CA2614353A1 - Management of plant pathogens - Google Patents
Management of plant pathogens Download PDFInfo
- Publication number
- CA2614353A1 CA2614353A1 CA002614353A CA2614353A CA2614353A1 CA 2614353 A1 CA2614353 A1 CA 2614353A1 CA 002614353 A CA002614353 A CA 002614353A CA 2614353 A CA2614353 A CA 2614353A CA 2614353 A1 CA2614353 A1 CA 2614353A1
- Authority
- CA
- Canada
- Prior art keywords
- plant
- weevil
- corn
- beetle
- pest
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 244000000003 plant pathogen Species 0.000 title abstract description 5
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 150
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 134
- 241000607479 Yersinia pestis Species 0.000 claims abstract description 127
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 86
- 230000000361 pesticidal effect Effects 0.000 claims abstract description 78
- 239000003001 serine protease inhibitor Substances 0.000 claims abstract description 62
- 101710187074 Serine proteinase inhibitor Proteins 0.000 claims abstract description 59
- 239000002243 precursor Substances 0.000 claims abstract description 47
- 230000002829 reductive effect Effects 0.000 claims abstract description 21
- 102000000541 Defensins Human genes 0.000 claims abstract description 20
- 108010002069 Defensins Proteins 0.000 claims abstract description 20
- 244000038559 crop plants Species 0.000 claims abstract description 9
- 241000196324 Embryophyta Species 0.000 claims description 322
- 239000000178 monomer Substances 0.000 claims description 56
- 240000008042 Zea mays Species 0.000 claims description 52
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 51
- 239000000203 mixture Substances 0.000 claims description 45
- 241000238631 Hexapoda Species 0.000 claims description 41
- 241000255967 Helicoverpa zea Species 0.000 claims description 37
- 241001629132 Blissus leucopterus Species 0.000 claims description 36
- 235000003222 Helianthus annuus Nutrition 0.000 claims description 34
- 229920000742 Cotton Polymers 0.000 claims description 33
- 241000219146 Gossypium Species 0.000 claims description 33
- 241001454293 Tetranychus urticae Species 0.000 claims description 32
- 244000068988 Glycine max Species 0.000 claims description 31
- 241000208818 Helianthus Species 0.000 claims description 31
- 241001478965 Melanoplus femurrubrum Species 0.000 claims description 31
- 241000256251 Spodoptera frugiperda Species 0.000 claims description 31
- 241001147398 Ostrinia nubilalis Species 0.000 claims description 29
- 241000566547 Agrotis ipsilon Species 0.000 claims description 28
- 241000400698 Elasmopalpus lignosellus Species 0.000 claims description 28
- 235000010469 Glycine max Nutrition 0.000 claims description 28
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 25
- 235000009973 maize Nutrition 0.000 claims description 25
- 241001415015 Melanoplus differentialis Species 0.000 claims description 24
- 240000007594 Oryza sativa Species 0.000 claims description 24
- 235000007164 Oryza sativa Nutrition 0.000 claims description 24
- 241001014341 Acrosternum hilare Species 0.000 claims description 23
- 235000011684 Sorghum saccharatum Nutrition 0.000 claims description 23
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 23
- 235000005822 corn Nutrition 0.000 claims description 23
- 240000006394 Sorghum bicolor Species 0.000 claims description 22
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims description 21
- 235000009566 rice Nutrition 0.000 claims description 21
- 241001609607 Delia platura Species 0.000 claims description 20
- 241000209140 Triticum Species 0.000 claims description 20
- 235000021307 Triticum Nutrition 0.000 claims description 20
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 18
- 244000061456 Solanum tuberosum Species 0.000 claims description 18
- 241000343781 Chaetocnema pulicaria Species 0.000 claims description 16
- 241000122106 Diatraea saccharalis Species 0.000 claims description 16
- 241001422926 Mayetiola hordei Species 0.000 claims description 16
- 241000922538 Melanoplus sanguinipes Species 0.000 claims description 16
- 241000208125 Nicotiana Species 0.000 claims description 16
- 241001160353 Oulema melanopus Species 0.000 claims description 16
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 16
- 241000286134 Phyllophaga crinita Species 0.000 claims description 16
- 241000167882 Rhopalosiphum maidis Species 0.000 claims description 16
- 241000722027 Schizaphis graminum Species 0.000 claims description 16
- 241000344246 Tetranychus cinnabarinus Species 0.000 claims description 16
- 241000339374 Thrips tabaci Species 0.000 claims description 16
- 239000002158 endotoxin Substances 0.000 claims description 16
- 238000012239 gene modification Methods 0.000 claims description 16
- 230000005017 genetic modification Effects 0.000 claims description 16
- 235000013617 genetically modified food Nutrition 0.000 claims description 16
- 241000489976 Diabrotica undecimpunctata howardi Species 0.000 claims description 15
- 241001147381 Helicoverpa armigera Species 0.000 claims description 15
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 15
- 235000010582 Pisum sativum Nutrition 0.000 claims description 15
- 240000004713 Pisum sativum Species 0.000 claims description 15
- 241000256247 Spodoptera exigua Species 0.000 claims description 15
- 241000254173 Coleoptera Species 0.000 claims description 14
- 235000007340 Hordeum vulgare Nutrition 0.000 claims description 14
- 240000005979 Hordeum vulgare Species 0.000 claims description 14
- 244000105624 Arachis hypogaea Species 0.000 claims description 13
- 241000258937 Hemiptera Species 0.000 claims description 13
- 241000219823 Medicago Species 0.000 claims description 13
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 claims description 13
- 244000062793 Sorghum vulgare Species 0.000 claims description 13
- 240000007241 Agrostis stolonifera Species 0.000 claims description 12
- 241000625764 Anticarsia gemmatalis Species 0.000 claims description 12
- 235000003255 Carthamus tinctorius Nutrition 0.000 claims description 12
- 244000020518 Carthamus tinctorius Species 0.000 claims description 12
- 241000254171 Curculionidae Species 0.000 claims description 12
- 241000879145 Diatraea grandiosella Species 0.000 claims description 12
- 241001619920 Euschistus servus Species 0.000 claims description 12
- 241000654868 Frankliniella fusca Species 0.000 claims description 12
- 241000255990 Helicoverpa Species 0.000 claims description 12
- 241000256244 Heliothis virescens Species 0.000 claims description 12
- 241000370523 Hypena scabra Species 0.000 claims description 12
- 241000501345 Lygus lineolaris Species 0.000 claims description 12
- 241000721451 Pectinophora gossypiella Species 0.000 claims description 12
- 241000517946 Phyllotreta nemorum Species 0.000 claims description 12
- 241000721694 Pseudatomoscelis seriatus Species 0.000 claims description 12
- 240000002791 Brassica napus Species 0.000 claims description 11
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims description 11
- 240000003768 Solanum lycopersicum Species 0.000 claims description 11
- 229940122618 Trypsin inhibitor Drugs 0.000 claims description 11
- 101710162629 Trypsin inhibitor Proteins 0.000 claims description 11
- 235000013339 cereals Nutrition 0.000 claims description 11
- 239000002753 trypsin inhibitor Substances 0.000 claims description 11
- 229940122644 Chymotrypsin inhibitor Drugs 0.000 claims description 10
- 101710137926 Chymotrypsin inhibitor Proteins 0.000 claims description 10
- 241000207199 Citrus Species 0.000 claims description 10
- 241000233866 Fungi Species 0.000 claims description 10
- 241000244206 Nematoda Species 0.000 claims description 10
- 241000208292 Solanaceae Species 0.000 claims description 10
- 239000003541 chymotrypsin inhibitor Substances 0.000 claims description 10
- 235000020232 peanut Nutrition 0.000 claims description 10
- 244000283070 Abies balsamea Species 0.000 claims description 9
- 235000007173 Abies balsamea Nutrition 0.000 claims description 9
- 244000075850 Avena orientalis Species 0.000 claims description 9
- 235000007319 Avena orientalis Nutrition 0.000 claims description 9
- 241000894006 Bacteria Species 0.000 claims description 9
- 235000011331 Brassica Nutrition 0.000 claims description 9
- 241000219198 Brassica Species 0.000 claims description 9
- 241000508723 Festuca rubra Species 0.000 claims description 9
- 241000482313 Globodera ellingtonae Species 0.000 claims description 9
- 235000003228 Lactuca sativa Nutrition 0.000 claims description 9
- 240000008415 Lactuca sativa Species 0.000 claims description 9
- 241000255777 Lepidoptera Species 0.000 claims description 9
- 240000006240 Linum usitatissimum Species 0.000 claims description 9
- 240000007817 Olea europaea Species 0.000 claims description 9
- 240000007377 Petunia x hybrida Species 0.000 claims description 9
- 235000010617 Phaseolus lunatus Nutrition 0.000 claims description 9
- 235000007238 Secale cereale Nutrition 0.000 claims description 9
- 102000012479 Serine Proteases Human genes 0.000 claims description 9
- 108010022999 Serine Proteases Proteins 0.000 claims description 9
- 241000044578 Stenotaphrum secundatum Species 0.000 claims description 9
- 235000020971 citrus fruits Nutrition 0.000 claims description 9
- 235000019713 millet Nutrition 0.000 claims description 9
- 241001143309 Acanthoscelides obtectus Species 0.000 claims description 8
- 241001136249 Agriotes lineatus Species 0.000 claims description 8
- 241000001996 Agrotis orthogonia Species 0.000 claims description 8
- 241001652650 Agrotis subterranea Species 0.000 claims description 8
- 241000254175 Anthonomus grandis Species 0.000 claims description 8
- 241000360281 Anthonomus quadrigibbus Species 0.000 claims description 8
- 241001600408 Aphis gossypii Species 0.000 claims description 8
- 241001414201 Bruchus pisorum Species 0.000 claims description 8
- 241001536086 Cephus cinctus Species 0.000 claims description 8
- 241001124201 Cerotoma trifurcata Species 0.000 claims description 8
- 241000877105 Chaetocnema confinis Species 0.000 claims description 8
- 241000256135 Chironomus thummi Species 0.000 claims description 8
- 241001367803 Chrysodeixis includens Species 0.000 claims description 8
- 244000060011 Cocos nucifera Species 0.000 claims description 8
- 235000013162 Cocos nucifera Nutrition 0.000 claims description 8
- 241001529599 Colaspis brunnea Species 0.000 claims description 8
- 241001587738 Cyclocephala borealis Species 0.000 claims description 8
- 241001090151 Cyrtopeltis Species 0.000 claims description 8
- 241001585354 Delia coarctata Species 0.000 claims description 8
- 241001529600 Diabrotica balteata Species 0.000 claims description 8
- 241000489947 Diabrotica virgifera virgifera Species 0.000 claims description 8
- 241000462639 Epilachna varivestis Species 0.000 claims description 8
- 241001183323 Epitrix cucumeris Species 0.000 claims description 8
- 241001442498 Globodera Species 0.000 claims description 8
- 241001442497 Globodera rostochiensis Species 0.000 claims description 8
- 241000498254 Heterodera glycines Species 0.000 claims description 8
- 241000379510 Heterodera schachtii Species 0.000 claims description 8
- 241001508566 Hypera postica Species 0.000 claims description 8
- 241001508564 Hypera punctata Species 0.000 claims description 8
- 241000258916 Leptinotarsa decemlineata Species 0.000 claims description 8
- 241000966204 Lissorhoptrus oryzophilus Species 0.000 claims description 8
- 241001130335 Maladera castanea Species 0.000 claims description 8
- 241000766511 Meligethes Species 0.000 claims description 8
- 241001477931 Mythimna unipuncta Species 0.000 claims description 8
- 241000721621 Myzus persicae Species 0.000 claims description 8
- 241000084931 Neohydatothrips variabilis Species 0.000 claims description 8
- 241000316608 Petrobia latens Species 0.000 claims description 8
- 241000275069 Phyllotreta cruciferae Species 0.000 claims description 8
- 241000254101 Popillia japonica Species 0.000 claims description 8
- 241001279786 Sipha flava Species 0.000 claims description 8
- 241000180219 Sitobion avenae Species 0.000 claims description 8
- 241000068648 Sitodiplosis mosellana Species 0.000 claims description 8
- 241000254152 Sitophilus oryzae Species 0.000 claims description 8
- 241000254154 Sitophilus zeamais Species 0.000 claims description 8
- 241001153341 Smicronyx sordidus Species 0.000 claims description 8
- 241000532885 Sphenophorus Species 0.000 claims description 8
- 241000254109 Tenebrio molitor Species 0.000 claims description 8
- 241001124191 Tenebrio obscurus Species 0.000 claims description 8
- 241000916142 Tetranychus turkestani Species 0.000 claims description 8
- 241000750338 Trialeurodes abutilonea Species 0.000 claims description 8
- 241000254113 Tribolium castaneum Species 0.000 claims description 8
- 241000254112 Tribolium confusum Species 0.000 claims description 8
- 241000532815 Zabrotes subfasciatus Species 0.000 claims description 8
- 241000314934 Zygogramma exclamationis Species 0.000 claims description 8
- 241001558877 Aceria tulipae Species 0.000 claims description 7
- 241000242266 Amphimallon majalis Species 0.000 claims description 7
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 7
- 235000006008 Brassica napus var napus Nutrition 0.000 claims description 7
- 235000004977 Brassica sinapistrum Nutrition 0.000 claims description 7
- 241000532642 Conotrachelus nenuphar Species 0.000 claims description 7
- 244000000626 Daucus carota Species 0.000 claims description 7
- 235000002767 Daucus carota Nutrition 0.000 claims description 7
- 241000489972 Diabrotica barberi Species 0.000 claims description 7
- 241000995027 Empoasca fabae Species 0.000 claims description 7
- 241000630740 Homoeosoma electellum Species 0.000 claims description 7
- 241000590524 Protaphis middletonii Species 0.000 claims description 7
- 241000209137 Agropyron cristatum Species 0.000 claims description 6
- 241000491617 Agropyron desertorum Species 0.000 claims description 6
- 241001184547 Agrostis capillaris Species 0.000 claims description 6
- 244000144725 Amygdalus communis Species 0.000 claims description 6
- 235000011437 Amygdalus communis Nutrition 0.000 claims description 6
- 244000226021 Anacardium occidentale Species 0.000 claims description 6
- 244000099147 Ananas comosus Species 0.000 claims description 6
- 235000007119 Ananas comosus Nutrition 0.000 claims description 6
- 241001674345 Callitropsis nootkatensis Species 0.000 claims description 6
- 244000045232 Canavalia ensiformis Species 0.000 claims description 6
- 235000009467 Carica papaya Nutrition 0.000 claims description 6
- 240000006432 Carica papaya Species 0.000 claims description 6
- 241000723377 Coffea Species 0.000 claims description 6
- 241000218631 Coniferophyta Species 0.000 claims description 6
- 235000009847 Cucumis melo var cantalupensis Nutrition 0.000 claims description 6
- 240000008067 Cucumis sativus Species 0.000 claims description 6
- 235000009355 Dianthus caryophyllus Nutrition 0.000 claims description 6
- 240000006497 Dianthus caryophyllus Species 0.000 claims description 6
- 241000255925 Diptera Species 0.000 claims description 6
- 244000078127 Eleusine coracana Species 0.000 claims description 6
- 241000025852 Eremochloa ophiuroides Species 0.000 claims description 6
- 240000002395 Euphorbia pulcherrima Species 0.000 claims description 6
- 241000234643 Festuca arundinacea Species 0.000 claims description 6
- 241000192306 Festuca longifolia Species 0.000 claims description 6
- 241000410074 Festuca ovina Species 0.000 claims description 6
- 241000448472 Gramma Species 0.000 claims description 6
- 235000005206 Hibiscus Nutrition 0.000 claims description 6
- 235000007185 Hibiscus lunariifolius Nutrition 0.000 claims description 6
- 244000267823 Hydrangea macrophylla Species 0.000 claims description 6
- 235000014486 Hydrangea macrophylla Nutrition 0.000 claims description 6
- 235000004431 Linum usitatissimum Nutrition 0.000 claims description 6
- 244000100545 Lolium multiflorum Species 0.000 claims description 6
- 235000014826 Mangifera indica Nutrition 0.000 claims description 6
- 240000007228 Mangifera indica Species 0.000 claims description 6
- 241000234479 Narcissus Species 0.000 claims description 6
- 241001668543 Pascopyrum smithii Species 0.000 claims description 6
- 241001330451 Paspalum notatum Species 0.000 claims description 6
- 241000044541 Paspalum vaginatum Species 0.000 claims description 6
- 235000007195 Pennisetum typhoides Nutrition 0.000 claims description 6
- 244000025272 Persea americana Species 0.000 claims description 6
- 235000008673 Persea americana Nutrition 0.000 claims description 6
- 241000218606 Pinus contorta Species 0.000 claims description 6
- 235000013267 Pinus ponderosa Nutrition 0.000 claims description 6
- 235000008577 Pinus radiata Nutrition 0.000 claims description 6
- 241000218621 Pinus radiata Species 0.000 claims description 6
- 235000008566 Pinus taeda Nutrition 0.000 claims description 6
- 241000218679 Pinus taeda Species 0.000 claims description 6
- 244000292693 Poa annua Species 0.000 claims description 6
- 241000136254 Poa compressa Species 0.000 claims description 6
- 241000209049 Poa pratensis Species 0.000 claims description 6
- 240000006597 Poa trivialis Species 0.000 claims description 6
- 240000001416 Pseudotsuga menziesii Species 0.000 claims description 6
- 241000736230 Puccinellia distans Species 0.000 claims description 6
- 241000208422 Rhododendron Species 0.000 claims description 6
- 241000109329 Rosa xanthina Species 0.000 claims description 6
- 235000004789 Rosa xanthina Nutrition 0.000 claims description 6
- 240000005498 Setaria italica Species 0.000 claims description 6
- 244000269722 Thea sinensis Species 0.000 claims description 6
- 244000299461 Theobroma cacao Species 0.000 claims description 6
- 235000009470 Theobroma cacao Nutrition 0.000 claims description 6
- 241000218638 Thuja plicata Species 0.000 claims description 6
- 235000010749 Vicia faba Nutrition 0.000 claims description 6
- 240000006677 Vicia faba Species 0.000 claims description 6
- 235000002098 Vicia faba var. major Nutrition 0.000 claims description 6
- 240000001102 Zoysia matrella Species 0.000 claims description 6
- 244000013123 dwarf bean Species 0.000 claims description 6
- 235000004426 flaxseed Nutrition 0.000 claims description 6
- 231100000331 toxic Toxicity 0.000 claims description 6
- 230000002588 toxic effect Effects 0.000 claims description 6
- 235000013311 vegetables Nutrition 0.000 claims description 6
- 241001626535 Agrostis canina Species 0.000 claims description 5
- 240000004585 Dactylis glomerata Species 0.000 claims description 5
- 240000004296 Lolium perenne Species 0.000 claims description 5
- 244000061176 Nicotiana tabacum Species 0.000 claims description 5
- 235000007199 Panicum miliaceum Nutrition 0.000 claims description 5
- 241000332477 Scutellonema bradys Species 0.000 claims description 5
- 244000082988 Secale cereale Species 0.000 claims description 5
- 241000256248 Spodoptera Species 0.000 claims description 5
- 244000022203 blackseeded proso millet Species 0.000 claims description 5
- 241000495828 Acleris gloverana Species 0.000 claims description 4
- 241000834107 Acleris variana Species 0.000 claims description 4
- 241000693815 Adelphocoris rapidus Species 0.000 claims description 4
- 241000175828 Adoxophyes orana Species 0.000 claims description 4
- 241000673185 Aeolus Species 0.000 claims description 4
- 241000993143 Agromyza Species 0.000 claims description 4
- 241000449794 Alabama argillacea Species 0.000 claims description 4
- 235000002732 Allium cepa var. cepa Nutrition 0.000 claims description 4
- 241001367806 Alsophila pometaria Species 0.000 claims description 4
- 241001259789 Amyelois transitella Species 0.000 claims description 4
- 241001673643 Anaphothrips obscurus Species 0.000 claims description 4
- 241001198505 Anarsia lineatella Species 0.000 claims description 4
- 241000153204 Anisota senatoria Species 0.000 claims description 4
- 241001427556 Anoplura Species 0.000 claims description 4
- 241000255978 Antheraea pernyi Species 0.000 claims description 4
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 4
- 235000018262 Arachis monticola Nutrition 0.000 claims description 4
- 241001332254 Araecerus fasciculatus Species 0.000 claims description 4
- 241001002469 Archips Species 0.000 claims description 4
- 241000384127 Argyrotaenia Species 0.000 claims description 4
- 241000335053 Beta vulgaris Species 0.000 claims description 4
- 241000255789 Bombyx mori Species 0.000 claims description 4
- 241001444260 Brassicogethes aeneus Species 0.000 claims description 4
- 241000982105 Brevicoryne brassicae Species 0.000 claims description 4
- 241001325378 Bruchus Species 0.000 claims description 4
- 241001388466 Bruchus rufimanus Species 0.000 claims description 4
- 241001517925 Bucculatrix Species 0.000 claims description 4
- 241000726760 Cadra cautella Species 0.000 claims description 4
- 241000906761 Calocoris Species 0.000 claims description 4
- 241000661337 Chilo partellus Species 0.000 claims description 4
- 241000255945 Choristoneura Species 0.000 claims description 4
- 241001414835 Cimicidae Species 0.000 claims description 4
- DBPRUZCKPFOVDV-UHFFFAOYSA-N Clorprenaline hydrochloride Chemical compound O.Cl.CC(C)NCC(O)C1=CC=CC=C1Cl DBPRUZCKPFOVDV-UHFFFAOYSA-N 0.000 claims description 4
- 241000143939 Colias eurytheme Species 0.000 claims description 4
- 241000683561 Conoderus Species 0.000 claims description 4
- 241001663470 Contarinia <gall midge> Species 0.000 claims description 4
- 241000993412 Corcyra cephalonica Species 0.000 claims description 4
- 241001114553 Coreidae Species 0.000 claims description 4
- 241001641310 Cunea Species 0.000 claims description 4
- 241000721021 Curculio Species 0.000 claims description 4
- 241001156075 Cyclocephala Species 0.000 claims description 4
- 241001634817 Cydia Species 0.000 claims description 4
- 241001635274 Cydia pomonella Species 0.000 claims description 4
- 241001351082 Datana integerrima Species 0.000 claims description 4
- 241001631715 Dendrolimus Species 0.000 claims description 4
- 241001124144 Dermaptera Species 0.000 claims description 4
- 241000489977 Diabrotica virgifera Species 0.000 claims description 4
- 241000381325 Diabrotica virgifera zeae Species 0.000 claims description 4
- 241001000394 Diaphania hyalinata Species 0.000 claims description 4
- 241001012951 Diaphania nitidalis Species 0.000 claims description 4
- 241001279823 Diuraphis noxia Species 0.000 claims description 4
- 241001517923 Douglasiidae Species 0.000 claims description 4
- 241001105160 Eleodes Species 0.000 claims description 4
- 241001608224 Ennomos subsignaria Species 0.000 claims description 4
- 241000661448 Eoreuma loftini Species 0.000 claims description 4
- 241000122098 Ephestia kuehniella Species 0.000 claims description 4
- 241001491718 Erannis Species 0.000 claims description 4
- 241000567412 Estigmene acrea Species 0.000 claims description 4
- 241001201696 Eulia Species 0.000 claims description 4
- 241000060469 Eupoecilia ambiguella Species 0.000 claims description 4
- 241000483001 Euproctis chrysorrhoea Species 0.000 claims description 4
- 241000515838 Eurygaster Species 0.000 claims description 4
- 241000167999 Euscepes Species 0.000 claims description 4
- 241000168001 Euscepes postfasciatus Species 0.000 claims description 4
- 241001368778 Euxoa messoria Species 0.000 claims description 4
- 241000242711 Fasciola hepatica Species 0.000 claims description 4
- 241000255896 Galleria mellonella Species 0.000 claims description 4
- 241001441330 Grapholita molesta Species 0.000 claims description 4
- 241001352371 Harrisina americana Species 0.000 claims description 4
- 241001201676 Hedya nubiferana Species 0.000 claims description 4
- 241000413128 Hemileuca oliviae Species 0.000 claims description 4
- 241001480224 Heterodera Species 0.000 claims description 4
- 241001481225 Heterodera avenae Species 0.000 claims description 4
- 244000284380 Hibiscus rosa sinensis Species 0.000 claims description 4
- 241001508558 Hypera Species 0.000 claims description 4
- 241001508570 Hypera brunneipennis Species 0.000 claims description 4
- 241001495069 Ischnocera Species 0.000 claims description 4
- 241000256602 Isoptera Species 0.000 claims description 4
- 241000400431 Keiferia lycopersicella Species 0.000 claims description 4
- 241001352367 Leucoma salicis Species 0.000 claims description 4
- 241001261104 Lobesia botrana Species 0.000 claims description 4
- 241000193981 Loxostege sticticalis Species 0.000 claims description 4
- 241000283636 Lygocoris pabulinus Species 0.000 claims description 4
- 241001414823 Lygus hesperus Species 0.000 claims description 4
- 241001492180 Lygus pratensis Species 0.000 claims description 4
- 241001048449 Lygus rugulipennis Species 0.000 claims description 4
- 241000721703 Lymantria dispar Species 0.000 claims description 4
- 241000081125 Macalla Species 0.000 claims description 4
- 241001155765 Macrodactylus Species 0.000 claims description 4
- 241000255676 Malacosoma Species 0.000 claims description 4
- 241000555303 Mamestra brassicae Species 0.000 claims description 4
- 241000732113 Mamestra configurata Species 0.000 claims description 4
- 241000369513 Manduca quinquemaculata Species 0.000 claims description 4
- 241000255908 Manduca sexta Species 0.000 claims description 4
- 241001232130 Maruca testulalis Species 0.000 claims description 4
- 241001367645 Melanchra picta Species 0.000 claims description 4
- 241001062280 Melanotus <basidiomycete fungus> Species 0.000 claims description 4
- 241000537142 Meligethes nigrescens Species 0.000 claims description 4
- 241000042079 Meligethes viridescens Species 0.000 claims description 4
- 241001143352 Meloidogyne Species 0.000 claims description 4
- 241000254043 Melolonthinae Species 0.000 claims description 4
- 241001477928 Mythimna Species 0.000 claims description 4
- 241000912288 Neolasioptera Species 0.000 claims description 4
- 241000615716 Nephotettix nigropictus Species 0.000 claims description 4
- 241001671709 Nezara viridula Species 0.000 claims description 4
- 244000061322 Nicotiana alata Species 0.000 claims description 4
- 241001548845 Nysius ericae Species 0.000 claims description 4
- 241001666448 Nysius raphanus Species 0.000 claims description 4
- 241001491877 Operophtera brumata Species 0.000 claims description 4
- 241001465800 Orgyia Species 0.000 claims description 4
- 241000906034 Orthops Species 0.000 claims description 4
- 241001585671 Paleacrita vernata Species 0.000 claims description 4
- 241001300993 Papilio cresphontes Species 0.000 claims description 4
- 241001517955 Phyllonorycter blancardella Species 0.000 claims description 4
- 241000437063 Phyllotreta striolata Species 0.000 claims description 4
- 241001313099 Pieris napi Species 0.000 claims description 4
- 241000907661 Pieris rapae Species 0.000 claims description 4
- 241001608845 Platynota Species 0.000 claims description 4
- 241001456328 Platynota stultana Species 0.000 claims description 4
- 241000495716 Platyptilia carduidactyla Species 0.000 claims description 4
- 241000595629 Plodia interpunctella Species 0.000 claims description 4
- 241000500437 Plutella xylostella Species 0.000 claims description 4
- 241001662912 Poecilocapsus lineatus Species 0.000 claims description 4
- 241000143945 Pontia protodice Species 0.000 claims description 4
- 241000193943 Pratylenchus Species 0.000 claims description 4
- 241001398044 Protaetia cuprea Species 0.000 claims description 4
- 241001657916 Proxenus mindara Species 0.000 claims description 4
- 241001510071 Pyrrhocoridae Species 0.000 claims description 4
- 241001124072 Reduviidae Species 0.000 claims description 4
- 241000004261 Sabulodes Species 0.000 claims description 4
- 241001351292 Schizura concinna Species 0.000 claims description 4
- 241000661450 Sesamia cretica Species 0.000 claims description 4
- 241000533293 Sesbania emerus Species 0.000 claims description 4
- 241000258242 Siphonaptera Species 0.000 claims description 4
- 241000254181 Sitophilus Species 0.000 claims description 4
- 241000254179 Sitophilus granarius Species 0.000 claims description 4
- 241000753145 Sitotroga cerealella Species 0.000 claims description 4
- 241001153355 Smicronyx Species 0.000 claims description 4
- 241001153342 Smicronyx fulvus Species 0.000 claims description 4
- 241001492664 Solenopsis <angiosperm> Species 0.000 claims description 4
- 241000779864 Solenopsis fugax Species 0.000 claims description 4
- 241000421631 Spanagonicus albofasciatus Species 0.000 claims description 4
- 241000950030 Sternechus Species 0.000 claims description 4
- 241001575047 Suleima Species 0.000 claims description 4
- 241001414989 Thysanoptera Species 0.000 claims description 4
- 241000255985 Trichoplusia Species 0.000 claims description 4
- 241001414983 Trichoptera Species 0.000 claims description 4
- 241001351286 Udea rubigalis Species 0.000 claims description 4
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 4
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 4
- 240000006365 Vitis vinifera Species 0.000 claims description 4
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 4
- 241000064240 Yponomeuta padellus Species 0.000 claims description 4
- 235000007244 Zea mays Nutrition 0.000 claims description 4
- 235000005489 dwarf bean Nutrition 0.000 claims description 4
- 208000006275 fascioliasis Diseases 0.000 claims description 4
- 244000079416 protozoan pathogen Species 0.000 claims description 4
- 235000020238 sunflower seed Nutrition 0.000 claims description 4
- 235000004507 Abies alba Nutrition 0.000 claims description 3
- 235000014081 Abies amabilis Nutrition 0.000 claims description 3
- 244000101408 Abies amabilis Species 0.000 claims description 3
- 244000178606 Abies grandis Species 0.000 claims description 3
- 235000017894 Abies grandis Nutrition 0.000 claims description 3
- 235000004710 Abies lasiocarpa Nutrition 0.000 claims description 3
- 240000005020 Acaciella glauca Species 0.000 claims description 3
- 241001133760 Acoelorraphe Species 0.000 claims description 3
- 235000005254 Allium ampeloprasum Nutrition 0.000 claims description 3
- 240000006108 Allium ampeloprasum Species 0.000 claims description 3
- 235000001274 Anacardium occidentale Nutrition 0.000 claims description 3
- 241000207875 Antirrhinum Species 0.000 claims description 3
- 241000047982 Axonopus Species 0.000 claims description 3
- 241000047987 Axonopus fissifolius Species 0.000 claims description 3
- 235000021533 Beta vulgaris Nutrition 0.000 claims description 3
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 claims description 3
- 241000145727 Bouteloua curtipendula Species 0.000 claims description 3
- 241000232315 Bouteloua gracilis Species 0.000 claims description 3
- 241000339490 Brachyachne Species 0.000 claims description 3
- 244000178993 Brassica juncea Species 0.000 claims description 3
- 235000014698 Brassica juncea var multisecta Nutrition 0.000 claims description 3
- 240000000385 Brassica napus var. napus Species 0.000 claims description 3
- 240000008100 Brassica rapa Species 0.000 claims description 3
- 235000006618 Brassica rapa subsp oleifera Nutrition 0.000 claims description 3
- 241000220243 Brassica sp. Species 0.000 claims description 3
- 241000743756 Bromus inermis Species 0.000 claims description 3
- 235000004936 Bromus mango Nutrition 0.000 claims description 3
- 241000544756 Bromus racemosus Species 0.000 claims description 3
- 241000320719 Buchloe Species 0.000 claims description 3
- 235000005881 Calendula officinalis Nutrition 0.000 claims description 3
- 240000001432 Calendula officinalis Species 0.000 claims description 3
- 241000218645 Cedrus Species 0.000 claims description 3
- 235000013912 Ceratonia siliqua Nutrition 0.000 claims description 3
- 240000008886 Ceratonia siliqua Species 0.000 claims description 3
- 235000007516 Chrysanthemum Nutrition 0.000 claims description 3
- 244000189548 Chrysanthemum x morifolium Species 0.000 claims description 3
- 235000010523 Cicer arietinum Nutrition 0.000 claims description 3
- 244000045195 Cicer arietinum Species 0.000 claims description 3
- 241000219112 Cucumis Species 0.000 claims description 3
- 235000010071 Cucumis prophetarum Nutrition 0.000 claims description 3
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims description 3
- 244000007835 Cyamopsis tetragonoloba Species 0.000 claims description 3
- 244000019459 Cynara cardunculus Species 0.000 claims description 3
- 235000019106 Cynara scolymus Nutrition 0.000 claims description 3
- 244000052363 Cynodon dactylon Species 0.000 claims description 3
- 235000014466 Douglas bleu Nutrition 0.000 claims description 3
- 235000007349 Eleusine coracana Nutrition 0.000 claims description 3
- 235000013499 Eleusine coracana subsp coracana Nutrition 0.000 claims description 3
- 241000234642 Festuca Species 0.000 claims description 3
- 241000218218 Ficus <angiosperm> Species 0.000 claims description 3
- 241000208150 Geraniaceae Species 0.000 claims description 3
- 240000000047 Gossypium barbadense Species 0.000 claims description 3
- 235000009429 Gossypium barbadense Nutrition 0.000 claims description 3
- 244000299507 Gossypium hirsutum Species 0.000 claims description 3
- 235000009432 Gossypium hirsutum Nutrition 0.000 claims description 3
- 244000020551 Helianthus annuus Species 0.000 claims description 3
- 244000308760 Helichrysum petiolatum Species 0.000 claims description 3
- 235000021506 Ipomoea Nutrition 0.000 claims description 3
- 241000207783 Ipomoea Species 0.000 claims description 3
- 244000017020 Ipomoea batatas Species 0.000 claims description 3
- 235000002678 Ipomoea batatas Nutrition 0.000 claims description 3
- 241000219729 Lathyrus Species 0.000 claims description 3
- 240000004322 Lens culinaris Species 0.000 claims description 3
- 235000014647 Lens culinaris subsp culinaris Nutrition 0.000 claims description 3
- 241000219745 Lupinus Species 0.000 claims description 3
- 241000208467 Macadamia Species 0.000 claims description 3
- 235000018330 Macadamia integrifolia Nutrition 0.000 claims description 3
- 240000007575 Macadamia integrifolia Species 0.000 claims description 3
- 240000003183 Manihot esculenta Species 0.000 claims description 3
- 235000004456 Manihot esculenta Nutrition 0.000 claims description 3
- 240000004658 Medicago sativa Species 0.000 claims description 3
- 235000010624 Medicago sativa Nutrition 0.000 claims description 3
- 241000234295 Musa Species 0.000 claims description 3
- 240000005561 Musa balbisiana Species 0.000 claims description 3
- 235000018290 Musa x paradisiaca Nutrition 0.000 claims description 3
- 235000002725 Olea europaea Nutrition 0.000 claims description 3
- 241000233855 Orchidaceae Species 0.000 claims description 3
- 241000209046 Pennisetum Species 0.000 claims description 3
- 244000026791 Pennisetum clandestinum Species 0.000 claims description 3
- 244000038248 Pennisetum spicatum Species 0.000 claims description 3
- 244000115721 Pennisetum typhoides Species 0.000 claims description 3
- 244000100170 Phaseolus lunatus Species 0.000 claims description 3
- 241000746983 Phleum pratense Species 0.000 claims description 3
- 235000001982 Physalis edulis Nutrition 0.000 claims description 3
- 240000004001 Physalis peruviana Species 0.000 claims description 3
- 240000000020 Picea glauca Species 0.000 claims description 3
- 235000008127 Picea glauca Nutrition 0.000 claims description 3
- 241000218595 Picea sitchensis Species 0.000 claims description 3
- 235000005205 Pinus Nutrition 0.000 claims description 3
- 241000218602 Pinus <genus> Species 0.000 claims description 3
- 235000008331 Pinus X rigitaeda Nutrition 0.000 claims description 3
- 235000011613 Pinus brutia Nutrition 0.000 claims description 3
- 241000018646 Pinus brutia Species 0.000 claims description 3
- 235000008593 Pinus contorta Nutrition 0.000 claims description 3
- 235000011334 Pinus elliottii Nutrition 0.000 claims description 3
- 241000142776 Pinus elliottii Species 0.000 claims description 3
- 244000019397 Pinus jeffreyi Species 0.000 claims description 3
- 241000555277 Pinus ponderosa Species 0.000 claims description 3
- 235000013269 Pinus ponderosa var ponderosa Nutrition 0.000 claims description 3
- 235000013268 Pinus ponderosa var scopulorum Nutrition 0.000 claims description 3
- 241001290151 Prunus avium subsp. avium Species 0.000 claims description 3
- 235000008572 Pseudotsuga menziesii Nutrition 0.000 claims description 3
- 235000005386 Pseudotsuga menziesii var menziesii Nutrition 0.000 claims description 3
- 241000508269 Psidium Species 0.000 claims description 3
- 240000001679 Psidium guajava Species 0.000 claims description 3
- 235000013929 Psidium pyriferum Nutrition 0.000 claims description 3
- 235000004443 Ricinus communis Nutrition 0.000 claims description 3
- 235000011449 Rosa Nutrition 0.000 claims description 3
- 241000209051 Saccharum Species 0.000 claims description 3
- 240000000111 Saccharum officinarum Species 0.000 claims description 3
- 235000007201 Saccharum officinarum Nutrition 0.000 claims description 3
- 241001138418 Sequoia sempervirens Species 0.000 claims description 3
- 235000008515 Setaria glauca Nutrition 0.000 claims description 3
- 235000007226 Setaria italica Nutrition 0.000 claims description 3
- 235000007230 Sorghum bicolor Nutrition 0.000 claims description 3
- 235000009184 Spondias indica Nutrition 0.000 claims description 3
- 235000021536 Sugar beet Nutrition 0.000 claims description 3
- 235000006468 Thea sinensis Nutrition 0.000 claims description 3
- 235000001484 Trigonella foenum graecum Nutrition 0.000 claims description 3
- 244000250129 Trigonella foenum graecum Species 0.000 claims description 3
- 235000007218 Tripsacum dactyloides Nutrition 0.000 claims description 3
- 235000019714 Triticale Nutrition 0.000 claims description 3
- 244000098338 Triticum aestivum Species 0.000 claims description 3
- 240000003021 Tsuga heterophylla Species 0.000 claims description 3
- 235000008554 Tsuga heterophylla Nutrition 0.000 claims description 3
- 241000722923 Tulipa Species 0.000 claims description 3
- 241000722921 Tulipa gesneriana Species 0.000 claims description 3
- 241000219977 Vigna Species 0.000 claims description 3
- 240000004922 Vigna radiata Species 0.000 claims description 3
- 235000010721 Vigna radiata var radiata Nutrition 0.000 claims description 3
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 claims description 3
- 235000010726 Vigna sinensis Nutrition 0.000 claims description 3
- 241000482268 Zea mays subsp. mays Species 0.000 claims description 3
- 235000020224 almond Nutrition 0.000 claims description 3
- 235000016520 artichoke thistle Nutrition 0.000 claims description 3
- 235000020226 cashew nut Nutrition 0.000 claims description 3
- 235000019693 cherries Nutrition 0.000 claims description 3
- 230000001055 chewing effect Effects 0.000 claims description 3
- 235000021331 green beans Nutrition 0.000 claims description 3
- 235000014684 lodgepole pine Nutrition 0.000 claims description 3
- 235000002252 panizo Nutrition 0.000 claims description 3
- 235000021018 plums Nutrition 0.000 claims description 3
- 235000003499 redwood Nutrition 0.000 claims description 3
- 235000000673 shore pine Nutrition 0.000 claims description 3
- 235000001019 trigonella foenum-graecum Nutrition 0.000 claims description 3
- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 3
- 241000228158 x Triticosecale Species 0.000 claims description 3
- 241001190490 Phryganidia Species 0.000 claims description 2
- 244000064895 Cucumis melo subsp melo Species 0.000 claims 6
- 241000209056 Secale Species 0.000 claims 4
- 241000318389 Anaphothrips Species 0.000 claims 3
- 241000257303 Hymenoptera Species 0.000 claims 3
- 241001658022 Lambdina fiscellaria fiscellaria Species 0.000 claims 3
- 241001658020 Lambdina fiscellaria lugubrosa Species 0.000 claims 3
- 241000088587 Meromyza Species 0.000 claims 3
- 241000234282 Allium Species 0.000 claims 2
- 241000219322 Dianthus Species 0.000 claims 2
- 241000218033 Hibiscus Species 0.000 claims 2
- 241001190492 Phryganidia californica Species 0.000 claims 2
- 235000016068 Berberis vulgaris Nutrition 0.000 claims 1
- 241001415013 Melanoplus Species 0.000 claims 1
- 238000013459 approach Methods 0.000 abstract description 8
- 210000004027 cell Anatomy 0.000 description 107
- 235000018102 proteins Nutrition 0.000 description 70
- 238000000034 method Methods 0.000 description 41
- 230000000694 effects Effects 0.000 description 36
- 230000014509 gene expression Effects 0.000 description 34
- 210000001519 tissue Anatomy 0.000 description 31
- 230000009466 transformation Effects 0.000 description 22
- 108091028043 Nucleic acid sequence Proteins 0.000 description 21
- 230000002068 genetic effect Effects 0.000 description 21
- 102000040430 polynucleotide Human genes 0.000 description 18
- 108091033319 polynucleotide Proteins 0.000 description 18
- 239000002157 polynucleotide Substances 0.000 description 18
- -1 but not limited to Chemical compound 0.000 description 17
- 210000000056 organ Anatomy 0.000 description 17
- 235000013601 eggs Nutrition 0.000 description 16
- 150000007523 nucleic acids Chemical class 0.000 description 16
- 239000003053 toxin Substances 0.000 description 15
- 231100000765 toxin Toxicity 0.000 description 15
- 108700012359 toxins Proteins 0.000 description 15
- 102000039446 nucleic acids Human genes 0.000 description 14
- 108020004707 nucleic acids Proteins 0.000 description 14
- 108090000765 processed proteins & peptides Proteins 0.000 description 13
- 239000000126 substance Substances 0.000 description 13
- 230000009261 transgenic effect Effects 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 12
- 102000004196 processed proteins & peptides Human genes 0.000 description 12
- 239000003550 marker Substances 0.000 description 11
- 239000000463 material Substances 0.000 description 11
- 102000035195 Peptidases Human genes 0.000 description 10
- 108091005804 Peptidases Proteins 0.000 description 10
- 229920001184 polypeptide Polymers 0.000 description 10
- 108020004414 DNA Proteins 0.000 description 9
- 235000005911 diet Nutrition 0.000 description 9
- 230000037213 diet Effects 0.000 description 9
- 230000001965 increasing effect Effects 0.000 description 9
- 239000003112 inhibitor Substances 0.000 description 9
- 230000000749 insecticidal effect Effects 0.000 description 9
- 238000000576 coating method Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 230000001404 mediated effect Effects 0.000 description 8
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 8
- 238000004166 bioassay Methods 0.000 description 7
- 239000011248 coating agent Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 230000001105 regulatory effect Effects 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 238000011282 treatment Methods 0.000 description 7
- 239000004365 Protease Substances 0.000 description 6
- 125000003275 alpha amino acid group Chemical group 0.000 description 6
- 239000004009 herbicide Substances 0.000 description 6
- 239000011859 microparticle Substances 0.000 description 6
- 239000004094 surface-active agent Substances 0.000 description 6
- 241000193388 Bacillus thuringiensis Species 0.000 description 5
- 108090000317 Chymotrypsin Proteins 0.000 description 5
- 206010061217 Infestation Diseases 0.000 description 5
- 108010025815 Kanamycin Kinase Proteins 0.000 description 5
- 102000004142 Trypsin Human genes 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 229920002494 Zein Polymers 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 239000000969 carrier Substances 0.000 description 5
- 229960002376 chymotrypsin Drugs 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- 230000015788 innate immune response Effects 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 230000008488 polyadenylation Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000005019 zein Substances 0.000 description 5
- 229940093612 zein Drugs 0.000 description 5
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- 241000701489 Cauliflower mosaic virus Species 0.000 description 4
- 108700010070 Codon Usage Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 108090000631 Trypsin Proteins 0.000 description 4
- 108010055615 Zein Proteins 0.000 description 4
- 238000007792 addition Methods 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 235000014113 dietary fatty acids Nutrition 0.000 description 4
- 229930195729 fatty acid Natural products 0.000 description 4
- 239000000194 fatty acid Substances 0.000 description 4
- 229910052738 indium Inorganic materials 0.000 description 4
- 230000001939 inductive effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 108010058731 nopaline synthase Proteins 0.000 description 4
- 244000052769 pathogen Species 0.000 description 4
- 235000019833 protease Nutrition 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000012552 review Methods 0.000 description 4
- 239000012588 trypsin Substances 0.000 description 4
- 108010000700 Acetolactate synthase Proteins 0.000 description 3
- 108700003918 Bacillus Thuringiensis insecticidal crystal Proteins 0.000 description 3
- 108020004705 Codon Proteins 0.000 description 3
- 101710151559 Crystal protein Proteins 0.000 description 3
- 244000241257 Cucumis melo Species 0.000 description 3
- 102000015833 Cystatin Human genes 0.000 description 3
- 108010066133 D-octopine dehydrogenase Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N Glutamine Chemical compound OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 3
- 240000002024 Gossypium herbaceum Species 0.000 description 3
- 235000004341 Gossypium herbaceum Nutrition 0.000 description 3
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 150000001412 amines Chemical class 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 210000003850 cellular structure Anatomy 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 239000007859 condensation product Substances 0.000 description 3
- 230000001276 controlling effect Effects 0.000 description 3
- 108050004038 cystatin Proteins 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000004520 electroporation Methods 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 210000005081 epithelial layer Anatomy 0.000 description 3
- 239000003337 fertilizer Substances 0.000 description 3
- 239000000417 fungicide Substances 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 210000000087 hemolymph Anatomy 0.000 description 3
- 230000002363 herbicidal effect Effects 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000002917 insecticide Substances 0.000 description 3
- 230000010354 integration Effects 0.000 description 3
- 101150066555 lacZ gene Proteins 0.000 description 3
- 230000001418 larval effect Effects 0.000 description 3
- 235000021374 legumes Nutrition 0.000 description 3
- 108010083942 mannopine synthase Proteins 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 229920000058 polyacrylate Polymers 0.000 description 3
- 235000019419 proteases Nutrition 0.000 description 3
- 210000001938 protoplast Anatomy 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 150000003871 sulfonates Chemical class 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 241000238876 Acari Species 0.000 description 2
- 244000291564 Allium cepa Species 0.000 description 2
- 101150078024 CRY2 gene Proteins 0.000 description 2
- 108010035563 Chloramphenicol O-acetyltransferase Proteins 0.000 description 2
- 241001057636 Dracaena deremensis Species 0.000 description 2
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 2
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 2
- 102100029880 Glycodelin Human genes 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 241001658021 Lambdina Species 0.000 description 2
- 241000209510 Liliopsida Species 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 240000009200 Macaranga tanarius Species 0.000 description 2
- 235000000487 Macaranga tanarius Nutrition 0.000 description 2
- SUZRRICLUFMAQD-UHFFFAOYSA-N N-Methyltaurine Chemical compound CNCCS(O)(=O)=O SUZRRICLUFMAQD-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 2
- 241001646398 Pseudomonas chlororaphis Species 0.000 description 2
- 101000611441 Solanum lycopersicum Pathogenesis-related leaf protein 6 Proteins 0.000 description 2
- 101001044900 Solanum tuberosum Proteinase inhibitor 1 Proteins 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 108700005078 Synthetic Genes Proteins 0.000 description 2
- 108010084455 Zeocin Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 210000004102 animal cell Anatomy 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 229940019748 antifibrinolytic proteinase inhibitors Drugs 0.000 description 2
- 239000003899 bactericide agent Substances 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000003542 behavioural effect Effects 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 230000000853 biopesticidal effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 150000007942 carboxylates Chemical class 0.000 description 2
- 230000002301 combined effect Effects 0.000 description 2
- 239000013065 commercial product Substances 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 2
- 239000002270 dispersing agent Substances 0.000 description 2
- 238000010410 dusting Methods 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 241001233957 eudicotyledons Species 0.000 description 2
- 210000001723 extracellular space Anatomy 0.000 description 2
- 150000002191 fatty alcohols Chemical class 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000004077 genetic alteration Effects 0.000 description 2
- 231100000118 genetic alteration Toxicity 0.000 description 2
- 102000005396 glutamine synthetase Human genes 0.000 description 2
- 108020002326 glutamine synthetase Proteins 0.000 description 2
- 150000002334 glycols Chemical class 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000003116 impacting effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 230000009571 larval growth Effects 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 2
- 238000000520 microinjection Methods 0.000 description 2
- 239000003750 molluscacide Substances 0.000 description 2
- 230000002013 molluscicidal effect Effects 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 239000005645 nematicide Substances 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 2
- 239000005014 poly(hydroxyalkanoate) Substances 0.000 description 2
- 229920000768 polyamine Polymers 0.000 description 2
- 229920000903 polyhydroxyalkanoate Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 230000000644 propagated effect Effects 0.000 description 2
- 230000001850 reproductive effect Effects 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000011426 transformation method Methods 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- 230000001052 transient effect Effects 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 230000009105 vegetative growth Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 230000004580 weight loss Effects 0.000 description 2
- 239000004563 wettable powder Substances 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- LDVVMCZRFWMZSG-OLQVQODUSA-N (3ar,7as)-2-(trichloromethylsulfanyl)-3a,4,7,7a-tetrahydroisoindole-1,3-dione Chemical compound C1C=CC[C@H]2C(=O)N(SC(Cl)(Cl)Cl)C(=O)[C@H]21 LDVVMCZRFWMZSG-OLQVQODUSA-N 0.000 description 1
- RLMLFADXHJLPSQ-NPPFTVEMSA-N (3s,6s,9s,12s,15s,18s,21s,24r,27s)-3,6-dibenzyl-12,24-bis[(2r)-butan-2-yl]-15-(2-hydroxypropan-2-yl)-4,10,16,22-tetramethyl-18-(2-methylpropyl)-9,21-di(propan-2-yl)-13-oxa-1,4,7,10,16,19,22,25-octazabicyclo[25.3.0]triacontane-2,5,8,11,14,17,20,23,26-nonon Chemical compound C([C@H]1C(=O)N2CCC[C@H]2C(=O)N[C@@H](C(N(C)[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N(C)[C@H](C(=O)O[C@H](C(=O)N(C)[C@@H](C(C)C)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N1C)[C@H](C)CC)C(C)(C)O)=O)[C@H](C)CC)C1=CC=CC=C1 RLMLFADXHJLPSQ-NPPFTVEMSA-N 0.000 description 1
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- SQAINHDHICKHLX-UHFFFAOYSA-N 1-naphthaldehyde Chemical class C1=CC=C2C(C=O)=CC=CC2=C1 SQAINHDHICKHLX-UHFFFAOYSA-N 0.000 description 1
- IBXNCJKFFQIKKY-UHFFFAOYSA-N 1-pentyne Chemical compound CCCC#C IBXNCJKFFQIKKY-UHFFFAOYSA-N 0.000 description 1
- ZBMRKNMTMPPMMK-UHFFFAOYSA-N 2-amino-4-[hydroxy(methyl)phosphoryl]butanoic acid;azane Chemical compound [NH4+].CP(O)(=O)CCC(N)C([O-])=O ZBMRKNMTMPPMMK-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- 102100026105 3-ketoacyl-CoA thiolase, mitochondrial Human genes 0.000 description 1
- XDRVGXCIPIURSL-UHFFFAOYSA-N 5,8-diethyl-3,10-dimethyldodec-6-yne-5,8-diol Chemical compound CCC(C)CC(O)(CC)C#CC(O)(CC)CC(C)CC XDRVGXCIPIURSL-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- WFPZSXYXPSUOPY-ROYWQJLOSA-N ADP alpha-D-glucoside Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H]1O)O)N1C=2N=CN=C(C=2N=C1)N)OP(O)(=O)OP(O)(=O)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O WFPZSXYXPSUOPY-ROYWQJLOSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 108010003902 Acetyl-CoA C-acyltransferase Proteins 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 241000589156 Agrobacterium rhizogenes Species 0.000 description 1
- 241000743339 Agrostis Species 0.000 description 1
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 241000256579 Anuraphis Species 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 101100080292 Aspergillus oryzae (strain ATCC 42149 / RIB 40) pltp gene Proteins 0.000 description 1
- 231100000699 Bacterial toxin Toxicity 0.000 description 1
- KHBQMWCZKVMBLN-UHFFFAOYSA-N Benzenesulfonamide Chemical compound NS(=O)(=O)C1=CC=CC=C1 KHBQMWCZKVMBLN-UHFFFAOYSA-N 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 229920002799 BoPET Polymers 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000589174 Bradyrhizobium japonicum Species 0.000 description 1
- 101100520142 Caenorhabditis elegans pin-2 gene Proteins 0.000 description 1
- 241000282461 Canis lupus Species 0.000 description 1
- 239000005745 Captan Substances 0.000 description 1
- 239000005746 Carboxin Substances 0.000 description 1
- 108010022172 Chitinases Proteins 0.000 description 1
- 102000012286 Chitinases Human genes 0.000 description 1
- 241000737241 Cocos Species 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 241000532667 Conotrachelus Species 0.000 description 1
- 101710190853 Cruciferin Proteins 0.000 description 1
- 206010011732 Cyst Diseases 0.000 description 1
- 102000005927 Cysteine Proteases Human genes 0.000 description 1
- 108010005843 Cysteine Proteases Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 101710096830 DNA-3-methyladenine glycosylase Proteins 0.000 description 1
- 102100039128 DNA-3-methyladenine glycosylase Human genes 0.000 description 1
- 241000209210 Dactylis Species 0.000 description 1
- 241000489975 Diabrotica Species 0.000 description 1
- 240000003421 Dianthus chinensis Species 0.000 description 1
- 241000122105 Diatraea Species 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 229920005682 EO-PO block copolymer Polymers 0.000 description 1
- 101150111720 EPSPS gene Proteins 0.000 description 1
- 241001631033 Eanus Species 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 241000220485 Fabaceae Species 0.000 description 1
- 108010087894 Fatty acid desaturases Proteins 0.000 description 1
- 241000233732 Fusarium verticillioides Species 0.000 description 1
- 101150062467 GAT gene Proteins 0.000 description 1
- 101710186901 Globulin 1 Proteins 0.000 description 1
- 239000005562 Glyphosate Substances 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 101000742054 Homo sapiens Protein phosphatase 1D Proteins 0.000 description 1
- 241000526466 Homoeosoma Species 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010021929 Infertility male Diseases 0.000 description 1
- 240000007049 Juglans regia Species 0.000 description 1
- 235000009496 Juglans regia Nutrition 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 229920001732 Lignosulfonate Polymers 0.000 description 1
- 241000234280 Liliaceae Species 0.000 description 1
- 241000209082 Lolium Species 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 101150050813 MPI gene Proteins 0.000 description 1
- 208000007466 Male Infertility Diseases 0.000 description 1
- 241000243785 Meloidogyne javanica Species 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 101100190814 Mus musculus Pltp gene Proteins 0.000 description 1
- 239000005041 Mylar™ Substances 0.000 description 1
- 102000018463 Myo-Inositol-1-Phosphate Synthase Human genes 0.000 description 1
- 108091000020 Myo-Inositol-1-Phosphate Synthase Proteins 0.000 description 1
- 101710202365 Napin Proteins 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 241000238814 Orthoptera Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000209117 Panicum Species 0.000 description 1
- 235000006443 Panicum miliaceum subsp. miliaceum Nutrition 0.000 description 1
- 235000009037 Panicum miliaceum subsp. ruderale Nutrition 0.000 description 1
- 241000218222 Parasponia andersonii Species 0.000 description 1
- 101710096342 Pathogenesis-related protein Proteins 0.000 description 1
- 101000870887 Phaseolus vulgaris Glycine-rich cell wall structural protein 1.8 Proteins 0.000 description 1
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N Phosphinothricin Natural products CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 description 1
- 241001148062 Photorhabdus Species 0.000 description 1
- 241001148064 Photorhabdus luminescens Species 0.000 description 1
- 241000233645 Phytophthora nicotianae Species 0.000 description 1
- 239000005924 Pirimiphos-methyl Substances 0.000 description 1
- 229920000331 Polyhydroxybutyrate Polymers 0.000 description 1
- 102100038675 Protein phosphatase 1D Human genes 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 108010003581 Ribulose-bisphosphate carboxylase Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010016634 Seed Storage Proteins Proteins 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 241000208122 Solanum peruvianum Species 0.000 description 1
- 235000002558 Solanum peruvianum Nutrition 0.000 description 1
- 244000194806 Solanum sisymbriifolium Species 0.000 description 1
- 235000018724 Solanum sisymbriifolium Nutrition 0.000 description 1
- 108010039811 Starch synthase Proteins 0.000 description 1
- ULUAUXLGCMPNKK-UHFFFAOYSA-N Sulfobutanedioic acid Chemical class OC(=O)CC(C(O)=O)S(O)(=O)=O ULUAUXLGCMPNKK-UHFFFAOYSA-N 0.000 description 1
- 102000002933 Thioredoxin Human genes 0.000 description 1
- 239000005843 Thiram Substances 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 241000218234 Trema tomentosa Species 0.000 description 1
- 241000219870 Trifolium subterraneum Species 0.000 description 1
- 108091068141 Type II family Proteins 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 241000607757 Xenorhabdus Species 0.000 description 1
- 101001036768 Zea mays Glucose-1-phosphate adenylyltransferase large subunit 1, chloroplastic/amyloplastic Proteins 0.000 description 1
- 101001040871 Zea mays Glutelin-2 Proteins 0.000 description 1
- 101000662549 Zea mays Sucrose synthase 1 Proteins 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 108091000039 acetoacetyl-CoA reductase Proteins 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 239000012872 agrochemical composition Substances 0.000 description 1
- 230000009418 agronomic effect Effects 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 150000003973 alkyl amines Chemical class 0.000 description 1
- 150000008055 alkyl aryl sulfonates Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000006229 amino acid addition Effects 0.000 description 1
- 150000001449 anionic compounds Chemical class 0.000 description 1
- 230000002547 anomalous effect Effects 0.000 description 1
- 230000001887 anti-feedant effect Effects 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229940009098 aspartate Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 239000005667 attractant Substances 0.000 description 1
- 108010008887 aureobasidin A Proteins 0.000 description 1
- 230000000680 avirulence Effects 0.000 description 1
- 229940097012 bacillus thuringiensis Drugs 0.000 description 1
- 239000000688 bacterial toxin Substances 0.000 description 1
- 239000013602 bacteriophage vector Substances 0.000 description 1
- 101150103518 bar gene Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012455 bioassay technique Methods 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 229920000704 biodegradable plastic Polymers 0.000 description 1
- 230000007321 biological mechanism Effects 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 230000009172 bursting Effects 0.000 description 1
- JIJAYWGYIDJVJI-UHFFFAOYSA-N butyl naphthalene-1-sulfonate Chemical compound C1=CC=C2C(S(=O)(=O)OCCCC)=CC=CC2=C1 JIJAYWGYIDJVJI-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229940117949 captan Drugs 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- GYSSRZJIHXQEHQ-UHFFFAOYSA-N carboxin Chemical compound S1CCOC(C)=C1C(=O)NC1=CC=CC=C1 GYSSRZJIHXQEHQ-UHFFFAOYSA-N 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000010307 cell transformation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000031902 chemoattractant activity Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 108010071959 class III chitinase Proteins 0.000 description 1
- 210000002777 columnar cell Anatomy 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 239000007799 cork Substances 0.000 description 1
- 238000009402 cross-breeding Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 208000031513 cyst Diseases 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 239000002852 cysteine proteinase inhibitor Substances 0.000 description 1
- 239000004062 cytokinin Substances 0.000 description 1
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 150000005690 diesters Chemical class 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- KWABLUYIOFEZOY-UHFFFAOYSA-N dioctyl butanedioate Chemical compound CCCCCCCCOC(=O)CCC(=O)OCCCCCCCC KWABLUYIOFEZOY-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 230000000856 effect on pests Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000005712 elicitor Substances 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 239000004495 emulsifiable concentrate Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000008393 encapsulating agent Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 150000002194 fatty esters Chemical class 0.000 description 1
- YYJNOYZRYGDPNH-MFKUBSTISA-N fenpyroximate Chemical compound C=1C=C(C(=O)OC(C)(C)C)C=CC=1CO/N=C/C=1C(C)=NN(C)C=1OC1=CC=CC=C1 YYJNOYZRYGDPNH-MFKUBSTISA-N 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- SLGWESQGEUXWJQ-UHFFFAOYSA-N formaldehyde;phenol Chemical class O=C.OC1=CC=CC=C1 SLGWESQGEUXWJQ-UHFFFAOYSA-N 0.000 description 1
- 239000003008 fumonisin Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000010363 gene targeting Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- IAJOBQBIJHVGMQ-BYPYZUCNSA-N glufosinate-P Chemical compound CP(O)(=O)CC[C@H](N)C(O)=O IAJOBQBIJHVGMQ-BYPYZUCNSA-N 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 description 1
- 229940097068 glyphosate Drugs 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 231100000001 growth retardation Toxicity 0.000 description 1
- 210000004209 hair Anatomy 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 239000010903 husk Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 230000000974 larvacidal effect Effects 0.000 description 1
- 230000007653 larval development Effects 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 150000004668 long chain fatty acids Chemical class 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 125000005609 naphthenate group Chemical group 0.000 description 1
- 230000001069 nematicidal effect Effects 0.000 description 1
- 230000031787 nutrient reservoir activity Effects 0.000 description 1
- 229940049964 oleate Drugs 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229910052615 phyllosilicate Inorganic materials 0.000 description 1
- 230000009894 physiological stress Effects 0.000 description 1
- QHOQHJPRIBSPCY-UHFFFAOYSA-N pirimiphos-methyl Chemical group CCN(CC)C1=NC(C)=CC(OP(=S)(OC)OC)=N1 QHOQHJPRIBSPCY-UHFFFAOYSA-N 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000005648 plant growth regulator Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 239000005015 poly(hydroxybutyrate) Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 244000062645 predators Species 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 238000002708 random mutagenesis Methods 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000008399 response to wounding Effects 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 230000008117 seed development Effects 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 238000012868 site-directed mutagenesis technique Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- FDEKNCBLOMYBBB-KJEVSKRMSA-M sodium (E)-3-[2-[1-[[2-(5-bromopyrimidin-2-yl)-3-cyclopentyl-1-methylindole-6-carbonyl]amino]cyclobutyl]-1-methylbenzimidazol-5-yl]prop-2-enoate Chemical compound [Na+].Cn1c(c(C2CCCC2)c2ccc(cc12)C(=O)NC1(CCC1)c1nc2cc(\C=C\C([O-])=O)ccc2n1C)-c1ncc(Br)cn1 FDEKNCBLOMYBBB-KJEVSKRMSA-M 0.000 description 1
- 229940080236 sodium cetyl sulfate Drugs 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- GGHPAKFFUZUEKL-UHFFFAOYSA-M sodium;hexadecyl sulfate Chemical compound [Na+].CCCCCCCCCCCCCCCCOS([O-])(=O)=O GGHPAKFFUZUEKL-UHFFFAOYSA-M 0.000 description 1
- NWZBFJYXRGSRGD-UHFFFAOYSA-M sodium;octadecyl sulfate Chemical compound [Na+].CCCCCCCCCCCCCCCCCCOS([O-])(=O)=O NWZBFJYXRGSRGD-UHFFFAOYSA-M 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 108010048090 soybean lectin Proteins 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- HOWHQWFXSLOJEF-MGZLOUMQSA-N systemin Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H]2N(CCC2)C(=O)[C@H]2N(CCC2)C(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)C(C)C)CCC1 HOWHQWFXSLOJEF-MGZLOUMQSA-N 0.000 description 1
- 108010050014 systemin Proteins 0.000 description 1
- 235000021195 test diet Nutrition 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 108060008226 thioredoxin Proteins 0.000 description 1
- 229960002447 thiram Drugs 0.000 description 1
- KUAZQDVKQLNFPE-UHFFFAOYSA-N thiram Chemical compound CN(C)C(=S)SSC(=S)N(C)C KUAZQDVKQLNFPE-UHFFFAOYSA-N 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/32—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Bacillus (G)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
- C07K14/8121—Serpins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8286—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for insect resistance
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
Abstract
The present invention relates generally to a multi-faceted approach to the control of plant pathogens including plant pests. More particularly, the present invention relates to plants such as crop plants genetically modified to produce at least two pesticidal or pestistatic agents which in at least one agent is a serine proteinase inhibitor or precursor thereof from the solanaceaefamily and at least one other is a non-serine proteinase inhibitor slected from the list a Bt protein, a member of the CRY family, a VIP and a defensin, and wherein said exhibits resistance or reduced susceptibility to a pest.
Description
MANAGEMENT OF PLANT PATHOGENS
BACKGROUND OF THE INVENTION
FIELD OF THE INVENTION
The present invention relates generally to a multi-faceted approach to the control of plant patliogens including plant pests. More particularly, the present invention relates to plants such as crop plants genetically modified to produce at least two pesticidal or pestistatic agents which in combination provide the plant with enhanced resistance or reduced susceptibility to plant pests. The present invention fu.rther provides genetic constructs encoding the pesticidal or pestistatic agents and compositions comprising the agents. An integrated plant pest management strategy is also part of the present invention.
DESCRIPTION OF THE PRIOR ART
Bibliographic details of the publications referred to in this specification are also collected at the end of the description.
Reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that this prior art forms part of the common general knowledge in any country.
Plant pathogens including insect pests and fungi are a major contributor to losses in agricultural crops. For example, the European corn borer alone is estimated to cost the corn growing industry up to a billion dollars a year in the United States in lost production and control measures. One particularly important agricultural crop is cotton.
The cotton industry is a rapidly growing cash crop in many countries and in Australia, for example, approximately 200,000 bales of cotton are spun each year. Of these, 94% are sent for export. However, cotton plants are susceptible to a wide range of pests and the cost of controlling pest infestation in cotton in Australia alone is in the tens of millions of dollars.
BACKGROUND OF THE INVENTION
FIELD OF THE INVENTION
The present invention relates generally to a multi-faceted approach to the control of plant patliogens including plant pests. More particularly, the present invention relates to plants such as crop plants genetically modified to produce at least two pesticidal or pestistatic agents which in combination provide the plant with enhanced resistance or reduced susceptibility to plant pests. The present invention fu.rther provides genetic constructs encoding the pesticidal or pestistatic agents and compositions comprising the agents. An integrated plant pest management strategy is also part of the present invention.
DESCRIPTION OF THE PRIOR ART
Bibliographic details of the publications referred to in this specification are also collected at the end of the description.
Reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that this prior art forms part of the common general knowledge in any country.
Plant pathogens including insect pests and fungi are a major contributor to losses in agricultural crops. For example, the European corn borer alone is estimated to cost the corn growing industry up to a billion dollars a year in the United States in lost production and control measures. One particularly important agricultural crop is cotton.
The cotton industry is a rapidly growing cash crop in many countries and in Australia, for example, approximately 200,000 bales of cotton are spun each year. Of these, 94% are sent for export. However, cotton plants are susceptible to a wide range of pests and the cost of controlling pest infestation in cotton in Australia alone is in the tens of millions of dollars.
Traditionally, the primary metliod of controlling plant pests such as insects is the use of varying spectrum chemical insecticides and fungicides. However, for environmental, regulatory and health reasons, the use of chemicals on crops is being discouraged. In fact, many broad spectrum chemicals have been banned from use or their use severely limited to particular crops or in particular agricultural regions. There is an urgent need to develop alternative strategies to the control and management of crop pests including insects and fungi.
One approach is the use of more natural agents. One such agent is a serine proteinase inhibitor.
Several members of the families Solanaceae and Fabaceae accumulate serine proteinase inhibitors in their storage organs and in leaves in response to wounding (Brown and Ryan, Biochemistry 23:3418-3422, 1984; Richard Phytochemistry 16:159-169, 1977). The inhibitory activities of these proteins are directed against a wide range of proteinases of microbial and animal origin, but rarely against plant proteinases (Richardson, 1977 supra).
It is believed that these inhibitors are involved in protection of the plants against pathogens and predators. In potato tubers and legume seeds, the inhibitors can comprise 10% or more of the stored proteins (Richardson, 1977 supra), while in leaves of tomato and potato (Green and Ryan, Science 776-777, 1972), and alfalfa (Brown and Ryan, 1984 supra), proteinase inhibitors can accumulate to levels of 2% of the soluble protein within 48 hours of insect attack, or other types of wounding (Brown and Ryan, 1984 supra;
Graham et al, Plants 169:399-405, 1986). High levels of these inhibitors (up to 50% of total soluble protein) are also present in unripe fruits of the wild tomato, Lycopersicon peruvianum (Pearce et al, Planta 175:527-531, 1988).
There are two families of serine proteinase inhibitors in tomato and potato (Brown and Ryan, 1984 supra). Type I inhibitors are small proteins (monomer Mr 8100) which inhibit chymotrypsin at a single reactive site (Melville and Ryan, Archives of Biochemistry and Biophysics 138:700-702, 1970; Plunkett et al, Arch Biochem Biophys 213:463-472, 1982).
Inhibitors of the type II family generally contain two reactive sites, one of which inhibits chymotrypsin and the other trypsin (Bryant et al, Biochemistry 15:3418-3424, 1976;
Plunkett et al, 1982 supra). The type II inhibitors have a monomer Mr of 12,300 (Plunkett et al, 1982 supra). Proteinase inhibitor I accumulates in etiolated tobacco (Nicotiana tabacum) leaves (Kuo et al, Arch Biochem Biophys 230:504-510, 1984), and elicitors from Phytophthora parasitica var. nicotianae were found to induce proteinase inhibitor I
accumulation in tobacco cell suspension cultures (Rickauer et al, Plant Physiol 9:1065-1070, 1989).
The serine proteinase inhibitor precursor from tobacco, NaPI, has been particularly well characterized and used in pest control (See United States Patent Nos.
6,031,087, 6,440,727, 6,451,573 and 6,261,821 all of which are incorporated herein by reference).
Certain species of microorganisms are also known to possess pathogenocidal activity against a broad range of pests including Lepidoptera, Diptera, Coleptera, Hemiptera and others. Bacillus thuringiensis and Bacillus papilliae are two particularly useful biological control agents. Pesticidal activity is concentrated in parasporal crystalline protein inclusions although pesticidal proteins have also been isolated from the vegetative growth stage of Bacillus. Several genes encoding pesticidal proteins have been cloned and characterised (See United States Patent Nos. 5,366,982 and 5,840,868).
Corn and cotton plants have been genetically engineered to produce pesticidal endotoxins and crytoxins from B. thuringiensis (Aronson Cell Mol Life Sci 59(3):417-425, 2002; Schnepf et al, Microbiol Mol Biol Rev 62(3):775-806, 1998).
Despite the success of biological control agents, there is always a danger of resistance developing. There is also some variability associated with the cytotoxicity of a single agent.
There is a need to develop alternative strategies for pest control in plants.
One approach is the use of more natural agents. One such agent is a serine proteinase inhibitor.
Several members of the families Solanaceae and Fabaceae accumulate serine proteinase inhibitors in their storage organs and in leaves in response to wounding (Brown and Ryan, Biochemistry 23:3418-3422, 1984; Richard Phytochemistry 16:159-169, 1977). The inhibitory activities of these proteins are directed against a wide range of proteinases of microbial and animal origin, but rarely against plant proteinases (Richardson, 1977 supra).
It is believed that these inhibitors are involved in protection of the plants against pathogens and predators. In potato tubers and legume seeds, the inhibitors can comprise 10% or more of the stored proteins (Richardson, 1977 supra), while in leaves of tomato and potato (Green and Ryan, Science 776-777, 1972), and alfalfa (Brown and Ryan, 1984 supra), proteinase inhibitors can accumulate to levels of 2% of the soluble protein within 48 hours of insect attack, or other types of wounding (Brown and Ryan, 1984 supra;
Graham et al, Plants 169:399-405, 1986). High levels of these inhibitors (up to 50% of total soluble protein) are also present in unripe fruits of the wild tomato, Lycopersicon peruvianum (Pearce et al, Planta 175:527-531, 1988).
There are two families of serine proteinase inhibitors in tomato and potato (Brown and Ryan, 1984 supra). Type I inhibitors are small proteins (monomer Mr 8100) which inhibit chymotrypsin at a single reactive site (Melville and Ryan, Archives of Biochemistry and Biophysics 138:700-702, 1970; Plunkett et al, Arch Biochem Biophys 213:463-472, 1982).
Inhibitors of the type II family generally contain two reactive sites, one of which inhibits chymotrypsin and the other trypsin (Bryant et al, Biochemistry 15:3418-3424, 1976;
Plunkett et al, 1982 supra). The type II inhibitors have a monomer Mr of 12,300 (Plunkett et al, 1982 supra). Proteinase inhibitor I accumulates in etiolated tobacco (Nicotiana tabacum) leaves (Kuo et al, Arch Biochem Biophys 230:504-510, 1984), and elicitors from Phytophthora parasitica var. nicotianae were found to induce proteinase inhibitor I
accumulation in tobacco cell suspension cultures (Rickauer et al, Plant Physiol 9:1065-1070, 1989).
The serine proteinase inhibitor precursor from tobacco, NaPI, has been particularly well characterized and used in pest control (See United States Patent Nos.
6,031,087, 6,440,727, 6,451,573 and 6,261,821 all of which are incorporated herein by reference).
Certain species of microorganisms are also known to possess pathogenocidal activity against a broad range of pests including Lepidoptera, Diptera, Coleptera, Hemiptera and others. Bacillus thuringiensis and Bacillus papilliae are two particularly useful biological control agents. Pesticidal activity is concentrated in parasporal crystalline protein inclusions although pesticidal proteins have also been isolated from the vegetative growth stage of Bacillus. Several genes encoding pesticidal proteins have been cloned and characterised (See United States Patent Nos. 5,366,982 and 5,840,868).
Corn and cotton plants have been genetically engineered to produce pesticidal endotoxins and crytoxins from B. thuringiensis (Aronson Cell Mol Life Sci 59(3):417-425, 2002; Schnepf et al, Microbiol Mol Biol Rev 62(3):775-806, 1998).
Despite the success of biological control agents, there is always a danger of resistance developing. There is also some variability associated with the cytotoxicity of a single agent.
There is a need to develop alternative strategies for pest control in plants.
SUMMARY OF THE INVENTION
Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.
In accordance with the present invention, a multi-faceted approach is provided for pest control in plants, and in particular crop plants. In this approach, modules of pesticidal or pestistatic agents derived from biological sources are combined in plants or in compositions. At least one agent is a serine proteinase inhibitor or a precursor thereof, and at least one other agent acts in a manner different to a proteinase inhibitor.
Generally, at least one agent is a Bt protein such as a Cry family protein a Vegetative Insecticidal Protein (VIP) or is a defensin like molecule. In a preferred embodiment, the serine proteinase inliibitor is from the Solanaceae family such as but not limited to NaPI from tobacco. Genetic material encoding the pesticidal or pestistatic agents is generally introduced into a desired plant which is then rendered genetically capable of producing the pesticidal or pestistatic agents. In combination, the two or more pesticidal or pestistatic modules confer enhanced, increased, more stable, more efficacious, greater pesticidal or pestistatic activity, broader spectrum and/or longer lasting resistance or reduced susceptibility to pest infestation. By "pest" includes any plant pathogen or insect or any agent which carriers a serine proteinase. Examples of plant pests include fungi, yeasts, and insects. Insects groups particular contemplated by the present invention include such as Helicoverpa, Leptiderpa, Coleoptera, Diptera, Orthoptera, Diatraea and Spocloptera.
Accordingly, one aspect of the present invention provides a plant genetically modified to produce at least two pesticidal or pestistatic agents wherein at least one agent is a serine proteinase inhibitor or precursor thereof wherein in precursor form, comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one other monomer is a chymotrypsin inhibitor and wherein at least one other agent is not a serine proteinase inhibitor.
Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.
In accordance with the present invention, a multi-faceted approach is provided for pest control in plants, and in particular crop plants. In this approach, modules of pesticidal or pestistatic agents derived from biological sources are combined in plants or in compositions. At least one agent is a serine proteinase inhibitor or a precursor thereof, and at least one other agent acts in a manner different to a proteinase inhibitor.
Generally, at least one agent is a Bt protein such as a Cry family protein a Vegetative Insecticidal Protein (VIP) or is a defensin like molecule. In a preferred embodiment, the serine proteinase inliibitor is from the Solanaceae family such as but not limited to NaPI from tobacco. Genetic material encoding the pesticidal or pestistatic agents is generally introduced into a desired plant which is then rendered genetically capable of producing the pesticidal or pestistatic agents. In combination, the two or more pesticidal or pestistatic modules confer enhanced, increased, more stable, more efficacious, greater pesticidal or pestistatic activity, broader spectrum and/or longer lasting resistance or reduced susceptibility to pest infestation. By "pest" includes any plant pathogen or insect or any agent which carriers a serine proteinase. Examples of plant pests include fungi, yeasts, and insects. Insects groups particular contemplated by the present invention include such as Helicoverpa, Leptiderpa, Coleoptera, Diptera, Orthoptera, Diatraea and Spocloptera.
Accordingly, one aspect of the present invention provides a plant genetically modified to produce at least two pesticidal or pestistatic agents wherein at least one agent is a serine proteinase inhibitor or precursor thereof wherein in precursor form, comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one other monomer is a chymotrypsin inhibitor and wherein at least one other agent is not a serine proteinase inhibitor.
More particularly, the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest.
In a preferred embodiment, the serine proteinase inhibitor or precursor is from tobacco and is referred to as NaPI. In another preferred embodiment, the other agent is an endotoxin such as a Bt protein including a Cry family protein, Cry, VIP or is a defensin or a component thereof. Reference to a defensin includes its N-terminal portion, acidic tail and/or a central portion alone or fused to a heterologous protein.
Accordingly, another aspect of the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is the NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest.
Another aspect of the present invention contemplates a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest except and wherein at least one other agent is a Bt family protein such as a Cry family protein or defensin. Examples of Cry proteins include Cry9, Cry2, Cry3, CrylAc, VIP.
More particularly, the present invention is directed to a genetically modified plant or its progeny resulting from self crossing, back crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest except wherein the least one other agent is selected from the list comprising a Bt protein and a defensin.
Examples of Bt crystal proteins as described in Hofte and Whiteley Microbial Reviews 53:242-255, 1989.
The present invention further provides two or more genetic constructs encoding the at least two pesticidal or pestistatic agents or a single construct encoding the at least two agents.
In addition, compositions capable of being applied to crops, leaves, seeds, flowers or roots such as by spray, paint, powder or micronized vapour and comprising the at least two agents are contemplated by the present invention.
Parts, reproductive parts, fruits, seeds, flowers and harvested portions of crops genetically modified to produce the at least two pesticidal or pestistatic agents also form part of the present invention.
In a preferred embodiment, the serine proteinase inhibitor or precursor is from tobacco and is referred to as NaPI. In another preferred embodiment, the other agent is an endotoxin such as a Bt protein including a Cry family protein, Cry, VIP or is a defensin or a component thereof. Reference to a defensin includes its N-terminal portion, acidic tail and/or a central portion alone or fused to a heterologous protein.
Accordingly, another aspect of the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is the NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest.
Another aspect of the present invention contemplates a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest except and wherein at least one other agent is a Bt family protein such as a Cry family protein or defensin. Examples of Cry proteins include Cry9, Cry2, Cry3, CrylAc, VIP.
More particularly, the present invention is directed to a genetically modified plant or its progeny resulting from self crossing, back crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest except wherein the least one other agent is selected from the list comprising a Bt protein and a defensin.
Examples of Bt crystal proteins as described in Hofte and Whiteley Microbial Reviews 53:242-255, 1989.
The present invention further provides two or more genetic constructs encoding the at least two pesticidal or pestistatic agents or a single construct encoding the at least two agents.
In addition, compositions capable of being applied to crops, leaves, seeds, flowers or roots such as by spray, paint, powder or micronized vapour and comprising the at least two agents are contemplated by the present invention.
Parts, reproductive parts, fruits, seeds, flowers and harvested portions of crops genetically modified to produce the at least two pesticidal or pestistatic agents also form part of the present invention.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 is a graphical representation of a mass of larvae surviving on a diet of cotton leaf discs coated with CrylAc. The control leaf discs were from cv Coker; the test leaf discs were from cv Coker homozygous for the NaPI gene. (Note: Leaves were selected to give only a small reduction in weight of larvae.) Figure 2 is a graphical representation of a mass of larvae surviving on a diet of cotton leaf discs coated with CrylAc. The control leaf discs were from cv Coker; the test leaf discs were from cv Coker homozygous for the NaPI gene.
Figure 3 is a graphical representation of mortality of larvae after feeding on a diet of cotton leaves coated with CrylAc. The control leaf discs were from cv Coker;
the test leaf discs were from cv Coker expressing the NaPI gene.
Figure 4 is a graphical representation showing the average weight of larvae from day 11 to day 12 (approximately 20 hours). Error bars represent standard error of the mean.
Figure 5 is a graphical representation showing the mean increase in larval mass from day 11 to day 12. Error bars represent standard error of the mean.
Figure 1 is a graphical representation of a mass of larvae surviving on a diet of cotton leaf discs coated with CrylAc. The control leaf discs were from cv Coker; the test leaf discs were from cv Coker homozygous for the NaPI gene. (Note: Leaves were selected to give only a small reduction in weight of larvae.) Figure 2 is a graphical representation of a mass of larvae surviving on a diet of cotton leaf discs coated with CrylAc. The control leaf discs were from cv Coker; the test leaf discs were from cv Coker homozygous for the NaPI gene.
Figure 3 is a graphical representation of mortality of larvae after feeding on a diet of cotton leaves coated with CrylAc. The control leaf discs were from cv Coker;
the test leaf discs were from cv Coker expressing the NaPI gene.
Figure 4 is a graphical representation showing the average weight of larvae from day 11 to day 12 (approximately 20 hours). Error bars represent standard error of the mean.
Figure 5 is a graphical representation showing the mean increase in larval mass from day 11 to day 12. Error bars represent standard error of the mean.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a multi-faceted approach to pest control in plants, and in particular crop plants.
Reference to a "multi-faceted approach" means the use of two or more agents in combination to effect a pesticidal or pestistatic outcome on a targets pest or group of pests on commencement of or during its infestation or attempted infestation of a plant. The multi-faceted approach is modular in design in that each pesticidal or pestistatic agent is considered a modular component. Different modular components are used in the overall pest management system.
By "pestistatic" is meant the reduction in growth or reproduction or colonization ability of a pest without necessitating death of the pest. Whilst pesticidal activity is preferred, a pestistatic effect is acceptable and forms part of the present invention.
As used herein" pesticidal activity" includes "insecticidal activity" when the target pest is an insect or "fungicidal activity" when the target pest is a fungus or "bacteriocidal activity"
when the target pest is a bacterium or "nematodicidal activity" when the target pest is a nematode. These terms refer to the activity of an agent, also referred to as a "pesticidal modular component" against a pest. The activity is conveniently measured in terms of the effect on pest mortality, pest weight loss, pest repellency and other behavioural, physiological and/or physical changes of the pest including ability to digest protein, the ability to grow, maintain itself and/or reproduce after exposure to the agent.
Accordingly, the agent has an adverse effect on pest fitness as determined by at least one measurable parameter. In accordance with the present invention, at least two agents have an effect on pest fitness which may be additive or synergistic or otherwise complementary.
Accordingly, the at least two agents may exhibit the same effect on the measurable parameter of pest fitness or the combined effect may differ to the individual effects of each agent. In addition, at least one agent may exhibit pesticidal activity and another agent may exhibit pestistatic activity. Preferably, however, both agents or the combined effect of the agents is pesticidal.
The agents are proteinaceous in nature when they are produced by plant cells but may be chemically modified when incorporated into a pesticidal or pestistatic composition. The agents may, therefore, be peptides, polypeptides or proteins as described herein or chemical analogs or chemical mimetics or functional equivalents.
As indicated above, the agents exhibit an effect on pests which effect may be referred to as an "impact". Examples of impacts including changes in feeding ability or habits or capacity, ability to undergo development changes, cytotoxicity, growth retardation, reduction in reproductivity, effects on eggs or larvae, and mortality or morbidity of the pest. In addition, the efficacy of the pesticidal or pestistatic activity can be assess by the effect on the crop plant e.g. growth yeild, fewer pests, reduced pathology.
The subject agents are provided to or in a plant in a "pesticidal effective amount" or in a "pestistatic effective amount". These amounts connotes the quantity of agents, separately or in combination, to provide individual or overall pesticidal or pestistatic activity or other appropriate impact.
Reference herein to a plant includes a monocotyledonous plant or a dicotyledonous plant and the plant may be generated from genetically transformed callus or tissue or it may be a progeny of the transformed plant or a cross between the transformed plant or its progeny and a non-transformed plant. As used herein, the term plant includes plant cells, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps, and plant cells that are intact in plants or parts of plants such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tips, anthers, and the like.
Examples of plants of interest include, but are not limited to, corn (Zea mays), Brassica sp.
(e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceuni), foxtail millet (Setaria italica), finger millet (Eleusine coracana), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanuni tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassaya (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifef a), cotton pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers.
Vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C.
cantalupensis), and musk melon (C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis). Plants of the present invention include crop plants (for example, cotton, corn, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants.
The present invention provides a multi-faceted approach to pest control in plants, and in particular crop plants.
Reference to a "multi-faceted approach" means the use of two or more agents in combination to effect a pesticidal or pestistatic outcome on a targets pest or group of pests on commencement of or during its infestation or attempted infestation of a plant. The multi-faceted approach is modular in design in that each pesticidal or pestistatic agent is considered a modular component. Different modular components are used in the overall pest management system.
By "pestistatic" is meant the reduction in growth or reproduction or colonization ability of a pest without necessitating death of the pest. Whilst pesticidal activity is preferred, a pestistatic effect is acceptable and forms part of the present invention.
As used herein" pesticidal activity" includes "insecticidal activity" when the target pest is an insect or "fungicidal activity" when the target pest is a fungus or "bacteriocidal activity"
when the target pest is a bacterium or "nematodicidal activity" when the target pest is a nematode. These terms refer to the activity of an agent, also referred to as a "pesticidal modular component" against a pest. The activity is conveniently measured in terms of the effect on pest mortality, pest weight loss, pest repellency and other behavioural, physiological and/or physical changes of the pest including ability to digest protein, the ability to grow, maintain itself and/or reproduce after exposure to the agent.
Accordingly, the agent has an adverse effect on pest fitness as determined by at least one measurable parameter. In accordance with the present invention, at least two agents have an effect on pest fitness which may be additive or synergistic or otherwise complementary.
Accordingly, the at least two agents may exhibit the same effect on the measurable parameter of pest fitness or the combined effect may differ to the individual effects of each agent. In addition, at least one agent may exhibit pesticidal activity and another agent may exhibit pestistatic activity. Preferably, however, both agents or the combined effect of the agents is pesticidal.
The agents are proteinaceous in nature when they are produced by plant cells but may be chemically modified when incorporated into a pesticidal or pestistatic composition. The agents may, therefore, be peptides, polypeptides or proteins as described herein or chemical analogs or chemical mimetics or functional equivalents.
As indicated above, the agents exhibit an effect on pests which effect may be referred to as an "impact". Examples of impacts including changes in feeding ability or habits or capacity, ability to undergo development changes, cytotoxicity, growth retardation, reduction in reproductivity, effects on eggs or larvae, and mortality or morbidity of the pest. In addition, the efficacy of the pesticidal or pestistatic activity can be assess by the effect on the crop plant e.g. growth yeild, fewer pests, reduced pathology.
The subject agents are provided to or in a plant in a "pesticidal effective amount" or in a "pestistatic effective amount". These amounts connotes the quantity of agents, separately or in combination, to provide individual or overall pesticidal or pestistatic activity or other appropriate impact.
Reference herein to a plant includes a monocotyledonous plant or a dicotyledonous plant and the plant may be generated from genetically transformed callus or tissue or it may be a progeny of the transformed plant or a cross between the transformed plant or its progeny and a non-transformed plant. As used herein, the term plant includes plant cells, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps, and plant cells that are intact in plants or parts of plants such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tips, anthers, and the like.
Examples of plants of interest include, but are not limited to, corn (Zea mays), Brassica sp.
(e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceuni), foxtail millet (Setaria italica), finger millet (Eleusine coracana), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanuni tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassaya (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifef a), cotton pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers.
Vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C.
cantalupensis), and musk melon (C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis). Plants of the present invention include crop plants (for example, cotton, corn, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants.
Turfgrasses include, but are not limited to: annual bluegrass (Poa annua);
annual ryegrass (Lolium multiflorum); Canada bluegrass (Poa compressa); Chewings fescue (Festuca rubra); colonial bentgrass (Agrostis tenuis); creeping bentgrass (Agrostis palustris);
crested wheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyron cristatum);
hard fescue (Festuca longifolia); Kentucky bluegrass (Poa pratensis);
orchardgrass (Dactylis glonaerata); perennial ryegrass (Lolium per=enne); red fescue (Festuca rubra);
redtop (Agrostis alba); rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina);
smooth bromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy (Phleum pratense); velvet bentgrass (Agrostis, canina); weeping alkaligrass (Puccinellia distans);
western wheatgrass (Agropyron smithii); Bermuda grass (Cynodon spp.); St.
Augustine grass (Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass (Paspalum notatum); carpet grass (Axonopus affinis); centipede grass (Eremochloa ophiuroides);
kikuyu grass (Pennisetum clandesinum); seashore paspalum (Paspalum vaginatum);
blue gramma (Bouteloua gracilis); buffalo grass (Buchloe dactyloids); sideoats gramma (Bouteloua curtipendula).
Plants of interest include grain plants that provide seeds of interest, oil-seed plants, and leguminous plants. Seeds of interest include grain seeds, such as corn, wheat, barley, rice, sorghum, rye, millet, etc. Oil-seed plants include cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, flax, castor, olive etc. Leguminous plants include beans and peas. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils, chickpea, etc.
Preferred plants contemplated herein include cotton, sweet corn, tomato, tobacco, piniento, potato, sunflower, citrus, plums, sorghum, leeks, soybean, alfalfa, beans, pidgeon peas, chick peas, artichokes, curcurbits, lettuce, Dianthus, geraniums, cape gooseberry, maize, flax and linseed, lupins, broad beans, garden peas, peanuts, canola, snapdragons, cherry, sunflower, pot marigolds, Helichrysum, wheat, barley, oats, triticale, carrots, onions, orchids, roses and petunias.
annual ryegrass (Lolium multiflorum); Canada bluegrass (Poa compressa); Chewings fescue (Festuca rubra); colonial bentgrass (Agrostis tenuis); creeping bentgrass (Agrostis palustris);
crested wheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyron cristatum);
hard fescue (Festuca longifolia); Kentucky bluegrass (Poa pratensis);
orchardgrass (Dactylis glonaerata); perennial ryegrass (Lolium per=enne); red fescue (Festuca rubra);
redtop (Agrostis alba); rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina);
smooth bromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy (Phleum pratense); velvet bentgrass (Agrostis, canina); weeping alkaligrass (Puccinellia distans);
western wheatgrass (Agropyron smithii); Bermuda grass (Cynodon spp.); St.
Augustine grass (Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass (Paspalum notatum); carpet grass (Axonopus affinis); centipede grass (Eremochloa ophiuroides);
kikuyu grass (Pennisetum clandesinum); seashore paspalum (Paspalum vaginatum);
blue gramma (Bouteloua gracilis); buffalo grass (Buchloe dactyloids); sideoats gramma (Bouteloua curtipendula).
Plants of interest include grain plants that provide seeds of interest, oil-seed plants, and leguminous plants. Seeds of interest include grain seeds, such as corn, wheat, barley, rice, sorghum, rye, millet, etc. Oil-seed plants include cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, flax, castor, olive etc. Leguminous plants include beans and peas. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils, chickpea, etc.
Preferred plants contemplated herein include cotton, sweet corn, tomato, tobacco, piniento, potato, sunflower, citrus, plums, sorghum, leeks, soybean, alfalfa, beans, pidgeon peas, chick peas, artichokes, curcurbits, lettuce, Dianthus, geraniums, cape gooseberry, maize, flax and linseed, lupins, broad beans, garden peas, peanuts, canola, snapdragons, cherry, sunflower, pot marigolds, Helichrysum, wheat, barley, oats, triticale, carrots, onions, orchids, roses and petunias.
The embodiments of the present invention may be effective against a variety of pests. For purposes of the present invention, pests include, but are not limited to, insects, fungi, bacteria, nematodes, acarids, protozoan pathogens, animal-parasitic liver flukes, and the like. Pests of particular interest are insect pests, particularly insect pests that cause significant damage to agricultural plants. The term "insect pests" as used herein refers to insects and other similar pests such as, for example, those of the order Acari including, but not limited to, mites and ticks. Insect pests of the present invention include, but are not limited to, insects of the order Lepidoptera, e.g. Achoroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura sp., Cochylls hospes, Colias eurytheme, Corcyra cephalonica, Cydia latifers eanus, Cydia pomonella, Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphania hyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraea saccharalis, Ennomos subsignaria, Eoreuma loftini, Esphestia elutella, Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoellia ambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoa messoria, Galleria mellonella, Grapholita molesta, Harrisina americana, Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemileuca oliviae, Homoeosoma electelluna, Hyphantia cunea, Keiferia lycopersicella, Lambdina fi.scellaria fiscellaria, Lambdina fi`scellaria lugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis, Lymantria dispar, Macalla thyrisalis, Malacosoma sp., Mamestra brassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta, Maruca testulalis, Melanchrapicta, Operophtera brumata, Orgyia sp., Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes, Pectinophora gossypiella, Phryganidia califoNnica, Phyllonorycter blancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella, Pontia protodice, Pseudaletia unipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizura concinna, Sitotroga cerealella, Spilonta ocellana, Spodoptera sp., Thaurnstopoea pityocampa, Tinsola bisselliella, Trichoplusia hi, Udea rubigalis, Xylomyges curiails, and Yponomeuta padella.
Also, the embodiments of the present invention may be effective against insect pests, including but not limited to insects selected from the orders Diptera, Hyrnenopter=a, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, Coleoptera, etc., particularly Lepidoptera. Insect pests of the invention for the major crops include, but are not limited to: Maize: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Helicoverpa zeae, corn earworm; Spodoptera frugiperda, fall armyworni;
Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer;
Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; nortllern corn rootworm, e.g., Diabrotica longicornis barberi;
southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis rnaidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; AnaphothNips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecirnpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplusfemurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meronayza anaericana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wlieat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera ftugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafliopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Also, the embodiments of the present invention may be effective against insect pests, including but not limited to insects selected from the orders Diptera, Hyrnenopter=a, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, Coleoptera, etc., particularly Lepidoptera. Insect pests of the invention for the major crops include, but are not limited to: Maize: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Helicoverpa zeae, corn earworm; Spodoptera frugiperda, fall armyworni;
Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer;
Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; nortllern corn rootworm, e.g., Diabrotica longicornis barberi;
southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis rnaidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; AnaphothNips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecirnpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplusfemurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meronayza anaericana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wlieat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera ftugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafliopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternuna hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle.
Furthermore, embodiments of the present invention may be effective against Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae. Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbean weevil;
Bruchus pisorum, pea weevil; Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio; Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil; Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphaN, plum curculio;
Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus naajalis, European chafer; Macrodactylus subspinosus, rose chafer;
Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor, yellow mealworm.
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternuna hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle.
Furthermore, embodiments of the present invention may be effective against Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae. Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbean weevil;
Bruchus pisorum, pea weevil; Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio; Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil; Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphaN, plum curculio;
Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus naajalis, European chafer; Macrodactylus subspinosus, rose chafer;
Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor, yellow mealworm.
Nematodes include parasitic nematodes such as root-knot, cyst, and lesion nematodes, including Heterodera spp., Meloidogyne spp., and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes).
Lesion nematodes include Pratylenchus spp.
The present invention is directed to a pest management system for plants which is multi-faceted in the sense that it involves two or more pesticidal and/or pestistatic agents generally designed to exhibit a combined impact on plant pests. At least one agent is a serine proteinase inhibitor and least one agent is a non-serine proteinase inhibitor (i.e. it acts via a mechanism different to the inhibition of serine proteinase activity). The multi-faceted system may employ both agents simultaneously or sequentially. In either event, this is described as the agents operating in "combination".
Accordingly, one aspect of the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest.
The serine proteinase inhibitor is preferably from tobacco and may comprise in precursor form at least three monomers, or four monomers, or five monomers, or six monomers, or seven monomers, or eight monomers wherein at least one monomer inhibits trypsin and at least one monomer inhibits chymotrypsin.
Lesion nematodes include Pratylenchus spp.
The present invention is directed to a pest management system for plants which is multi-faceted in the sense that it involves two or more pesticidal and/or pestistatic agents generally designed to exhibit a combined impact on plant pests. At least one agent is a serine proteinase inhibitor and least one agent is a non-serine proteinase inhibitor (i.e. it acts via a mechanism different to the inhibition of serine proteinase activity). The multi-faceted system may employ both agents simultaneously or sequentially. In either event, this is described as the agents operating in "combination".
Accordingly, one aspect of the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest.
The serine proteinase inhibitor is preferably from tobacco and may comprise in precursor form at least three monomers, or four monomers, or five monomers, or six monomers, or seven monomers, or eight monomers wherein at least one monomer inhibits trypsin and at least one monomer inhibits chymotrypsin.
The term "precursor" as used herein is not intended to place any limitation on the utility of the precursor molecule itself or a requirement that the molecule first be processed into monomers before proteinase inhibitory activity is expressed. The precursor molecule has proteinase inhibitory activity and the present invention is directed to the precursor and to the individual monomers of the precursor.
In a most preferred embodiment, the proteinase inhibitor or its precursor is from Nicotiana alata (NaPI) as described in International Patent Application No.
PCT/AU93/00659 [WO
94/13 8 10] which is incorporated herein by reference.
Accordingly, another aspect of the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is the NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest.
The preferred at least one other agent is a toxin including an endotoxin such as derived from a microorganism or fungus. Particularly useful examples include toxins derived from Bacillus species. Reference to a toxin includes a peptide, polypeptide or protein exhibiting pesticidal or pestistatic activity but which does not inhibit solely a serine proteinase. It may inhibit a serine proteinase only if it also exhibits another mode of pesticidal or pestistatic activity. Particularly useful toxins are Bt-related toxins from B.
thuringiensis such as the Cry family of endotoxins. Examples of these endotxins include Cry9, Cry2s, Cry3s or CrylAc family of endotoxins.
The term "Cry9 family" is used herein to refer to nucleotide or amino acid sequences which share a high degree of sequence identity or similarity to previously described wild type Cry9 or Cry9D sequences. Bt toxin or endotoxin is intended to include the broader class of Cry toxins found in various status of B. thuringiensis and includes such toxins, as CrylAc, Cryls, Cry2s and Cry3s. Examples of suitable toxins can be found in US
Patent Application Nos. 2005/0138685, 2004/0016020 and 2004/0221333 which are each incorporated herein by reference.
Other suitable toxins include vegetative insecticidal protein (VIP) and a defensin. Suitable defensins are described in International Patent Application No.
which is incorporated herein by reference.
According, another aspect of the present invention contemplates a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest except and wherein at least one other agent is selected from the list comprising a Bt protein, a Cry9 family protein, a Cry2 family protein or Cry3 family protein, Cry1AC, a VIP and a defensin.
More particularly, the present invention is directed to a genetically modified plant or its progeny resulting from self crossing, back crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest except wherein the least one other agent is selected from the list comprising a Bt protein including a Cry family protein, a VIP and a defensin.
In a most preferred embodiment, the proteinase inhibitor or its precursor is from Nicotiana alata (NaPI) as described in International Patent Application No.
PCT/AU93/00659 [WO
94/13 8 10] which is incorporated herein by reference.
Accordingly, another aspect of the present invention provides a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is the NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest.
The preferred at least one other agent is a toxin including an endotoxin such as derived from a microorganism or fungus. Particularly useful examples include toxins derived from Bacillus species. Reference to a toxin includes a peptide, polypeptide or protein exhibiting pesticidal or pestistatic activity but which does not inhibit solely a serine proteinase. It may inhibit a serine proteinase only if it also exhibits another mode of pesticidal or pestistatic activity. Particularly useful toxins are Bt-related toxins from B.
thuringiensis such as the Cry family of endotoxins. Examples of these endotxins include Cry9, Cry2s, Cry3s or CrylAc family of endotoxins.
The term "Cry9 family" is used herein to refer to nucleotide or amino acid sequences which share a high degree of sequence identity or similarity to previously described wild type Cry9 or Cry9D sequences. Bt toxin or endotoxin is intended to include the broader class of Cry toxins found in various status of B. thuringiensis and includes such toxins, as CrylAc, Cryls, Cry2s and Cry3s. Examples of suitable toxins can be found in US
Patent Application Nos. 2005/0138685, 2004/0016020 and 2004/0221333 which are each incorporated herein by reference.
Other suitable toxins include vegetative insecticidal protein (VIP) and a defensin. Suitable defensins are described in International Patent Application No.
which is incorporated herein by reference.
According, another aspect of the present invention contemplates a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest except and wherein at least one other agent is selected from the list comprising a Bt protein, a Cry9 family protein, a Cry2 family protein or Cry3 family protein, Cry1AC, a VIP and a defensin.
More particularly, the present invention is directed to a genetically modified plant or its progeny resulting from self crossing, back crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest except wherein the least one other agent is selected from the list comprising a Bt protein including a Cry family protein, a VIP and a defensin.
Reference herein to NaPI, proteinase inhibitors, Bt, Cry9, CrylAc, Cryls, Cry2s, Cry3s, VIP and defensin and the like include derivatives and fragments and otherwise modified forms. Examples of modified forms include mutagenized forms such as mutants with altered substrate specificity or which exhibit greater stability or which direct the agent to a plant cell organelle such as a vacuole.
Serine Protease inhibitors such as NaPIs are known to act directly on the gut proteases of the digestive system of lepidopteran pests. A complex is formed between the specific inhibitor (either a trypsin or chymotrypsin inhibitor) and the protease (either a trypsin or chymotrypsin). These complexes are then excreted in the frass. The effect is to diminish the digestive capacity of the insect for protein and to deplete the protein reserves of the insect larvae.
Protease inhibitors may also affect the innate immunity system of insects. For this effect to occur, the inhibitors must move into the haemolymph where they can complex with the proteases in the cascades of the innate immunity system. Normally there is very restricted access of foreign molecules to this fundamental immune system of insects. An aspect of our invention is to provide conditions which allow enhanced access of PIs to the innate immunity system.
As indicated above, the present invention extends to parts of the genetically modified plants or,their progeny which include seeds, pollen, leaves, flowers, stigmas, stems, roots, root hairs, bark and apical meristems. Parts also include root stock or other form of commercially exploitable plant parts.
The present invention further provides nucleic acid molecules encoding the pesticidal or pestistatic agents. The nucleic acid molecules may be in a single construct, in multiple constructs or may be in situ in a plant cell genome. Conveniently, the nucleic acid molecules form a genetic composition. The nucleic acid molecules may also be part of beads such as beads used in biolistic DNA transfer.
Serine Protease inhibitors such as NaPIs are known to act directly on the gut proteases of the digestive system of lepidopteran pests. A complex is formed between the specific inhibitor (either a trypsin or chymotrypsin inhibitor) and the protease (either a trypsin or chymotrypsin). These complexes are then excreted in the frass. The effect is to diminish the digestive capacity of the insect for protein and to deplete the protein reserves of the insect larvae.
Protease inhibitors may also affect the innate immunity system of insects. For this effect to occur, the inhibitors must move into the haemolymph where they can complex with the proteases in the cascades of the innate immunity system. Normally there is very restricted access of foreign molecules to this fundamental immune system of insects. An aspect of our invention is to provide conditions which allow enhanced access of PIs to the innate immunity system.
As indicated above, the present invention extends to parts of the genetically modified plants or,their progeny which include seeds, pollen, leaves, flowers, stigmas, stems, roots, root hairs, bark and apical meristems. Parts also include root stock or other form of commercially exploitable plant parts.
The present invention further provides nucleic acid molecules encoding the pesticidal or pestistatic agents. The nucleic acid molecules may be in a single construct, in multiple constructs or may be in situ in a plant cell genome. Conveniently, the nucleic acid molecules form a genetic composition. The nucleic acid molecules may also be part of beads such as beads used in biolistic DNA transfer.
Accordingly, another aspect of the present invention provides a genetic composition comprising at least one construct encoding a serine proteinase inhibitor or a precursor form thereof wherein in precursor form it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one other monomer inhibits chymotrypsin and at least one construct encodes a non-serine proteinase inllibitor.
Generally, the nucleic acid molecules are operably linked to one or more promoters.
The promoter may regulate the expression of the nucleotide sequence encoding the agent, or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, or pathogens, or metal ions, amongst others.
Preferably, the promoter is capable of regulating expression of a nucleic acid molecule in a plant cell, tissue or organ, at least during the period of time over which the nucleotide sequence encoding the agent is expressed therein.
Plant-operable promoters are particularly preferred for use in the constructs of the present invention. Examples of suitable promoters include pCaMV 35S (Fang et al, Plant Cell 1:141-150, 1989), PGEL1 (Hajdukiewicz et al, Plant Mol Biol 25:989-994, 1994), class III
chitinase (Samac and Shah, Plant Cell 3:1063-1072, 1991), pin2 (Keil et al, EMBO J
8:1323-1330, 1989), PEP carboxylase (Pathirana et al, Plant J 12:293-304, 1997; MAP
kinase (Schoenbeck et al, Molec Plant-Microbe Interact, 1999), MSV (Legavre et al, In:
Vth International Congress of Plant Molecular Biology, Singapore, 1997), pltp (Hsu et al, Plant Sci 143:63-70, 1999), pmpi (Cordero et al, In: General Meeting of the International Program on Rice Biotechnology of the Rockefeller Foundation, Malacca, Malaysia, 1997) or glutamin synthase (Pujade-Renaud et al, Plant Physiol Biochem 35:85-93, 1997).
In the present context, the terms "in operable connection with" or "operably under the control" or similar shall be taken to indicate that expression of the nucleic acid molecule is under the control of the promoter sequence with which it is spatially connected; in a cell, tissue, organ or whole plant.
A number of promoters can be used in the practice of the invention. The promoters can be selected based on the desired outcome. The nucleic acids can be combined with constitutive, tissue-preferred, inducible, or other promoters for expression in the host organism. Suitable constitutive promoters for use in a plant host cell include, for example, the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO
99/43838 and U.S. Pat. No. 6,072,050; the core CaMV 35S promoter (Odell et al, Nature 313:810-812, 1985); rice actin McElroy et al, Plant Cell 2:163-171, 1990);
ubiquitin (Christensen et al, Plant Mol Biol 12:619-632, 1989 and Christensen et al, Plant Mol Biol 18:675-689, 1992); pEMU (Last et al, TheoN Appl Genet 81:581-588, 1991); MAS
(Velten et al, EMBO J 3:2723-2730, 1984); ALS promoter (U.S. Pat. No. 5,659,026), and the like.
Other constitutive promoters include, for example, those discussed in U.S.
Pat. Nos.
5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463;
5,608,142;
and 6,177,611.
Depending on the desired outcome, it may be beneficial to express the gene from an inducible promoter. Of particular interest for regulating the expression of the nucleotide sequences of the present invention in plants are wound-inducible promoters.
Such wound-inducible promoters, may respond to damage caused by insect feeding, and include potato proteinase inhibitor (pin II) gene (Ryan Ann Rev Phytopath 28:425-449, 1990;
Duan et al, Nature Biotechnology 14:494-498, 1996); wunl and wun2, U.S. Pat. No.
5,428,148; winl and win2 (Stanford et al, Mol Gen Genet 215:200-208, 1989); systemin (McGurl et al, Science 225:1570-1573, 1992); WIP1 (Rohmeier et al, Plant Mol Biol 22:783-792, 1993;
Eckelkamp et al, FEBS Letters 323:73-76, 1993); MPI gene (Corderok et al, Plant J
6(2):141-150, 1994); and the like, herein incorporated by reference.
Additionally, pathogen-inducible promoters may be employed in the methods and nucleotide constructs of the present invention. Such pathogen-inducible promoters include those from pathogenesis-related proteins (PR proteins), which are induced following infection by a pathogen; e.g., PR proteins, SAR proteins, beta-l,3-glucanase, chitinase, etc.
See, for example, Redolfi et al, Meth JPlant Pathol 89:245-254, 1983; Uknes et al, Plant Cell 4:645-656, 1992; and Van Loon Plant Mol Virol 4:111-116, 1985. See also WO
99/43819, herein incorporated by reference.
Of interest are promoters that are expressed locally at or near the site of pathogen infection. See, for example, Marineau et al, Plant Mol Biol 9:335-342, 1987;
Matton et al, Molecular Plant-Microbe Interactions 2:325-331, 1989; Somsisch et al, Proc Natl Acad Sci USA 83:2427-2430, 1986; Somsisch et al, Mol Gen Genet 2:93-98, 1988; and Yang Proc Natl Acad Sci USA 93:14972-14977, 1996. See also, Chen et al, Plant J
10:955-966, 1996; Zhang et al, Proc Natl Acad Sci USA 91:2507-2511, 1994; Warner et al, Plant J
3:191-201, 1993; Siebertz et al, Plant Cell 1:961-968, 1989; U.S. Pat. No.
5,750,386 (nematode-inducible); and the references cited therein. Of particular interest is the inducible promoter for the maize PRms gene, whose expression is induced by the pathogen Fusarium moniliforme (see, for example, Cordero et al, Physiol Mol Plant Path 41:189-200, 1992).
Chemical-regulated promoters can be used to modulate the expression of a gene in a plant through the application of an exogenous chemical regulator. Depending upon the objective, the promoter may be a chemical-inducible promoter, where application of the chemical induces gene expression, or a chemical-repressible promoter, where application of the chemical represses gene expression. Chemical-inducible promoters are known in the art and include, but are not limited to, the maize In2-2 promoter, which is activated by benzenesulfonamide herbicide safeners, the maize GST promoter, which is activated by hydrophobic electrophilic compounds that are used as pre-emergent herbicides, and the tobacco PR-la promoter, which is activated by salicylic acid. Other chemical-regulated promoters of interest include steroid-responsive promoters (see, for example, the glucocorticoid-inducible promoter in Schena et al, Proc Natl Acad Sci USA
88:10421-10425, 1991 and McNellis et al, Plant J 14(2):247-257, 1998 and tetracycline-inducible and tetracycline-repressible promoters (see, for example, Gatz et al, Mol Gen Genet 227:229-237, 1991, and U.S. Pat. Nos. 5,814,618 and 5,789,156), herein incorporated by reference.
Tissue-preferred promoters can be utilized to target enhanced pesticidal protein expression within a particular plant tissue. Tissue-preferred promoters include those discussed in Yamamoto et al, Plant J 12(2):255-265, 1997; Kawamata et al, Plant Cell Physiol 38(7):792-803, 1997; Hansen et al, Mol Gen Genet 254(3):337-343, 1997; Russell et al, Transgenic Res 6(2):157-168, 1997; Rinehart et al, Plant Physiol 112(3):1331-1341, 1996;
Van Camp et al, Plant Physiol 112(2):525-535, 1996; Canevascini et al, Plant Physiol 112(2):513-524, 1996; Yamamoto et al, Plant Cell Physiol 35(5):773-778, 1994;
Lam Results Probl Cell Differ 20:181-196, 1994; Orozco et al, Plant Mol Biol 23(6):1129-1138, 1993; Matsuoka et al, Proc Natl Acad Sci USA 90(20):9586-9590, 1993; and Guevara-Garcia et al, Plant J 4(3):495-505, 1993. Such promoters can be modified, if necessary, for weak expression.
Leaf-preferred promoters are known in the art. See, for example, Yamamoto et al, 1997 supra; Kwon et al, Plant Physiol 105:357-67, 1994; Yamamoto et al, 1994 supra;
Gotor et al, Plant J 3:509-18, 1993; Orozco et al, 1993 supra and Matsuoka et al, 1993 supra.
Root-preferred or root-specific promoters are known and can be selected from the many available from the literature or isolated de novo from various compatible species. See, for example, Hire et al, Plant Mol Biol 20(2):207-218, 1992 (soybean root-specific glutainine synthetase gene); Keller and Baumgartner Plant Cell 3(10):1051-1061, 1991 (root-specific control element in the GRP 1.8 gene of French bean); Sanger et al, Plant Mol Biol 14(3):433-443, 1990 (root-specific promoter of the mannopine synthase (MAS) gene of Agrobacterium tumefaciens); and Miao et al, Plant Cell 3(1):11-22, 1991 (full-length cDNA clone encoding cytosolic glutamine synthetase (GS), which is expressed in roots and root nodules of soybean). See also Bogusz et al, Plant Cell 2(7):633-641, 1990, where two root-specific promoters isolated from hemoglobin genes from the nitrogen-fixing nonlegume Parasponia andersonii and the related non-nitrogen-fixing nonlegume Trema tomentosa are described. The promoters of these genes were linked to a(3-glucuronidase reporter gene and introduced into both the nonlegume Nicotiana tabacum and the legume Lotus cof-niculatus, and in both instances root-specific promoter activity was preserved.
Promoters of ro1C and rolD root-inducing genes of Agrobacterium rhizogenes rnay also be used (see Limerick Plant Science 79(1):69-76). Teeri et al, EMBO J 8(2):343-350, 1989) describe gene fusion to lacZ to show that the Agrobacteriuna T-DNA gene encoding octopine synthase is especially active in the epidermis of the root tip and that the TR2' gene is root specific in the intact plant and stimulated by wounding in leaf tissue, an especially desirable combination of characteristics for use with an insecticidal or larvicidal gene. The TR1' gene fused to nptII (neomycin phosphotransferase II) showed similar characteristics. Additional root-preferred promoters include the VfENOD-GRP3 gene promoter (Kuster et al, Plant Mol Biol 29(4):759-772, 1995); and rolb promoter (Capana et al, Plant Mol Biol 25(4):681-691, 1994. See also U.S. Pat. Nos. 5,837,876;
5,750,386;
5,633,363; 5,459,252; 5,401,836; 5,110,732; and 5,023,179.
"Seed-preferred" promoters include both "seed-specific" promoters (those promoters active during seed development such as promoters of seed storage proteins) as well as "seed-germinating" promoters (those promoters active during seed germination). See Thompson et al, BioEssays 10:108, 1989, herein incorporated by reference. Such seed-preferred promoters include, but are not limited to, Ciml (cytokinin-induced message);
cZ19B1 (maize 19 kDa zein); and milps (myo-inositol-1-phosphate synthase) (see U.S.
Pat. No.
6,225,529, herein incorporated by reference). Gamma-zein and Glob-1 are endosperm-specific promoters. For dicots, seed-specific promoters include, but are not limited to, bean 0-phaseolin, napin, (3-conglycinin, soybean lectin, cruciferin, and the like.
For monocots, seed-specific promoters include, but are not limited to, maize 15 kDa zein, 22 kDa zein, 27 kDa zein, g-zein, waxy, shrunken 1, shrunken 2, globulin 1, etc. See also WO
00/12733, where seed-preferred promoters from endl and end2 genes are disclosed; herein incorporated by reference. A promoter that has "preferred" expression in a particular tissue is expressed in that tissue to a greater degree than in at least one other plant tissue. Some tissue-preferred promoters show expression almost exclusively in the particular tissue.
Where low level expression is desired, weak promoters will be used. Generally, the term "weak promoter" as used herein refers to a promoter that drives expression of a coding sequence at a low level. By low level expression at levels of about 1/1000 transcripts to about 1/100,000 transcripts to about 1/500,000 transcripts is intended.
Alternatively, it is recognized that the term "weak promoters" also encompasses promoters that drive expression in only a few cells and not in others to give a total low level of expression.
Where a promoter drives expression at unacceptably high levels, portions of the promoter sequence can be deleted or modified to decrease expression levels.
Such weak constitutive promoters include, for example the core promoter of the Rsyn7 promoter (WO 99/43838 and U.S. Pat. No. 6,072,050), the core 35S CaMV
promoter, and the like. Other constitutive promoters include, for example, those disclosed in U.S. Pat.
Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680;
5,268,463;
5,608,142; and 6,177,611; herein incorporated by reference.
The construct preferably contains additional regulatory elements for efficient transcription, for example, a transcription termination (or terminators) sequence.
The term "terminator" refers to a DNA sequence at the end of a transcriptional unit which signals termination of transcription. Terminators are 3'-non-translated DNA
sequences generally containing a polyadenylation signal, which facilitates the addition of polyadenylate sequences to the 3'-end of a primary transcript. Terminators active in plant cells are known and described in the literature. They may be isolated from bacteria, fungi, viruses, animals and/or plants or synthesized de novo.
As with promoter sequences, the terminator may be any termination sequence which is operable in the cells, tissues or organs in which it is intended to be used.
Examples of terminators particularly suitable for use in the synthetic genes of the present invention include the SV40 polyadenylation signal, the HSV TK polyadenylation signal, the CYCl terminator, ADH terminator, SPA terminator, nopaline synthase (NOS) gene terminator of Agrobacterium tumefaciens, the terminator of the cauliflower mosaic virus (CaMV) 35S gene, the zein gene terminator from Zea mays, the Rubisco small subunit gene (SSU) gene terminator sequences, subclover stunt virus (SCSV) gene sequence terminators, any rho-independent E. coli terminator, or the lacZ alpha terminator, amongst others.
In a particularly preferred embodiment, the terminator is the SV40 polyadenylation signal or the HSV TK polyadenylation signal which are operable in animal cells, tissues and organs, octopine synthase (OCS) or nopaline synthase (NOS) terminator active in plant cells, tissue or organs, or the lacZ alpha terminator which is active in prokaryotic cells.
Those skilled in the art will be aware of additional terminator sequences which may be suitable for use in performing the subject invention. Such sequences may readily be used without any undue experimentation.
Means for introducing (i.e. transfecting or transforming) cells with the constructs are well-known to those skilled in the art.
The constructs described supra are capable of being modified further, for example, by the inclusion of marker nucleotide sequences encoding a detectable marker enzyme or a functional analogue or derivative thereof, to facilitate detection of the synthetic gene in a cell, tissue or organ in which it is expressed. According to this embodiment, the marker nucleotide sequences will be present in a translatable format and be expressed. In addition, transport sequences may be included to direct one or more agents to particular plant organnelles.
Those skilled in the art will be aware of how to produce the constructs described herein and of the requirements for obtaining the expression thereof, when so desired, in a specific cell or cell-type under the conditions desired. In particular, it will be known to those skilled in the art that the genetic manipulations required to perform the present invention may require the propagation of a genetic construct described herein or a derivative thereof in a prokaryotic cell such as an E. coli cell or a plant cell or an animal cell.
Generally, the nucleic acid molecules are operably linked to one or more promoters.
The promoter may regulate the expression of the nucleotide sequence encoding the agent, or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, or pathogens, or metal ions, amongst others.
Preferably, the promoter is capable of regulating expression of a nucleic acid molecule in a plant cell, tissue or organ, at least during the period of time over which the nucleotide sequence encoding the agent is expressed therein.
Plant-operable promoters are particularly preferred for use in the constructs of the present invention. Examples of suitable promoters include pCaMV 35S (Fang et al, Plant Cell 1:141-150, 1989), PGEL1 (Hajdukiewicz et al, Plant Mol Biol 25:989-994, 1994), class III
chitinase (Samac and Shah, Plant Cell 3:1063-1072, 1991), pin2 (Keil et al, EMBO J
8:1323-1330, 1989), PEP carboxylase (Pathirana et al, Plant J 12:293-304, 1997; MAP
kinase (Schoenbeck et al, Molec Plant-Microbe Interact, 1999), MSV (Legavre et al, In:
Vth International Congress of Plant Molecular Biology, Singapore, 1997), pltp (Hsu et al, Plant Sci 143:63-70, 1999), pmpi (Cordero et al, In: General Meeting of the International Program on Rice Biotechnology of the Rockefeller Foundation, Malacca, Malaysia, 1997) or glutamin synthase (Pujade-Renaud et al, Plant Physiol Biochem 35:85-93, 1997).
In the present context, the terms "in operable connection with" or "operably under the control" or similar shall be taken to indicate that expression of the nucleic acid molecule is under the control of the promoter sequence with which it is spatially connected; in a cell, tissue, organ or whole plant.
A number of promoters can be used in the practice of the invention. The promoters can be selected based on the desired outcome. The nucleic acids can be combined with constitutive, tissue-preferred, inducible, or other promoters for expression in the host organism. Suitable constitutive promoters for use in a plant host cell include, for example, the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO
99/43838 and U.S. Pat. No. 6,072,050; the core CaMV 35S promoter (Odell et al, Nature 313:810-812, 1985); rice actin McElroy et al, Plant Cell 2:163-171, 1990);
ubiquitin (Christensen et al, Plant Mol Biol 12:619-632, 1989 and Christensen et al, Plant Mol Biol 18:675-689, 1992); pEMU (Last et al, TheoN Appl Genet 81:581-588, 1991); MAS
(Velten et al, EMBO J 3:2723-2730, 1984); ALS promoter (U.S. Pat. No. 5,659,026), and the like.
Other constitutive promoters include, for example, those discussed in U.S.
Pat. Nos.
5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463;
5,608,142;
and 6,177,611.
Depending on the desired outcome, it may be beneficial to express the gene from an inducible promoter. Of particular interest for regulating the expression of the nucleotide sequences of the present invention in plants are wound-inducible promoters.
Such wound-inducible promoters, may respond to damage caused by insect feeding, and include potato proteinase inhibitor (pin II) gene (Ryan Ann Rev Phytopath 28:425-449, 1990;
Duan et al, Nature Biotechnology 14:494-498, 1996); wunl and wun2, U.S. Pat. No.
5,428,148; winl and win2 (Stanford et al, Mol Gen Genet 215:200-208, 1989); systemin (McGurl et al, Science 225:1570-1573, 1992); WIP1 (Rohmeier et al, Plant Mol Biol 22:783-792, 1993;
Eckelkamp et al, FEBS Letters 323:73-76, 1993); MPI gene (Corderok et al, Plant J
6(2):141-150, 1994); and the like, herein incorporated by reference.
Additionally, pathogen-inducible promoters may be employed in the methods and nucleotide constructs of the present invention. Such pathogen-inducible promoters include those from pathogenesis-related proteins (PR proteins), which are induced following infection by a pathogen; e.g., PR proteins, SAR proteins, beta-l,3-glucanase, chitinase, etc.
See, for example, Redolfi et al, Meth JPlant Pathol 89:245-254, 1983; Uknes et al, Plant Cell 4:645-656, 1992; and Van Loon Plant Mol Virol 4:111-116, 1985. See also WO
99/43819, herein incorporated by reference.
Of interest are promoters that are expressed locally at or near the site of pathogen infection. See, for example, Marineau et al, Plant Mol Biol 9:335-342, 1987;
Matton et al, Molecular Plant-Microbe Interactions 2:325-331, 1989; Somsisch et al, Proc Natl Acad Sci USA 83:2427-2430, 1986; Somsisch et al, Mol Gen Genet 2:93-98, 1988; and Yang Proc Natl Acad Sci USA 93:14972-14977, 1996. See also, Chen et al, Plant J
10:955-966, 1996; Zhang et al, Proc Natl Acad Sci USA 91:2507-2511, 1994; Warner et al, Plant J
3:191-201, 1993; Siebertz et al, Plant Cell 1:961-968, 1989; U.S. Pat. No.
5,750,386 (nematode-inducible); and the references cited therein. Of particular interest is the inducible promoter for the maize PRms gene, whose expression is induced by the pathogen Fusarium moniliforme (see, for example, Cordero et al, Physiol Mol Plant Path 41:189-200, 1992).
Chemical-regulated promoters can be used to modulate the expression of a gene in a plant through the application of an exogenous chemical regulator. Depending upon the objective, the promoter may be a chemical-inducible promoter, where application of the chemical induces gene expression, or a chemical-repressible promoter, where application of the chemical represses gene expression. Chemical-inducible promoters are known in the art and include, but are not limited to, the maize In2-2 promoter, which is activated by benzenesulfonamide herbicide safeners, the maize GST promoter, which is activated by hydrophobic electrophilic compounds that are used as pre-emergent herbicides, and the tobacco PR-la promoter, which is activated by salicylic acid. Other chemical-regulated promoters of interest include steroid-responsive promoters (see, for example, the glucocorticoid-inducible promoter in Schena et al, Proc Natl Acad Sci USA
88:10421-10425, 1991 and McNellis et al, Plant J 14(2):247-257, 1998 and tetracycline-inducible and tetracycline-repressible promoters (see, for example, Gatz et al, Mol Gen Genet 227:229-237, 1991, and U.S. Pat. Nos. 5,814,618 and 5,789,156), herein incorporated by reference.
Tissue-preferred promoters can be utilized to target enhanced pesticidal protein expression within a particular plant tissue. Tissue-preferred promoters include those discussed in Yamamoto et al, Plant J 12(2):255-265, 1997; Kawamata et al, Plant Cell Physiol 38(7):792-803, 1997; Hansen et al, Mol Gen Genet 254(3):337-343, 1997; Russell et al, Transgenic Res 6(2):157-168, 1997; Rinehart et al, Plant Physiol 112(3):1331-1341, 1996;
Van Camp et al, Plant Physiol 112(2):525-535, 1996; Canevascini et al, Plant Physiol 112(2):513-524, 1996; Yamamoto et al, Plant Cell Physiol 35(5):773-778, 1994;
Lam Results Probl Cell Differ 20:181-196, 1994; Orozco et al, Plant Mol Biol 23(6):1129-1138, 1993; Matsuoka et al, Proc Natl Acad Sci USA 90(20):9586-9590, 1993; and Guevara-Garcia et al, Plant J 4(3):495-505, 1993. Such promoters can be modified, if necessary, for weak expression.
Leaf-preferred promoters are known in the art. See, for example, Yamamoto et al, 1997 supra; Kwon et al, Plant Physiol 105:357-67, 1994; Yamamoto et al, 1994 supra;
Gotor et al, Plant J 3:509-18, 1993; Orozco et al, 1993 supra and Matsuoka et al, 1993 supra.
Root-preferred or root-specific promoters are known and can be selected from the many available from the literature or isolated de novo from various compatible species. See, for example, Hire et al, Plant Mol Biol 20(2):207-218, 1992 (soybean root-specific glutainine synthetase gene); Keller and Baumgartner Plant Cell 3(10):1051-1061, 1991 (root-specific control element in the GRP 1.8 gene of French bean); Sanger et al, Plant Mol Biol 14(3):433-443, 1990 (root-specific promoter of the mannopine synthase (MAS) gene of Agrobacterium tumefaciens); and Miao et al, Plant Cell 3(1):11-22, 1991 (full-length cDNA clone encoding cytosolic glutamine synthetase (GS), which is expressed in roots and root nodules of soybean). See also Bogusz et al, Plant Cell 2(7):633-641, 1990, where two root-specific promoters isolated from hemoglobin genes from the nitrogen-fixing nonlegume Parasponia andersonii and the related non-nitrogen-fixing nonlegume Trema tomentosa are described. The promoters of these genes were linked to a(3-glucuronidase reporter gene and introduced into both the nonlegume Nicotiana tabacum and the legume Lotus cof-niculatus, and in both instances root-specific promoter activity was preserved.
Promoters of ro1C and rolD root-inducing genes of Agrobacterium rhizogenes rnay also be used (see Limerick Plant Science 79(1):69-76). Teeri et al, EMBO J 8(2):343-350, 1989) describe gene fusion to lacZ to show that the Agrobacteriuna T-DNA gene encoding octopine synthase is especially active in the epidermis of the root tip and that the TR2' gene is root specific in the intact plant and stimulated by wounding in leaf tissue, an especially desirable combination of characteristics for use with an insecticidal or larvicidal gene. The TR1' gene fused to nptII (neomycin phosphotransferase II) showed similar characteristics. Additional root-preferred promoters include the VfENOD-GRP3 gene promoter (Kuster et al, Plant Mol Biol 29(4):759-772, 1995); and rolb promoter (Capana et al, Plant Mol Biol 25(4):681-691, 1994. See also U.S. Pat. Nos. 5,837,876;
5,750,386;
5,633,363; 5,459,252; 5,401,836; 5,110,732; and 5,023,179.
"Seed-preferred" promoters include both "seed-specific" promoters (those promoters active during seed development such as promoters of seed storage proteins) as well as "seed-germinating" promoters (those promoters active during seed germination). See Thompson et al, BioEssays 10:108, 1989, herein incorporated by reference. Such seed-preferred promoters include, but are not limited to, Ciml (cytokinin-induced message);
cZ19B1 (maize 19 kDa zein); and milps (myo-inositol-1-phosphate synthase) (see U.S.
Pat. No.
6,225,529, herein incorporated by reference). Gamma-zein and Glob-1 are endosperm-specific promoters. For dicots, seed-specific promoters include, but are not limited to, bean 0-phaseolin, napin, (3-conglycinin, soybean lectin, cruciferin, and the like.
For monocots, seed-specific promoters include, but are not limited to, maize 15 kDa zein, 22 kDa zein, 27 kDa zein, g-zein, waxy, shrunken 1, shrunken 2, globulin 1, etc. See also WO
00/12733, where seed-preferred promoters from endl and end2 genes are disclosed; herein incorporated by reference. A promoter that has "preferred" expression in a particular tissue is expressed in that tissue to a greater degree than in at least one other plant tissue. Some tissue-preferred promoters show expression almost exclusively in the particular tissue.
Where low level expression is desired, weak promoters will be used. Generally, the term "weak promoter" as used herein refers to a promoter that drives expression of a coding sequence at a low level. By low level expression at levels of about 1/1000 transcripts to about 1/100,000 transcripts to about 1/500,000 transcripts is intended.
Alternatively, it is recognized that the term "weak promoters" also encompasses promoters that drive expression in only a few cells and not in others to give a total low level of expression.
Where a promoter drives expression at unacceptably high levels, portions of the promoter sequence can be deleted or modified to decrease expression levels.
Such weak constitutive promoters include, for example the core promoter of the Rsyn7 promoter (WO 99/43838 and U.S. Pat. No. 6,072,050), the core 35S CaMV
promoter, and the like. Other constitutive promoters include, for example, those disclosed in U.S. Pat.
Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680;
5,268,463;
5,608,142; and 6,177,611; herein incorporated by reference.
The construct preferably contains additional regulatory elements for efficient transcription, for example, a transcription termination (or terminators) sequence.
The term "terminator" refers to a DNA sequence at the end of a transcriptional unit which signals termination of transcription. Terminators are 3'-non-translated DNA
sequences generally containing a polyadenylation signal, which facilitates the addition of polyadenylate sequences to the 3'-end of a primary transcript. Terminators active in plant cells are known and described in the literature. They may be isolated from bacteria, fungi, viruses, animals and/or plants or synthesized de novo.
As with promoter sequences, the terminator may be any termination sequence which is operable in the cells, tissues or organs in which it is intended to be used.
Examples of terminators particularly suitable for use in the synthetic genes of the present invention include the SV40 polyadenylation signal, the HSV TK polyadenylation signal, the CYCl terminator, ADH terminator, SPA terminator, nopaline synthase (NOS) gene terminator of Agrobacterium tumefaciens, the terminator of the cauliflower mosaic virus (CaMV) 35S gene, the zein gene terminator from Zea mays, the Rubisco small subunit gene (SSU) gene terminator sequences, subclover stunt virus (SCSV) gene sequence terminators, any rho-independent E. coli terminator, or the lacZ alpha terminator, amongst others.
In a particularly preferred embodiment, the terminator is the SV40 polyadenylation signal or the HSV TK polyadenylation signal which are operable in animal cells, tissues and organs, octopine synthase (OCS) or nopaline synthase (NOS) terminator active in plant cells, tissue or organs, or the lacZ alpha terminator which is active in prokaryotic cells.
Those skilled in the art will be aware of additional terminator sequences which may be suitable for use in performing the subject invention. Such sequences may readily be used without any undue experimentation.
Means for introducing (i.e. transfecting or transforming) cells with the constructs are well-known to those skilled in the art.
The constructs described supra are capable of being modified further, for example, by the inclusion of marker nucleotide sequences encoding a detectable marker enzyme or a functional analogue or derivative thereof, to facilitate detection of the synthetic gene in a cell, tissue or organ in which it is expressed. According to this embodiment, the marker nucleotide sequences will be present in a translatable format and be expressed. In addition, transport sequences may be included to direct one or more agents to particular plant organnelles.
Those skilled in the art will be aware of how to produce the constructs described herein and of the requirements for obtaining the expression thereof, when so desired, in a specific cell or cell-type under the conditions desired. In particular, it will be known to those skilled in the art that the genetic manipulations required to perform the present invention may require the propagation of a genetic construct described herein or a derivative thereof in a prokaryotic cell such as an E. coli cell or a plant cell or an animal cell.
The constructs of the present invention may be introduced to a suitable cell, tissue or organ without modification as linear DNA, optionally contained within a suitable carrier, such as a cell, virus particle or liposome, amongst others. To produce a genetic construct, a nucleic acid is inserted into a suitable vector or episome molecule, such as a bacteriophage vector, viral vector or a plasmid, cosmid or artificial chromosome vector which is capable of being maintained and/or replicated and/or expressed in the host cell, tissue or organ into which it is subsequently introduced.
Accordingly, a further aspect of the present invention provides a genetic construct which at least comprises a genetic element encoding one or both pesticidal or pestistatic agents as herein described and one or more origins of replication and/or selectable marker gene sequences.
Usually, an origin of replication or a selectable marker gene suitable for use in bacteria is physically-separated from those genetic sequences contained in the genetic construct which are intended to be expressed or transferred to a plant cell, or integrated into the genome of a plant cell.
As used herein, the term "selectable marker gene" includes any gene which confers a phenotype on a cell on which it is expressed to facilitate the identification and/or selection of cells which are transfected or transformed with a genetic construct of the invention or a derivative thereof.
Suitable selectable marker genes contemplated herein include the ampicillin-resistance gene (Amp'), tetracycline-resistance gene (Tc% bacterial kanamycin-resistance gene (Kan'), the zeocin resistance gene (Zeocin is a drug of the bleomycin family which is trade mark of InVitrogen Corporation), the AURI-C gene which confers resistance to the antibiotic aureobasidin A, phosphinothricin-resistance gene, neomycin phosphotransferase gen (nptII), hygromycin-resistance gene, (3-glucuronidase (GUS) gene, chloramphenicol acetyltransferase (CAT) gene, green fluorescent protein-encoding gene or the luciferase gene, amongst others.
Accordingly, a further aspect of the present invention provides a genetic construct which at least comprises a genetic element encoding one or both pesticidal or pestistatic agents as herein described and one or more origins of replication and/or selectable marker gene sequences.
Usually, an origin of replication or a selectable marker gene suitable for use in bacteria is physically-separated from those genetic sequences contained in the genetic construct which are intended to be expressed or transferred to a plant cell, or integrated into the genome of a plant cell.
As used herein, the term "selectable marker gene" includes any gene which confers a phenotype on a cell on which it is expressed to facilitate the identification and/or selection of cells which are transfected or transformed with a genetic construct of the invention or a derivative thereof.
Suitable selectable marker genes contemplated herein include the ampicillin-resistance gene (Amp'), tetracycline-resistance gene (Tc% bacterial kanamycin-resistance gene (Kan'), the zeocin resistance gene (Zeocin is a drug of the bleomycin family which is trade mark of InVitrogen Corporation), the AURI-C gene which confers resistance to the antibiotic aureobasidin A, phosphinothricin-resistance gene, neomycin phosphotransferase gen (nptII), hygromycin-resistance gene, (3-glucuronidase (GUS) gene, chloramphenicol acetyltransferase (CAT) gene, green fluorescent protein-encoding gene or the luciferase gene, amongst others.
Preferably, the selectable marker gene is the nptII gene or Kan' gene or green fluorescent protein (GFP)-encoding gene.
Those skilled in the art will be aware of other selectable marker genes useful in the performance of the present invention and the subject invention is not limited by the nature of the selectable marker gene.
The present invention extends to all genetic constructs essentially as described herein, which include further genetic sequences intended for the maintenance and/or replication of said genetic construct in prokaryotes or eukaryotes and/or the integration of said genetic construct or a part thereof into the genome of a eukaryotic cell or organism.
Standard methods may be used to introduce the constructs into the cell, tissue or organ, for example, liposome-mediated transfection or transformation, transformation of cells with attenuated virus particles or bacterial cells, cell mating, transformation or transfection procedures known to those skilled in the art.
Additional means for introducing recombinant DNA into plant tissue or cells include, but are not limited to, transformation using CaC12 and variations thereof, direct DNA uptake into protoplasts, PEG-mediated uptake to protoplasts, microparticle bombardment, electroporation, microinjection of DNA, microparticle bombardment of tissue explant or cells, vacuum-infiltration of tissue with nucleic acid, or in the case of plants, T-DNA-mediated transfer from 4grobactef ium to the plant tissue.
For microparticle bombardment of cells, a microparticle is propelled into a cell to produce a transformed cell. Any suitable ballistic cell transformation methodology and apparatus can be used in performing the present invention. Exemplary apparatus and procedures are disclosed by Stomp et al, (U.S. Patent No. 5,122,466) and Sanford and Wolf (U.S. Patent No. 4,945,050). When using ballistic transformation procedures, the genetic construct may incorporate a plasmid capable of replicating in the cell to be transformed.
Those skilled in the art will be aware of other selectable marker genes useful in the performance of the present invention and the subject invention is not limited by the nature of the selectable marker gene.
The present invention extends to all genetic constructs essentially as described herein, which include further genetic sequences intended for the maintenance and/or replication of said genetic construct in prokaryotes or eukaryotes and/or the integration of said genetic construct or a part thereof into the genome of a eukaryotic cell or organism.
Standard methods may be used to introduce the constructs into the cell, tissue or organ, for example, liposome-mediated transfection or transformation, transformation of cells with attenuated virus particles or bacterial cells, cell mating, transformation or transfection procedures known to those skilled in the art.
Additional means for introducing recombinant DNA into plant tissue or cells include, but are not limited to, transformation using CaC12 and variations thereof, direct DNA uptake into protoplasts, PEG-mediated uptake to protoplasts, microparticle bombardment, electroporation, microinjection of DNA, microparticle bombardment of tissue explant or cells, vacuum-infiltration of tissue with nucleic acid, or in the case of plants, T-DNA-mediated transfer from 4grobactef ium to the plant tissue.
For microparticle bombardment of cells, a microparticle is propelled into a cell to produce a transformed cell. Any suitable ballistic cell transformation methodology and apparatus can be used in performing the present invention. Exemplary apparatus and procedures are disclosed by Stomp et al, (U.S. Patent No. 5,122,466) and Sanford and Wolf (U.S. Patent No. 4,945,050). When using ballistic transformation procedures, the genetic construct may incorporate a plasmid capable of replicating in the cell to be transformed.
Examples of microparticles suitable for use in such systems include 1 to 5 m gold spheres. The DNA construct may be deposited on the microparticle by any suitable technique, such as by precipitation.
The methods of the invention involve introducing a polypeptide or polynucleotide into a plant. "Introducing" is intended to mean presenting to the plant the polynucleotide or polypeptide in such a manner that the sequence gains access to the interior of a cell of the plant. The methods of the invention do not depend on a particular method for introducing a polynucleotide or polypeptide into a plant, only that the polynucleotide or polypeptides gains access to the interior of at least one cell of the plant. Methods for introducing polynucleotide or polypeptides into plants are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods.
"Stable transformation" is intended to mean that the nucleotide construct introduced into a plant integrates into the genome of the plant and is capable of being inherited by the progeny thereof. "Transient transformation" is intended to mean that a polynucleotide is introduced into the plant and does not integrate into the genome of the plant or a polypeptide is introduced into a plant.
Transformation protocols as well as protocols for introducing nucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation. Suitable methods of introducing nucleotide sequences into plant cells and subsequent insertion into the plant genome include microinjection (Crossway et al, Biotechniques 4:320-334, 1986), electroporation (Riggs et al, Proc Natl Acad Sci USA 83:5602-5606, 1986), Agrobacterium-mediated transformation (U.S.
Pat.
Nos. 5,563,055 and 5,981,840), direct gene transfer (Paszkowski et al, EMBO J
3:2717-2722, 1984), and ballistic particle acceleration (see, for example, U.S. Pat.
Nos. 4,945,050;
5,879,918; 5,886,244; and 5,932,782; Tomes et al, Plant Cell, Tissue, and Organ Culture:
Fundamental Methods, 1995; and McCabe et al, Biotechnology 6:923-926, 1988);
and Lecl transformation (WO 00/28058). For potato transformation see Tu et al, Plant Molecular Biology 37:829-838, 1998 and Chong et al, Transgenic Research 9:71-78, 2000.
Additional transformation procedures can be found in Weissinger et al, Ann Rev Genet 22:421-477, 1988; Sanford et al, Particulate Science and Technology 5:27-37, (onion); Christou et al, Plant Physiol 87:671-674, 1988 (soybean); McCabe et al, Bio/Technology 6:923-926, 1988 (soybean); Finer and McMullen In Vitro Cell Dev Biol 27P:175-182, 1991 (soybean); Singh et al, Theor Appl Genet 96:319-324, 1998 (soybean);
Datta et al, Biotechnology 8:736-740, 1990 (rice); Klein et al, Proc Natl Acad Sci USA
85:4305-4309, 1988 (maize); Klein et al, Biotechnology 6:559-563, 1988 (maize); U.S.
Pat. Nos. 5,240,855; 5,322,783 and 5,324,646; Klein et al, Plant Physiol 91:440-444, 1988 (maize); Fromm et al, Biotechnology 8:833-839, 1990 (maize); Hooykaas-Van Slogteren et al, Nature (London) 311:763-764, 1984; U.S. Pat. No. 5,736,369 (cereals);
Bytebier et al, Proc Natl Acad Sci USA 84:5345-5349, 1987 (Liliaceae); De Wet et al, The Experimental Manipulation of Ovule Tissues, 197-209, 1985 (pollen); Kaeppler et al, Plant Cell Reports 9:415-418, 1990 and Kaeppler et al, Theor Appl Genet 84:560-566, 1992 (whisker-mediated transformation); D'Halluin et al, Plant Cell 4:1495-1505, 1992 (electroporation);
Li et al, Plant Cell Reports 12:250-255, 1993 and Christou and Ford Annals of Botany 75:407-413, 1995 (rice); Osjoda et al, Nature Biotechnology 14:745-750, 1996 (maize via Agrobacterium tumefaciens); all of which are herein incorporated by reference.
In a further embodiment of the present invention, the genetic constructs described herein are adapted for integration into the genome of a cell in which it is expressed. Those skilled in the art will be aware that, in order to achieve integration of a genetic sequence or genetic construct into the genome of a host cell, certain additional genetic sequences may be required. In the case of plants, left and right border sequences from the T-DNA of the Agrobacterium tumefaciens Ti plasmid will generally be required.
The present invention further extends to an isolated cell, tissue or organ comprising the constructs or parts thereof. The present invention extends further to regenerated tissues, organs and whole organisms derived from said cells, tissues and organs and to propagules and progeny thereof as well as seeds and other reproductive material.
The methods of the invention involve introducing a polypeptide or polynucleotide into a plant. "Introducing" is intended to mean presenting to the plant the polynucleotide or polypeptide in such a manner that the sequence gains access to the interior of a cell of the plant. The methods of the invention do not depend on a particular method for introducing a polynucleotide or polypeptide into a plant, only that the polynucleotide or polypeptides gains access to the interior of at least one cell of the plant. Methods for introducing polynucleotide or polypeptides into plants are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods.
"Stable transformation" is intended to mean that the nucleotide construct introduced into a plant integrates into the genome of the plant and is capable of being inherited by the progeny thereof. "Transient transformation" is intended to mean that a polynucleotide is introduced into the plant and does not integrate into the genome of the plant or a polypeptide is introduced into a plant.
Transformation protocols as well as protocols for introducing nucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation. Suitable methods of introducing nucleotide sequences into plant cells and subsequent insertion into the plant genome include microinjection (Crossway et al, Biotechniques 4:320-334, 1986), electroporation (Riggs et al, Proc Natl Acad Sci USA 83:5602-5606, 1986), Agrobacterium-mediated transformation (U.S.
Pat.
Nos. 5,563,055 and 5,981,840), direct gene transfer (Paszkowski et al, EMBO J
3:2717-2722, 1984), and ballistic particle acceleration (see, for example, U.S. Pat.
Nos. 4,945,050;
5,879,918; 5,886,244; and 5,932,782; Tomes et al, Plant Cell, Tissue, and Organ Culture:
Fundamental Methods, 1995; and McCabe et al, Biotechnology 6:923-926, 1988);
and Lecl transformation (WO 00/28058). For potato transformation see Tu et al, Plant Molecular Biology 37:829-838, 1998 and Chong et al, Transgenic Research 9:71-78, 2000.
Additional transformation procedures can be found in Weissinger et al, Ann Rev Genet 22:421-477, 1988; Sanford et al, Particulate Science and Technology 5:27-37, (onion); Christou et al, Plant Physiol 87:671-674, 1988 (soybean); McCabe et al, Bio/Technology 6:923-926, 1988 (soybean); Finer and McMullen In Vitro Cell Dev Biol 27P:175-182, 1991 (soybean); Singh et al, Theor Appl Genet 96:319-324, 1998 (soybean);
Datta et al, Biotechnology 8:736-740, 1990 (rice); Klein et al, Proc Natl Acad Sci USA
85:4305-4309, 1988 (maize); Klein et al, Biotechnology 6:559-563, 1988 (maize); U.S.
Pat. Nos. 5,240,855; 5,322,783 and 5,324,646; Klein et al, Plant Physiol 91:440-444, 1988 (maize); Fromm et al, Biotechnology 8:833-839, 1990 (maize); Hooykaas-Van Slogteren et al, Nature (London) 311:763-764, 1984; U.S. Pat. No. 5,736,369 (cereals);
Bytebier et al, Proc Natl Acad Sci USA 84:5345-5349, 1987 (Liliaceae); De Wet et al, The Experimental Manipulation of Ovule Tissues, 197-209, 1985 (pollen); Kaeppler et al, Plant Cell Reports 9:415-418, 1990 and Kaeppler et al, Theor Appl Genet 84:560-566, 1992 (whisker-mediated transformation); D'Halluin et al, Plant Cell 4:1495-1505, 1992 (electroporation);
Li et al, Plant Cell Reports 12:250-255, 1993 and Christou and Ford Annals of Botany 75:407-413, 1995 (rice); Osjoda et al, Nature Biotechnology 14:745-750, 1996 (maize via Agrobacterium tumefaciens); all of which are herein incorporated by reference.
In a further embodiment of the present invention, the genetic constructs described herein are adapted for integration into the genome of a cell in which it is expressed. Those skilled in the art will be aware that, in order to achieve integration of a genetic sequence or genetic construct into the genome of a host cell, certain additional genetic sequences may be required. In the case of plants, left and right border sequences from the T-DNA of the Agrobacterium tumefaciens Ti plasmid will generally be required.
The present invention further extends to an isolated cell, tissue or organ comprising the constructs or parts thereof. The present invention extends further to regenerated tissues, organs and whole organisms derived from said cells, tissues and organs and to propagules and progeny thereof as well as seeds and other reproductive material.
For example, plants may be regenerated from transformed plant cells or tissues or organs on hormone-containing media and the regenerated plants may take a variety of forms, such as chimeras of transformed cells and non-transformed cells; clonal transformants (e.g. all cells transformed to contain the expression cassette); grafts of transformed and untransformed tissue (e.g. a transformed root stock grafted to an untransformed scion in citrus species). Transformed plants may be propagated by a variety of means, such as by clonal propagation or classical breeding techniques. For example, first generation (or Tl) transformed plants may be selfed to give homozygous second generation (or T2) transformed plants, and the T2 plants further propagated through classical breeding techniques.
The present invention contemplates any other DNA sequence differing in its codon usage but encoding the same protein or a similar protein with substantially the same pesticidal or pestistatic activity, can be constructed, depending on the particular purpose.
It has been described in some prokaryotic and eucaryotic expression systems that changing the codon usage to that of the host cell is desired for gene expression in foreign hosts (Bennetzen &
Hall, J Biol Chem 257:3026, 1982; Itakura, Science 198:1056-1063, 1977). Codon usage tables are available in the literature (Wada et al, Nucl Acids Res 18:2367-1411, 1990;
Murray et al, Nucleic Acids Research 17:477-498, 1989) and in the major DNA
sequence databases. Accordingly, synthetic DNA sequences can be constructed so that the same or substantially the same proteins are produced. It is evident that several DNA
sequences can be devised once the amino acid sequence of the instant agents proteins of this invention.
Such other DNA sequences include synthetic or semi-synthetic DNA sequences that have been changed in order to inactivate certain sites in the gene, e.g. by selectively inactivating certain cryptic regulatory or processing elements present in the native sequence, or by adapting the overall codon usage to that of a more related host organism, preferably that of the host organism in which expression is desired. Synthetic DNA sequences could also be made following the procedures described in EP 0 385 962, EP 0 618 967, or EP 0 682 115.
Small modifications to a DNA sequence such as described above can be routinely made by PCR-mediated mutagenesis (Ho et al, Gene 77:51-59, 1989; White et al, Trends in Genet 5:185-189, 1989). New synthetic or semi-synthetic genes can be made by automated DNA
synthesis and ligation of the resulting DNA fragments.
To prevent or delay the development of resistance by pests, it is preferred to also express in the same plant host, preferably a transgenic plant, the other protein or protein complex, which has a different mode of action, and a high toxicity to the same pest targeted by the first agent (e.g. serine proteinase inhibitor) when produced in a transgenic host, preferably a plant. Suitable candidates to be combined with the serine proteinase inhibitor include the mature VIP1Aa protein when combined with the mature VIP2Aa or VIP2Ab protein of PCT publication WO 96/10083 in case these VIP proteins have a different mode of action compared to the serine proteinase inhibitors; the corn rootworm toxins of Photorhabdus or Xenorhabdus spp., e.g., the insecticidal proteins of Photorhabdus luminescens W-14 (Guo et al, J Biol Chem 274:9836-9842, 1999); the CryET70 protein of WO 00/26378;
the insecticidal proteins produced by Bt strains PS80JJ1, PS149B1 and PS167H2 as described in WO 97/40162, particularly the about 14 kD and about 44 kD proteins of Bt strain PS149B1; the Cry3Bb protein of U.S. Pat. No. 6,023,013; protease inhibitors such as the N2 and Rl cysteine proteinase inhibitors of soybean (Zhao et al, Plant Physiol 111:1299-1306, 1996) or oryzastatine such as rice cystatin (Genbank entry S49967), corn cystatin (Genbank entries D38130, D10622, D63342) such as the corn cystatin expressed in plants as described by Irie et al, Plant Mol Biol 30:149-157, 1996). Also included herein are all equivalents and variants, such as truncated proteins retaining insecticidal activity, of any of the above proteins.
DNA of the encoding serine proteinase inhibitor genes of the subject invention, can be ligated in suitable expression vectors and transformed in E. coli, and the clones can then be screened by conventional colony immunoprobing methods (French et al, Anal Biochem 156:417-423, 1986) for expression of the toxin with monoclonal or polyclonal antibodies raised against proteinase inhibitor. Also, the DNA can be ligated in suitable Bt shuttle vectors (Lereclus et al, Bio/Technology 10:418, 1992) and transformed in a crystal minus Bt-mutant. The clones are then screened for production of ISP proteins (by SDS-PAGE, Western blot and/or insect assay).
The genes encoding the invention can be sequenced in a conventional manner (Maxam and Gilbert Methods in Enzymol 65:499-560, 1980; Sanger Proc Natl Acad Sci USA
74:5463-5467, 1977) to obtain the DNA sequence. Sequence comparisons indicated that the genes are different from previously described genes encoding proteins secreted during the vegetative growth phase of Bacillus or other bacterial species and Bacillus thuringiensis crystal proteins with activity against Coleoptera (Crickmore, et al, Microbiology and Molecular Biology Reviews 62:807-813, 1998; WO 98/44137, WO 94/21795, WO
96/10083, WO 00/09697, WO 9957282, and WO 9746105).
A pesticidal or pestistatic composition of the subject invention can also be formulated in a conventional manner using the microorganisms transformed with the genes, or preferably their respective proteins or pesticidally or pestistatically effective portions thereof as an active ingredient, together with suitable carriers, diluents, emulsifiers and/or dispersants (e.g., as described by Bernhard and Utz, An Environmental Biopesticide: Theory and Practice 255-267, 1993). This pesticidal or pestistatic composition can be formulated as a wettable powder, pellets, granules or dust or as a liquid formulation with aqueous or non-aqueous solvents as a foam, gel, suspension, concentrate, etc. Known microorganisms include cells of Pseudomonas or other bacteria that serve to encapsulate the proteins in a stable environment prior to application to the insects. Also included in the invention is a product comprising the agents described herein as a combined preparation for simultaneous, separate or sequential use to protect corn plants against corn rootworms, particularly such product is an insecticidal composition or a transgenic corn plant.
A method for controlling pests in accordance with the subject invention can comprise applying (e.g., spraying), to a locus (area) to be protected, an pesticidal or pestistatic effective amount of the agents or host cells transformed with the gene of the subject invention. The locus to be protected can include, for example, the habitat of the insect pests or growing vegetation or an area where vegetation is to be grown.
The present invention contemplates any other DNA sequence differing in its codon usage but encoding the same protein or a similar protein with substantially the same pesticidal or pestistatic activity, can be constructed, depending on the particular purpose.
It has been described in some prokaryotic and eucaryotic expression systems that changing the codon usage to that of the host cell is desired for gene expression in foreign hosts (Bennetzen &
Hall, J Biol Chem 257:3026, 1982; Itakura, Science 198:1056-1063, 1977). Codon usage tables are available in the literature (Wada et al, Nucl Acids Res 18:2367-1411, 1990;
Murray et al, Nucleic Acids Research 17:477-498, 1989) and in the major DNA
sequence databases. Accordingly, synthetic DNA sequences can be constructed so that the same or substantially the same proteins are produced. It is evident that several DNA
sequences can be devised once the amino acid sequence of the instant agents proteins of this invention.
Such other DNA sequences include synthetic or semi-synthetic DNA sequences that have been changed in order to inactivate certain sites in the gene, e.g. by selectively inactivating certain cryptic regulatory or processing elements present in the native sequence, or by adapting the overall codon usage to that of a more related host organism, preferably that of the host organism in which expression is desired. Synthetic DNA sequences could also be made following the procedures described in EP 0 385 962, EP 0 618 967, or EP 0 682 115.
Small modifications to a DNA sequence such as described above can be routinely made by PCR-mediated mutagenesis (Ho et al, Gene 77:51-59, 1989; White et al, Trends in Genet 5:185-189, 1989). New synthetic or semi-synthetic genes can be made by automated DNA
synthesis and ligation of the resulting DNA fragments.
To prevent or delay the development of resistance by pests, it is preferred to also express in the same plant host, preferably a transgenic plant, the other protein or protein complex, which has a different mode of action, and a high toxicity to the same pest targeted by the first agent (e.g. serine proteinase inhibitor) when produced in a transgenic host, preferably a plant. Suitable candidates to be combined with the serine proteinase inhibitor include the mature VIP1Aa protein when combined with the mature VIP2Aa or VIP2Ab protein of PCT publication WO 96/10083 in case these VIP proteins have a different mode of action compared to the serine proteinase inhibitors; the corn rootworm toxins of Photorhabdus or Xenorhabdus spp., e.g., the insecticidal proteins of Photorhabdus luminescens W-14 (Guo et al, J Biol Chem 274:9836-9842, 1999); the CryET70 protein of WO 00/26378;
the insecticidal proteins produced by Bt strains PS80JJ1, PS149B1 and PS167H2 as described in WO 97/40162, particularly the about 14 kD and about 44 kD proteins of Bt strain PS149B1; the Cry3Bb protein of U.S. Pat. No. 6,023,013; protease inhibitors such as the N2 and Rl cysteine proteinase inhibitors of soybean (Zhao et al, Plant Physiol 111:1299-1306, 1996) or oryzastatine such as rice cystatin (Genbank entry S49967), corn cystatin (Genbank entries D38130, D10622, D63342) such as the corn cystatin expressed in plants as described by Irie et al, Plant Mol Biol 30:149-157, 1996). Also included herein are all equivalents and variants, such as truncated proteins retaining insecticidal activity, of any of the above proteins.
DNA of the encoding serine proteinase inhibitor genes of the subject invention, can be ligated in suitable expression vectors and transformed in E. coli, and the clones can then be screened by conventional colony immunoprobing methods (French et al, Anal Biochem 156:417-423, 1986) for expression of the toxin with monoclonal or polyclonal antibodies raised against proteinase inhibitor. Also, the DNA can be ligated in suitable Bt shuttle vectors (Lereclus et al, Bio/Technology 10:418, 1992) and transformed in a crystal minus Bt-mutant. The clones are then screened for production of ISP proteins (by SDS-PAGE, Western blot and/or insect assay).
The genes encoding the invention can be sequenced in a conventional manner (Maxam and Gilbert Methods in Enzymol 65:499-560, 1980; Sanger Proc Natl Acad Sci USA
74:5463-5467, 1977) to obtain the DNA sequence. Sequence comparisons indicated that the genes are different from previously described genes encoding proteins secreted during the vegetative growth phase of Bacillus or other bacterial species and Bacillus thuringiensis crystal proteins with activity against Coleoptera (Crickmore, et al, Microbiology and Molecular Biology Reviews 62:807-813, 1998; WO 98/44137, WO 94/21795, WO
96/10083, WO 00/09697, WO 9957282, and WO 9746105).
A pesticidal or pestistatic composition of the subject invention can also be formulated in a conventional manner using the microorganisms transformed with the genes, or preferably their respective proteins or pesticidally or pestistatically effective portions thereof as an active ingredient, together with suitable carriers, diluents, emulsifiers and/or dispersants (e.g., as described by Bernhard and Utz, An Environmental Biopesticide: Theory and Practice 255-267, 1993). This pesticidal or pestistatic composition can be formulated as a wettable powder, pellets, granules or dust or as a liquid formulation with aqueous or non-aqueous solvents as a foam, gel, suspension, concentrate, etc. Known microorganisms include cells of Pseudomonas or other bacteria that serve to encapsulate the proteins in a stable environment prior to application to the insects. Also included in the invention is a product comprising the agents described herein as a combined preparation for simultaneous, separate or sequential use to protect corn plants against corn rootworms, particularly such product is an insecticidal composition or a transgenic corn plant.
A method for controlling pests in accordance with the subject invention can comprise applying (e.g., spraying), to a locus (area) to be protected, an pesticidal or pestistatic effective amount of the agents or host cells transformed with the gene of the subject invention. The locus to be protected can include, for example, the habitat of the insect pests or growing vegetation or an area where vegetation is to be grown.
To obtain the agents , cells of the recombinant hosts expressing the agents can be grown in a conventional manner on a suitable culture medium and the protein can then be obtained from the medium using conventional means. The agent can then be separated and purified by standard techniques such as chromatography, extraction, electrophoresis, or the like.
The protease-resistant toxin form can then be obtained by protease, e.g.
cysteine or serine protease, digestion of the protein.
While the invention does not depend on a particular biological mechanism for increasing the resistance of a plant to a plant pest, expression of the nucleotide sequences of the invention in a plant can result in the production of the pesticidal or pestistatic proteins of the invention and in an increase in the resistance of the plant to a plant pest. The plants of the invention find use in agriculture in methods for impacting plant pests.
Certain embodiments of the invention provide transformed crop plants, such as, for example, cotton plants, which find use in methods for impacting insect pests of the plant.
A "subject plant or plant cell" is one in which genetic alteration, such as transformation, has been effected as to a gene of interest, or is a plant or plant cell which is descended from a plant or cell so altered and which comprises the alteration. A
"control" or "control plant" or "control plant cell" provides a reference point for measuring changes in phenotype of the subject plant or plant cell.
A control plant or plant cell may comprise, for example: (a) a wild-type plant or cell, i.e., of the same genotype as the starting material for the genetic alteration which resulted in the subject plant or cell; (b) a plant or plant cell of the same genotype as the starting material but which has been transformed with a null construct (i.e., with a construct which has no known effect on the trait of interest, such as a construct comprising a marker gene); (c) a plant or plant cell which is a non-transformed segregant among progeny of a subject plant or plant cell; (d) a plant or plant cell genetically identical to the subject plant or plant cell but which is not exposed to conditions or stimuli that would induce expression of the gene of interest; or (e) the subject plant or plant cell itself, under conditions in which the gene of interest is not expressed.
The protease-resistant toxin form can then be obtained by protease, e.g.
cysteine or serine protease, digestion of the protein.
While the invention does not depend on a particular biological mechanism for increasing the resistance of a plant to a plant pest, expression of the nucleotide sequences of the invention in a plant can result in the production of the pesticidal or pestistatic proteins of the invention and in an increase in the resistance of the plant to a plant pest. The plants of the invention find use in agriculture in methods for impacting plant pests.
Certain embodiments of the invention provide transformed crop plants, such as, for example, cotton plants, which find use in methods for impacting insect pests of the plant.
A "subject plant or plant cell" is one in which genetic alteration, such as transformation, has been effected as to a gene of interest, or is a plant or plant cell which is descended from a plant or cell so altered and which comprises the alteration. A
"control" or "control plant" or "control plant cell" provides a reference point for measuring changes in phenotype of the subject plant or plant cell.
A control plant or plant cell may comprise, for example: (a) a wild-type plant or cell, i.e., of the same genotype as the starting material for the genetic alteration which resulted in the subject plant or cell; (b) a plant or plant cell of the same genotype as the starting material but which has been transformed with a null construct (i.e., with a construct which has no known effect on the trait of interest, such as a construct comprising a marker gene); (c) a plant or plant cell which is a non-transformed segregant among progeny of a subject plant or plant cell; (d) a plant or plant cell genetically identical to the subject plant or plant cell but which is not exposed to conditions or stimuli that would induce expression of the gene of interest; or (e) the subject plant or plant cell itself, under conditions in which the gene of interest is not expressed.
As indicated above, one of skill in the art will readily acknowledge that advances in the field of molecular biology such as site-specific and random mutagenesis, polymerase chain reaction methodologies, and protein engineering techniques provide an extensive collection of tools and protocols suitable for use to alter or engineer both the amino acid sequence and underlying genetic sequences of proteins of agricultural interest.
Thus, the Cry9 family proteins of the invention may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of the pesticidal or pestistatic proteins can be prepared by introducing mutations into a synthetic nucleic acid (e.g., DNA molecule). Methods for mutagenesis and nucleic acid alterations are well known in the art. For exainple, designed changes can be introduced using an oligonucleotide-mediated site-directed mutagenesis technique. See, for example, Kunkel Proc Natl Acad Sci USA 82:488-492, 1985; Kunkel et al, Methods in Enzymol 154:367-382, 1987; U.S. Pat. No. 4,873,192; Walker and Gaastra, Techniques in Molecular Biology 1983, and the references cited therein.
The mutagenized Cry9 family nucleotide sequences of the invention may be modified so as to change about 1, 2, 3, 4, 5, 6, 8, 10, 12 or more of the amino acids present in the primary sequence of the encoded polypeptide. Alternatively, even more changes from the native sequence may be introduced such that the encoded protein may have at least about 1% or 2%, or about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, or even about 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, 21%, 22%, 23%, 24%, or 25%, 30%, 35%, or 40%
or more of the codons altered, or otherwise modified compared to the corresponding wild-type protein. In the same manner, the encoded protein may have at least about 1% or 2%, or about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, or even about 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, 21%, 22%, 23%, 24%, or 25%, 30%, 35%, or 40% or more additional codons compared to the corresponding wild-type protein. It should be understood that the mutagenized Cry9 family nucleotide sequences of the present invention are intended to encompass biologically functional, equivalent peptides which have pesticidal activity, such as an improved pesticidal activity as determined by antifeedant properties against fall armyworm larvae. Such sequences may arise as a consequence of codon redundancy and functional equivalency that are known to occur naturally within nucleic acid sequences and the proteins thus encoded.
One of skill in the art would recognize that amino acid additions and/or substitutions are generally based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, charge, size, and the like. Exemplary amino acid substitution groups that take various of the foregoing characteristics into consideration are well known to those of skill in the art and include: arginine and lysine;
glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine, and isoleucine.
In certain embodiments the nucleic acid sequences of the present invention can be stacked with any combination of polynucleotide sequences of interest in order to create plants with a desired phenotype. For example, the polynucleotides of the present invention may be stacked with any other polynucleotides encoding polypeptides having pesticidal and/or insecticidal activity, such as other B. thuringiensis toxic proteins (described in U.S. Pat.
Nos. 5,366,892; 5,747,450; 5,736,514; 5,723,756; 5,593,881; and Geiser et al, Gene 48:109, 1986), pentin (described in U.S. Pat. No. 5,981,722) and the like. The combinations generated can also include multiple copies of any one of the polynucleotides of interest. The polynucleotides of the present invention can also be stacked with any other gene or combination of genes to produce plants with a variety of desired trait combinations including but not limited to traits desirable for animal feed such as high oil genes (e.g., U.S. Pat. No. 6,232,529); balanced amino acids (e.g. hordothionins (U.S. Pat.
Nos.
5,990,389; 5,885,801; 5,885,802; and 5,703,049); barley high lysine (Williamson et al, Eur J Biochem 165:99-106,1987; and WO 98/20122) and high methionine proteins (Pedersen et al, JBiol Chem 261:6279, 1986; Kirihara et al, Gene 71:359, 1988; and Musumura et al, Plant Mol Biol 12:123, 1989); increased digestibility (e.g., modified storage proteins (U.S.
Application Ser. No. 10/053,410, filed Nov. 7, 2001); and thioredoxins (U.S.
Application Ser. No. 10/005,429, filed Dec. 3, 2001)), the disclosures of which are herein incorporated by reference.
The polynucleotides of the present invention can also be stacked with traits desirable for disease or herbicide resistance (e.g., fumonisin detoxification genes (U.S.
Pat. No.
5,792,931); avirulence and disease resistance genes (Jones et al, Science 266:789, 1994;
Martin et al, Science 262:1432, 1993; and Mindrinos et al, Cell 78:1089, 1994);
acetolactate synthase (ALS) mutants that lead to herbicide resistance such as the S4 and/or Hra mutations; inhibitors of glutainine synthase such as phosphinothricin or basta (e.g., bar gene); and glyphosate resistance (EPSPS gene and GAT gene as disclosed in U.S.
application Ser. No. 10/004,357; and 10/427,692); and traits desirable for processing or process products such as high oil (e.g., U.S. Pat. No. 6,232,529); modified oils (e.g., fatty acid desaturase genes (U.S. Pat. No. 5,952,544; WO 94/11516)); modified starches (e.g., ADPG pyrophosphorylases (AGPase), starch synthases (SS), starch branching enzymes (SBE) and starch debranching enzymes (SDBE)); and polymers or bioplastics (e.g., U.S.
Pat. No. 5,602,321; beta-ketothiolase, polyhydroxybutyrate synthase, and acetoacetyl-CoA
reductase (Schubert et al, J Bacteriol 170:5837-5847, 1988) facilitate expression of polyhydroxyalkanoates (PHAs)), the disclosures of which are herein incorporated by reference. One could also combine the polynucleotides of the present invention with polynucleotides providing agronomic traits such as male sterility (e.g., see U.S. Pat. No.
5,583,210), stalk strength, flowering time, or transformation technology traits such as cell cycle regulation or gene targeting (e.g. WO 99/61619; WO 00/17364; WO
99/25821), the disclosures of which are herein incorporated by reference.
These stacked combinations can be created by any method including but not limited to cross breeding plants by any conventional or TopCrosse methodology, or genetic transformation. If the traits are stacked by genetically transforming the plants, the polynucleotide sequences of interest can be combined at any time and in any order. For example, a transgenic plant comprising one or more desired traits can be used as the target to introduce furtller traits by subsequent transformation. The traits can be introduced simultaneously in a co-transformation protocol with the polynucleotides of interest provided by any combination of transformation cassettes. For example, if two sequences will be introduced, the two sequences can be contained in separate transformation cassettes (trans) or contained on the same transformation cassette (cis). Expression of the sequences can be driven by the same promoter or by different promoters. In certain cases, it may be desirable to introduce a transformation cassette that will suppress the expression of the polynucleotide of interest. This may be combined with any combination of other suppression cassettes or overexpression cassettes to generate the desired combination of traits in the plant. It is further recognized that polynucleotide sequences can be stacked at a desired genomic location using a site-specific recombination system. See, for example, W099/25821, W099/25854, W099/25840, W099/25855, and W099/25853, all of which are herein incorporated by reference.
Compositions of the invention find use in protecting plants, seeds, and plant products in a variety of ways. For example, the compositions can be used in a method that involves placing an effective ainount of the pesticidal or pestistatic composition in the environment of the pest by a procedure selected from the group consisting of spraying, dusting, broadcasting, or seed coating.
Before plant propagation material (fruit, tuber, bulb, corm, grains, seed), but especially seed, is sold as a commercial product, it is customarily treated with a protectant coating comprising herbicides, insecticides, fungicides, bactericides, nematicides, molluscicides, or mixtures of several of these preparations, if desired together with further carriers, surfactants, or application-promoting adjuvants customarily employed in the art of formulation to provide protection against damage caused by bacterial, fungal, or animal pests. In order to treat the seed, the protectant coating may be applied to the seeds either by impregnating the tubers or grains with a liquid fomlulation or by coating them with a combined wet or dry formulation. In addition, in special cases, other methods of application to plants are possible, e.g., treatment directed at the buds or the fruit.
The plant seed of the invention comprising the nucleotide sequences encoding the agents may be treated with a seed protectant coating comprising a seed treatment compound, such as, for example, captan, carboxin, thiram, methalaxyl, pirimiphos-methyl, and others that are commonly used in seed treatment. In one embodiment within the scope of the invention, a seed protectant coating comprising a pesticidal composition of the invention is used alone or in combination with one of the seed protectant coatings customarily used in seed treatment.
In the present invention, a composition includes whole organisms, cells, spore(s), pesticidal or pestistatic protein(s), pesticidal or pestistatic component(s), pest-impacting component(s), mutant(s), living or dead cells and cell components, including mixtures of living and dead cells and cell components, and including broken cells and cell components or an isolated pesticidal or pestistatic protein can be formulated with an acceptable carrier into a pesticidal composition(s) that is, for example, a suspension, a solution, an emulsion, a dusting powder, a dispersible granule, a wettable powder, and an emulsifiable concentrate, an aerosol, an impregnated granule, an adjuvant, a coatable paste, and also encapsulations in, for example, polymer substances.
Such compositions disclosed above may be obtained by the addition of a surface-active agent, an inert carrier, a preservative, a humectant, a feeding stimulant, an attractant, an encapsulating agent, a binder, an emulsifier, a dye, a UV protectant, a buffer, a flow agent or fertilizers, micronutrient donors, or other preparations that influence plant growth. One or more agrochemicals including, but not limited to, herbicides, insecticides, fungicides, bactericides, nematicides, molluscicides, acaracides, plant growth regulators, harvest aids, and fertilizers, can be combined with carriers, surfactants or adjuvants customarily employed in the art of formulation or other components to facilitate product handling and application for particular target pests. Suitable carriers and adjuvants can be solid or liquid and correspond to the substances ordinarily employed in formulation technology, e.g., natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, binders, or fertilizers. The active ingredients of the present invention are normally applied in the form of compositions and can be applied to the crop area, plant, or seed to be treated.
For example, the compositions of the present invention may be applied to grain in preparation for or during storage in a grain bin or silo, etc. The compositions of the present invention may be applied simultaneously or in succession with other compounds.
Methods of applying an active ingredient of the present invention or an agrochemical composition of the present invention that contains at least one of the pesticidal proteins produced by the bacterial strains of the present invention include, but are not limited to, foliar application, seed coating, and soil application. The number of applications and the rate of application depend on the intensity of infestation by the corresponding pest.
Suitable surface-active agents include, but are not limited to, anionic compounds such as a carboxylate of, for example, a metal; a carboxylate of a long chain fatty acid; an N-acylsarcosinate; mono or di-esters of phosphoric acid with fatty alcohol ethoxylates or salts of such esters; fatty alcohol sulfates such as sodium dodecyl sulfate, sodium octadecyl sulfate or sodium cetyl sulfate; etlioxylated fatty alcohol sulfates;
ethoxylated alkylphenol sulfates; lignin sulfonates; petroleum sulfonates; alkyl aryl sulfonates such as alkyl-benzene sulfonates or lower alkylnaphtalene sulfonates, e.g., butyl-naphthalene sulfonate;
salts of sulfonated naphthalene-formaldehyde condensates; salts of sulfonated phenol-formaldehyde condensates; more complex sulfonates such as the amide sulfonates, e.g., the sulfonated condensation product of oleic acid and N-methyl taurine; or the dialkyl sulfosuccinates, e.g., the sodium sulfonate of dioctyl succinate. Non-ionic agents include condensation products of fatty acid esters, fatty alcohols, fatty acid amides or fatty-alkyl-or alkenyl-substituted phenols with ethylene oxide, fatty esters of polyhydric alcohol ethers, e.g., sorbitan fatty acid esters, condensation products of such esters with ethylene oxide, e.g., polyoxyethylene sorbitar fatty acid esters, block copolymers of ethylene oxide and propylene oxide, acetylenic glycols such as 2,4,7,9-tetraethyl-5-decyn-4,7-diol, or ethoxylated acetylenic glycols. Examples of a cationic surface-active agent include, for instance, an aliphatic mono-, di-, or polyamine such as an acetate, naphthenate or oleate; or oxygen-containing amine such as an amine oxide of polyoxyethylene alkylamine;
an amide-linked amine prepared by the condensation of a carboxylic acid with a di-or polyamine; or a quaternary ammonium salt.
Examples of inert materials include but are not limited to inorganic minerals such as kaolin, phyllosilicates, carbonates, sulfates, phosphates, or botanical materials such as cork, powdered corncobs, peanut hulls, rice hulls, and walnut shells.
Thus, the Cry9 family proteins of the invention may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of the pesticidal or pestistatic proteins can be prepared by introducing mutations into a synthetic nucleic acid (e.g., DNA molecule). Methods for mutagenesis and nucleic acid alterations are well known in the art. For exainple, designed changes can be introduced using an oligonucleotide-mediated site-directed mutagenesis technique. See, for example, Kunkel Proc Natl Acad Sci USA 82:488-492, 1985; Kunkel et al, Methods in Enzymol 154:367-382, 1987; U.S. Pat. No. 4,873,192; Walker and Gaastra, Techniques in Molecular Biology 1983, and the references cited therein.
The mutagenized Cry9 family nucleotide sequences of the invention may be modified so as to change about 1, 2, 3, 4, 5, 6, 8, 10, 12 or more of the amino acids present in the primary sequence of the encoded polypeptide. Alternatively, even more changes from the native sequence may be introduced such that the encoded protein may have at least about 1% or 2%, or about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, or even about 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, 21%, 22%, 23%, 24%, or 25%, 30%, 35%, or 40%
or more of the codons altered, or otherwise modified compared to the corresponding wild-type protein. In the same manner, the encoded protein may have at least about 1% or 2%, or about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, or even about 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, 21%, 22%, 23%, 24%, or 25%, 30%, 35%, or 40% or more additional codons compared to the corresponding wild-type protein. It should be understood that the mutagenized Cry9 family nucleotide sequences of the present invention are intended to encompass biologically functional, equivalent peptides which have pesticidal activity, such as an improved pesticidal activity as determined by antifeedant properties against fall armyworm larvae. Such sequences may arise as a consequence of codon redundancy and functional equivalency that are known to occur naturally within nucleic acid sequences and the proteins thus encoded.
One of skill in the art would recognize that amino acid additions and/or substitutions are generally based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, charge, size, and the like. Exemplary amino acid substitution groups that take various of the foregoing characteristics into consideration are well known to those of skill in the art and include: arginine and lysine;
glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine, and isoleucine.
In certain embodiments the nucleic acid sequences of the present invention can be stacked with any combination of polynucleotide sequences of interest in order to create plants with a desired phenotype. For example, the polynucleotides of the present invention may be stacked with any other polynucleotides encoding polypeptides having pesticidal and/or insecticidal activity, such as other B. thuringiensis toxic proteins (described in U.S. Pat.
Nos. 5,366,892; 5,747,450; 5,736,514; 5,723,756; 5,593,881; and Geiser et al, Gene 48:109, 1986), pentin (described in U.S. Pat. No. 5,981,722) and the like. The combinations generated can also include multiple copies of any one of the polynucleotides of interest. The polynucleotides of the present invention can also be stacked with any other gene or combination of genes to produce plants with a variety of desired trait combinations including but not limited to traits desirable for animal feed such as high oil genes (e.g., U.S. Pat. No. 6,232,529); balanced amino acids (e.g. hordothionins (U.S. Pat.
Nos.
5,990,389; 5,885,801; 5,885,802; and 5,703,049); barley high lysine (Williamson et al, Eur J Biochem 165:99-106,1987; and WO 98/20122) and high methionine proteins (Pedersen et al, JBiol Chem 261:6279, 1986; Kirihara et al, Gene 71:359, 1988; and Musumura et al, Plant Mol Biol 12:123, 1989); increased digestibility (e.g., modified storage proteins (U.S.
Application Ser. No. 10/053,410, filed Nov. 7, 2001); and thioredoxins (U.S.
Application Ser. No. 10/005,429, filed Dec. 3, 2001)), the disclosures of which are herein incorporated by reference.
The polynucleotides of the present invention can also be stacked with traits desirable for disease or herbicide resistance (e.g., fumonisin detoxification genes (U.S.
Pat. No.
5,792,931); avirulence and disease resistance genes (Jones et al, Science 266:789, 1994;
Martin et al, Science 262:1432, 1993; and Mindrinos et al, Cell 78:1089, 1994);
acetolactate synthase (ALS) mutants that lead to herbicide resistance such as the S4 and/or Hra mutations; inhibitors of glutainine synthase such as phosphinothricin or basta (e.g., bar gene); and glyphosate resistance (EPSPS gene and GAT gene as disclosed in U.S.
application Ser. No. 10/004,357; and 10/427,692); and traits desirable for processing or process products such as high oil (e.g., U.S. Pat. No. 6,232,529); modified oils (e.g., fatty acid desaturase genes (U.S. Pat. No. 5,952,544; WO 94/11516)); modified starches (e.g., ADPG pyrophosphorylases (AGPase), starch synthases (SS), starch branching enzymes (SBE) and starch debranching enzymes (SDBE)); and polymers or bioplastics (e.g., U.S.
Pat. No. 5,602,321; beta-ketothiolase, polyhydroxybutyrate synthase, and acetoacetyl-CoA
reductase (Schubert et al, J Bacteriol 170:5837-5847, 1988) facilitate expression of polyhydroxyalkanoates (PHAs)), the disclosures of which are herein incorporated by reference. One could also combine the polynucleotides of the present invention with polynucleotides providing agronomic traits such as male sterility (e.g., see U.S. Pat. No.
5,583,210), stalk strength, flowering time, or transformation technology traits such as cell cycle regulation or gene targeting (e.g. WO 99/61619; WO 00/17364; WO
99/25821), the disclosures of which are herein incorporated by reference.
These stacked combinations can be created by any method including but not limited to cross breeding plants by any conventional or TopCrosse methodology, or genetic transformation. If the traits are stacked by genetically transforming the plants, the polynucleotide sequences of interest can be combined at any time and in any order. For example, a transgenic plant comprising one or more desired traits can be used as the target to introduce furtller traits by subsequent transformation. The traits can be introduced simultaneously in a co-transformation protocol with the polynucleotides of interest provided by any combination of transformation cassettes. For example, if two sequences will be introduced, the two sequences can be contained in separate transformation cassettes (trans) or contained on the same transformation cassette (cis). Expression of the sequences can be driven by the same promoter or by different promoters. In certain cases, it may be desirable to introduce a transformation cassette that will suppress the expression of the polynucleotide of interest. This may be combined with any combination of other suppression cassettes or overexpression cassettes to generate the desired combination of traits in the plant. It is further recognized that polynucleotide sequences can be stacked at a desired genomic location using a site-specific recombination system. See, for example, W099/25821, W099/25854, W099/25840, W099/25855, and W099/25853, all of which are herein incorporated by reference.
Compositions of the invention find use in protecting plants, seeds, and plant products in a variety of ways. For example, the compositions can be used in a method that involves placing an effective ainount of the pesticidal or pestistatic composition in the environment of the pest by a procedure selected from the group consisting of spraying, dusting, broadcasting, or seed coating.
Before plant propagation material (fruit, tuber, bulb, corm, grains, seed), but especially seed, is sold as a commercial product, it is customarily treated with a protectant coating comprising herbicides, insecticides, fungicides, bactericides, nematicides, molluscicides, or mixtures of several of these preparations, if desired together with further carriers, surfactants, or application-promoting adjuvants customarily employed in the art of formulation to provide protection against damage caused by bacterial, fungal, or animal pests. In order to treat the seed, the protectant coating may be applied to the seeds either by impregnating the tubers or grains with a liquid fomlulation or by coating them with a combined wet or dry formulation. In addition, in special cases, other methods of application to plants are possible, e.g., treatment directed at the buds or the fruit.
The plant seed of the invention comprising the nucleotide sequences encoding the agents may be treated with a seed protectant coating comprising a seed treatment compound, such as, for example, captan, carboxin, thiram, methalaxyl, pirimiphos-methyl, and others that are commonly used in seed treatment. In one embodiment within the scope of the invention, a seed protectant coating comprising a pesticidal composition of the invention is used alone or in combination with one of the seed protectant coatings customarily used in seed treatment.
In the present invention, a composition includes whole organisms, cells, spore(s), pesticidal or pestistatic protein(s), pesticidal or pestistatic component(s), pest-impacting component(s), mutant(s), living or dead cells and cell components, including mixtures of living and dead cells and cell components, and including broken cells and cell components or an isolated pesticidal or pestistatic protein can be formulated with an acceptable carrier into a pesticidal composition(s) that is, for example, a suspension, a solution, an emulsion, a dusting powder, a dispersible granule, a wettable powder, and an emulsifiable concentrate, an aerosol, an impregnated granule, an adjuvant, a coatable paste, and also encapsulations in, for example, polymer substances.
Such compositions disclosed above may be obtained by the addition of a surface-active agent, an inert carrier, a preservative, a humectant, a feeding stimulant, an attractant, an encapsulating agent, a binder, an emulsifier, a dye, a UV protectant, a buffer, a flow agent or fertilizers, micronutrient donors, or other preparations that influence plant growth. One or more agrochemicals including, but not limited to, herbicides, insecticides, fungicides, bactericides, nematicides, molluscicides, acaracides, plant growth regulators, harvest aids, and fertilizers, can be combined with carriers, surfactants or adjuvants customarily employed in the art of formulation or other components to facilitate product handling and application for particular target pests. Suitable carriers and adjuvants can be solid or liquid and correspond to the substances ordinarily employed in formulation technology, e.g., natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, binders, or fertilizers. The active ingredients of the present invention are normally applied in the form of compositions and can be applied to the crop area, plant, or seed to be treated.
For example, the compositions of the present invention may be applied to grain in preparation for or during storage in a grain bin or silo, etc. The compositions of the present invention may be applied simultaneously or in succession with other compounds.
Methods of applying an active ingredient of the present invention or an agrochemical composition of the present invention that contains at least one of the pesticidal proteins produced by the bacterial strains of the present invention include, but are not limited to, foliar application, seed coating, and soil application. The number of applications and the rate of application depend on the intensity of infestation by the corresponding pest.
Suitable surface-active agents include, but are not limited to, anionic compounds such as a carboxylate of, for example, a metal; a carboxylate of a long chain fatty acid; an N-acylsarcosinate; mono or di-esters of phosphoric acid with fatty alcohol ethoxylates or salts of such esters; fatty alcohol sulfates such as sodium dodecyl sulfate, sodium octadecyl sulfate or sodium cetyl sulfate; etlioxylated fatty alcohol sulfates;
ethoxylated alkylphenol sulfates; lignin sulfonates; petroleum sulfonates; alkyl aryl sulfonates such as alkyl-benzene sulfonates or lower alkylnaphtalene sulfonates, e.g., butyl-naphthalene sulfonate;
salts of sulfonated naphthalene-formaldehyde condensates; salts of sulfonated phenol-formaldehyde condensates; more complex sulfonates such as the amide sulfonates, e.g., the sulfonated condensation product of oleic acid and N-methyl taurine; or the dialkyl sulfosuccinates, e.g., the sodium sulfonate of dioctyl succinate. Non-ionic agents include condensation products of fatty acid esters, fatty alcohols, fatty acid amides or fatty-alkyl-or alkenyl-substituted phenols with ethylene oxide, fatty esters of polyhydric alcohol ethers, e.g., sorbitan fatty acid esters, condensation products of such esters with ethylene oxide, e.g., polyoxyethylene sorbitar fatty acid esters, block copolymers of ethylene oxide and propylene oxide, acetylenic glycols such as 2,4,7,9-tetraethyl-5-decyn-4,7-diol, or ethoxylated acetylenic glycols. Examples of a cationic surface-active agent include, for instance, an aliphatic mono-, di-, or polyamine such as an acetate, naphthenate or oleate; or oxygen-containing amine such as an amine oxide of polyoxyethylene alkylamine;
an amide-linked amine prepared by the condensation of a carboxylic acid with a di-or polyamine; or a quaternary ammonium salt.
Examples of inert materials include but are not limited to inorganic minerals such as kaolin, phyllosilicates, carbonates, sulfates, phosphates, or botanical materials such as cork, powdered corncobs, peanut hulls, rice hulls, and walnut shells.
The compositions of the present invention can be in a suitable form for direct application or as a concentrate of primary composition that requires dilution with a suitable quantity of water or other diluant before application. The pesticidal concentration will vary depending upon the nature of the particular formulation, specifically, whether it is a concentrate or to be used directly. The composition contains 1 to 98% of a solid or liquid inert carrier, and 0 to 50% or 0.1 to 50% of a surfactant. These compositions will be administered at the labeled rate for the commercial product, for example, about 0.01 lb-5.0 lb.
per acre when in dry form and at about 0.01 pts.-10 pts. per acre when in liquid form.
Insect pests may be tested for pesticidal or pestistatic activity of compositions of the instant invention in early developmental stages, e.g., as larvae or other immature forms.
The insects may be reared in total darkness at from about 20 C. to about 30 C.
and from about 30% to about 70% relative humidity. Bioassays may be performed as described in Czapla and Lang JEcon Entomol 83(6):2480-2485, 1990. Methods of rearing insect larvae and performing bioassays are well known to one of ordinary skill in the art.
A wide variety of bioassay techniques are known to one skilled in the art.
General procedures include addition of the experimental compound or organism to the diet source in an enclosed container. Pesticidal activity can be measured by, but is not limited to, changes in mortality, weight loss, attraction, repellency and other behavioral and physical changes after feeding and exposure for an appropriate length of time.
Bioassays described herein can be used with any feeding insect pest in the larval or adult stage.
The present invention is further described by the following non-limiting Examples.
per acre when in dry form and at about 0.01 pts.-10 pts. per acre when in liquid form.
Insect pests may be tested for pesticidal or pestistatic activity of compositions of the instant invention in early developmental stages, e.g., as larvae or other immature forms.
The insects may be reared in total darkness at from about 20 C. to about 30 C.
and from about 30% to about 70% relative humidity. Bioassays may be performed as described in Czapla and Lang JEcon Entomol 83(6):2480-2485, 1990. Methods of rearing insect larvae and performing bioassays are well known to one of ordinary skill in the art.
A wide variety of bioassay techniques are known to one skilled in the art.
General procedures include addition of the experimental compound or organism to the diet source in an enclosed container. Pesticidal activity can be measured by, but is not limited to, changes in mortality, weight loss, attraction, repellency and other behavioral and physical changes after feeding and exposure for an appropriate length of time.
Bioassays described herein can be used with any feeding insect pest in the larval or adult stage.
The present invention is further described by the following non-limiting Examples.
NaPI
A bioassay was developed based on feeding larvae of H. arnaigera on cotton leaves homozygous for NaPI and coated with varying concentrations of CrylAc. The NaPI
leaves were specifically selected to give a small reduction in weight of the larvae. (In previous bioassays, without CrylAc, leaves have been used which give a greater effect on larval growth and development.) Preparation of Leaves The leaves were cut into discs using a hole puncher and coated with CrylAc solutions by immersing in the test solution for 5 minutes. The discs were removed from the solution with tweezers and then air dried on plastic mesh (approx. 30 min). When there was no visible moisture on the leaf disc surface, the discs were transferred to wells of a 24 well plate.
The wells contained filter paper moistened with water. Untransformed leaf material of cv Coker was used in control experiments.
Handling of Eggs H. armigera eggs (supplied by Department of Primary Industry & Fisheries, Indooroopilly, Queensland, Australia) were delivered by air freight to the laboratory and incubated (18-20 C depending on developmental stage) to the brown egg stage (pre hatching).
Eggs were suspended in polyacrylate (Aquakeep polyacrylate, 1 mg/ml H20) and one egg placed in each well containing a leaf disc. Single eggs were then transferred to individual wells.
The wells were covered with perforated Mylar film and incubated at 25 in a controlled temperature cabinet with lights (16 hr light, 8 hr dark). The larvae were removed at day 6 or 7 and individually weighed.
A bioassay was developed based on feeding larvae of H. arnaigera on cotton leaves homozygous for NaPI and coated with varying concentrations of CrylAc. The NaPI
leaves were specifically selected to give a small reduction in weight of the larvae. (In previous bioassays, without CrylAc, leaves have been used which give a greater effect on larval growth and development.) Preparation of Leaves The leaves were cut into discs using a hole puncher and coated with CrylAc solutions by immersing in the test solution for 5 minutes. The discs were removed from the solution with tweezers and then air dried on plastic mesh (approx. 30 min). When there was no visible moisture on the leaf disc surface, the discs were transferred to wells of a 24 well plate.
The wells contained filter paper moistened with water. Untransformed leaf material of cv Coker was used in control experiments.
Handling of Eggs H. armigera eggs (supplied by Department of Primary Industry & Fisheries, Indooroopilly, Queensland, Australia) were delivered by air freight to the laboratory and incubated (18-20 C depending on developmental stage) to the brown egg stage (pre hatching).
Eggs were suspended in polyacrylate (Aquakeep polyacrylate, 1 mg/ml H20) and one egg placed in each well containing a leaf disc. Single eggs were then transferred to individual wells.
The wells were covered with perforated Mylar film and incubated at 25 in a controlled temperature cabinet with lights (16 hr light, 8 hr dark). The larvae were removed at day 6 or 7 and individually weighed.
Preparation of CrylAc solution CrylAc crystal protein/spore mix (10 mg from CSIRO, Canberra, Australia) was dissolved in 0.9 ml of 50mM Na2 C03, (pH 10.5). Bovine Pancreatic Trypsin (Sigma T-8642 Type 8) stock 0.75 mg/ml was prepared in the same buffer and 0.1 ml of stock added to the crystal protein spore mix and incubated overnight at room temperature. The solution was centrifuged (14,000 rpm) for 5 minutes and the supernatant collected and stored at -70 C.
The concentration of activated CrylAc protein was estimated as 2.5 mg/ml on the basis of 50% starting material being CrylAc, and 50% of this being the final activated material.
Estimated molecular weights protoxin 133 kD: activated toxin 66 kD. Dilutions were prepared in 0.03% v/v Triton Ag 98 as wetting agent.
A suitable assay for CrylAc is described in Liao et al, Jlnvertebrate Pathol 80:55-63.
Experiment 1- Examination of the effect on mass of larvae In this experiment, sub-lethal concentrations of CrylAc were selected. The focus was to examine the effect on mass of the larvae. The second fully expanded leaf (from growing tip) of glasshouse grown plants was used for disc preparation. Experiment duration was 7 days.
Results The egg hatch was 70-100%; 12 eggs per test (1 egg/well). The mass of surviving larvae is shown in Figure 1. There was insignificant mortality.
It appears that both CrylAc and NaPI contribute to the decrease in weight of the larvae with increasing concentrations of CrylAc. (The result for cv Coker at 10 ug Cry 1 Ac seems anomalous.
The concentration of activated CrylAc protein was estimated as 2.5 mg/ml on the basis of 50% starting material being CrylAc, and 50% of this being the final activated material.
Estimated molecular weights protoxin 133 kD: activated toxin 66 kD. Dilutions were prepared in 0.03% v/v Triton Ag 98 as wetting agent.
A suitable assay for CrylAc is described in Liao et al, Jlnvertebrate Pathol 80:55-63.
Experiment 1- Examination of the effect on mass of larvae In this experiment, sub-lethal concentrations of CrylAc were selected. The focus was to examine the effect on mass of the larvae. The second fully expanded leaf (from growing tip) of glasshouse grown plants was used for disc preparation. Experiment duration was 7 days.
Results The egg hatch was 70-100%; 12 eggs per test (1 egg/well). The mass of surviving larvae is shown in Figure 1. There was insignificant mortality.
It appears that both CrylAc and NaPI contribute to the decrease in weight of the larvae with increasing concentrations of CrylAc. (The result for cv Coker at 10 ug Cry 1 Ac seems anomalous.
Experiment 2 - Effects of high concentration of CryIAc This experiment was designed to achieve mortality by using higher concentrations of CrylAc. Higher numbers of eggs were used. For Day 1, the third fully expanded leaf was used. For Day 4, additions of leaf material of the second fully expanded leaf was used.
Experiment duration was 6 days.
Results The egg hatch was 50-83%; 40 eggs per test (1 egg/well). The mass of surviving larvae is shown in Figure 2. The mortality is shown in Figure 3.
This experiment is essentially the same as experiment 1 except that the concentrations of CrylAc are increased over the range 50 to 250 ug/ml. These concentrations result in significant mortality, and thus the data presented refers to a subset of the original sample that have survived at each CrylAc concentration.
The difference in mass between the larvae feeding on Coker or NaPI leaves is essentially the same at each concentration of CrylAc over the range 50-250 ug/ml.
With no added CrylAc, there is not significant difference in mortality. At each concentration of CrylAc there is a significant enhancement of mortality in leaves expressing NaPI over that obtained with Coker control material.
In no case, in any of the experiments, did expression of the NaPI gene diminish the effect of CrylAc.
Experiment duration was 6 days.
Results The egg hatch was 50-83%; 40 eggs per test (1 egg/well). The mass of surviving larvae is shown in Figure 2. The mortality is shown in Figure 3.
This experiment is essentially the same as experiment 1 except that the concentrations of CrylAc are increased over the range 50 to 250 ug/ml. These concentrations result in significant mortality, and thus the data presented refers to a subset of the original sample that have survived at each CrylAc concentration.
The difference in mass between the larvae feeding on Coker or NaPI leaves is essentially the same at each concentration of CrylAc over the range 50-250 ug/ml.
With no added CrylAc, there is not significant difference in mortality. At each concentration of CrylAc there is a significant enhancement of mortality in leaves expressing NaPI over that obtained with Coker control material.
In no case, in any of the experiments, did expression of the NaPI gene diminish the effect of CrylAc.
Experiment 3 - Bioassay with third instar larvae The aim of this experiment was to determine the effect of Bt toxin (CrylAc) and NaPI on 3rd to 4th instar H. armigera larvae.
Larvae (48) were grown for 11 days on cotton leaves cv Coker before being transferred to new plates containing fresh leaf discs. The 48 larvae were separated into 4 treatments of 12 larvae.
The four treatments were as follows:
- Untransformed Coker 315 leaves - Transgenic NaPI leaves - Untransformed Coker leaves coated with 100 ug/mL CrylAc - Transgenic NaPI leaves coated with 100 ug/mL CrylAc The larvae were grown for a further 20 hours at 25 C. Larval weight was recorded at days 7, 11 and 12.
Experimental details Two, 24 well plates were used. Single H. armigera eggs, suspended in Polyacrylate (lg/L), were placed in each well. Initially one leaf disc was placed in each well.
Further discs were added when required.
Coker leaves at positions 3, 4 and 5 were used for the first 11 days. The leaves were harvested, cut into discs and pooled before they were added to the wells. Only leaves from position 2 or 3 were used for the treatments.
Larvae (48) were grown for 11 days on cotton leaves cv Coker before being transferred to new plates containing fresh leaf discs. The 48 larvae were separated into 4 treatments of 12 larvae.
The four treatments were as follows:
- Untransformed Coker 315 leaves - Transgenic NaPI leaves - Untransformed Coker leaves coated with 100 ug/mL CrylAc - Transgenic NaPI leaves coated with 100 ug/mL CrylAc The larvae were grown for a further 20 hours at 25 C. Larval weight was recorded at days 7, 11 and 12.
Experimental details Two, 24 well plates were used. Single H. armigera eggs, suspended in Polyacrylate (lg/L), were placed in each well. Initially one leaf disc was placed in each well.
Further discs were added when required.
Coker leaves at positions 3, 4 and 5 were used for the first 11 days. The leaves were harvested, cut into discs and pooled before they were added to the wells. Only leaves from position 2 or 3 were used for the treatments.
Results All H. armigera eggs used in the assay hatched (Table 1).At day 11 the larvae were weighed and then fed the test diets. About 20 hours later (day 12) the larvae were weighed again (Table 1, Figure 4).
Table 1. Results of H. arnaigera bioassay using 3ra instar larvae CrylAc concentration 0 ug/mL 100ug/mL
C N C N
No eggs 12 12 12 12 Hatched larvae 12 12 12 12 Surviving larvae 11 11 12 12 Average weight (mg) day 11 64.6 65.3 67.1 69.0 Average weight (mg) day 12 95.2 91.0 81.4 74.5 1o increase in mass day 11-12 47% 39% 21% 8%
C- untransforined Coker leaves, N- transgenic leaves expressing NaPI
There was a small difference in average weight gained between the larvae fed on control Coker leaves and the transgenic NaPI expressing leaves over the 20 hours. The control larvae increased their weight by an average of 48% while the NaPI fed larvae increased their weight by an average of 41% (Table 1, Figure 5). Note that the average weight of larvae at day 11 was slightly different for each test group (Table 1) and this must be taken into account in the calculations. Since the larvae were all fed the same diet, this difference is due to natural variation of the larvae.
There was a greater difference in weight when the larvae were fed CrylAc. The CrylAc fed larvae only increased their weight by an average of 21% (Table 1, Figure 5).
Table 1. Results of H. arnaigera bioassay using 3ra instar larvae CrylAc concentration 0 ug/mL 100ug/mL
C N C N
No eggs 12 12 12 12 Hatched larvae 12 12 12 12 Surviving larvae 11 11 12 12 Average weight (mg) day 11 64.6 65.3 67.1 69.0 Average weight (mg) day 12 95.2 91.0 81.4 74.5 1o increase in mass day 11-12 47% 39% 21% 8%
C- untransforined Coker leaves, N- transgenic leaves expressing NaPI
There was a small difference in average weight gained between the larvae fed on control Coker leaves and the transgenic NaPI expressing leaves over the 20 hours. The control larvae increased their weight by an average of 48% while the NaPI fed larvae increased their weight by an average of 41% (Table 1, Figure 5). Note that the average weight of larvae at day 11 was slightly different for each test group (Table 1) and this must be taken into account in the calculations. Since the larvae were all fed the same diet, this difference is due to natural variation of the larvae.
There was a greater difference in weight when the larvae were fed CrylAc. The CrylAc fed larvae only increased their weight by an average of 21% (Table 1, Figure 5).
Finally, larvae fed transgenic leaves coated with Cry1Ac only increased their weight by an average of 7%. It is interesting to note that 3 out of the 121arvae actually lost weight or did not increase in weight (Table 1, Figure 5).
These results suggest that the combination of NaPI and CrylAc increases the effect in diminishing larval growth. These results are in line with those obtained from Examples 1 and 2 in which freshly hatched larvae were allowed to feed for 7 days on test leaves.
These results suggest that the combination of NaPI and CrylAc increases the effect in diminishing larval growth. These results are in line with those obtained from Examples 1 and 2 in which freshly hatched larvae were allowed to feed for 7 days on test leaves.
Properties of midgut membrane When H arnaigera larvae are fed a diet containing NaPI or Bt proteins, such as CrylAc, there are different effects on the gut epithelial cells.
The presence of Bt in the diet of lepidopteran pests results in a swelling of the columnar cells and a proliferation of stem cells at the base of the epithelial layer.
Ultimately cells in the epithelial layer burst when exposed to Bt and related bacterial toxins. In contrast, the presence of NaPIs in the diet causes a limited swelling of cells in the epithelial layer without bursting. There is some associated movement of material into the intercellular spaces. When the diet contains both Bt and NaPI, there is an extensive and more rapid breakdown of the epithelial cell structures. The intercellular spaces are more distended and there is enhanced access of material to the haemolymph. Material which penetrates through to the haemolyinph may then come into contact with proteases involved in the innate immunity response. In particular, when PIs gain access to the haemolymph they can interfere with the protein cascades essential to this innate immunity response. The ability of the insect gut to regenerate after damage from either toxin is reduced.
Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to, or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.
The presence of Bt in the diet of lepidopteran pests results in a swelling of the columnar cells and a proliferation of stem cells at the base of the epithelial layer.
Ultimately cells in the epithelial layer burst when exposed to Bt and related bacterial toxins. In contrast, the presence of NaPIs in the diet causes a limited swelling of cells in the epithelial layer without bursting. There is some associated movement of material into the intercellular spaces. When the diet contains both Bt and NaPI, there is an extensive and more rapid breakdown of the epithelial cell structures. The intercellular spaces are more distended and there is enhanced access of material to the haemolymph. Material which penetrates through to the haemolyinph may then come into contact with proteases involved in the innate immunity response. In particular, when PIs gain access to the haemolymph they can interfere with the protein cascades essential to this innate immunity response. The ability of the insect gut to regenerate after damage from either toxin is reduced.
Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to, or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.
BIBLIOGRAPHY
Aronson Cell Mol Life Sci 59(3):417-425, 2002 Bennetzen & Hall, JBiol Chem 257:3026, 1982 Bernhard and Utz, An Environmental Biopesticide: Theory and Practice 255-267, Bogusz et al, Plant Cell 2(7):633-641, 1990 Brown and Ryan, Biochemistry 23:3418-3422, 1984 Bryant et al, Biochemistry 15:3418-3424, 1976 Bytebier et al, Proc Natl Acad Sci USA 84:5345-5349, 1987 Canevascini et al, Plant Physiol 112(2):513-524, 1996 Capana et al, Plant Mol Biol 25(4):681-691, 1994 Chen et al, Plant J 10:955-966, 1996 Chong et al, Transgenic Research 9:71-78, 2000 Christensen et al, Plant Mol Biol 12:619-632, 1989 Christensen et al, Plant Mol Biol 18:675-689, 1992 Christou et al, Plant Physiol 87:671-674, 1988 Christou and Ford Annals of Botany 75:407-413, 1995 Cordero et al, In: General Meeting of the International Program on Rice Biotechnology of the Rockefeller Foundation, Malacca, Malaysia, 1997 Cordero et al, Physiol Mol Plant Path 41:189-200, 1992 Corderok et al, Plant J 6(2) :141-150, 1994 Crickmore, et al, Microbiology and Molecular Biology Reviews 62:807-813, 1998 Crossway et al, Biotechniques 4:320-334, 1986 Czapla and Lang JEcon Entomol 83(6):2480-2485, 1990 Datta et al, Biotechnology 8:736-740, 1990 De Wet et al, The Experimental Manipulation of Ovule Tissues:197-209, 1985 D'Halluin et al, Plant Cell 4:1495-1505, 1992 Duan et al, Nature Biotechnology 14:494-498, 1996 Eckelkamp et al, FEBS Letters 323:73-76, 1993 Fang et al, Plant Cell 1:141-150, 1989 Finer and McMullen In Vitro Cell Dev Biol 27P:175-182, 1991 French et al, Anal Biochem 156:417-423, 1986 Fromm et al, Biotechnology 8:833-839, 1990 Gatz et al, Mol Gen Genet 227:229-237, 1991 Geiser et al, Gene 48:109, 1986 Gotor et al, Plant J 3:509-18, 1993 Graham et al, Plants 169:399-405, 1986 Green and Ryan, Science 776-777, 1972 Guevara-Garcia et al, Plant J 4(3):495-505, 1993 Guo et al, JBiol Chem 274:9836-9842, 1999 Hajdukiewicz et al, Plant Mol Biol 25:989-994, 1994 Hansen et al, Mol Gen Genet 254(3):337-343, 1997 Hire et al, Plant Mol Biol 20(2):207-218, 1992 Ho et al, Gene 77:51-59, 1989 Hofte and Whiteley Microbial Reviews 53:242-255, 1989 Hooykaas-Van Slogteren et al, Natzire (London) 311:763-764, 1984 Hsu et al, Plant Sci 143:63-70, 1999 Irie et al, Plant Mol Biol 30:149-157, 1996 Itakura, Science 198:1056-1063, 1977 Jones et al, Science 266:789, 1994 Kaeppler et al, Plant Cell Reports 9:415-418, 1990 Kaeppler et al, Theor Appl Genet 84:560-566, 1992 Kawamata et al, Plant Cell Physiol 38(7):792-803, 1997 Keil et al, EMBO J8:1323-1330, 1989 Keller and Baumgartner Plant Cell 3(10):1051-1061, 1991 Kirihara et al, Gene 71:359, 1988 Klein et al, Biotechnology 6:559-563, 1988 Klein et al, Plant Physiol 91:440-444, 1988 Klein et al, Proc Natl Acad Sci USA 85:4305-4309, 1988 Kunkel et al, Methods in Enzymol 154:367-3 82, 1987 Kunkel Proc Natl Acad Sci USA 82:488-492, 1985 Kuo et al, Arch Biochem Biophys 230:504-510, 1984 Kuster et al, Plant Mol Biol 29(4):759-772, 1995 Kwon et al, Plant Physiol 105:357-67, 1994 Lam Results Probl Cell Differ 20:181-196, 1994 Last et al, TheoN Appl Genet 81:581-588, 1991 Leach and Aoyagi (1991) Legavre et al, In: Vth International Congress of Plant Molecular Biology, Singapore, 1997 Lereclus et al, Bio/Technology 10:418, 1992 Li et al, Plant Cell Reports 12:250-255, 1993 Liao et al, Jlnvertebrate Pathol 80:55-63 Limerick Plant Science 79(1):69-76 McCabe et al, Bio/Technology 6:923-926, 1988 McCabe et al, Biotechnology 6:923-926, 1988 McElroy et al, Plant Cell 2:163-171, 1990 McGurl et al, Science 225:1570-1573, 1992 McNellis et al, Plant J 14(2):247-257, 1998 Marineau et al, Plant Mol Biol 9:335-342, 1987 Martin et al, Science 262:1432, 1993 Matsuoka et al, Proc Natl Acad Sci USA 90(20):9586-9590, 1993 Matton et al, Molecular Plant-Microbe Interactions 2:325-331, 1989 Maxam and Gilbert Methods in Enzymol 65:499-560, 1980 Melville and Ryan, Archives of Biochemistry and Biophysics 138:700-702, 1970 Miao et al, Plant Cell 3(1):11-22, 1991 Mindrinos et al, Cell 78:1089, 1994 Murray et al, Nucleic Acids Research 17:477-498, 1989 Musumura et al, Plant Mol Biol 12:123, 1989 Odell et al, Nature 313:810-812, 1985 Orozco et al, Plant Mol Biol 23(6):1129-1138, 1993 Osjoda et al, Nature Biotechnology 14:745-750, 1996 Paszkowski et al, EMBO J 3:2717-2722, 1984 Pathirana et al, Plant J 12:293-304, 1997 Pearce et al, Planta 175:527-531, 1988 Pedersen et al, JBiol Chem 261:6279, 1986 Plunkett et al, Arch Biochem Biophys 213:463-472, 1982 Pujade-Renaud et al, Plant Physiol Biochem 35:85-93, 1997).
Redolfi et al, Meth JPlant Pathol 89:245-254, 1983 Richard Phytochemistry 16:159-169, 1977 Rickauer et al, Plant Physiol 9:1065-1070, 1989 Riggs et al, Proc Natl Acad Sci USA 83:5602-5606, 1986 Rinehart et al, Plant Physiol 112(3):1331-1341, 1996 Rohmeier et al, Plant Mol Biol 22:783-792, 1993 Russell et al, Transgenic Res 6(2):157-168, 1997 Ryan Ann Rev Phytopath 28:425-449, 1990 Samac and Shah, Plant Cell 3:1063-1072, 1991 Sanford et al, Particulate Science and Technology 5:27-37, 1987 Sanger et al, Plant Mol Biol 14(3):433-443, 1990 Sanger Proc Natl Acad Sci USA 74:5463-5467, 1977 Schena et al, Proc Natl Acad Sci USA 88:10421-10425, 1991 Schnepf et al, Microbiol Mol Biol Rev 62(3):775-806, 1998 Schoenbeck et al, Molec Plant-Microbe Interact, 1999 Schubert et al, JBacteriol 170:5837-5847, 1988 Siebertz et al, Plant Cell 1:961-968, 1989 Singh et al, Theor Appl Genet 96:319-324, 1998 Somsisch et al, Mol Gen Genet 2:93-98, 1988 Somsisch et al, Proc Natl Acad Sci USA 83:2427-2430, 1986 Stanford et al, Mol Gen Genet 215:200-208, 1989 Teeri et al, EMBO J8(2):343-350, 1989 Thompson et al, BioEssays 10:108, 1989 Tomes et al, Plant Cell, Tissue, and Organ Culture: Fundamental Methods, 1995 Tu et al, Plant Molecular Biology 37:829-838, 1998 Uknes et al, Plant Cell 4:645-656, 1992 Van Camp et al, Plant Physiol 112(2):525-535, 1996 Van Loon Plant Mol Virol 4:111-116, 1985 Velten et al, EMBO J 3:2723-2730, 1984 Wada et al, Nucl Acids Res 18:2367-1411, 1990 Walker and Gaastra, Techniques in Molecular Biology 1983 Warner et al, Plant J 3:191-201, 1993 Weissinger et al, Ann Rev Genet 22:421-477, 1988 White et al, Trends in Genet 5:185-189, 1989 Williamson et al, Eur JBiochem 165:99-106,1987 Yainamoto et al, Plant Cell Physiol 35(5):773-778, 1994 Yamamoto et al, Plant J 12(2):255-265, 1997 Yang Proc Natl Acad Sci USA 93:14972-14977, 1996 Zhang et al, Proc Natl Acad Sci USA 91:2507-2511, 1994 Zhao et al, Plant Physiol 111:1299-1306, 1996
Aronson Cell Mol Life Sci 59(3):417-425, 2002 Bennetzen & Hall, JBiol Chem 257:3026, 1982 Bernhard and Utz, An Environmental Biopesticide: Theory and Practice 255-267, Bogusz et al, Plant Cell 2(7):633-641, 1990 Brown and Ryan, Biochemistry 23:3418-3422, 1984 Bryant et al, Biochemistry 15:3418-3424, 1976 Bytebier et al, Proc Natl Acad Sci USA 84:5345-5349, 1987 Canevascini et al, Plant Physiol 112(2):513-524, 1996 Capana et al, Plant Mol Biol 25(4):681-691, 1994 Chen et al, Plant J 10:955-966, 1996 Chong et al, Transgenic Research 9:71-78, 2000 Christensen et al, Plant Mol Biol 12:619-632, 1989 Christensen et al, Plant Mol Biol 18:675-689, 1992 Christou et al, Plant Physiol 87:671-674, 1988 Christou and Ford Annals of Botany 75:407-413, 1995 Cordero et al, In: General Meeting of the International Program on Rice Biotechnology of the Rockefeller Foundation, Malacca, Malaysia, 1997 Cordero et al, Physiol Mol Plant Path 41:189-200, 1992 Corderok et al, Plant J 6(2) :141-150, 1994 Crickmore, et al, Microbiology and Molecular Biology Reviews 62:807-813, 1998 Crossway et al, Biotechniques 4:320-334, 1986 Czapla and Lang JEcon Entomol 83(6):2480-2485, 1990 Datta et al, Biotechnology 8:736-740, 1990 De Wet et al, The Experimental Manipulation of Ovule Tissues:197-209, 1985 D'Halluin et al, Plant Cell 4:1495-1505, 1992 Duan et al, Nature Biotechnology 14:494-498, 1996 Eckelkamp et al, FEBS Letters 323:73-76, 1993 Fang et al, Plant Cell 1:141-150, 1989 Finer and McMullen In Vitro Cell Dev Biol 27P:175-182, 1991 French et al, Anal Biochem 156:417-423, 1986 Fromm et al, Biotechnology 8:833-839, 1990 Gatz et al, Mol Gen Genet 227:229-237, 1991 Geiser et al, Gene 48:109, 1986 Gotor et al, Plant J 3:509-18, 1993 Graham et al, Plants 169:399-405, 1986 Green and Ryan, Science 776-777, 1972 Guevara-Garcia et al, Plant J 4(3):495-505, 1993 Guo et al, JBiol Chem 274:9836-9842, 1999 Hajdukiewicz et al, Plant Mol Biol 25:989-994, 1994 Hansen et al, Mol Gen Genet 254(3):337-343, 1997 Hire et al, Plant Mol Biol 20(2):207-218, 1992 Ho et al, Gene 77:51-59, 1989 Hofte and Whiteley Microbial Reviews 53:242-255, 1989 Hooykaas-Van Slogteren et al, Natzire (London) 311:763-764, 1984 Hsu et al, Plant Sci 143:63-70, 1999 Irie et al, Plant Mol Biol 30:149-157, 1996 Itakura, Science 198:1056-1063, 1977 Jones et al, Science 266:789, 1994 Kaeppler et al, Plant Cell Reports 9:415-418, 1990 Kaeppler et al, Theor Appl Genet 84:560-566, 1992 Kawamata et al, Plant Cell Physiol 38(7):792-803, 1997 Keil et al, EMBO J8:1323-1330, 1989 Keller and Baumgartner Plant Cell 3(10):1051-1061, 1991 Kirihara et al, Gene 71:359, 1988 Klein et al, Biotechnology 6:559-563, 1988 Klein et al, Plant Physiol 91:440-444, 1988 Klein et al, Proc Natl Acad Sci USA 85:4305-4309, 1988 Kunkel et al, Methods in Enzymol 154:367-3 82, 1987 Kunkel Proc Natl Acad Sci USA 82:488-492, 1985 Kuo et al, Arch Biochem Biophys 230:504-510, 1984 Kuster et al, Plant Mol Biol 29(4):759-772, 1995 Kwon et al, Plant Physiol 105:357-67, 1994 Lam Results Probl Cell Differ 20:181-196, 1994 Last et al, TheoN Appl Genet 81:581-588, 1991 Leach and Aoyagi (1991) Legavre et al, In: Vth International Congress of Plant Molecular Biology, Singapore, 1997 Lereclus et al, Bio/Technology 10:418, 1992 Li et al, Plant Cell Reports 12:250-255, 1993 Liao et al, Jlnvertebrate Pathol 80:55-63 Limerick Plant Science 79(1):69-76 McCabe et al, Bio/Technology 6:923-926, 1988 McCabe et al, Biotechnology 6:923-926, 1988 McElroy et al, Plant Cell 2:163-171, 1990 McGurl et al, Science 225:1570-1573, 1992 McNellis et al, Plant J 14(2):247-257, 1998 Marineau et al, Plant Mol Biol 9:335-342, 1987 Martin et al, Science 262:1432, 1993 Matsuoka et al, Proc Natl Acad Sci USA 90(20):9586-9590, 1993 Matton et al, Molecular Plant-Microbe Interactions 2:325-331, 1989 Maxam and Gilbert Methods in Enzymol 65:499-560, 1980 Melville and Ryan, Archives of Biochemistry and Biophysics 138:700-702, 1970 Miao et al, Plant Cell 3(1):11-22, 1991 Mindrinos et al, Cell 78:1089, 1994 Murray et al, Nucleic Acids Research 17:477-498, 1989 Musumura et al, Plant Mol Biol 12:123, 1989 Odell et al, Nature 313:810-812, 1985 Orozco et al, Plant Mol Biol 23(6):1129-1138, 1993 Osjoda et al, Nature Biotechnology 14:745-750, 1996 Paszkowski et al, EMBO J 3:2717-2722, 1984 Pathirana et al, Plant J 12:293-304, 1997 Pearce et al, Planta 175:527-531, 1988 Pedersen et al, JBiol Chem 261:6279, 1986 Plunkett et al, Arch Biochem Biophys 213:463-472, 1982 Pujade-Renaud et al, Plant Physiol Biochem 35:85-93, 1997).
Redolfi et al, Meth JPlant Pathol 89:245-254, 1983 Richard Phytochemistry 16:159-169, 1977 Rickauer et al, Plant Physiol 9:1065-1070, 1989 Riggs et al, Proc Natl Acad Sci USA 83:5602-5606, 1986 Rinehart et al, Plant Physiol 112(3):1331-1341, 1996 Rohmeier et al, Plant Mol Biol 22:783-792, 1993 Russell et al, Transgenic Res 6(2):157-168, 1997 Ryan Ann Rev Phytopath 28:425-449, 1990 Samac and Shah, Plant Cell 3:1063-1072, 1991 Sanford et al, Particulate Science and Technology 5:27-37, 1987 Sanger et al, Plant Mol Biol 14(3):433-443, 1990 Sanger Proc Natl Acad Sci USA 74:5463-5467, 1977 Schena et al, Proc Natl Acad Sci USA 88:10421-10425, 1991 Schnepf et al, Microbiol Mol Biol Rev 62(3):775-806, 1998 Schoenbeck et al, Molec Plant-Microbe Interact, 1999 Schubert et al, JBacteriol 170:5837-5847, 1988 Siebertz et al, Plant Cell 1:961-968, 1989 Singh et al, Theor Appl Genet 96:319-324, 1998 Somsisch et al, Mol Gen Genet 2:93-98, 1988 Somsisch et al, Proc Natl Acad Sci USA 83:2427-2430, 1986 Stanford et al, Mol Gen Genet 215:200-208, 1989 Teeri et al, EMBO J8(2):343-350, 1989 Thompson et al, BioEssays 10:108, 1989 Tomes et al, Plant Cell, Tissue, and Organ Culture: Fundamental Methods, 1995 Tu et al, Plant Molecular Biology 37:829-838, 1998 Uknes et al, Plant Cell 4:645-656, 1992 Van Camp et al, Plant Physiol 112(2):525-535, 1996 Van Loon Plant Mol Virol 4:111-116, 1985 Velten et al, EMBO J 3:2723-2730, 1984 Wada et al, Nucl Acids Res 18:2367-1411, 1990 Walker and Gaastra, Techniques in Molecular Biology 1983 Warner et al, Plant J 3:191-201, 1993 Weissinger et al, Ann Rev Genet 22:421-477, 1988 White et al, Trends in Genet 5:185-189, 1989 Williamson et al, Eur JBiochem 165:99-106,1987 Yainamoto et al, Plant Cell Physiol 35(5):773-778, 1994 Yamamoto et al, Plant J 12(2):255-265, 1997 Yang Proc Natl Acad Sci USA 93:14972-14977, 1996 Zhang et al, Proc Natl Acad Sci USA 91:2507-2511, 1994 Zhao et al, Plant Physiol 111:1299-1306, 1996
Claims (61)
1. A genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest.
2. The genetically modified plant of Claim 1 wherein the serine proteinase inhibitor is provided in precursor form.
3. The genetically modified plant of Claim 2 wherein the precursor comprises at least four monomers.
4. The genetically modified plant of Claim 2 wherein the precursor comprises at least five monomers.
5. The genetically modified plant of Claim 2 wherein the precursor comprises at least six monomers.
6. The genetically modified plant of Claim 5 wherein the serine proteinase inhibitor is NaPI from Nicotiana alata.
7. The genetically modified plant of any one of Claims 1 to 6 wherein the non-serine proteinase inhibitor is an endotoxin.
8. The genetically modified plant of Claim 7 wherein the non-serine proteinase inhibitor is a Bt protein.
9. The genetically modified plant of Claim 7 or 8 wherein the endotoxin is selected from the list conistsing of a member of the Cry family and VIP.
10. The genetically modified plant of any one of Claims 1 to 6 wherein the non-serine proteinase inhibitor is a defensin molecule or a pest toxic part thereof.
11. A genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is the NaPI precursor or a monomer thereof or a recombinant or derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest.
12. The genetically modified plant of any one of Claim 11 wherein the non-serine proteinase inhibitor is an endotoxin.
13. The genetically modified plant of Claim 12 wherein the non-serine proteinase inhibitor is a Bt protein.
14. The genetically modified plant of Claim 12 or 13 wherein the endotoxin is selected from the list conistsing of a member of the Cry familyand VIP.
15. The genetically modified plant of Claim 11 wherein the non-serine proteinase inhibitor is a defensin molecule or a pest toxic part thereof.
16. A genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest except and wherein at least one other agent is selected from the list comprising a Bt protein, a Cry family protein, a VIP and a defensin.
17. A genetically modified plant or its progeny resulting from self crossing, back crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is NaPI precursor or a monomer thereof or a recombinant or modular derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest except wherein the least one other agent is selected from the list comprising a Bt protein, a member of the Cry family, a VIP and a defensin.
18. The genetically modified plant of Claim 1 or 11 or 16 or 17 wherein the plant is selected from the list consisting of corn (Zea mays), Brassica sp. (e.g., B.
napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceum), foxtail millet (Setaria italica), finger millet (Eleusine coracana), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassaya (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera), cotton pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers, vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C, cantalupensis), and musk melon (C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis). Plants of the present invention include crop plants (for example, cotton, corn, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants, turfgrasses include, but are not limited to: annual bluegrass (Poa annua);
annual ryegrass (Lolium multiflorum); Canada bluegrass (Poa compressa); Chewings fescue (Festuca rubra); colonial bentgrass (Agrostis tenuis); creeping bentgrass (Agrostis palustris);
crested wheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyron cristatum);
hard fescue (Festuca longifolia); Kentucky bluegrass (Poa pratensis);
orchardgrass (Dactylis glomerata); perennial ryegrass (Lolium perenne); red fescue (Festuca rubra);
redtop (Agrostis alba); rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina);
smooth bromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy (Phleum pratense); velvet bentgrass (Agrostis canina); weeping alkaligrass (Puccinellia distans);
western wheatgrass (Agropyron smithii); Bermuda grass (Cynodon spp.); St.
Augustine grass (Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass (Paspalum notatum); carpet grass (Axonopus affinis); centipede grass (Eremochloa ophiuroides);
kikuyu grass (Pennisetum clandesinum); seashore paspalum (Paspalum vaginatum);
blue gramma (Bouteloua gracilis); buffalo grass (Buchloe dactyloids); sideoats gramma (Bouteloua curtipendula), oil-seed plants, leguminous plants, corn, wheat, barley, rice, sorghum, rye, millet, cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, flax, castor, olive beans, peas, guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils and chickpea.
napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceum), foxtail millet (Setaria italica), finger millet (Eleusine coracana), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassaya (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera), cotton pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers, vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C, cantalupensis), and musk melon (C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis). Plants of the present invention include crop plants (for example, cotton, corn, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants, turfgrasses include, but are not limited to: annual bluegrass (Poa annua);
annual ryegrass (Lolium multiflorum); Canada bluegrass (Poa compressa); Chewings fescue (Festuca rubra); colonial bentgrass (Agrostis tenuis); creeping bentgrass (Agrostis palustris);
crested wheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyron cristatum);
hard fescue (Festuca longifolia); Kentucky bluegrass (Poa pratensis);
orchardgrass (Dactylis glomerata); perennial ryegrass (Lolium perenne); red fescue (Festuca rubra);
redtop (Agrostis alba); rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina);
smooth bromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy (Phleum pratense); velvet bentgrass (Agrostis canina); weeping alkaligrass (Puccinellia distans);
western wheatgrass (Agropyron smithii); Bermuda grass (Cynodon spp.); St.
Augustine grass (Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass (Paspalum notatum); carpet grass (Axonopus affinis); centipede grass (Eremochloa ophiuroides);
kikuyu grass (Pennisetum clandesinum); seashore paspalum (Paspalum vaginatum);
blue gramma (Bouteloua gracilis); buffalo grass (Buchloe dactyloids); sideoats gramma (Bouteloua curtipendula), oil-seed plants, leguminous plants, corn, wheat, barley, rice, sorghum, rye, millet, cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, flax, castor, olive beans, peas, guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils and chickpea.
19. The genetically modified plant of Claim 18 wherein the plant is a cotton, sweet corn, tomato, tobacco, piniento, potato, sunflower, citrus, plums, sorghum, leeks, soybean, alfalfa, beans, pidgeon peas, chick peas, artichokes, curcurbits, lettuce, Dianthus, geraniums, cape gooseberry, maize, flax and linseed, lupins, broad beans, garden peas, peanuts, canola, snapdragons, cherry, sunflower, pot marigolds, Helichrysum, wheat, barley, oats, triticale, carrots, onions, orchids, roses or petunias.
20. The genetically modified plant of Claim 1 or 11 or 16 or 17 wherein the plant is cotton.
21. The genetically modified plant of Claim 1 or 11 or 16 or 17 wherein the plant pest is selected from the listing consisting of insects, fungi, bacteria, nematodes, acarids, protozoan pathogens, animal-parasitic liver flukes, and the like.
22. The genetically modified plant of Claim 21 wherein the pest is an insect.
23. The genetically modified plant of Claim 22 wherein the insect is selected from the list consisting of Helicoverpa, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, Coleoptera.
24. The genetically modified plant of Claim 22 wherein the pest is selected from the list consisting of Achoroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura sp., Cochylls hospes, Colias eurytheme, Corcyra cephalonica, Cydia latiferreanus, Cydia pomonella, Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphania hyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraea saccharalis, Ennomos subsignaria, Eoreuma loftini, Esphestia elutella, Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoellia ambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoa messoria, Galleria mellonella, Grapholita molesta, Harrisina americana, Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemileuca oliviae, Homoeosoma electellum, Hyphantia cunea, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis, Lymantria dispar, Macalla thyrisalis, Malacosoma sp., Mamestra brassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta, Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp., Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes, Pectinophora gossypiella, Phryganidia californica, Phyllonorycter blancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella, Pontia protodice, Pseudaletia unipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizura concinna, Sitotroga cerealella, Spilonta ocellana, Spodoptera sp., Thaurnstopoea pityocampa, Tinsola bisselliella, Trichoplusia hi, Udea rubigalis, Xylomyges curiails, and Yponomeuta padella, Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Helicoverpa zeae, corn earworm; Spodoptera frugiperda, fall armyworm; Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer;
Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; northern corn rootworm, e.g., Diabrotica longicornis barberi;
southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubruna, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle, Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae.
Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbean weevil; Bruchus pisorum, pea weevil;
Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio;
Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil;
Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus majalis, European chafer; Macrodactylus subspinosus, rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor and yellow mealworm.
Helicoverpa zeae, corn earworm; Spodoptera frugiperda, fall armyworm; Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer;
Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; northern corn rootworm, e.g., Diabrotica longicornis barberi;
southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubruna, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle, Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae.
Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbean weevil; Bruchus pisorum, pea weevil;
Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio;
Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil;
Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus majalis, European chafer; Macrodactylus subspinosus, rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor and yellow mealworm.
25. The genetically modified plant of Claim 22 wherein the pest is a nematode selected from the list consisting of Heterodera spp., Meloidogyne spp., and Globodera spp.;
particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode);
Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes) and lesion nematodes including Pratylenchus spp.
particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode);
Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes) and lesion nematodes including Pratylenchus spp.
26. A pest management control system comprising generating a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest.
27. The pest management control system of Claim 26 wherein the serine proteinase inhibitor is provided in precursor form.
28. The pest management control system of Claim 27 wherein the precursor comprises at least four monomers.
29. The pest management control system of Claim 27 wherein the precursor comprises at least five monomers.
30. The pest management control system of Claim 27 wherein the precursor comprises at least six monomers.
31. The pest management control system of Claim 30 wherein the serine proteinase inhibitor is NaPI from Nicotiana alata.
32. The pest management control system of any one of Claims 26 to 31 wherein the non-serine proteinase ibhibitor is an endotoxin.
33. The pest management control system of Claim 32 wherein the non-serine proteinase inhibitor is a Bt protein.
34. The pest management control system of Claim 32 or 33 wherein the endotoxin is selected from the list conistsing of a member of Cry family and VIP.
35. The pest management control system of any one of Claims 26 to 31 wherein the non-serine proteinase inhibitor is a defensin molecule or a pest toxic part thereof.
36. A pest management control system comprising generating a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is the NaPI precursor or a monomer thereof or a recombinant or modular derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest.
37. The pest management control system of Claim 36 wherein the non-serine proteinase inhibitor is an endotoxin.
38. The pest management control system of Claim 37 wherein the non-serine proteinase inhibitor is a Bt protein.
39. The pest management control system of Claim 37 or 38 wherein the endotoxin is selected from the list conistsing of a member of the Cry family and VIP.
40. The pest management control system of Claim 36 wherein the non-serine proteinase inhibitor is a defensin molecule or a pest toxic part thereof.
41. A pest management control system comprising generating a genetically modified plant or its progeny resulting from self-crossing, back-crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is a serine proteinase inhibitor or precursor thereof from the Solanaceae family wherein in precursor form, it comprises at least three monomers wherein at least one monomer is a trypsin inhibitor and at least one monomer is a chymotrypsin inhibitor and wherein at least one other agent is a non-serine proteinase inhibitor, wherein said plant exhibits resistance or reduced susceptibility to a pest except and wherein at least one other agent is selected from the list comprising a Bt protein, a Cry family protein, a VIP and a defensin.
42. A pest management control system comprising generating a genetically modified plant or its progeny resulting from self crossing, back crossing or crossing with another plant, said plant comprising at least two pesticidal and/or pestistatic agents wherein at least one of said agents results from the genetic modification or crossing wherein at least one agent is NaPI precursor or a monomer thereof or a recombinant or modular derivative thereof and wherein at least one other agent is a non-serine proteinase inhibitor wherein said plant exhibits resistance or reduced susceptibility to a pest except wherein the least one other agent is selected from the list comprising a Bt protein, a Cry family protein, a VIP and a defensin.
43. A pest management control system of Claim 26 or 36 or 41 or 42 wherein the plant is selected from the list consisting of corn (Zea mays), Brassica sp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet (Pennisetum glaucum), proso millet (Panicum miliaceum), foxtail millet (Setaria italica), finger millet (Eleusine coracana), sunflower (Helianthus annuus), safflower (Carthamus tinctorius), wheat (Triticum aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium barbadense, Gossypium hirsutum), sweet potato (Ipomoea batatus), cassaya (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera), cotton pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidentale), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers, vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C.
cantalupensis), and musk melon (C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis). Plants of the present invention include crop plants (for example, cotton, corn, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants, turfgrasses include, but are not limited to: annual bluegrass (Poa annua);
annual ryegrass (Lolium multiflorum); Canada bluegrass (Poa compressa); Chewings fescue (Festuca rubra); colonial bentgrass (Agrostis tenuis); creeping bentgrass (Agrostis palustris);
crested wheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyron cristatum);
hard fescue (Festuca longifolia); Kentucky bluegrass (Poa pratensis);
orchardgrass (Dactylis glomerata); perennial ryegrass (Lolium perenne); red fescue (Festuca rubra);
redtop (Agrostis alba); rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina);
smooth bromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy (Phleum pratense); velvet bentgrass (Agrostis canina); weeping alkaligrass (Puccinellia distans);
western wheatgrass (Agropyron smithii); Bermuda grass (Cynodon spp.); St.
Augustine grass (Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass (Paspalum notatum); carpet grass (Axonopus affinis); centipede grass (Eremochloa ophiuroides);
kikuyu grass (Pennisetum clandesinum); seashore paspalum (Paspalum vaginatum);
blue gramma (Bouteloua gracilis); buffalo grass (Buchloe dactyloids); sideoats gramma (Bouteloua curtipendula), oil-seed plants, leguminous plants, corn, wheat, barley, rice, sorghum, rye, millet, cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, flax, castor, olive beans, peas, guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils and chickpea.
cantalupensis), and musk melon (C. melo). Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii); Western hemlock (Tsuga canadensis); Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis). Plants of the present invention include crop plants (for example, cotton, corn, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.), such as corn and soybean plants, turfgrasses include, but are not limited to: annual bluegrass (Poa annua);
annual ryegrass (Lolium multiflorum); Canada bluegrass (Poa compressa); Chewings fescue (Festuca rubra); colonial bentgrass (Agrostis tenuis); creeping bentgrass (Agrostis palustris);
crested wheatgrass (Agropyron desertorum); fairway wheatgrass (Agropyron cristatum);
hard fescue (Festuca longifolia); Kentucky bluegrass (Poa pratensis);
orchardgrass (Dactylis glomerata); perennial ryegrass (Lolium perenne); red fescue (Festuca rubra);
redtop (Agrostis alba); rough bluegrass (Poa trivialis); sheep fescue (Festuca ovina);
smooth bromegrass (Bromus inermis); tall fescue (Festuca arundinacea); timothy (Phleum pratense); velvet bentgrass (Agrostis canina); weeping alkaligrass (Puccinellia distans);
western wheatgrass (Agropyron smithii); Bermuda grass (Cynodon spp.); St.
Augustine grass (Stenotaphrum secundatum); zoysia grass (Zoysia spp.); Bahia grass (Paspalum notatum); carpet grass (Axonopus affinis); centipede grass (Eremochloa ophiuroides);
kikuyu grass (Pennisetum clandesinum); seashore paspalum (Paspalum vaginatum);
blue gramma (Bouteloua gracilis); buffalo grass (Buchloe dactyloids); sideoats gramma (Bouteloua curtipendula), oil-seed plants, leguminous plants, corn, wheat, barley, rice, sorghum, rye, millet, cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, flax, castor, olive beans, peas, guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils and chickpea.
44. The pest management control system of Claim 43 wherein the plant is a cotton, sweet corn, tomato, tobacco, piniento, potato, sunflower, citrus, plums, sorghum, leeks, soybean, alfalfa, beans, pidgeon peas, chick peas, artichokes, curcurbits, lettuce, Dianthus, geraniums, cape gooseberry, maize, flax and linseed, lupins, broad beans, garden peas, peanuts, canola, snapdragons, cherry, sunflower, pot marigolds, Helichrysum, wheat, barley, oats, triticale, carrots, onions, orchids, roses and petunias.
45. The pest management control system of Claim 26 or 36 or 41 or 42 wherein the plant is cotton.
46. The pest management control system of Claim 26 or 36 or 41 or 42 wherein the plant pest is selected from the listing consisting of insects, fungi, bacteria, nematodes, acarids, protozoan pathogens, animal-parasitic liver flukes, and the like.
47. The pest management control system of Claim 46 wherein the pest is an insect.
48. The pest management control system of Claim 47 wherein the insect is selected from the list consisting of Helicoverpa, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, Coleoptera.
49. The pest management control system of Claim 47 wherein the pest is selected from the list consisting of Lepidoptera, e.g. Achoroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura sp., Cochylls hospes, Colias eurytheme, Corcyra cephalonica, Cydia latiferreanus, Cydia pomonella, Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphania hyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraea saccharalis, Ennomos subsignaria, Eoreuma loftini, Esphestia elutella, Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoellia ambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoa messoria, Galleria mellonella, Grapholita molesta, Harrisina americana, Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemileuca oliviae, Homoeosoma electellum, Hyphantia cunea, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis, Lymantria dispar, Macalla thyrisalis, Malacosoma sp., Mamestra brassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta, Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp., Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes, Pectinophora gossypiella, Phryganidia calfornica, Phyllonorycter blancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella, Pontia protodice, Pseudaletia unipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizura concinna, Sitotroga cerealella, Spilonta ocellana, Spodoptera sp., Thaurnstopoea pityocampa, Tinsola bisselliella, Trichoplusia hi, Udea rubigalis, Xylomyges curiails, and Yponomeuta padella, Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Helicoverpa zeae, corn earworm; Spodoptera frugiperda, fall armyworm;
Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; northern corn rootworm, e.g., Diabrotica longicornis barberi; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle, Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae.
Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbean weevil; Bruchus pisorum, pea weevil;
Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio;
Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil;
Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus majalis, European chafer; Macrodactylus subspinosus, rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor and yellow mealworm.
Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; northern corn rootworm, e.g., Diabrotica longicornis barberi; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet armyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle, Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae.
Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus, broadbean weevil; Bruchus pisorum, pea weevil;
Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio;
Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil;
Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus majalis, European chafer; Macrodactylus subspinosus, rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor and yellow mealworm.
50. The pest management control system of Claim 47 wherein the pest is a nematode selected from the list consisting of Heterodera spp., Meloidogyne spp., and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode);
Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes) and lesion nematodes including Pratylenchus spp.
Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes) and lesion nematodes including Pratylenchus spp.
51. A pesticidal or pestistatic composition comprising a serine proteinase inhibitor and at least one non-serine proteinase inhibitor.
52. The pesticidal or pestistatic composition of Claim 51 wherein the serine proteinase inhibitor is NaPI from Nicotiana alata.
53. The pesticidal or pestistatic composition of Claim 51 or 52 wherein the non-serine proteinase ibhibitor is an endotoxin.
54. The pesticidal or pestistatic composition of Claim 53 wherein the non-serine proteinase inhibitor is a Bt protein.
55. The pesticidal or pestistatic composition of Claim 53 or 54 wherein the endotoxin is selected from the list conistsing of a member of the Cry family and VIP.
56. The pesticidal or pestistatic composition of Claim 51 wherein the non-serine proteinase inhibitor is a defensin molecule or a pest toxic part thereof.
57. The pesticidal or pestistatic composition of Claim 51 wherein the plant pest is selected from the listing consisting of insects, fungi, bacteria, nematodes, acarids, protozoan pathogens, animal-parasitic liver flukes, and the like.
58. The pesticidal or pestistatic composition of Claim 57 wherein the pest is an insect.
59. The pesticidal or pestistatic composition of Claim 58 wherein the insect is selected from the list consisting of Helicoverpa, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera and Coleoptera.
60. The pesticidal or pestistatic composition of Claim 58 wherein the pest is selected from the list consisting of Lepidoptera, e.g. Achoroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura sp., Cochylls hospes, Colias eurytheme, Corcyra cephalonica, Cydia latiferreanus, Cydia pomonella, Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphania hyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraea saccharalis, Ennomos subsignaria, Eoreuma loftini, Esphestia elutella, Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoellia ambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoa messoria, Galleria mellonella, Grapholita molesta, Harrisina americana, Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemileuca oliviae, Homoeosoma electellum, Hyphantia cunea, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis, Lymantria dispar, Macalla thyrisalis, Malacosoma sp., Mamestra brassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta, Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp., Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes, Pectinophora gossypiella, Phryganidia californica, Phyllonorycter blancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella, Pontia protodice, Pseudaletia unipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizura concinna, Sitotroga cerealella, Spilonta ocellana, Spodoptera sp., Thaurnstopoea pityocampa, Tinsola bisselliella, Trichoplusia hi, Udea rubigalis, Xylomyges curiails, and Yponomeuta padella, Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Helicoverpa zeae, corn earworm; Spodoptera frugiperda, fall armyworm;
Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; northern corn rootworm, e.g., Diabrotica longicornis barberi; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet annyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle, Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae.
Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufinanus, broadbean weevil; Bruchus pisorum, pea weevil;
Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio;
Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil;
Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus majalis, European chafer; Macrodactylus subspinosus, rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor and yellow mealworm.
Diatraea grandiosella, southwestern corn borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera; northern corn rootworm, e.g., Diabrotica longicornis barberi; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms;
Cyclocephala borealis, northern masked chafer (white grub); Cyclocephala immaculata, southern masked chafer (white grub); Popillia japonica, Japanese beetle;
Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis, corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes, migratory grasshopper; Hylemya platura, seedcorn maggot; Agromyza parvicornis, corn blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant;
Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer;
Spodoptera frugiperda, fall armyworm; Helicoverpa zea, corn earworm;
Elasmopalpus lignosellus, leser cornstalk borer; Feltia subterranea, granulate cutworm;
Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis, maize billbug; Rhopalosiphum maidis; corn leaf aphid; Sipha flava, yellow sugarcane aphid;
chinch bug, e.g., Blissus leucopterus leucopterus; Contarinia sorghicola, sorghum midge;
Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly;
Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil;
Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
Neolasioptera murtfeldtiana, sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Spodoptera exigua, beet annyworm;
Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper;
Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug;
Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper;
Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice:
Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm;
Helicoverpa zea, corn earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil; Sitophilus oryzae, rice weevil; Nephotettix nigropictus, rice leafhoper; chinch bug, e.g., Blissus leucopterus leucopterus; Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticarsia gemmatalis, velvetbean caterpillar; Plathypena scabra, green cloverworm; Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm; Spodoptera exigua, beet armyworm;
Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips;
Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European corn borer; Agrotis ipsilon, black cutworm;
Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus;
Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug;
Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite;
Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens;
Potato:
Leptinotarsa decemlineata, Colorado potato beetle, Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus, Poecilocapsus lineatus, Blissus leucopterus, Nysius ericae, Nysius raphanus, Euschistus servus, Nezara viridula, Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, and Cimicidae.
Pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufinanus, broadbean weevil; Bruchus pisorum, pea weevil;
Zabrotes subfasciatus, Mexican bean weevil; Diabrotica balteata, banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus quadrigibbus, apple curculio;
Sternechus paludatus, bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil;
Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio;
Sitophilus zeamais, maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus, West Indian sweet potato weevil; Maladera castanea, Asiatic garden beetle;
Rhizotrogus majalis, European chafer; Macrodactylus subspinosus, rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor and yellow mealworm.
61. The pesticidal and pestistatic composition of Claim 58 wherein the pest is a nematode selected from the list consisting of Heterodera spp., Meloidogyne spp., and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode);
Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes) and lesion nematodes including Pratylenchus spp.
Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes) and lesion nematodes including Pratylenchus spp.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US69777605P | 2005-07-08 | 2005-07-08 | |
US60/697,776 | 2005-07-08 | ||
PCT/AU2006/000952 WO2007006079A1 (en) | 2005-07-08 | 2006-07-06 | Management of plant pathogens |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2614353A1 true CA2614353A1 (en) | 2007-01-18 |
Family
ID=37636655
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002614353A Abandoned CA2614353A1 (en) | 2005-07-08 | 2006-07-06 | Management of plant pathogens |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP1909559A4 (en) |
CN (1) | CN101257792A (en) |
AU (1) | AU2006269808A1 (en) |
BR (1) | BRPI0612634A2 (en) |
CA (1) | CA2614353A1 (en) |
WO (1) | WO2007006079A1 (en) |
Families Citing this family (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BRPI0916867A2 (en) * | 2008-08-05 | 2017-05-23 | Hexima Ltd | methods for protecting a plant from a disease associated with pathogen infection, and for identifying a defensive, use of a plant defensin and a proteinase inhibitor or a precursor form, genetically modified plant or progeny thereof, and defensive |
US9889184B2 (en) | 2008-08-05 | 2018-02-13 | Hexima Limited | Anti-pathogen systems |
NZ603561A (en) * | 2010-04-23 | 2015-06-26 | Dow Agrosciences Llc | Combinations including cry34ab/35ab and cry3aa proteins to prevent development of resistance in corn rootworms (diabrotica spp.) |
CN102037964B (en) * | 2011-01-06 | 2014-01-22 | 兰州大学 | Application of farrerol in preparing bactericides |
CN104244705B (en) | 2012-01-27 | 2018-05-22 | 德克萨斯A&M大学系统 | The citrus composition of pathogen-resistance, organism, system and method |
CN105145648A (en) * | 2015-07-20 | 2015-12-16 | 邱永力 | Agent for killing cotton aphids and manufacturing method for agent |
CN105850897B (en) * | 2016-05-09 | 2018-10-26 | 漯河市森林病虫害防治检疫站 | A kind of artificial fecundation method of Tetrastichus septentrionalis |
KR101929388B1 (en) | 2017-11-10 | 2018-12-17 | 대한민국 | Composition for prevention, treatment or improvement of liver disease comprising italian ryegrass extraction fraction as an active ingredient |
CN108314728B (en) * | 2018-02-07 | 2020-01-17 | 中国农业科学院植物保护研究所 | Alfalfa trypsin inhibitor MT-mth2-89i19, and coding gene and application thereof |
CN108059671B (en) * | 2018-02-07 | 2020-01-17 | 中国农业科学院植物保护研究所 | Alfalfa trypsin inhibitor MT-mth2-36p5, and coding gene and application thereof |
CN111513065A (en) * | 2020-05-15 | 2020-08-11 | 中国农业科学院植物保护研究所 | Coffee bean weevil attractant composition and application thereof |
CN112175946B (en) * | 2020-06-30 | 2022-10-21 | 华南农业大学 | miR-283-3p of targeted Trypsin-9 gene and application thereof |
CN112471079B (en) * | 2020-12-11 | 2022-02-01 | 兰州大学 | Indoor artificial feeding method for Qinghai-Tibet plateau grassland caterpillars |
CN114532175B (en) * | 2022-02-18 | 2023-10-27 | 新疆农垦科学院 | Identification method for physiological aphid resistance based on fuzz characteristics of cotton leaves |
CN117481146B (en) * | 2023-12-29 | 2024-03-08 | 西北农林科技大学深圳研究院 | Agricultural composition containing synergistic agent |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
TR24186A (en) * | 1988-04-11 | 1991-05-30 | Monsanto Co | YOENTEM TO INCREASE THE ACTIVITY OF POISONOUS POISONS |
DE69333782T2 (en) * | 1992-12-16 | 2006-01-26 | Hexima Ltd., Melbourne | A PROTEINASE INHIBITOR, PRESENTER, AND GENETIC SEQUENCES THEREFOR |
EP1263281B1 (en) * | 2000-02-29 | 2012-04-11 | Auburn University | Multiple gene expression for engineering novel pathways and hyperexpression of foreign proteins in plants |
CA2437606A1 (en) * | 2001-02-08 | 2002-08-15 | Marilyn Anne Anderson | Plant-derived molecules and genetic sequences encoding same and uses therefor |
-
2006
- 2006-07-06 CA CA002614353A patent/CA2614353A1/en not_active Abandoned
- 2006-07-06 BR BRPI0612634A patent/BRPI0612634A2/en not_active IP Right Cessation
- 2006-07-06 EP EP06752677A patent/EP1909559A4/en not_active Withdrawn
- 2006-07-06 AU AU2006269808A patent/AU2006269808A1/en not_active Abandoned
- 2006-07-06 WO PCT/AU2006/000952 patent/WO2007006079A1/en active Application Filing
- 2006-07-06 CN CNA2006800326476A patent/CN101257792A/en active Pending
Also Published As
Publication number | Publication date |
---|---|
CN101257792A (en) | 2008-09-03 |
EP1909559A4 (en) | 2009-01-21 |
WO2007006079A1 (en) | 2007-01-18 |
EP1909559A1 (en) | 2008-04-16 |
AU2006269808A1 (en) | 2007-01-18 |
BRPI0612634A2 (en) | 2017-06-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US7629504B2 (en) | Bacillus thuringiensis cry9 nucleic acids | |
CA2551102C (en) | Genes encoding proteins with pesticidal activity | |
US7696412B2 (en) | Genes encoding novel Bacillus thuringiensis proteins with pesticidal activity against Coleopterans | |
US7378499B2 (en) | Genes encoding proteins with pesticidal activity | |
CA2611539C (en) | Insect-specific protease recognition sequences | |
CA2614353A1 (en) | Management of plant pathogens | |
US7629449B2 (en) | Nucleic acid molecules encoding cysteine proteases | |
ABAD et al. | Patent 2551102 Summary | |
MXPA06007245A (en) | Genes encoding proteins with pesticidal activity | |
MXPA06007246A (en) | Plant activation of insect toxin |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
FZDE | Discontinued | ||
FZDE | Discontinued |
Effective date: 20100706 |