OA20372A - Dominikia Sp. strain, compositions comprising it and uses. - Google Patents
Dominikia Sp. strain, compositions comprising it and uses. Download PDFInfo
- Publication number
- OA20372A OA20372A OA1202100107 OA20372A OA 20372 A OA20372 A OA 20372A OA 1202100107 OA1202100107 OA 1202100107 OA 20372 A OA20372 A OA 20372A
- Authority
- OA
- OAPI
- Prior art keywords
- composition
- dominikia
- mycorrhizal
- strain
- seeds
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 139
- 241001158850 Dominikia sp. Species 0.000 title claims abstract description 85
- 235000013339 cereals Nutrition 0.000 claims abstract description 39
- 238000000034 method Methods 0.000 claims abstract description 36
- 239000002054 inoculum Substances 0.000 claims description 49
- 241000233866 Fungi Species 0.000 claims description 47
- 239000000758 substrate Substances 0.000 claims description 37
- 239000000126 substance Substances 0.000 claims description 33
- 239000011248 coating agent Substances 0.000 claims description 31
- 238000000576 coating method Methods 0.000 claims description 31
- 239000000853 adhesive Substances 0.000 claims description 20
- 230000001070 adhesive Effects 0.000 claims description 20
- 239000000417 fungicide Substances 0.000 claims description 11
- 239000002917 insecticide Substances 0.000 claims description 10
- 239000012141 concentrate Substances 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- 230000000855 fungicidal Effects 0.000 claims description 8
- 239000008187 granular material Substances 0.000 claims description 7
- 230000000749 insecticidal Effects 0.000 claims description 7
- 238000003801 milling Methods 0.000 claims description 7
- 238000010899 nucleation Methods 0.000 claims description 7
- 239000002245 particle Substances 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000000021 stimulant Substances 0.000 claims description 5
- 230000002363 herbicidal Effects 0.000 claims description 4
- 239000004009 herbicide Substances 0.000 claims description 4
- 235000021073 macronutrients Nutrition 0.000 claims description 4
- 239000011785 micronutrient Substances 0.000 claims description 4
- 235000013369 micronutrients Nutrition 0.000 claims description 4
- 239000005645 nematicide Substances 0.000 claims description 4
- 239000000049 pigment Substances 0.000 claims description 4
- 238000007873 sieving Methods 0.000 claims description 4
- 235000013619 trace mineral Nutrition 0.000 claims description 4
- 238000004040 coloring Methods 0.000 claims description 3
- 239000008247 solid mixture Substances 0.000 claims description 3
- 239000004495 emulsifiable concentrate Substances 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 description 87
- 239000002689 soil Substances 0.000 description 77
- 210000004215 spores Anatomy 0.000 description 59
- 239000000047 product Substances 0.000 description 35
- 230000000694 effects Effects 0.000 description 30
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 30
- 230000018109 developmental process Effects 0.000 description 27
- 230000012010 growth Effects 0.000 description 25
- 230000002262 irrigation Effects 0.000 description 23
- 238000003973 irrigation Methods 0.000 description 23
- 239000004927 clay Substances 0.000 description 22
- 229910052570 clay Inorganic materials 0.000 description 22
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 21
- 229910052757 nitrogen Inorganic materials 0.000 description 20
- 240000008042 Zea mays Species 0.000 description 19
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 18
- 229910052698 phosphorus Inorganic materials 0.000 description 18
- 238000004519 manufacturing process Methods 0.000 description 17
- 241000209140 Triticum Species 0.000 description 16
- 238000004458 analytical method Methods 0.000 description 15
- 235000021307 wheat Nutrition 0.000 description 15
- OAICVXFJPJFONN-UHFFFAOYSA-N phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 14
- 239000011574 phosphorus Substances 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 238000003306 harvesting Methods 0.000 description 13
- 238000011081 inoculation Methods 0.000 description 13
- 235000015097 nutrients Nutrition 0.000 description 13
- 230000002538 fungal Effects 0.000 description 12
- 229910052700 potassium Inorganic materials 0.000 description 12
- 240000007594 Oryza sativa Species 0.000 description 11
- 235000005822 corn Nutrition 0.000 description 11
- 235000005824 corn Nutrition 0.000 description 11
- 239000011780 sodium chloride Substances 0.000 description 11
- 230000006399 behavior Effects 0.000 description 10
- 230000004720 fertilization Effects 0.000 description 10
- 241000894007 species Species 0.000 description 10
- 238000010186 staining Methods 0.000 description 10
- 230000000007 visual effect Effects 0.000 description 10
- 239000011734 sodium Substances 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- 241000045080 Dominikia minuta Species 0.000 description 8
- 210000004276 Hyalin Anatomy 0.000 description 8
- 201000009910 diseases by infectious agent Diseases 0.000 description 8
- 239000005416 organic matter Substances 0.000 description 8
- 241001609260 Dominikia Species 0.000 description 7
- 235000007164 Oryza sativa Nutrition 0.000 description 7
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 7
- 239000011575 calcium Substances 0.000 description 7
- 235000009973 maize Nutrition 0.000 description 7
- 241000209082 Lolium Species 0.000 description 6
- 240000004296 Lolium perenne Species 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 230000035784 germination Effects 0.000 description 6
- 230000003071 parasitic Effects 0.000 description 6
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 6
- 239000011591 potassium Substances 0.000 description 6
- 235000009566 rice Nutrition 0.000 description 6
- 229910052708 sodium Inorganic materials 0.000 description 6
- 241001583499 Glomeromycotina Species 0.000 description 5
- 240000005979 Hordeum vulgare Species 0.000 description 5
- 235000007340 Hordeum vulgare Nutrition 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000003337 fertilizer Substances 0.000 description 5
- 238000011049 filling Methods 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 5
- 238000004806 packaging method and process Methods 0.000 description 5
- 229920003023 plastic Polymers 0.000 description 5
- 239000004033 plastic Substances 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 238000005070 sampling Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000009331 sowing Methods 0.000 description 5
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 4
- 241001609258 Dominikia aurea Species 0.000 description 4
- 240000006394 Sorghum bicolor Species 0.000 description 4
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 4
- 235000007264 Triticum durum Nutrition 0.000 description 4
- 241000209143 Triticum turgidum subsp. durum Species 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 229910052791 calcium Inorganic materials 0.000 description 4
- 238000002856 computational phylogenetic analysis Methods 0.000 description 4
- 229920003013 deoxyribonucleic acid Polymers 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000005747 Chlorothalonil Substances 0.000 description 3
- CRQQGFGUEAVUIL-UHFFFAOYSA-N Chlorothalonil Chemical compound ClC1=C(Cl)C(C#N)=C(Cl)C(C#N)=C1Cl CRQQGFGUEAVUIL-UHFFFAOYSA-N 0.000 description 3
- 241000045073 Dominikia indica Species 0.000 description 3
- 241001354220 Rhizophagus iranicus Species 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 230000035558 fertility Effects 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000001717 pathogenic Effects 0.000 description 3
- 244000052769 pathogens Species 0.000 description 3
- 235000020004 porter Nutrition 0.000 description 3
- 238000003908 quality control method Methods 0.000 description 3
- 230000007226 seed germination Effects 0.000 description 3
- 239000007921 spray Substances 0.000 description 3
- NINIDFKCEFEMDL-UHFFFAOYSA-N sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 3
- 239000011701 zinc Substances 0.000 description 3
- UCSJYZPVAKXKNQ-HZYVHMACSA-N 1-[(1S,2R,3R,4S,5R,6R)-3-carbamimidamido-6-{[(2R,3R,4R,5S)-3-{[(2S,3S,4S,5R,6S)-4,5-dihydroxy-6-(hydroxymethyl)-3-(methylamino)oxan-2-yl]oxy}-4-formyl-4-hydroxy-5-methyloxolan-2-yl]oxy}-2,4,5-trihydroxycyclohexyl]guanidine Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 229920002483 18S ribosomal RNA Polymers 0.000 description 2
- UFNOUKDBUJZYDE-UHFFFAOYSA-N 2-(4-chlorophenyl)-3-cyclopropyl-1-(1H-1,2,4-triazol-1-yl)butan-2-ol Chemical compound C1=NC=NN1CC(O)(C=1C=CC(Cl)=CC=1)C(C)C1CC1 UFNOUKDBUJZYDE-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 229960005069 Calcium Drugs 0.000 description 2
- 239000005757 Cyproconazole Substances 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N D-sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 241000045082 Dominikia iranica Species 0.000 description 2
- 235000019749 Dry matter Nutrition 0.000 description 2
- 240000005149 Epigaea repens Species 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 210000004209 Hair Anatomy 0.000 description 2
- GUBGYTABKSRVRQ-UUNJERMWSA-N Lactose Natural products O([C@@H]1[C@H](O)[C@H](O)[C@H](O)O[C@@H]1CO)[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 GUBGYTABKSRVRQ-UUNJERMWSA-N 0.000 description 2
- 240000003183 Manihot esculenta Species 0.000 description 2
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 2
- 241000539716 Mea Species 0.000 description 2
- -1 Metalaxyl-Ridomil Chemical compound 0.000 description 2
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 2
- 239000005811 Myclobutanil Substances 0.000 description 2
- HZJKXKUJVSEEFU-UHFFFAOYSA-N Myclobutanil Chemical compound C=1C=C(Cl)C=CC=1C(CCCC)(C#N)CN1C=NC=N1 HZJKXKUJVSEEFU-UHFFFAOYSA-N 0.000 description 2
- 235000011205 Ocimum Nutrition 0.000 description 2
- 241001529734 Ocimum Species 0.000 description 2
- 235000010120 Ocimum sp Nutrition 0.000 description 2
- CZMRCDWAGMRECN-GDQSFJPYSA-N Sucrose Natural products O([C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1)[C@@]1(CO)[C@H](O)[C@@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-GDQSFJPYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- 241000223259 Trichoderma Species 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 2
- 238000004166 bioassay Methods 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M buffer Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- KXDHJXZQYSOELW-UHFFFAOYSA-M carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 210000004027 cells Anatomy 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 238000003926 complexometric titration Methods 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 230000000875 corresponding Effects 0.000 description 2
- 238000010192 crystallographic characterization Methods 0.000 description 2
- 230000000249 desinfective Effects 0.000 description 2
- 239000003651 drinking water Substances 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- KQTVWCSONPJJPE-UHFFFAOYSA-N etridiazole Chemical compound CCOC1=NC(C(Cl)(Cl)Cl)=NS1 KQTVWCSONPJJPE-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000005048 flame photometry Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 238000005469 granulation Methods 0.000 description 2
- 230000003179 granulation Effects 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 229920001903 high density polyethylene Polymers 0.000 description 2
- 230000001524 infective Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- 230000005541 medical transmission Effects 0.000 description 2
- 230000002503 metabolic Effects 0.000 description 2
- 230000000813 microbial Effects 0.000 description 2
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 2
- 239000008108 microcrystalline cellulose Substances 0.000 description 2
- 229940016286 microcrystalline cellulose Drugs 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 235000021231 nutrient uptake Nutrition 0.000 description 2
- 238000001543 one-way ANOVA Methods 0.000 description 2
- 230000000737 periodic Effects 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- DNIAPMSPPWPWGF-UHFFFAOYSA-N propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- KEAYESYHFKHZAL-UHFFFAOYSA-N sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 230000004083 survival Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229920002554 vinyl polymer Polymers 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- 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
- BOTWFXYSPFMFNR-PYDDKJGSSA-N (E)-Phytol Chemical compound CC(C)CCC[C@@H](C)CCC[C@@H](C)CCC\C(C)=C\CO BOTWFXYSPFMFNR-PYDDKJGSSA-N 0.000 description 1
- 239000001707 (E,7R,11R)-3,7,11,15-tetramethylhexadec-2-en-1-ol Substances 0.000 description 1
- 229920000160 (ribonucleotides)n+m Polymers 0.000 description 1
- NVKAWKQGWWIWPM-ABEVXSGRSA-N 17-β-hydroxy-5-α-Androstan-3-one Chemical compound C1C(=O)CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 NVKAWKQGWWIWPM-ABEVXSGRSA-N 0.000 description 1
- MNHVNIJQQRJYDH-UHFFFAOYSA-N 2-[2-(1-chlorocyclopropyl)-3-(2-chlorophenyl)-2-hydroxypropyl]-1,2-dihydro-1,2,4-triazole-3-thione Chemical compound N1=CNC(=S)N1CC(C1(Cl)CC1)(O)CC1=CC=CC=C1Cl MNHVNIJQQRJYDH-UHFFFAOYSA-N 0.000 description 1
- YJISHJVIRFPGGN-UHFFFAOYSA-N 5-[5-[3,4-dihydroxy-6-(hydroxymethyl)-5-methoxyoxan-2-yl]oxy-6-[[3,4-dihydroxy-6-(hydroxymethyl)-5-methoxyoxan-2-yl]oxymethyl]-3,4-dihydroxyoxan-2-yl]oxy-6-(hydroxymethyl)-2-methyloxane-3,4-diol Chemical compound O1C(CO)C(OC)C(O)C(O)C1OCC1C(OC2C(C(O)C(OC)C(CO)O2)O)C(O)C(O)C(OC2C(OC(C)C(O)C2O)CO)O1 YJISHJVIRFPGGN-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 235000016626 Agrimonia eupatoria Nutrition 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 241000416162 Astragalus gummifer Species 0.000 description 1
- 239000005730 Azoxystrobin Substances 0.000 description 1
- WFDXOXNFNRHQEC-GHRIWEEISA-N Azoxystrobin Chemical compound CO\C=C(\C(=O)OC)C1=CC=CC=C1OC1=CC(OC=2C(=CC=CC=2)C#N)=NC=N1 WFDXOXNFNRHQEC-GHRIWEEISA-N 0.000 description 1
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 241000194103 Bacillus pumilus Species 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 240000008371 Bacillus subtilis Species 0.000 description 1
- 229940075615 Bacillus subtilis Drugs 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000827797 Beauveria sp. Species 0.000 description 1
- 241000219430 Betula pendula Species 0.000 description 1
- 229960003563 Calcium Carbonate Drugs 0.000 description 1
- 239000005745 Captan Substances 0.000 description 1
- 229940117949 Captan Drugs 0.000 description 1
- 239000005746 Carboxin Substances 0.000 description 1
- DJHJJVWPFGHIPH-OODMECLYSA-N Chitin Chemical compound O[C@@H]1C(NC(=O)C)[C@H](O)OC(CO)[C@H]1COC[C@H]1C(NC(C)=O)[C@@H](O)[C@H](COC[C@H]2C([C@@H](O)[C@H](O)C(CO)O2)NC(C)=O)C(CO)O1 DJHJJVWPFGHIPH-OODMECLYSA-N 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 241000941525 Chromobacterium sp. Species 0.000 description 1
- 241000317178 Claroideoglomus etunicatum Species 0.000 description 1
- 101700011961 DPOM Proteins 0.000 description 1
- 241000045065 Dominikia achra Species 0.000 description 1
- 241000235494 Endogonaceae Species 0.000 description 1
- 241000235496 Endogone Species 0.000 description 1
- 210000002615 Epidermis Anatomy 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 239000005769 Etridiazole Substances 0.000 description 1
- IWDQPCIQCXRBQP-UHFFFAOYSA-M Fenaminosulf Chemical compound [Na+].CN(C)C1=CC=C(N=NS([O-])(=O)=O)C=C1 IWDQPCIQCXRBQP-UHFFFAOYSA-M 0.000 description 1
- NHOWDZOIZKMVAI-UHFFFAOYSA-N Fenarimol Chemical compound C=1N=CN=CC=1C(C=1C(=CC=CC=1)Cl)(O)C1=CC=C(Cl)C=C1 NHOWDZOIZKMVAI-UHFFFAOYSA-N 0.000 description 1
- 239000005786 Flutolanil Substances 0.000 description 1
- 239000005789 Folpet Substances 0.000 description 1
- 241001123597 Funneliformis mosseae Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000235500 Gigaspora Species 0.000 description 1
- 240000007842 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000976924 Inca Species 0.000 description 1
- 239000005867 Iprodione Substances 0.000 description 1
- ONUFESLQCSAYKA-UHFFFAOYSA-N Iprodione Chemical compound O=C1N(C(=O)NC(C)C)CC(=O)N1C1=CC(Cl)=CC(Cl)=C1 ONUFESLQCSAYKA-UHFFFAOYSA-N 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 235000003228 Lactuca sativa Nutrition 0.000 description 1
- 241000209510 Liliopsida Species 0.000 description 1
- 235000019738 Limestone Nutrition 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 101710029649 MDV043 Proteins 0.000 description 1
- 239000005802 Mancozeb Substances 0.000 description 1
- YKSNLCVSTHTHJA-UHFFFAOYSA-L Maneb Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S YKSNLCVSTHTHJA-UHFFFAOYSA-L 0.000 description 1
- 241000196323 Marchantiophyta Species 0.000 description 1
- 238000007476 Maximum Likelihood Methods 0.000 description 1
- 239000005807 Metalaxyl Substances 0.000 description 1
- ZQEIXNIJLIKNTD-UHFFFAOYSA-N Metalaxyl Chemical compound COCC(=O)N(C(C)C(=O)OC)C1=C(C)C=CC=C1C ZQEIXNIJLIKNTD-UHFFFAOYSA-N 0.000 description 1
- 241000488289 Metarhizium sp. Species 0.000 description 1
- 229910003110 Mg K Inorganic materials 0.000 description 1
- 241000244206 Nematoda Species 0.000 description 1
- 101700061424 POLB Proteins 0.000 description 1
- 241000887182 Paraphaeosphaeria minitans Species 0.000 description 1
- 241001668579 Pasteuria Species 0.000 description 1
- LKPLKUMXSAEKID-UHFFFAOYSA-N Pentachloronitrobenzene Chemical compound [O-][N+](=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1Cl LKPLKUMXSAEKID-UHFFFAOYSA-N 0.000 description 1
- BOTWFXYSPFMFNR-QYLFUYDXSA-N Phytol Natural products CC(C)CCC[C@@H](C)CCC[C@@H](C)CCC\C(C)=C/CO BOTWFXYSPFMFNR-QYLFUYDXSA-N 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- 239000005821 Propamocarb Substances 0.000 description 1
- WZZLDXDUQPOXNW-UHFFFAOYSA-N Propamocarb Chemical compound CCCOC(=O)NCCCN(C)C WZZLDXDUQPOXNW-UHFFFAOYSA-N 0.000 description 1
- 239000005825 Prothioconazole Substances 0.000 description 1
- 101700054624 RF1 Proteins 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 229960005322 Streptomycin Drugs 0.000 description 1
- 239000005842 Thiophanate-methyl Substances 0.000 description 1
- 239000005843 Thiram Substances 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
- 229920001615 Tragacanth Polymers 0.000 description 1
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Tris Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000010162 Tukey test Methods 0.000 description 1
- FSCWZHGZWWDELK-UHFFFAOYSA-N Vinclozolin Chemical compound O=C1C(C)(C=C)OC(=O)N1C1=CC(Cl)=CC(Cl)=C1 FSCWZHGZWWDELK-UHFFFAOYSA-N 0.000 description 1
- 235000007244 Zea mays Nutrition 0.000 description 1
- 239000005870 Ziram Substances 0.000 description 1
- 229930000028 abscisic acids Natural products 0.000 description 1
- JLIDBLDQVAYHNE-OAHLLOKOSA-N abscisin II Chemical compound OC(=O)C=C(C)C=C[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-OAHLLOKOSA-N 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000001476 alcoholic Effects 0.000 description 1
- 239000011717 all-trans-retinol Substances 0.000 description 1
- 235000019169 all-trans-retinol Nutrition 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 229960000892 attapulgite Drugs 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- OIPMQULDKWSNGX-UHFFFAOYSA-N bis[[ethoxy(oxo)phosphaniumyl]oxy]alumanyloxy-ethoxy-oxophosphanium Chemical compound [Al+3].CCO[P+]([O-])=O.CCO[P+]([O-])=O.CCO[P+]([O-])=O OIPMQULDKWSNGX-UHFFFAOYSA-N 0.000 description 1
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 1
- 229910052796 boron Inorganic materials 0.000 description 1
- 150000001647 brassinosteroids Chemical class 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 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
- 239000005018 casein Substances 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 150000003841 chloride salts Chemical class 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- PFIADAMVCJPXSF-UHFFFAOYSA-N chloroneb Chemical compound COC1=CC(Cl)=C(OC)C=C1Cl PFIADAMVCJPXSF-UHFFFAOYSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N edta Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing Effects 0.000 description 1
- 235000021112 essential micronutrients Nutrition 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable Effects 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 229930003935 flavonoids Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- PTCGDEVVHUXTMP-UHFFFAOYSA-N flutolanil Chemical compound CC(C)OC1=CC=CC(NC(=O)C=2C(=CC=CC=2)C(F)(F)F)=C1 PTCGDEVVHUXTMP-UHFFFAOYSA-N 0.000 description 1
- HKIOYBQGHSTUDB-UHFFFAOYSA-N folpet Chemical compound C1=CC=C2C(=O)N(SC(Cl)(Cl)Cl)C(=O)C2=C1 HKIOYBQGHSTUDB-UHFFFAOYSA-N 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000001295 genetical Effects 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000003621 irrigation water Substances 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N iso-propanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- 239000006028 limestone Substances 0.000 description 1
- 235000021388 linseed oil Nutrition 0.000 description 1
- 239000000944 linseed oil Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000003050 macronutrient Effects 0.000 description 1
- 229920000940 maneb Polymers 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- 239000010445 mica Substances 0.000 description 1
- 229910052618 mica group Inorganic materials 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 230000000877 morphologic Effects 0.000 description 1
- 230000001069 nematicidal Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 229910052625 palygorskite Inorganic materials 0.000 description 1
- 230000024241 parasitism Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 238000005453 pelletization Methods 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 230000000243 photosynthetic Effects 0.000 description 1
- 230000001863 plant nutrition Effects 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 239000011118 polyvinyl acetate Substances 0.000 description 1
- 229920002689 polyvinyl acetate Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 235000019422 polyvinyl alcohol Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000000750 progressive Effects 0.000 description 1
- 235000013772 propylene glycol Nutrition 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920002973 ribosomal RNA Polymers 0.000 description 1
- 230000035040 seed growth Effects 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 235000010956 sodium stearoyl-2-lactylate Nutrition 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 230000028070 sporulation Effects 0.000 description 1
- 230000003019 stabilising Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- QGHREAKMXXNCOA-UHFFFAOYSA-N thiophanate-methyl Chemical compound COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC QGHREAKMXXNCOA-UHFFFAOYSA-N 0.000 description 1
- 229960002447 thiram Drugs 0.000 description 1
- 238000003971 tillage Methods 0.000 description 1
- 210000001519 tissues Anatomy 0.000 description 1
- RROQIUMZODEXOR-UHFFFAOYSA-N triforine Chemical compound O=CNC(C(Cl)(Cl)Cl)N1CCN(C(NC=O)C(Cl)(Cl)Cl)CC1 RROQIUMZODEXOR-UHFFFAOYSA-N 0.000 description 1
- 238000009966 trimming Methods 0.000 description 1
- 238000004450 types of analysis Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
- XLOMVQKBTHCTTD-UHFFFAOYSA-N zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
- 235000014692 zinc oxide Nutrition 0.000 description 1
- CHNQZRKUZPNOOH-UHFFFAOYSA-J zinc;manganese(2+);N-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[Zn+2].[S-]C(=S)NCCNC([S-])=S.[S-]C(=S)NCCNC([S-])=S CHNQZRKUZPNOOH-UHFFFAOYSA-J 0.000 description 1
- DUBNHZYBDBBJHD-UHFFFAOYSA-L ziram Chemical compound [Zn+2].CN(C)C([S-])=S.CN(C)C([S-])=S DUBNHZYBDBBJHD-UHFFFAOYSA-L 0.000 description 1
Abstract
Object of the invention is the strain of fungus Dominikia sp. deposited under accession number MUCL 57072, that can be included into compositions; said composition being suitable to be used as bio-stimulant and bio-nematicidal in plants, preferably in cereals. Also object of the invention is a process for obtaining said compositions.
Description
“Dominikia sp. STRAIN, COMPOSITIONS COMPRISING IT AND USES”
FIELD OFTHE INVENTION
The présent invention relates to the agronomie field. Specifically, it relates to a Dominikia sp. strain, compositions comprising it and uses of said compositions, such as the use as bio-stimulant and bio-nematicidal. Compositions comprising a Dominikia sp. strain were found to be particularly useful in cereal crops.
BACKGROUND ART
Currently approximately 300 species in 30 généra are described in the Glomeromycota. However, nucleic acid (DNA, RNA) based biodiversity studies point towards a much higher diversity in Glomeromycota and several new species and généra hâve been described recently.
Most Glomeromycota are arbuscular mycorrhizal fungi (AMF) which are mutualistic symbionts of approximately 80% of ail vascular land plants. Nutrient uptake of plants which are colonised by AMF occurs directly via the root epidermis and the root hairs, but also via the fungus-root-interface which has the characteristic form of arbuscules or intraradical hyphal coils. Besides their rôle in enhancing nutrient uptake in their host, AMF play important rôles in soil aggregation and in protecting plants against drought stress and soil born plant pathogens. Because of this highly bénéficiai nature, several mycorrhizal compositions are known in the art, and they hâve been developed to provide bénéficiai effects on the crops they are provided to.
WO2015/000612 and WO2015/000613 disclose compositions comprising Glomus iranicum var. tenuihypharum var. nov. strain and their uses.
In particular, WO2015/000612 discloses a composition comprising Glomus iranicum var. tenuihypharum var. nov. strain and 2:1 smectite clays. The composition of WO2015/000612 is disclosed to provide a positive impact on crop yield, i.e. that can be used as bio-stimulant, in lettuce crops.
Similarly, WO2015/000613 discloses a composition comprising Glomus iranicum var. tenuihypharum var. nov. strain and 2:1 smectite clays, métal ions and chitin. WO2015/000613 discloses that said composition acts as bio-nematicidal in tomato crops.
However, the compositions disclosed in the prior art do not provide an acceptable positive impact on cereal crops, both in terms of bio-stimulant and bio-nematicidal effect.
SUMMARY OF THE INVENTION
An aim of the présent invention is thus to provide a strain of fungus that is suitable to be included into compositions, namely compositions suitable to provide a positive impact on crops, in particular on cereal crops.
Another aim of the invention is to provide a composition that is effective as bio-stimulant on cereal crops.
Still another aim of the présent invention is to provide a composition that is effective as bio-nematicidal on cereal crops.
A further aim of the présent invention is to provide a process to for obtaining compositions.
DETA1LED DESCRIPTION
An object of the présent invention is thus a strain of Glomeromycota, i.e. a Dominikia sp. strain, deposited under accession number MUCL 57072.
The strain of the invention was isolated from a very hydromorphic, highly compacted sodium saline soil type (Solonetz Gley) with a lot of sait deposit at the surface, in the locality of Fortuna, Murcia (Spain).
Dominikia sp. strain of the invention was deposited on 21/03/2018 at the International Depositary Authority Belgian Coordinated Collections of Micro-organisms (BCCM), Université catholique de Louvain, Mycothequèque de l’Université catholique de Louvain (MUCL), Croix du Sud 2, box L7.05.06, 1348 Louvain-la-Neuve, Belgium, by Symborg S.L., with address Campus de Espinardo, 7, edificio CEEIM, 30100 Murcia, Spain.
The strain Dominikia sp. of the invention has been identified by the depositor with the reference SMB01, and received the deposit number MUCL 57072 by the international depositary authority.
The Sporocarps of the strain of the invention are unknown. In the strain of the invention, spores occurs in loose clusters in the soil, and may be terminal or intercalary; spores may be also formed within the roots. Spores are hyaline to light ochraceous, with a subglobose to globose structure (rarely irregularly), relatively small, i.e. from 24.0 to 42 pm in diameter, with an average value of 30.7 ± 3.7 pm in diameter . Spores présent a composite three-layered spore wall (1-4 pm thick). Particularly, an outer layer, a middle layer and an inner layer can be recognized.
The outer layer of the spore wall is substantially mucilagous and evanescent, thus giving young spores a rough appearance, while older spores show a somewhat shaggy appearance. The outer layer of the spore wall exhibits a dextrinoid reaction, when stained with Melzer’s reagent, giving young spores a brownish red color. The middle layer of the spore wall is substantially permanent, and is 0.5 - 2.0 pm thick.
The inner layer of the spore wall is substantially laminar, with a thickness of 0.5-1.5 pm. The contents of the spores hâve a pale and guttulate appearance. The hyphae that hold the spores hâve a hyaline to pale ochraceous color, and are straight or wavy of 2.5 4.5 pm in diameter (with an average value of 3.0 pm) The hyphae hâve a cylindrical to slightly funnel shape, which merges with the open-pored layers of the spore wall, at least in mature spores. The germination structure comprises a germ tube grown and develops back through the union of the hyphae with the spores. It forms vesicular arbuscular mycorrhizae.
The extraradical mycélium forms an extensive network.
A phylogenetic analysis of 134 18S rDNA sequences, including environmental and referenced sequences, was generated by the Applicant. From most Dominikia species no or relatively short partial 18S sequences are available. Dominikia indica and D. iranica sequences can be included in this phylogenetic analysis. The sequences of these two species were shorter than those of the rest of the dataset. Trimming the dataset to the length of these sequences resulted in rather incompletely resolved trees. Using the full length sequences, trees with branches representing the higher taxonomie levels could be generated. However, a robust, monophyletic clade consisting of sequences from Dominikia spp. could not be generated with this dataset. Dominikia sp. forms one, well supported clade together with Dominikia iranica and anonymous, environmental glomeralean sequences from different host plants and different locations. Phylogenetic analysis of an SSU-ITS1 dataset including sequences from the majority of described Dominikia species indicates that Dominikia sp. branches next to D. aurea.
Small spores (up to 65-70 pm of diameter), usually aggregated in loose to compact clusters, are characteristic for the genus Dominikia. The spore wall of the members of the genus Dominikia consists of two or three layers. The outer layer, forming the spore surface, is mucilaginous, short-lived and stains in Melzer’s reagent or is unit (not divided into sublayers), permanent and does not react in Melzer’s reagent. The hyphae which hold the spores are cylindrical to funnel-shaped with a pore that may be open or occluded by a septum. Dominikia minuta (Basidionym: Glomus minutum) was designated as type-species. The strain of the invention, i.e. Dominikia sp., has hyaline spores of substantially the same size than those of Dominikia minuta but, in contrast to Dominikia sp., the pore of. G. minutum (i.e. Dominikia minuta) is occluded by a septum.
D. sp. also differs from Dominikia minuta by its three-layers spore wall, as opposed to the two-layered spore wall in D. minuta, and because of the presence of the dextrinoid reaction, which is absent in Dominikia minuta.
Dominikia sp. also differs from the other described Dominikia species and strains. The 5 outer layer and the inner layer of spores of Dominikia achra stain deeply red in Melzer's reagent, while in Dominikia sp. only the outer layer shows a dextrinoid reaction. Dominikia indica differs from Dominikia sp. by forming small, hyaline spores in hypogeous aggregates. The spore wall of D. indica consists of two hyaline layers: a mucilaginous, short-lived, thin outer layer staining pinkish to pink in Melzer's reagent 10 and a laminate, smooth, permanent, thicker inner layer as opposed to the three layers in
Dominikia sp.
Based on the 18S-ITS1 phylogeny, Dominikia aurea is the closest relative of the Dominikia sp. strain of the invention. The two species differ in many morphologically characters (see Table 1), the most évident différence being the mostly ovoid spores of 15 Dominikia aurea which are aggregated in irregular sporocarps.
Table 1 | ||
Dominikia aurea (Oehl et al. 2003) | Dominikia sp. | |
spore aggregation | irregular sporocarps without peridium | loose clusters |
intraradical spores | Not described | présent |
intercalary spores | Not described | présent |
colour | Light orange to orange | ochraceous |
form | Ovoid, rarely globose | globose to subglobose |
size | Ovoid spores: (36-)55 - 65 x (30)45- 52 pm Globose spores: (27-) 40-60 pm | (24,0) 30,7 ± 3,7 (42) pm |
spore wall | two layered | three layered |
Outermost layer | evanescent, hyaline up to 1.5 pm, deteriorated in mature spores, dextrinoid | mucilaginous, roughened, hyaline (0.4-) 1.0(1.5) pm, |
deteriorated in mature spores, dextrinoid | ||
Middle layer | permanent, slightly rough 0.5 - 2.0 pm thick | |
Innermost layer | laminate, light orange, 1.5-3 (-4) pm | laminate, smooth (0.5-1.5 pm) |
Subtending hypae | Light orange to orange, straight or curved, cylindrical or slightly funnel shaped; 6 -10 pm | hyaline to pale ochraceous, straight or undulating 2.5 - 5 pm |
Germination | Unknown | Through the subtending hypha |
Many highly similar sequences to the Dominikia sp. of the invention sequences could be found when blasted against gene bank (e-value 0.00, Identity 99%). These sequences occur from wide range of different hosts (e.g. liverworts, monocotyledons) from different countries and continents, indicating a world-wide distribution. These findings indicate that Dominikia sp. is widely distributed.
As above mentioned, Dominikia sp. was found to be particularly useful to be included into compositions, in particular compositions to be provided to cereal crops.
Another object of the invention is a composition comprising Dominikia sp., deposited on 21/03/2018 under accession number MUCL 57072, as above mentioned.
Without being bound to a spécifie explanation, it has been surprisingly observed that Dominikia sp., and composition comprising the same, are particularly effective in providing bénéficiai effects to crops, in particular to cereal crops.
In particular, it has been observed that, by providing a composition comprising Dominikia sp. to a crop, e.g. to a cereal crop (for example, a maize crop, a wheat crop, a barley crop, or a rice crop) an increase in the uptake of the nutrient and an improvement in the yield of the cereal crop can be obtained, with respect to crops that were not provided with Dominikia sp..
Preferably, the concentration of the Dominikia sp. in the composition is from 4.0 % to 1.0% by weight, more preferably from 3.0% to 2.0% by weight, even more preferably 5
2.5% to 2.3 % by weight.
According to embodiments, the composition of the invention, is a liquid, a solid or a gel composition.
Preferably, the composition of the invention, is a solid composition.
According to embodiments, the composition of the invention may be in the form of powder, emulsifiable concentrate, granules, or microgranules.
The composition of the invention is a solid composition, and the concentration of the propagules of Dominikia sp. in the composition, is measured according to the “Most Probable Number Method” (Porter, Aust. J. Soil Res., 1979, 17, 515-19) from 180 to 120 propagules per gram of composition, preferably from 150 to 120 propagules per gram of composition, more preferably from 125 to 120 propagules per gram of composition.
The concentration of the propagules is referred to the concentration of the propagules in the final product.
According to a preferred embodiment, the composition of the invention is in the form of microgranules.
According to embodiments, said microgranules hâve a size ranging from 500 pm to 2000 pm, preferably from 800 pm to 1500 pm, more preferably from 900 pm to 1200 pm.
Advantageously, the concentration of the Dominikia sp. in the composition of the invention, as well as the form of présentation of the composition, e.g. microgranules, may be selected according to the predetermined final application.
According to embodiments, the composition of the invention further comprises at least one fungicide and/or at least one bio-fungicide and/or at least one insecticide and/or at least one bio-insecticide and/or at least one nematicide and/or at least one biostimulant.
According to a preferred embodiment, the composition of the invention is in the form for seeds coating.
For example, said fungicide is selected from the group consisting of Maneb, Mancozeb, Metalaxyl-Ridomil, Myclobutanil, Olpisan, Propamocarb, Quintozene, Streptomycin, Sulfur, Thiophanate-methyl, Thiram, triforine, vinclozolin, Zinc white, Zineb, Ziram, Banrot, Fixed copper, Chlorothalonil, Chlorothalonil, Captan, Chloroneb, Cyproconazole, Zinc ethelene, bisdithiocarbamate, Etridiazole, Fenaminosulf, Fenarimol, Flutolanil, Folpet, Fosetyl-AL and Iprodione.
Exemplary bio-fungicides are: Trichodermas sp, Bacillus subtilis, Bacillus licheniformis, Bacillus pumilus, Bacillus amyloliquefaciens, Streptomyces sp, Coniothyrium minitans and Pythïum oligandrum.
According to embodiments, the insecticide is selected from the group consisting of organophosphate, carbamate and neonicotinoid.
According to embodiments, said bio-insecticide is selected from the group consisting of Bacillus sp., Chromobacterium sp., Beauveria sp. and Metarhizium sp.
According to embodiments, said nematicide is organophosphate or carbamate. According to embodiments, said bio -nematicide is Pasteuria sp.
Another object of the présent invention is a process for the production of a composition comprising the Dominikia sp. strain of the invention.
The process of the invention comprises the following steps:
a. Providing a substrate;
b. Providing said substrate with the seeds of a host plant and with Dominikia sp. strain deposited under accession number MUCL 57072;
c. Cultivating said host plant and watering to maintain said substrate with a moisture level of at least 75 % of field capacity;
d. Discontinuing said watering for a period of at least 7 days;
e. Removing the aerial part of said host plant and the substrate;
f. Drying the removed substrate;
g. Milling the dried substrate to obtain granules having a particle size of below 100 pm.
Preferably, the substrate comprises a clay.
According to embodiments, the Dominikia sp. provided in step b. is an inoculum comprising Dominikia sp. propagules. Preferably, said inoculum is obtained by providing the root System of a host plant with a substantially pure Dominikia sp., cultivating said host plant on a substrate preferably comprising a clay (preferably a stérile smectite clay), preferably for a life cycle of the host plant, and subsequently extracting the root System, to obtain an inoculum. In this case, the inoculum comprises an amount of the substrate, rootlets, and Dominikia sp. propagules.
According to a preferred embodiment, the process of the invention further comprises a step h1. of microgranulation.
Preferably, by microgranulation, a composition comprising the Dominikia sp. strain of the invention in the form of microgranules may be obtained.
As above mentioned, said microgranules hâve a size ranging from 500 pm to 2000 pm, preferably from 800 pm to 1500 pm, more preferably from 900 pm to 1200 pm.
According to a preferred embodiment, the process of the invention further comprises a step h2. of préparation of a concentrate biological inoculant for seeds coating of cereals, namely concentrate biological inoculant for seeds coating of cereals of the milled product obtained according to step g. It has to be noted that, in this context, the step h2. of concentrate biological inoculant for seeds coating of cereals is meant as a step wherein the concentration of the Dominikia sp. (e.g., the Dominikia sp. propagules) in the composition increases.
For example, said step h2. of concentration may be carried out by sieving the milled product obtained according to step g. to select the granules having a particle size above a predetermined value, e.g., above 35 pm.
Advantageously, the concentration step h2. allows to increase the concentration of Dominikia sp. (e.g., the Dominikia sp. propagules) up to 10 times the initial concentration, preferably up to 50 times the initial concentration (i.e. the concentration of Dominikia sp. in the product before step h2), more preferably up to 100 times the initial concentration, i.e. 100 times with respect to the concentration of the composition before step h2..
According to embodiments, it is possible to increase the concentration of Dominikia sp. (e.g., the Dominikia sp. propagules) more than 100 times the initial concentration (i.e. the concentration of Dominikia sp. in the product before step h2).
According to a preferred embodiment, the process of the invention further comprises a step i. of seeds coating, namely seeds coating of the concentrate product obtained according to step h2.
Said step i. of seed coating comprises the following steps:
il. Covering the seeds are with an adhesive substance i2. Adding the concentrate biological inoculant, i3. Optionally provide one or more compatible treatments selected from:
i3.1 Treatments with fungicides, insecticides and/or herbicides compatible with mycorrhizal forming fungi, i3.2 Treatment with bénéficiai microorganisms,
Î3.3 Treatment with macro and/or micronutrients, i3.4 Treatment with stimulants,
Î3.5 Treatment with colouring pigments compatible with mycorrhizal fungi;
i4. Drying the seeds.
Another object of the présent invention is the use of a composition comprising Dominikia sp., deposited under accession number MUCL 57072 as bio- stimulant.
As used herein, the term “bio-stimulation” refer to the stimulation exerted by a composition according to the invention in plants that are provided with said composition. Without being bound to a spécifie scientific explanation, it can be hypothesized that the Dominikia sp. strain of the invention exerts the function of translocation of nutrients, taking these nutrients from the soil or substrate and using said nutrients in its metabolic System, it translocates said nutrients from its mycélium network and it subsequently exchanges them in the root cells.
According to a preferred embodiment, the composition of the présent invention is used as a bio-stimulant on cereals.
Another object of the présent invention is the use of a composition comprising Dominikia sp., deposited under accession number MUCL 57072 as bio- nematicidal.
In other words, the composition of the présent invention can be used to protect plants from nematodes.
According to a preferred embodiment, the composition of the présent invention is used as a bio-nematicidal on cereals.
Advantageously, the composition of the invention can be provided to plants, i.e. to a crop, for example to a cereal crop, in several ways.
According to embodiments, the composition of the invention can be applied to the plant by seed treatment (i.e. seed coating), root treatment, roots embedded in an émulsion, addition to irrigation water, irrigation, application of powder to the root System or application of émulsion injected into the root System.
Preferably, in particular when it has to be provided to cereals, the composition of the invention is provided by seed coating or in conjunction with the seeds at the time of seeding, or in the form of microganulate and in conjunction with the seeds at the time of seeding.
BRIEF DESCRIPTION OF THE FIGURES
- Figure 1: Dynamics of spore production and fungal occupation, expressed through Visual density (%), in plants treated with Dominikia sp..
Figure 2: Behaviour of the arbuscular external mycélium and the arbuscular endophyte in the treatment with Dominikia sp..
- Figure 3: Relationship between the external arbuscular mycélium and the arbuscular endophyte in the treatment with the Dominikia sp..
EXPERIMENTAL SECTION
Example 1. Molecular analyses
DNA- Extraction
Isolated hyphae and spores were transferred into 1,5 mL Eppendorf Tubes with 0.2 g of glass-beads (2 mm diameter) and 100 pL CTAB-buffer (2 % CTAB = cetyltrimethylammoniumbromide, 1.4 M NaCI, 0.1 M Tris-HCI pH 7.5, 0.2 M Na-EDTA). This mixture was homogenized using a Retsch MM301 Bail Mill at 50 Hz for 30 seconds. Another 400 pL of CTAB-buffer were added and the mixture was incubated at 65 °C for one hour. 400 pL Cholroform-lsoamylalkohol (24 : 1) were added to the suspension and mixed by inverting the reaction tubes and subsequently centrifuged for 5 min at 15000 x g and the top layer recovered into a clean Eppendorf tube. This step was repeated twice. To this suspension 200 pL 5 M Ammonium acetate were added; this mixture was incubated at 4 °C for at least 30 minutes followed by 20 minutes spinning at 4 °C and 15000 x g. DNA was precipitated with 700 pL Isopropanol at -20 °C overnight. The DNA pellet obtained after Isopropanol-precipitation was washed with ice-cold 70 % éthanol, air dried and re-dissolved in 50 pL TE buffer (10 mM Tris,10 mM EDTA, pH 8) + 4.5 U RNase/mL.
PCR- conditons
The 18S rDNA was amplified using the primers GEOA2, GEO11 (Schwarzott D, Schüssler A (2001) A simple and reliable method for SSL) rRNA gene DNA extraction, amplification, and cloning from single AM fungal spores. Mycorrhiza 10: 203-207), Primers used for PCR amplification and for sequencing of the 18S rDNA- internai transcribed spacer région were Glom1310 and ITS4i (Redecker D. (2000) Spécifie PCR primers to identify arbuscular mycorrhizal fungi within colonized roots. Mycorrhiza 10: 73-80). Amplifications were performed in 0.2 mM dNTP-mix, 1 mM of each primer, 10 % of PCR reaction buffer and stérile, molecular grade water. GoTaq® DNA polymerase (Promega, Mannheim, Germany) was added at 4 u / 100 pL of reaction mix; 2 pL of genomic DNA template was used in each 20 pL reaction. Amplifications were carried out in a Primus 96 advanced- thermocycler (peqLab Biotechnology) in 200 pL reaction tubes using the following PCR conditions: 96 °C, 180 s initial dénaturation; followed by 35 cycles: 96 °C 30 s, 58 °C, 30 s, 72 °C, 90 s; and a final extention at 72 °C for 10 minutes.
Sequence data and Glomeromycota taxonomy
The reference alignment published by Krüger et al. (Krüger M, Krüger C, Walker C, Stockinger H, Schüβler A (2012) Phylogenetic reference data for systematics and phylotaxonomy of arbuscular mycorrhizal fungi from phylum to species level. New Phytol 193: 970-984; downloadable at www.amf-phylogeny.com) was used as basis for an 18S phylogeny. To identify similar environmental sequences the sequence of Dominikia sp. was blasted against GenBank and highly similar sequences were included in the alignment forming the basis of the 18S tree. Published partial 18S-ITS1partial 5.8S sequences were used for the Dominikia phylogeny .
Data analyses
Alignments were initially carried out by Clustal W. A maximum likelihood phylogenetic analysis was computed through the CIPRES web-portal (Miller MA, Pfeiffer W, Schwartz T. (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic très. In Proceedings of the Gateway Computing Environments Workshop (GCE), 14 Nov. 2010, New Orléans, LA pp 1 - 8; http://www.phylo.org/) with RAxML version 8.0 (Stamatakis et al. 2014) using 100 bootstrap replicates and the GTRGAMMA model. A Bayesian consensus tree was constructed using MrBayes version 2.0.5. Two separated MC3 runs with randomly generated starting trees were carried out for 2M générations each with one cold and three heated chains using the GTR+I model. Ail parameters were estimated from the data. Trees were sampled every 1000 générations. 200000 générations were discarded as “burn-in” and consensus trees constructed from the returning sample.
Example 2. Production Procedure of an embodiment ofthe composition ofthe invention - Concentrated composition for seeds coating First Phase - Greenhouse conditions
Substrate: Smectite clays are selected with pH between 7.8 and 8 and are sterilized in alternating cycles of 3 days before use as substrate.
AMF strain: Dominikia sp. propagules in pure conditions are used as started inoculum. This type of inoculant is always in continuous playback in cycles of 90-180 days, either in growth chamber or in greenhouses with controlled host plant conditions. Different plants are used to reproduce the inoculum in successive cycles to avoid disease transmission in the same host.
Summer- Fall: Sorghum vulgare and Ocimum sp.
Winter - Spring: Lolium perenne.
Culture Pots: Pots with 15 liter volume is used.
Growth conditions of host plants:
In growth chamber or greenhouse, cultivation begins with the straight inoculation of the root System of the host plant with pure inoculum of the selected strain of arbuscularmycorrhizal fungus (Dominikia sp.) in a stérile Smectite clay substrate. These plants are grown through their complété life cycle, according to the type of plant, taking between 90 and 180 days. The plants are always kept well hydrated, with a daily irrigation supply (Stérile water) with a température range between 25 and 28 °C and a relative humidity of 65%. Once completed, the root System is extracted, which contains smectite substrate, rootlets and pure AMF propagules, to be subsequently used to scale up in the second phase.
To détermine the quality of the inoculum, the following minimum spécifications are used: Total Spores: 50-225 spores/g.
Extramatrical mycélium: > 70 mg/kg substrate.
Rootlets colonized percentage: Sorghum > 50 %, Lolium > 45 %, Ocimum > 40 %.
MPN concentration: > 1x104 propagules per 100 ml of substrate.
Second Phase - Scale Up
Step 1 : Préparation of beds
The beds are constructed with a plastic liner material so that the bed is isolated from surrounding soil; the beds are constructed so that the drainage occurs and the growth of undesired végétation is prevented, preferably with a plastic covering.
Beds should be filled with the selected Smectite clay (Arcilla Roja Galve). The humidity of the clay should be approximately 15% to facilitate handling during bed filling. After filling irrigation should be provided to saturation to improve the structure.
- Beds are located on well-drained sites.
- Beds can hâve any dimensions, taking into considération access required to facilitate the transit of people and equipment necessary for bed care.
- Beds hâve irrigation System as dictated based on local needs. The preferred System setup may be either a drip irrigation or sprinkler and should be automated and allow independent watering of selected areas of the bed.
Step 2: Host plant species
Détermination of host plant species and mycorrhizal fungi that can be set in the System: The sélection and identification of the host plant and fungal species will correspond to the spécifie site conditions and objectives of the production. Rye grass (Lolium perenne) and the AMF previously produced at First Phase is used.
Step 3: Seedinq the host plant and inoculation
Prior to planting of the host plant, seed germination is tested. Based on the results of this test, the appropriate seeding rate is determined. In the case of perennial ryegrass seed is sown broadeast at 80 kg/ha of seed previously certified using pelleted seed. Also in conjunction with the seed 20 g of AMF inoculum m2 bed is applied directly to the Smectite clay.
Immediately after sowing irrigation is applied as a fine spray to prevent the redistribution of seed and inoculum.
The water used for irrigation should preferably hâve the following characteristics:
- pH values of > 6 to < 7.5
- Electrical conductivity: < 1.6 mS/m
- Soluble Total Salts: < 1000 ppm
- Sodium Absorption Ratio (SAR) <10
- free of heavy éléments and pathogens. It is preferred that the water used for irrigation is drinking water.
Step 4: Cultural activities and irrigation
Irrigation applied should be sufficient to achieve 100% of field capacity, but avoid applying too much water and causing ponding or standing water. Beds should be irrigated again when clay moisture drops to 75 - 80% of field capacity.
Step 5: Managing the establishment of mycorrhizal symbiosis and knowledge of the dynamics of development of the colonization of mycorrhizal fungus.
During the growth and development of the host plant, AMF root colonization occurs and establishes the symbiosis between plant and fungus. To evaluate the development of this relationship, periodic sampling of roots System is conducted to evaluate the mycorrhizal development. Methods used to assess colonization include Gerdeman & Nicolson (1963) (Gerdemann J.W., Nicolson T.H. 1963. Spores of mycorrhizal endogen species extracted from soil by wet sieving and decanting. Transactions of the British Mycological Society 46(2):235-44.), McGonigle (1990) (McGonigle, T.P., Miller M.H., Evans D.G., Fairchild G.L., Swan J.A. 1990. A new method which gives an objective measure of colonization of roots by vesicular arbuscular mycorrhizal fungi. New Phytologist 115 (3):495-501), and Phillips & Hayman (1970) (Phillips, J.M., Hayman D.S. 1970. Improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection. Transactions of the British Mycological Society 55:158-161.).
Sampling begins at two months after planting and continues monthly until the end of the growing season. With the information obtained from these samples the dynamics of development of mycorrhizal symbiosis within the inoculant can be determined.
Production process assessment is based on mycorrhizal root colonization, extrametrical concentration of mycélium, and spore content clay samples that are taken periodically.
Knowing the dynamics of mycorrhizal production development allows détermination of the optimum harvest time and makes the most of the process of symbiosis on host plant and mycorrhizal fungus.
Step 6: Harvest
If the host plant that is used is the perennial Lolium, then the optimal harvest time normally occurs between 6 and 7 months after seeding, because in that period the plants mature, complété their life cycle and show a tendency to loss of vigor and become yellow.
Fifteen days before the scheduled harvest date the irrigation supply is eliminated and the foliage is maintained to allow the clay to lose moisture slowly ensuring completion of inoculation process. If this activity coïncides with the rainy season, it will be necessary to protect the bed from the rain to allow it to dry in a timely manner, by covering the bed with waterproof plastic.
Above ground foliage from host plants is first manually removed. Harvesting is done by removing clay from the bed. Remove the substrate by dividing the mass of clay on portions as thinly as possible to facilitate mixing their content throughout the depth of the profile and place it in the bags to transport.
Step 7: Dry and millinq of inoculum
Drying: The harvested substrate and mycorrhizal propagules are subject to solarization and thermal disinfection for 30 days at 50 °C. The drying period may be extended until a moisture content below 5%, in order to facilitate the milling process.
Milling: The product is ground in an industrial mill, cooled to 2 °C to prevent overheating mycorrhizal propagules. Grinding continues until a particle size of below 100 microns is achieved.
Third Phase - Concentration
After Step 7, the grounded biomass, it is corne to the product concentration using a sieve of 35 microns. The particle obtained below of this measure (between 60- 70% of the initial product) is discarded and stayed with the and 25 to 30% of the ultraconcentrated product does not pass through that size and will be at the end, a concentrated composition. With the use of this technology we passed from 1.2 x104 propagules per 100 ml to 1.2 x 106 propagules per 100 ml.
The outlet relative humidity of the product is below 5 %.
Quality Control: The final product purity and concentration is determined following the most probable number method of Porter (1979) (Porter, W.M. 1979. The most probable number method for enumerating infective propagules of vesicular arbuscular mycorrhizal fungi in soils. Aust. J. Soil Res. 17:515-519.).
Packaging: Finished product is packaged and labeled for shipment.
The final concentration of the concentrate composition is:
MPN concentration: > 1.2 x 106 propagules per 100 ml of product. Fourth Phase - Seed Coatinq
Coating the seeds with mycorrhizal inoculant can be performed in a spécial machine for coating seeds, in a conventional concrète mixer or manually in a mixing vessel. Said coating requires various steps that are described below in the order in which they are performed.
1. As an initial step, the seeds are coated with an adhesive substance. The adhesive substances that can possibly be used in addition to water include organic adhesives (gelatin, ethyl cellulose, propylene glycol, etc.) and inorganic adhesives (minerai oils, polyvinyls, plastic resins, etc.). The preferred adhesives used are polymeric or copolymeric of the polyvinyl group such as polyvinylpyrrolidone and polyvinyl acetate. The adhesive is added to an aqueous or alcoholic solution to its optimal solubility, varying the amount of adhesive used between 0.1% and 15%, preferably between 0.5% and 10% and still more preferably between 1.0% and 5% of the total weight of the seeds to be coated. The amount of adhesive used will dépend on its Chemical properties as well as the type of seed to be treated. The time of treatment of the seeds with the adhesive may vary between 1 and 60 seconds, preferably between 5 and 50 seconds and still more preferably between 10 and 40 seconds per 100 kg of seeds used.
2. After coating with adhesive, the mycorrhizal inoculant is then added. The proportion of the inoculant added to the seeds may be selected in the range between 0.1% and 15%, preferably between 0.5% and 10% and still more preferably between 1% and 5% of the weight of seeds, which will dépend on the type and variety. The time of treatment of the seeds with the mycorrhizal inoculant may vary between 1 and 50 seconds, preferably between 5 and 40 seconds and still more preferably between 10 and 30 seconds per 100 kg of seeds used.
The mycorrhizal treatment may be combined with other treatments that are described in sections 2a to 2e. These treatments may also be performed independently of the mycorrhizal inoculant treatment, obtaining a multi-layer coating. The steps to be performed in the coating of each layer are the same as those in the case of the mycorrhizal inoculant. These layers may be separated by coating the seeds with innocuous substances of calcareous (calcium carbonate and similar), clay or polymeric origins. The pelletizing or encrusting substance must not exceed 50%, preferably 40% and still more preferably 30% of the weight of the seed and is added to the seed with an 15 adhesive substance as mentioned in section 1.
2a. Treatments with fungicides, insecticides and/or herbicides compatible with mycorrhizal forming fungi. In general terms, ail commercial herbicides and insecticides are compatible with mycorrhizal forming fungi. However, not ail fungicides are compatible with the survival of mycorrhizal fungi. The main fungicides to be used include azoxystrobin, carboxin, cyproconazole, chlorothalonil, metalaxyl, myclobutanil and prothioconazole. The amount of pesticide used will dépend on the manufacturées recommendations and various pesticides can be used depending on the need.
2b. Treatment with bénéficiai microorganisms including Trichoderma spp., Rhizobium bacteria and/or a combination of microorganisms bénéficiai to the rhizosphere such as Aspergillus, Pénicillium and nitrogen fixing bacteria.
2c. Treatment with macro and/or micronutrients, where nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnésium (Mg) and sulphur (S) are the essential macronutrients and iron (Fe), zinc (Zn), manganèse (Mn), boron (B), copper (Cu), molybdenum (Mo) and chlorine (Cl) are the essential micronutrients. The dose of nutrients for coating may vary between 0.01% and 15%, preferably between 0.05% and 10%, and still more preferably between 0.1% and 5% of the total weight of the seeds to be coated.
2d. Treatment with stimulants. Coating the seeds with stimulants that induce seed germination and growth and plant defence and sporulation and mycorrhizal fungal growth. The stimulants can be phytohormones (abscisic acid, strigolactones, brassinosteroids, etc.) and their inducers and dérivatives, secondary métabolites (flavonoids, terpenoids, etc.) and cofactors (métal ions, etc.).
2e. Treatment with colouring pigments: these pigments must be compatible with the survival of the mycorrhizal fungus and be a clear differentiator between treated and nontreated seeds.
3. Steps 1 and 2 mean that a complété coating of the mycorrhizal inoculant by the adhesive substance is performed in a time of from 1 to 40 seconds, preferably from 5 to 30 seconds and still more preferably from 10 to 20 seconds, performing a complété coating of the mycorrhizal inoculant by the adhesive substance.
4. After performing the mycorrhizal treatment, the seeds are dried for a time that can vary between 1 and 50 seconds, preferably between 5 and 40 seconds and still more preferably between 10 and 30 seconds. This step may be performed after step 5.
5. Finally the coated seeds are discharged into containers from the coating component. The duration of the discharge may vary from 5 to 30 seconds, preferably from 10 to 25 seconds and still more preferably from 15 to 20 seconds.
Example 3. Production Procedure of an embodiment ofthe composition ofthe invention - Microgranulated composition
First Phase - Greenhouse conditions
Substrate: Smectite clays are selected with pH between 7.8 and 8 and are sterilized in alternating cycles of 3 days before use as substrate.
AMF strain: Dominikia sp. propagules in pure conditions are used as started inoculum. This type of inoculant is always in continuous playback in cycles of 90-180 days, either in growth chamber or in greenhouses with controlled host plant conditions. Different plants are used to reproduce the inoculum in successive cycles to avoid disease transmission in the same host.
Summer- Fall: Sorghum vulgare and Ocimum sp.
Winter - Spring: Lolium perenne.
Culture Pots: Pots with 15 liter volume is used.
Growth conditions of host plants:
In growth chamber or greenhouse, cultivation begins with the straight inoculation of the root System of the host plant with pure inoculum of the selected strain of arbuscularmycorrhizal fungus (Dominikia sp.) in a stérile smectite clay substrate. These plants are grown through their complété life cycle, according to the type of plant, taking between 90 and 180 days. The plants are always kept well hydrated, with a daily irrigation supply (Stérile water) with a température range between 25 and 28 °C and a relative humidity of 65%. Once completed, the root System is extracted, which contains smectite substrate, rootlets and pure AMF propagules, to be subsequently used to scale up in the second phase.
To détermine the quality of the inoculum, the following minimum spécifications are used: Total Spores: 50-225 spores/g.
Extramatrical mycélium: > 70 mg/kg substrate.
Rootlets colonized percentage: Sorghum > 50 %, Lolium > 45 %, Ocimum > 40 %.
MPN concentration: > 1x104 propagules per 100 ml of substrate.
Second Phase - Scale Up
Step 1 : Préparation of beds
The beds are constructed with a plastic liner material so that the bed is isolated from surrounding soil; the beds are constructed so that the drainage occurs and the growth of undesired végétation is prevented, preferably with a plastic covering.
Beds should be filled with the selected Smectite clay (Arcilla Roja Galve). The humidity of the clay should be approximately 15% to facilitate handling during bed filling. After filling irrigation should be provided to saturation to improve the structure.
- Beds are located on well-drained sites.
- Beds can hâve any dimensions, taking into considération access required to facilitate the transit of people and equipment necessary for bed care.
- Beds hâve irrigation System as dictated based on local needs. The preferred System setup may be either a drip irrigation or sprinkler and should be automated and allow independent watering of selected areas of the bed.
Step 2: Host plant species
Détermination of host plant species and mycorrhizal fungi that can be set in the System: The sélection and identification of the host plant and fungal species will correspond to the spécifie site conditions and objectives of the production. Rye grass (Lolium perenne) and the AMF previously produced at First Phase is used.
Step 3: Seedinq the host plant and inoculation
Prior to planting of the host plant, seed germination is tested. Based on the results of this test, the appropriate seeding rate is determined. In the case of perennial ryegrass seed is sown broadeast at 80 kg/ha of seed previously certified using pelleted seed. Also in conjunction with the seed 20 g of AMF inoculum m2 bed is applied directly to the Smectite clay.
Immediately after sowing irrigation is applied as a fine spray to prevent the redistribution of seed and inoculum.
The water used for irrigation should preferably hâve the following characteristics: - pH values of > 6 to < 7.5
- Electrical conductivity: < 1.6 mS/m
- Soluble Total Salts: < 1000 ppm
- Sodium Absorption Ratio (SAR) <10
- free of heavy éléments and pathogens. It is preferred that the water used for irrigation is drinking water.
Step 4: Cultural activities and irrigation
Irrigation applied should be sufficient to achieve 100% of field capacity, but avoid applying too much water and causing ponding or standing water. Beds should be irrigated again when clay moisture drops to 75 - 80% of field capacity.
Step 5: Managing the establishment of mycorrhizal symbiosis and knowledge of the dynamics of development of the colonization of mycorrhizal fungus.
During the growth and development of the host plant, AMF root colonization occurs and establishes the symbiosis between plant and fungus. To evaluate the development of this relationship, periodic sampling of roots System is conducted to evaluate the mycorrhizal development. Methods used to assess colonization include Gerdeman & Nicolson (1963) (Gerdemann J.W., Nicolson T.H. 1963. Spores of mycorrhizal endogone species extracted from soil by wet sieving and decanting. Transactions of the British Mycological Society 46(2):235-44.), McGonigle (1990) (McGonigle, T.P., Miller M.H., Evans D.G., Fairchild G.L., Swan J.A. 1990. A new method which gives an objective measure of colonization of roots by vesicular arbuscular mycorrhizal fungi. New Phytologist 115 (3):495-501.), and Phillips & Hayman (1970) (Phillips, J.M., Hayman D.S. 1970. Improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection. Transactions of the British Mycological Society 55:158-161.).
Sampling begins at two months after planting and continues monthly until the end of the growing season. With the information obtained from these samples the dynamics of development of mycorrhizal symbiosis within the inoculant can be determined.
Production process assessment is based on mycorrhizal root colonization, extrametrical concentration of mycélium, and spore content clay samples that are taken periodically.
Knowing the dynamics of mycorrhizal production development allows détermination of the optimum harvest time and makes the most of the process of symbiosis on host plant and mycorrhizal fungus.
Step 6: Harvest
The harvest is the most critical step in the production process. If the host plant that is used is the perennial Lolium, then the optimal harvest time normally occurs between 6 and 7 months after seeding, because in that period the plants mature, complété their life cycle and show a tendency to loss of vigor and become yellow.
Fifteen days before the scheduled harvest date the irrigation supply is eliminated and the foliage is maintained to allow the clay to lose moisture slowly ensuring completion of inoculation process. If this activity coïncides with the rainy season, it will be necessary to protect the bed from the rain to allow it to dry in a timely manner, by covering the bed with waterproof plastic.
Above ground foliage from host plants is first manually removed. Harvesting is done by removing clay from the bed. Remove the substrate by dividing the mass of clay on portions as thinly as possible to facilitate mixing their content throughout the depth of the profile and place it in the bags to transport.
Step 7: Dry and milling of inoculum
Drying: The harvested substrate and mycorrhizal propagules are subject to solarization and thermal disinfection for 30 days at 50 °C. The drying period may be extended until a moisture content below 5%, in order to facilitate the milling process.
Milling: The product is ground in an industrial mill, cooled to 2 °C to prevent overheating mycorrhizal propagules. Grinding continues until a particle size of below 100 microns is achieved.
Third Phase - Micro granulation
The micro granulation process is divided in three steps:
- Addition of granular support into the rotating biconical mixer. To produce the support was used: Mix of mica, attapulgite and limestone in a range between 40-90% by weight, more preferably between 50-80% by weight, even more preferably between 60-70 by weight.
- Addition of mycorrhizae (arbuscular mycorrhizal fungus): the AMF is dosed between 10-60% by weight, more preferably between 20-50% by weight, even more preferably between 30-40% by weight of Dominikia sp. previously produced (Step 7) along with a binder (wax, linseed oil, gum arabic, gum tragacanth, methyl cellulose, polyvinyl alcohol, tapioca flour, lactose, sucrose, microcrystalline cellulose, poly vinyl pyrrolidone, lactose powder, sucrose powder, tapioca starch (cassava flour) and microcrystalline cellulose, gums, or protein such as egg white or casein) in a range of 1 to 25% by weight, more preferably of 5 to 20% by weight, even more preferably of 10 to 15 % by weight inside the rotating biconical mixer by a screw. During this operation, the mixer is kept moving until total granule homogenization.
- Packaging: once the formulated product is totally homogenized it is discharged onto a sieve to remove clumps and dust produced. Finally, the product is collected in a packaging hopper from which automatically proceeds to give the corresponding filling containers.
Quality control and packaging
Quality Control: The final product purity and concentration is determined following the most probable number method of Porter (1979) (Porter, W.M. 1979. The most probable number method for enumerating infective propagules of vesicular arbuscular mycorrhizal fungi in soils. Aust. J. Soil Res. 17:515-519.).
Packaging: Finished product is packaged and labeled for shipment.
The final concentration of the microgranulated composition is: MPN concentration: > 1x104 propagules per 100 ml of product.
Example 4. Effect of the composition of the invention on the Development of Wheat (Triticum durum)
In order to demonstrate the effectiveness of the composition of the présent invention, which comprises the arbuscular mycorrhizal fungus (AMF) Dominikia sp., deposited under accession number MUCL 57072 on wheat, a trial was conducted in the experimental field “Très caminos”, property of CEBAS-CSIC in Murcia.
The experimental design was randomized block with four réplications. The experimental plots were 3 m long per 4.2 m wide.
The following treatments were applied:
- Control: no Chemical fertilization and no fungus applied
- Composition of the invention: provided in microgranulated form, with a rate of 10 kg/ha, in combination with Chemical fertilization (Guerra, B. E. Micorriza arbuscular. Recurso microbiolôglco en la agricultura sostenible. Tecnologia en Marcha, 2008, vol. 21, no. 1, p. 191-201).
- Standard product: Chemical fertilization (Guerra, B. E. Micorriza arbuscular. Recurso microbiolôgico en la agricultura sostenible. Tecnologia en Marcha, 2008, vol. 21, no. 1, p. 191-201): 150 kg/ha of nitrogen, 54 kg/ha of phosphorus, 100 kg/ha of potassium, 15 kg/ha of calcium, 15 kg/ha of magnésium, 23 kg/ha of sulphur and other micronutrients.
Planting was done on 1 November 2013 and the harvesttook place on June 1,2014, for a total of 240 days. The type of sowing and inoculation used was mechanical and was applied by a grain drill stroke and the composition of the invention, in microgranulated form, was applied at 10 kg/ha along with the seed.
The soil used was classified as Nitisol (IUSS Working Group. WRB. World reference base for soil resources 2006. World Soil Resources Reports. Rome: FAO, 2006). For Chemical characterization of the soil the following analytical methods were used:
- pH: soil-solution ratio 1:2.5.
- Organic matter (MO): Walkley and Black.
- P2O5: Oniani.
- Exchangeable cations: extraction with NH4 AC at 1 mol/L at pH 7 and complexometric titration (Ca and Mg) and flame photometry (Na and K).
These methods are described in the manual of analytical techniques for the analysis of soil, foliar, organic fertilizers and Chemical fertilizers (Instituto Nacional de Ciencias Agricoles. Manual de técnicas Analiticas para anâlisis de suelo, foliar, abonos orgânicos y fertilizantes quimicos: La Habana. 1989).
The results of Chemical soil characteristics (Table 2) show a medium to high fertility, noting an average content of organic matter, which respond to that described for this type of soil (Hernandez, A.; Morell, F.; Ascanio, Μ. O.; Borges, Y.; Morales, M. y Yong, A. Cambios globales de los suelos Ferraliticos Rojos Lixiviados (Nitisoles Rôdicos Éutricos) de la provincia La Habana. Cultivos Tropicales, 2006, vol. 27. no. 2, p. 4150.).
Table 2: Chemical characteristics of soil.
Table 2 | |
Organic matter (%) | 3.21 |
PH | 7.02 |
p2o5 | 397.25 |
K (cmol/kg soil) | 0.96 |
Ca (cmol/kg soil) | 14.26 |
Mg (cmol/kg soil) | 3.70 |
Na (cmol/kg soil) | 0.11 |
The fungal strain of the invention of Dominikia sp. was isolated from a saline soil, this fungus is tolérant to high sait concentrations.
The variables analyzed were: the number of spores of AMF per gram of rhizospheric soil (Gederman, J. W. y Trappe, J. M. The endogonaceae in the pacifie northwest. Mycologia Memoir No 5. The New York Botanic Garden. 1974, no. 5), the percentage of mycorrhizal colonization by the technique of staining of the roots (Phillips, J. M. y Hayman, D. S. Improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection. Tranfer. Britanic: Mycology Society, 1970,vol. 55, p. 158-161), the percentage of Visual density through the intercepts method (Giovannetti, M. y Mosse, B. An évaluation of techniques to measure vesicular-arbuscular infection in roots. New Phytology, 1980, vol. 84, p. 489-500) and the total glomalin (glycoprotein) content which was obtained by the pressure cooker method (Wright, S. E.; Nichols, K. A. y Schmidt, W. F. Comparison of efficacy of three extradants to solubilize glomalin on hyphae and in soil. Chemosphere, 22
2006, vol. 64, no. 7, p. 1219-1224).
Foliar nutrients content (% N, P and K) and the total protein content were determined by the methods described in the laboratory manual of analytical techniques of INCA (Instituto Nacional de Ciencias Agricolas. Manual de técnicas Analiticas para anâlisis de suelo, foliar, abonos orgânicos y fertilizantes quimicos: La Habana. 1989).
The yield was evaluated as follows: the number of grains per spike were evaluated; number of spike per m2; mass per 1000 grains and agricultural yields (T/ha).
The statistical Processing of the experimental results was done by analysis of one way ANOVA and test of Duncan (Duncan, D. B. Multiple range and multiple F tests. Biométries, 1955, vol. 11, no. 1) was used, when there were différences between treatment means statistical processor SPSS 11.5 was used in ail cases.
Results and Discussion
Table 3 shows the results of the different treatments on the mycorrhizal variables. These values clearly showed an increased number of spores where the composition of the invention was applied in relation with the control (no Chemical fertilization) and with the standard product.
Table 3: Effect of treatments on mycorrhizal variables studied.
Table 3 | ||||
Treatments | Spores/gram of soil | Colonizati on (%) | VD (%) | Glomalin (mg/g soil) |
Control | 42 b | 12.3 b | 0.12 b | 1.8 b |
Composition of the invention | 156 a | 65.3 a | 3.25 a | 9.27 a |
Standard product | 36 b | 11.2 b | 0.11 b | 1.78 b |
P | 0.014** | 0.04*** | 0.021 | 0.017** |
F | 8.5 | 9.2 | 7.1 | 6.5 |
Different letters in the same column are significantly different at p < 0.05.
The above it may be due to a strong colonization activity, making difficult the root colonization by other species of arbuscular mycorrhizal fungi. On the other hand, it shows that mycorrhizal colonization was also more effective with the composition of the invention reflecting higher values compared to other treatments.
It should be noted that native strains in soil showed low values of colonization. By 23 observing the values of Visual density (variable measuring the intensity of mycorrhizal colonization), shows that the highest percentage was obtained with the composition of the invention .
Another variable to highlight was total glycoprotein content which was higher with the composition of the invention. This phenomenon reaffirms a greater mycorrhizal symbiosis when the composition of the invention is applied and the possible influence of the inoculant to increase the formation of aggregates in the soil. Moreover, treatments where mycorrhizal inoculant was not applied had low values of this variable, which may be attributable to the low effectivity of native mycorrhizae.
Table 4 shows the foliar minerai contents on each treatment. For nitrogen and phosphorus the content was higher where the composition of the invention was applied while the sodium content was higher for the Chemical fertilization.
Table 4: Foliar nitrogen, phosphorus, potassium and total protein.
Table 4 | ||||
Treatments | % N | % P | % K | Total protein (%) |
Control | 1.25 b | 0.26 b | 1.12 | 8.04 b |
Composition of the invention | 1.45 a | 0.40 a | 1.18 | 10.1 a |
Standard product | 1.41 a | 0.36 a | 1.23 | 9.89 a |
P | 0.001** | 0.023** | 1.23 ns | 0.013* |
F | 7.2 | 5.6 | 8.8 | 6.5 |
Different letters in the same column are significantly different at p < 0.05.
It should be noted that reported values (Lôpez-Bellido, L. Cultivos herbâceos. Cereales. Ed. Mundi-Prensa, 1991. p. 151-158) for this variable (% N foliar) are above the critical index of wheat yields between 4 and 5 T/ha.
Similar results (Cornejo, P.; Borie, F.; Rubio, R. y Azcon, R. Influence of nitrogen source on the viability, functionality and persistence of Glomus etunicatum fungal propagules in an Andisol. Applied Soil Ecology, 2007, vol. 35, no. 2, p. 423-431; Echeverria, E. y Stiddert, G. A. El contenido de nitrôgeno en la hoja bandera del trigo como predictivo del incremento de proteina en el grano por aplicaciones de nitrôgeno en la espigazôn (The nitrogen content in the wheat flag leaf as prédictive of the increase in protein in the grain by nitrogen applications to the ear). Revista de la Facultad de Agronomia, 1998, vol. 103, no. 1, p. 10) indicate the presence of this nutrient in leaf tissue, due to mycorrhizal symbiosis process which allows the absorption and transport of nutrients through the mycelia. The content of foliar phosphorus did not show significant différences for any of the treatments, which may be due to the content of this element 24 on soil (Table 4), which is taken up by plants from the soil solution through its radical and System, in this particular case by the tripartite interaction plant-arbuscular mycorrhizal (mycorrhizal symbiosis) and soil, pointing out a higher value than the foliar critical index of this nutrient in durum wheat (Lôpez-Bellido, L. Cultivos herbâceos.
Cereales. Ed. Mundi-Prensa, 1991. p. 151-158) and indicating a good crop development. As well as this value was higher trend where the composition of the invention was applied.
Another variable that is relevant is the foliar protein content, which is higher in the treatment with the composition of the invention (even higher than treated with Chemical 10 fertilization). Finally, the control treatment had a significant lower percentage.
The results of yield and its components (Table 4) reflect clearly the response of durum wheat to the application of the composition of the invention and standard product, both with high contents of N.
Table 5: Effect of treatments on yield and its variables.
Table 5 | |||||
T reatments | Grains/spike | N° spikes/m 2 | Mass of 1000 grains (g) | Yield (T/ha) | Increase (%) |
Control | 25.00 b | 576 | 29.0 b | 3.25 b | |
Composition of the invention | 33.00 a | 641 | 33.4 a | 5.22 a | 8.8 |
Standard product | 30.00 a | 635 | 32.0 a | 4.60 a | — |
P | 0.02** | 11.25 ns | 0.015* | 0.009** | |
F | 11.2 | 10.2 | 6.1 | 7.8 |
* Different letters in the same column are significantly different at p < 0.05.
Treatments with the composition of the invention and standard product gave the highest values on each variable with respect to the control. It could be explained by greater effectiveness of the solid inoculant and the application of high doses of nitrogen.
It is good to note that wheat cultivation associated with the composition of the invention, allowed to achieve adéquate productive response with acceptable indicators of mycorrhizal operation, not only to improve soil biological activity, but promoted a significant increase production of 8.8% in relation to the control and the standard 25 product.
Example 5. Effect of the composition of the invention on the Development of Corn (Zea mays).
In order to demonstrate the effectiveness of the composition of the invention, which comprises the arbuscular mycorrhizal fungus (AMF) Dominikia sp., deposited under accession number MUCL 57072, on maize, a trial was conducted on a field close to the town Egea de los Caballeros. A microgranular form of the composition of the invention was tested. A standard product was used to compare the effect of both products on the cultivation of maize, var DKC 6717, Monsanto; on a Cambisol soil.
Planting was done on 12 April and the harvest took place on December 14, 2014, for a total of 230 days. The type of sowing and inoculation used was mechanical and was applied by a grain drill stroke and the microgranulate composition of the invention was applied at 10 kg/ha along with the seed. The planting used was 0.7 m x 0.16 m for a total of 80 plants per square meter. The spray System used was 18 x 18 m.
The experimental design was randomized block with three réplications. The experimental plots were 20 m long x 20 m wide.
The following treatments were applied:
- Composition of the invention: provided at 10 kg/ha and Chemical fertilization (Guerra, B. E. Micorriza arbuscular. Recurso microbiolôgico en la agricultura sostenible. Tecnologia en Marcha, 2008, vol. 21, no. 1, p. 191-201).
- Standard product: Chemical fertilization (Guerra, B. E. Micorriza arbuscular. Recurso microbiolôgico en la agricultura sostenible. Tecnologia en Marcha, 2008, vol. 21, no. 1, p. 191-201): 325 kg/ha of nitrogen, 80 kg/ha of phosphorus and 200 kg/ha of potassium, with an initial contribution of organic matter in the form of slurry 10 T/ha.
The soil used was classified as Cambisol (IUSS Working Group. WRB. World reference base for soil resources 2006. World Soil Resources Reports. Rome:FAO, 2006). For Chemical characterization of the soil the following analytical methods were used:
- pH: soil-solution ratio 1: 2.5.
- Organic matter (MO): Walkley and Black.
- P2O5: Oniani.
- Exchangeable cations: extraction with NH4 AC at 1 mol/L at pH 7 and complexometric titration (Ca and Mg) and flame photometry (Na and K).
These methods are described in the manual of analytical techniques for the analysis of soil, foliar, organic fertilizers and Chemical fertilizers (Instituto Nacional de Ciencias Agricolas. Manual de técnicas Analiticas para anâlisis de suelo, foliar, abonos orgânicos y fertilizantes quimicos: La Habana. 1989).
The results of Chemical soil characteristics (Table 6) show a medium to high fertility, noting an average content of organic matter, which respond to that described for this type of soil (Hernandez, A.; Morell, F.; Ascanio, Μ. O.; Borges, Y.; Morales, M. y Yong, A. Cambios globales de los suelos Ferraliticos Rojos Lixiviados (Nitisoles Rôdicos Éutricos) de la provincia La Habana. Cultives Tropicales, 2006, vol. 27. no. 2, p. 41-50). Table 6: Chemical characteristics of soil
Table 6 | |
Organic matter (%) | 2.51 |
pH | 8.02 |
P2O5 | 258.60 |
K (cmol/kg soil) | 1.40 |
Ca (cmol/kg soil) | 15.44 |
Mg (cmol/kg soil) | 2.90 |
Na (cmol/kg soil) | 0.30 |
The fungal strain of the invention of Dominikia sp. was isolated from a saline soil, this fungus is tolérant to high sait concentrations.
The variables analyzed were: percentage of mycorrhizal colonization by the technique of staining of the roots (Phillips, J. M. y Hayman, D. S. Improved procedures for clearing roots and staining parasitic and vesiculararbuscular mycorrhizal fungi for rapid assessment of infection. Tranfer. Britanic: Mycology Society, 1970,vol. 55, p. 158-161) and percentage of Visual density (VD) through the intercepts method (Giovannetti, M. y Mosse, B. An évaluation of techniques to measure vesicular-arbuscular infection in roots. New Phytology, 1980, vol. 84, p. 489-500). Foliar nutrients content (% N, P and K) were determined by the methods described in the laboratory manual of analytical techniques (Instituto Nacional de Ciencias Agricolas. Manual de técnicas Analiticas para anâlisis de suelo, foliar, abonos orgânicos y fertilizantes quimicos: La Habana. 1989).
The yield was evaluated as follows: the number of grains per spike were evaluated; number of spike per square meter; mass per 1000 grains and agricultural yields (T/ha). The statistical processing of the experimental results was done by analysis of one way ANOVA and test of Duncan (Duncan, D. B. Multiple range and multiple F tests.
Biométries, 1955, vol. 11, no. 1) was used, when there were différences between treatment means statistical processor SPSS 11.5 was used in ail cases.
Results and Discussion
Table 7 shows the results of the mycorrhizal activity évolution at 45 and 120 days after planting. These values clearly showed an increased activity where the composition of the invention was applied, reaching not only a greater mycorrhizal colonization percentage, but also a greater intensity of colonization, reflected through higher Visual density.
Table 7: Effect of treatments on mycorrhizal variables studied.
Table 7 | ||||
Colonization (%) 45 days | Colonization (%) 120 days | VD (%) 45 days | VD (%) 120 days | |
Composition of the invention | 36.2 a | 64.12 a | 3.4 a | 4.28 a |
Standard Product | 10.1 b | 11.36 b | 0.3 b | 0.85 b |
P | 0.011** | 0.02*** | 0.01* | 0.006* |
F | 7.2 | 8.2 | 8.1 | 9.4 |
* Different letters in the same column are significantly different at p < 0.05.
This assay showed that the most effective inoculant was the composition of the invention, which reflected higher values compared to other treatments.
The values of Visual density (variable which measures the intensity of the mycorrhizal colonization), shows that the highest percentage was also related to the composition of the invention, closely related to the effectiveness.
Foliar minerai contents were measured (Table 8). The results showed no significant différences between both treatments, the composition of the invention and standard product. However there was a trend to a greater nutritional supply, especially in the case of nitrogen content in the presence of Dominikia sp. (composition of the invention). Table 8: Foliar nitrogen, phosphorus, potassium at 120 days on maize.
Table 8 | |||
Treatments | % N | % P | % K |
Composition of the invention | 1.48 | 0.41 | 1.81 |
Standard product | 1.40 | 0.35 | 1.9 |
P | 0.12 ns | 0.1 ns | 0.22 ns |
F | 8.5 | 8.2 | 7.9 |
* Different letters in the same column are significantly different at p < 0.05.
It should be noted that nitrogen content values obtained with the treatment with the composition of the invention are above the critical index of wheat for production yields between 4 and 5 T/ha (Lôpez-Bellido, L. Cultivos herbâceos. Cereales. Ed. MundiPrensa, 1991. p. 151-158).
The content of foliar phosphorus did not show significant différences for any of the treatments, which may be due to the content of this element on the soil used (Table 8). Phosphorus is taken up by plants through its radical System, in this particular case by the tripartite interaction plant-arbuscular mycorrhizal (mycorrhizal symbiosis) and soil. The foliar phosphorus content was higher than the critical index of this nutrient in durum wheat (Lôpez-Bellido, L. Cultivos herbâceos. Cereales. Ed. Mundi-Prensa, 1991. p. 151-158) which indicate a good crop development. However this value was higher trend in the treatment treated with the composition of the invention.
Similar results in wheat and maize demonstrate the effect of minerai fertilization on the percentage of length of colonized roots, where low doses of phosphorus were applied. This support the importance and usefulness of the composition of the invention for corn and wheat plants to increase the absorption of phosphorus from the soil in either presence or absence of nitrogen and phosphorus, which allows to minimize the dose fertilizer to apply.
Foliar potassium contents were similar for the different treatments under study, showing satisfactory levels for this crop, which coïncides with the statements for cereals about this macro element (Lôpez-Bellido, L. Cultivos herbâceos. Cereales. Ed. Mundi-Prensa, 1991. p. 151-158). Moreover, high concentrations of this element are found in both mycorrhizal plants and those that are not (Bolleta, A. y Krugger, H. Fertilizaciôn e inoculation con hongos micorrizicos arbusculares en trigo. Buenos Airesdnstituto National de Tecnologia Agropecuaria. 2004., Saleque, M. A.; Timsina, J.; Panaullah, G. M.; Ishaque, M.; Pathan, D. J.; Saha, P. K.; Quayyum, M. A.; Humphreys, E. y Meisner, C. A. Nutrient uptake and apparent balances for rice-wheat sequences. II. Phosphorus. Journal of Plant Nutrition, 2006, vol. 29, no. 1, p. 157-172), which may be due to thefact that this element moves more easily in the soil solution.
The following table sum up the results obtained (Table 9), which clearly reflect the maize response to the application of the composition of the invention and to the standard product.
Table 9. Effect of treatments on yield and its components in the culture.
Table 9 | |||
Treatments | Yield (kg/ha) | Grain humidity (%) | Increase (%) |
Composition of the invention | 14700.2 a | 22 | 9.18 |
Standard Product | 13500.3 b | 20 | — |
P | 0.042** | ||
F | 29.40 |
* Different letters in the same column are significantly different at p< 0.05.
Plants treated with the composition of the invention reached the highest values in comparison with the standard product, which could be explained by a greater effectiveness of the solid inoculant to establish a symbiosis with the plant and thus a better assimilation of high doses of nitrogen.
Finally and in conclusion is good to point out that maize cultivation associated with the composition of the invention, allowed to achieve a positive production response from the point of view of performance, reaching a production increase of 9.18% compared to minerai fertilization. As well as, nutritional indicators showed a higher trend and the use of arbuscular mycorrhizal fungus of the invention not only improve soil biological activity, but promoted a significant increase in production based on a more sustainable management.
Example 6. Effectiveness of coating corn seed with Dominikia sp. and an adhesive substance on the mycorrhizal activity.
Objective: To learn about the efficacy of coating corn with Dominikia sp. and an adhesive substance on the mycorrhizal activity.
To achieve this objective, three planters (each considered a répétition) were planted with corn seeds coated with Dominikia sp. deposited under accession number MUCL 57072 and an adhesive substance.
The seeds were planted on 8/3/2016 and mycorrhizal colonisation assays were performed on the root using the Phillips and Hayman, 1970, staining technique at 21 and 35 days after planting.
Table 10. Percentage of mycorrhizal colonisation (%MC) of the rootlets from corn seeds coated with the combination of Dominikia sp. and an adhesive substance at 21 and 35 days post planting (dpp) respectively.
Table 10 | |||||
Date évaluation | of | Planter 1 (% MC) | Planter 2 (% MC) | Planter 3 (% MC) | Mean (X) |
28/3/2016 dpp) | (21 | 7 | 9 | 10 | 8.6 |
10/4/2016 dpp) | (35 | 12 | 11 | 15 | 12.6 |
Table 10 shows that in each of the samples analysed, there was a positive mycorrhizal colonisation percentage, which increased as the plants were growing. At this stage, only incipient colonisation points on the corn were detected, formed by the network of extramatrical mycélium that had just begun to form.
Conclusions:
The coating of the corn seed with the concentration of inoculant containing Dominikia sp. and an adhesive substance was effective because it generated mycorrhizal structures in the root as extramatrical mycélium in the first stages of the plant. Spores of the germinated species were seen and formed an internai germination and colonisation network. The values of colonisation depending on the times of corn growth are also shown.
Example 7. Results obtained in the field with the application of the composition of the invention in microoranulated form in wheat and barley.
Results obtained in the field with the application of composition of the invention, comprising Dominikia sp. deposited under accession number MUCL 57072, in microgranulated form in wheat and barley are reported in following Table 11.
Table 11
Crop | Variety | Irrigation | Treated (kg. ha1) | Not Treated (kg. ha1) | Improvement (%) |
Barley | Voley | Rainfed Cultivation | 2600 | 3000 | 15 |
Barley | Signoro | Irrigated Cultivation | 7100 | 7700 | 8.5 |
Wheat | Bonifacio | Irrigated Cultivation | 6350 | 7000 | 10.2 |
Wheat | Badré | Irrigated Cultivation | 4981 | 5463 | 9.7 |
Wheat | Chambo | Rainfed Cultivation | 3350 | 3700 | 10.4 |
Wheat | durum | Rainfed Cultivation | 4300 | 4900 | 14 |
Wheat | durum | Rainfed Cultivation | 4570 | 5110 | 11.8 |
Wheat | durum | Irrigated Cultivation | 6345 | 7135 | 12 |
Example 8. Results obtained in the field with the application of the composition of the invention in microgranulated form in corn.
Results obtained in the field with the application of the composition of the invention, 5 comprising Dominikia sp. deposited under accession number MUCL 57072, in microgranulated form in corn are reported in following table 12.
Table 12 | |||||
VarietyCycle | Irrigation | Not T reated (dry Matter) | T reated (dry Matter) | IMPROVEMENT (kg/ha) | |
Zone 1 | Lg 30369 - 365 | Irrigated | 7.87 | 8.25 | 809.091 |
Zone 2 | Lg 30369 - 365 | Irrigated | 4.24 | 5.27 | 2408.655 |
Zone 3 | Lg 30369 - 335 | Irrigated | 4.85 | 5.26 | 963.080 |
Zone 4 | Lg 30369 365 | Irrigated | 6.84 | 7.89 | 2085.287 |
Zone 5 | Maisadour -350 | Irrigated | 7.40 | 8.09 | 1379.649 |
Example 9. Micorrhizal activity of Dominikia sp. in the cultivation ofrice (Oriza sativa) in flooded soil.
An experiment was carried out to define the mycorrhizal activity of the composition of the invention containing Dominikia sp., deposited under accession number MUCL 57072, in concentrated form, in flooded saline soil conditions. The composition used had a concentration between 1-4 x106 propagules/g of substrate. Conversely, according to embodiments, the concentration of the composition may be between 1-2 x104, preferably between 1,2-1,8 x104 propagules/g of substrate.
Materials and Methods:
The experiment was performed at the Très Caminos experimental farm owned by CEBAS-CSIC and located in the Matanza area, municipality of Santomera (Murcia). The plants were grown in a monolayer tunnel-type experimental greenhouse with an approximate surface area of 60 m2, covered with polycarbonate and top window protected with antitrips mesh. It was equipped with a cooling System and an aluminised screen shade System. The trial was performed with rice plants (Oriza sativa), variety J 104, subjected to two mycorrhizal inoculant treatments and the corresponding control without mycorrizal treatment. The mycorrizal treatment was the composition of the invention containing Dominikia sp., deposited under accession number MUCL 57072, in concentrated form at a dose of 1 kg/ha coating the rice seed.
When the inoculation was assured in individual shoots, at 15 days after germination, the plants were transplanted at a concentration of 10 plants per concrète channel, of 2 m2 surface area, using Hidromôrfico Gley Nodular Salinizado soil, according to the UNESCO soil classification (Hernâdez, A; Pérez, J.M; Bosch, D; Rivera, L: Nueva Version de Clasificaciôn Genetica de los Suelos de Cuba. Soil Institute. AGRINFOR, La Habana, 1999. 64p) as a substrate in both containers. The main features are shown in Table 1. The cultivation work was performed and a layer of water was added after 18 days of sowing the seeds in ail treatments.
Table 13. Some Chemical properties and number of spores.50 g soil'1 of Hidromôrfico Gley Nodular Salinizado soil used in the experiment.
Table 13 | ||||||||
M.O | P | P | Ca | Mg | K | Na | C.E | No |
(%) | H | (cmol.kg· 1) | cmol.kg'1 | (qS.cm 1) | spores.g soil'1 | |||
2.3 | 7. 5 | 13.2 | 10.2 | 5.6 | 0.9 | 2.2 | 2876 | 1.2 |
Characteristics of the composition of the invention in concentrated form
In the présent example, the mycorrhizal inoculant composition of the invention in concentrated form, contains the arbuscular mycorrhizal forming fungus Dominikia sp. deposited under accession number MUCL 57072. The composition used had a concentration between 1-4 x106 propagules/g of substrate.
Déterminations performed
The dynamics of growth for 90 days post transplanting (dpt) was determined, where the plant height and depth of the root System was measured and also the harvest yield and some of its components.
Mycorrhizal action was determined during crop development by performing measurements of percentage colonisation (%), Visual density (%), arbuscular extramatrical mycélium and arbuscular endophytes using a stéréo microscope (Zeiss, West Germany -5-) and an Axiostar compound microscope (Zeiss, West Germany). The ratio between arbuscular external and endophyte mycélium was calculated (MEA:EA).
Mycorrhizal évaluation of the samples was performed using the root staining technique (Phillips, D.M and Hayman, D.S. Improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection. Trans. Br. Mycol. Soc. 55. 158-161. 1970) and the percentage colonisation was determined by the method of intercepts (Giovannetti, M. and Mosse, B. (1980): An évaluation of techniques to measure vesicular-arbuscular infection in roots. New Phytology., 84:489-500). The mathematical calculation of visual density, arbuscular endophytes and mycorrhizal activity was determined according to proposed protocols (Trouvelot, A., Kough, J. and Gianinazzi Pearson, V. (1986). Mesure du Taux de Mycorhization VA d'un Système Radiculaire. Recherche de Méthodes d'Estimation ayant une Signification Fonctionnelle. Proceedings of the 1st European Symposium on Mycorrhizae: Physiological and Genetical Aspects of Mycorrhizae, Dijon, 15 July, 1985. À(eds. V. Gianinazzi Pearson and S. Gianinazzi). INRA, Paris, pp. 217 222; Herrera
Peraza, R. Eduardo Furrazola, Roberto L. Ferrer, Rigel Fernandez Valle and Yamir Terres Arias. 2004. Functional strategies of root hairs and arbuscular mycorrhizae in an evergreen tropical forest, Sierra del Rosario, Cuba. Revista CENIC Ciencias Biolôgicas, Vol. 35, No. 2, 2004). The total spore populations, g soil'1 were also determined.
Statistical Analysis
The statistical Processing of the results were performed by simple analysis of variance of classification and Tukey's test was used when there were significant différences between means, using the program Statgraphics® Plus, 4.1. For drawing graphs (i.e., Figures 1,2 and 3), the SigmaPlot 4 program was used.
The percentage values of mycorrhizal colonisation were transformed using the expression 2arcsenVx.
Results and Discussion
Table 13, above reported, shows some of the Chemical properties of the soil used in the experiment. The soil had a slightly alkaline pH, average levels of organic matter, P and values of Ca2+ of the order of 10 cmol.kg’1. Regarding the saline characteristics, there were high contents of Na and high electrical conductivity, indicating strongly saline characteristics, although fertility was acceptable for development of a rice crop.
The number of spores found in this substrate was very low, a feature of heavily used agricultural soils where the diversity and intensity of arbuscular mycorrhizal fungi (AMF) is reduced, given intensive tillage, overexploitation and typical chemicalization and salinization processes, etc. (Rao, D.L.N., 1998. Biological amelioration of salt-affected soils. In: Microbial Interactions in Agriculture and Forestry, vol. 1. Science Publishers, Enfield, USA, pp. 21-238)
The analysis of the growth dynamics of rice plants under these conditions showed distinct behaviour with the treatments in the variables studied. The height of the plants increased steadily, accelerating their growth after day 27 (Table 14).
Table 14. Height (cm), depth of root System (cm) and mycorrhizal colonisation (%MC) in plants treated with Dominikia sp. (D.t) and control (C) plants during 90 dpt in saline soil conditions.
Table 14
DPT | 1 | 7 | 10 | 13 | 18 | 20 | 23 | 27 | 32 | 39 | 60 | 90 |
Heig ht | ||||||||||||
D.t | 10.5 b | 10.5 b | 10.9 b | 12.4 b | 13.0 b | 17.1 a | 17.0 a | 19.8 | 23.8 a | 32.8 a | 63.5 a | 74.1 a |
C | 11.1 a | 11.2 a | 12.2 a | 14.4 a | 15.6 a | 15.6 b | 16.7 b | 18.9 | 20.7 b | 25.7 b | 58.6 b | 68.1 b |
St. Sig. | 0.2* | 0.12 | 0.1** | 0.2** | 0.3* ** | 0.2* ** | 0.4* ** | 0.9 ns | 0.1* | 0.2* ** | 0.6* ** | 0.2* |
DRS | ||||||||||||
D.t | 0 | 0.6 | 1.4 b | 2.6 | 4.3 | 6.18 | 6.48 | 6.82 | 7.9 a | 7.08 | 13.9 a | 16.9 a |
C | 0 | 0.6 | 2.1 a | 2.1 | 4.7 | 6.29 | 6.32 | 6.4 | 6.4 b | 7.23 | 10.4 b | 13.1 b |
St. Sig. | 0.0 ns | 0.3 ns | 0.2** | 0.6 n.s | 0.8 ns | 0.3 ns | 0.5 ns | 0.4 ns | 0.2* ** | 0.3 ns | 0.1* | 0.2* |
(% MC) | ||||||||||||
D.t | 2 a | 6 a | 13 a | 14a | 14 a | 17a | 18 a | 24 a | 35 a | 38 a | 38 a | 41 a |
C | 0 b | 3 b | 3 b | 6 b | 10 b | 12 b | 13 b | 15 b | 20 b | 21 b | 19 b | 22 b |
St. Sig. | 0.3* | 0.2** | 0.13 | 0.4** | 0.6* | 0.1* ** | 0.2* ** | 0.1* ** | 0.2* ** | 0.3* | 0.2* | 0.4* ** |
Legend: DRS: Depth of root System, DPT: Days post treatment, St. Sig.: Standard Déviation.
Same letters in the same column do not differ significantly at p < 0.05.
Both treatments show significant différences during height gain. The highest values were obtained in the control treatment up to 18 days, after which there was a change in behaviour and highest values for plant height were obtained in the inoculant treatment with the efficient arbuscular mycorrhizal fungus.
The analysis of the growth dynamics of rice plants under these conditions revealed distinct behaviour of the variables with the treatment. The height of the plants increased steadily, accelerating their growth after day 27.
Both treatments show significant différences during height gain. The highest values were obtained in the control treatment up to 18 days, after which there was a change in behaviour and highest values for plant height were obtained in the inoculation treatment with the efficient arbuscular mycorrhizal fungus.
A similar behaviour of the height variable was observed in the root System depth, which was greater in treated than control plants after 32 dpt. In this case, the behaviour was accentuated because this is the part of the plant where the fungus is established.
The study of mycorrhizal colonisation showed distinct behaviour to that found in the previously analysed variables. In this case and in both treatments, colonisation was progressive and always reached highest values in the inoculated treatment compared to the control treatment, which although was not inoculated showed natural levels of mycorrhizal colonisation, a typical resuit in experiments conducted in natural conditions.
The development was well marked in the case of treatment inoculated with AMF, reaching high values if taking into account that the experiment was conducted in flooded conditions or with a layer of water after 20 days. At the end of the cultivation, root colonisation reached a value of 44%, which is considered high compared to other studies of mycorrhizae in rice, where the colonisation maximums do not exceed 25% with inoculation on a solid base. (Fernandez, F.; Ortiz, R.; Martinez, M.A.; Costales, A.; Llonin, D. The effect of commercial arbuscular mycorrhizal fungi (AMF) inoculants on rice (Oryza sativa) in different types of soils. Cultivos Tropicales 18 (1): 5-9, 1997).
Figure 1 shows the development of two very important fungal variables in mycorrizal activity in the treatment with the inoculant with the composition of the invention in concentrated form, the population of spores and fungal occupation, expressed through the Visual density percentage, which is nothing other than the intensity with which the mycélium colonises the interior root.
The Visual density showed a typical microbial behaviour with a well-defined latency phase where the fungus slowly colonised the root interior from 0 to 20 days and subsequently showed exponential growth to 40 days, the time when it reached the stationary phase and the end of the cultivation.
In the case of the spore population, values in the soil were detected in the first days, derived from the inoculation (up to 15 days in the stems), which then gradually disappeared over time, product of the germination in favourable conditions of humidity and high température until their population in the soil fell. After 30 days, production of new spores started, derived from the development of external fungal biomass and development of symbiosis with the plant. In this case, the population continued to grow to values close to 12 spores, g’1 de soil.
The analysis of the arbuscular external mycélium and the arbuscular endophytes variables (Fig. 2) is very interesting as it shows how the internai and external behaviours of the mycorrhizal symbiont occur as symbiosis develops in annual cultivation cycles.
In this case high values of external mycélium were observed during the first stages of symbiosis development, caused by the fungal growth at the expense of that of the plant, expressed after some years (Bethlenfalvay, G.J., Brown, M.S., Franson, R.L, Mihara, K.L., 1989. The glycine-glomus-bradyrhizobium symbiosis. IX. Nutritional, morphological and physiological response of nodulated soybean to géographie isolâtes of the mycorrhizal fungus of Glomus mosseae. Physiol. Plant. 76, 226-232), as a frank parasitic process, derived from exubérant growth of the mycélium in early stages of mycorrhizal colonisation in plants (hours) with a low photosynthetic phase and a high metabolic cost. The development of arbuscular endophytes follows the opposite tendency. There were very low values during the first few days, not achieving significant growth until after 25 days of growth, a stage considered to be transitional in arbuscular mycorrhizal symbiosis.
After 30 days there is a réduction and stabilisation of the external mycélium and a graduai increase of the endophyte related to plant growth and development of symbiosis.
Figure 3 shows the relationship established between the mycorrhizal components in the rice crop (The ratio between arbuscular external mycélium and endophyte mycélium (MEA:EA)). This correspondence between the two main components of symbiosis, the external mycélium and the endophyte, expresses the activity of this association, which passes through different stages of development. (Hirrel, M.C., 1981. The effect of sodium and chloride salts on the germination of Gigaspora margaria. Mycology 43, 610-617).
An initial stage where there are high values of external mycélium in correspondence with very low values of endophytes, which propitiates a frank parasitism, expressed not only by these variables but also in a réduction of plant growth compared to the noninoculated or inefficiently mycorrhized control, which does not cause substantial changes in plant development (Table 2).
An intermediate or transition phase where both parts start to balance and a mutualist phase of exchange, when the components equilibrate with a value of 1 and even less, so that there is a notable increase inside the root that ensures proper exchange of nutrients at the level of the arbuscules inside the cells.
For this crop in particular, two phases can be well defined; an initial-transition, up to 2025 days and a mutualist phase after this time. Parallel to this, an analysis of plant height shows a take-off in plant growth in relation to the non-inoculated control after 27 days of planting, which coïncides with the mutualist phase of the mycorrhizal symbiosis under these salinity conditions.
This effect can be subsequently shown in the analysis of the yield and of its components (Table 15).
Table 15. Number of panicles per plant (n), weight of panicles (g), weight of 100 grains (g) and yield (g. plant'1) in rice plants treated with Dominikia sp. (D.t) and control untreated plants (C) in saline soil.
Table 15 | ||||
Treatments | NPP | PP | P 100 g | R |
D.t | 8.33 a | 2.70 a | 3.69 a | 21.66 a |
C | 5.40 b | 1.87 b | 2.70 b | 15.40 b |
St. Sig. | 0.12 *** | 0.05*** | 0.001*** | 1.34*** |
C.V (%) | 11.2 | 9.2 | 6.5 | 14.6 |
Legend: NPP: number of panicles per plant (n), PP: Weight of panicles (g), P 100 G: Weight of 100 grains (g), R: yield (g.plant'1), St. Sig.: Standard Déviation. Same letters in the same column are not significantly different at p < 0.05.
There was an increase in ail the components of the yield measured in plants treated with Dominikia sp. compared to control non-inoculated plants. This was particularly interesting in the salinity conditions of the work.
By way of conclusion, the use of this strain of mycorrhizal fungus was effective for these soil conditions. This is very interesting as a viable and sustainable alternative for the adverse conditions caused by saline stress.
Claims (23)
1. Dominikia sp. strain, deposited under accession number MUCL 57072.
2. Composition comprising Dominikia sp. strain, deposited under accession number MUCL 57072, wherein the concentration of the Dominikia sp. in said composition is from 4.0% to 1.0% by weight.
3. Composition according to claim 2, wherein the concentration of the Dominikia sp. in said composition is from 3.0% to 2.0% by weight.
4. Composition according to claim 3, wherein the concentration of the Dominikia sp. in said composition is from 2.5% to 2.3% by weight
5. Composition according to claim 2, wherein said composition is a solid composition.
6. Composition according to claim 5, wherein said composition is in the form of powder, emulsifiable concentrate, granules, or microgranules.
7. Composition according to claim 6, wherein said composition is in the form of microgranules.
8. Composition according to claim 7, wherein said microgranules hâve a size ranging from 500 pm to 2000 pm.
9. Composition according to claim 8, wherein said microgranules hâve a size ranging from 800 pm to 1500 pm.
10. Composition according to claim 9, wherein said microgranules hâve a size ranging from 900 pm to 1200 pm
11. Composition according to any of one of daims 2 to 10, further comprising at least one fungicide and/or at least one bio-fungicide and/or at least one insecticide and/or at least one bio-insecticide and/or at least one nematicide and/or at least one biostimulant.
12. Process for the préparation of a composition comprising Dominikia sp. strain deposited under accession number MUCL 57072 comprising the following steps: a. Providing a substrate;
b. Providing said substrate with the seeds of a host plant and with Dominikia sp. strain deposited under accession number MUCL 57072;
c. Cultivating said host plant and watering to maintain said substrate with a moisture level of at least 75% of field capacity;
d. Discontinuing said watering for a period of at least 7days;
e. Removing the aerial part of said host plant and the substrate;
f. Drying the removed substrate;
g. milling the dried substrate to obtain granules having a particle size of below 100 pm.
13. Process according to claim 12, wherein said Dominikia sp. strain of step b. is an inoculum comprising Dominikia sp. strain propagules.
14. Process according to claim 12 or 13, further comprising a step h1. of microgranulation.
15. Process according to claim 12 or 13, further comprising a step h2. of préparation of a concentrate biological inoculant for seeds coating of cereals.
16. Process according to claim 15, wherein said step h2. of préparation of a concentrate biological inoculant for seeds coating of cereals is carried out by sieving the milled product obtained in step g. to select the granules having a particle size above a predetermined value.
17. Process according to claim 15, further comprising a step i. of seed coating.
18. Process according to claim 17, wherein said step i. of seed coating comprises the steps:
il. Covering the seeds with an adhesive substance;
i2. Adding said concentrate biological inoculant;
i3. Optionally provide one or more compatible treatments selected from:
Î3.1 Treatments with fungicides, insecticides and/or herbicides, compatible with mycorrhizal forming fungi,
Î3.2 Treatment with bénéficiai microorganisms, i3.3 Treatment with macro and/or micronutrients, i3.4 Treatment with stimulants, i3.5 Treatment with colouring pigments compatible with mycorrhizal fungi;
i4. Drying the seeds.
19. Use of a composition comprising Dominikia sp. strain, deposited under accession number MUCL 57072 as bio- stimulant in plants.
20. Use of a composition comprising Dominikia sp. strain deposited under accession number MUCL 57072 as bio-nematicidal in plants.
21. Use according to claim 19 or 20, wherein said plants are cereals.
22. Composition according to claim 2 characterized by being provided by seed coating or in conjunction with seeds at the time of seeding.
23. Cereal seed coated with a composition according to claim 2.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP18382653.6 | 2018-09-12 |
Publications (1)
Publication Number | Publication Date |
---|---|
OA20372A true OA20372A (en) | 2022-07-01 |
Family
ID=
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105579573B (en) | Compositions and methods involving isolated endophytes | |
US10470465B2 (en) | Glomus iranicum var. tenuihypharum var. nov. strain and use thereof as biostimulant | |
Dilbo et al. | Integrated management of garlic white rot (Sclerotium cepivorum Berk) using some fungicides and antifungal Trichoderma species | |
Talbi et al. | Effect of double inoculation with endomycorrhizae and Trichoderma harzianum on the growth of carob plants | |
Khokhar et al. | Effect of Penicillium species culture filtrate on seedling growth of wheat | |
AU2021290228A1 (en) | A method for improving the mean dry shoot weight, mean dry grain weight, and suppressing seed-borne infection in a cereal crop | |
Vitorino et al. | Growth promotion mediated by endophytic fungi in cloned seedlings of Eucalyptus grandis x Eucalyptus urophylla hybrids | |
US11896013B2 (en) | Dominikia sp. strain, compositions comprising it and uses | |
JP5909695B1 (en) | Microbial control agent and seed coating agent for bacterial disease of plant, and seed coated with said seed coating agent | |
US10201164B2 (en) | Method for increasing the production of flowers, seeds and/or fruit of a plant | |
OA20372A (en) | Dominikia Sp. strain, compositions comprising it and uses. | |
Gaston et al. | Impact of previous legumes on millet mycorrhization and yields in sandy soil of West African Sahel | |
EA045371B1 (en) | DOMINIKIA SP. STRAIN, COMPOSITIONS CONTAINING IT AND APPLICATION OPTIONS | |
JP6384087B2 (en) | Plant cultivation method | |
Tetarwal et al. | Sustainable and eco-friendly management of root rot of soybean caused by Rhizoctonia solani and Fusarium solani | |
Marieska et al. | Effectiveness of Lettuce Seed Encapsulation Containing Trichoderma Sp. in Control of Damping-off Disease | |
JP5052873B2 (en) | Control agent and method for biological control of soybean stem blight | |
Webster | Managing foliar blights on specialty crops |