KR20230105465A - Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects - Google Patents
Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects Download PDFInfo
- Publication number
- KR20230105465A KR20230105465A KR1020220000878A KR20220000878A KR20230105465A KR 20230105465 A KR20230105465 A KR 20230105465A KR 1020220000878 A KR1020220000878 A KR 1020220000878A KR 20220000878 A KR20220000878 A KR 20220000878A KR 20230105465 A KR20230105465 A KR 20230105465A
- Authority
- KR
- South Korea
- Prior art keywords
- strain
- pepper
- polymyxa
- plant
- disease
- Prior art date
Links
- 230000008635 plant growth Effects 0.000 title claims description 12
- 238000011161 development Methods 0.000 title description 10
- 208000035143 Bacterial infection Diseases 0.000 title description 5
- 230000000853 biopesticidal effect Effects 0.000 title description 2
- 230000004936 stimulating effect Effects 0.000 title description 2
- 235000002566 Capsicum Nutrition 0.000 claims abstract description 65
- 241000722363 Piper Species 0.000 claims abstract description 61
- 239000006002 Pepper Substances 0.000 claims abstract description 60
- 235000016761 Piper aduncum Nutrition 0.000 claims abstract description 60
- 235000017804 Piper guineense Nutrition 0.000 claims abstract description 60
- 235000008184 Piper nigrum Nutrition 0.000 claims abstract description 60
- 201000010099 disease Diseases 0.000 claims abstract description 53
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 53
- 241000196324 Embryophyta Species 0.000 claims abstract description 49
- 230000036039 immunity Effects 0.000 claims abstract description 12
- 239000011814 protection agent Substances 0.000 claims abstract description 12
- 244000005700 microbiome Species 0.000 claims description 33
- 239000003795 chemical substances by application Substances 0.000 claims description 26
- 244000052769 pathogen Species 0.000 claims description 25
- 230000000813 microbial effect Effects 0.000 claims description 23
- 241000194105 Paenibacillus polymyxa Species 0.000 claims description 17
- 241000193738 Bacillus anthracis Species 0.000 claims description 16
- 241000132152 Polymyxa Species 0.000 claims description 14
- 230000001737 promoting effect Effects 0.000 claims description 14
- 230000001717 pathogenic effect Effects 0.000 claims description 13
- 235000013399 edible fruits Nutrition 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 9
- 235000006040 Prunus persica var persica Nutrition 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 9
- 244000144730 Amygdalus persica Species 0.000 claims description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 7
- 230000003042 antagnostic effect Effects 0.000 claims description 6
- 230000001939 inductive effect Effects 0.000 claims description 6
- 241001123536 Colletotrichum acutatum Species 0.000 claims description 5
- 241000589771 Ralstonia solanacearum Species 0.000 claims description 5
- 239000002773 nucleotide Substances 0.000 claims description 5
- 125000003729 nucleotide group Chemical group 0.000 claims description 5
- 241000815873 Xanthomonas euvesicatoria Species 0.000 claims description 4
- 239000003337 fertilizer Substances 0.000 claims description 4
- 241000588701 Pectobacterium carotovorum Species 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 206010027146 Melanoderma Diseases 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 33
- 230000012010 growth Effects 0.000 abstract description 25
- 230000001580 bacterial effect Effects 0.000 abstract description 19
- 241000894006 Bacteria Species 0.000 abstract description 18
- 239000002689 soil Substances 0.000 abstract description 13
- 244000052616 bacterial pathogen Species 0.000 abstract description 10
- 238000000034 method Methods 0.000 abstract description 9
- 238000012258 culturing Methods 0.000 abstract description 5
- 244000053095 fungal pathogen Species 0.000 abstract description 4
- 230000002538 fungal effect Effects 0.000 abstract description 3
- 238000011282 treatment Methods 0.000 description 38
- 238000012360 testing method Methods 0.000 description 22
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 description 12
- 239000000417 fungicide Substances 0.000 description 10
- 230000002401 inhibitory effect Effects 0.000 description 10
- 239000000575 pesticide Substances 0.000 description 10
- 230000018109 developmental process Effects 0.000 description 9
- 239000001974 tryptic soy broth Substances 0.000 description 9
- 240000004160 Capsicum annuum Species 0.000 description 8
- 238000011835 investigation Methods 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 235000007862 Capsicum baccatum Nutrition 0.000 description 7
- 241000607479 Yersinia pestis Species 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 239000001728 capsicum frutescens Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 230000002906 microbiologic effect Effects 0.000 description 7
- 240000007594 Oryza sativa Species 0.000 description 6
- 235000007164 Oryza sativa Nutrition 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 230000006378 damage Effects 0.000 description 6
- 230000000855 fungicidal effect Effects 0.000 description 6
- 229920001690 polydopamine Polymers 0.000 description 6
- 235000009566 rice Nutrition 0.000 description 6
- 235000013311 vegetables Nutrition 0.000 description 6
- 241000179039 Paenibacillus Species 0.000 description 5
- 241000758706 Piperaceae Species 0.000 description 5
- 239000003242 anti bacterial agent Substances 0.000 description 5
- 229940088710 antibiotic agent Drugs 0.000 description 5
- 241000894007 species Species 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000563903 Bacillus velezensis Species 0.000 description 4
- 240000008384 Capsicum annuum var. annuum Species 0.000 description 4
- ABUGVBRDFWGJRD-CHOYNLESSA-N [9-[(2r,3r,4s,5r)-3,4-dihydroxy-5-methyloxolan-2-yl]-2-(2,4-dinitrophenyl)sulfanylpurin-6-yl] [hydroxy(phosphonooxy)phosphoryl] hydrogen phosphate Chemical compound O[C@@H]1[C@H](O)[C@@H](C)O[C@H]1N1C2=NC(SC=3C(=CC(=CC=3)[N+]([O-])=O)[N+]([O-])=O)=NC(OP(O)(=O)OP(O)(=O)OP(O)(O)=O)=C2N=C1 ABUGVBRDFWGJRD-CHOYNLESSA-N 0.000 description 4
- 230000003698 anagen phase Effects 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 241001123534 Colletotrichum coccodes Species 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 3
- 108010028921 Lipopeptides Proteins 0.000 description 3
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 240000003768 Solanum lycopersicum Species 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 239000012773 agricultural material Substances 0.000 description 3
- 208000022362 bacterial infectious disease Diseases 0.000 description 3
- 244000000005 bacterial plant pathogen Species 0.000 description 3
- 238000009709 capacitor discharge sintering Methods 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- 239000002734 clay mineral Substances 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 238000012252 genetic analysis Methods 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 230000010534 mechanism of action Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 2
- 240000007124 Brassica oleracea Species 0.000 description 2
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 description 2
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 2
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 description 2
- 108010076119 Caseins Proteins 0.000 description 2
- 241001529387 Colletotrichum gloeosporioides Species 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 241000768412 Paenibacillus polymyxa E681 Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 241001646398 Pseudomonas chlororaphis Species 0.000 description 2
- 238000012300 Sequence Analysis Methods 0.000 description 2
- 239000000589 Siderophore Substances 0.000 description 2
- 244000061456 Solanum tuberosum Species 0.000 description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 description 2
- 229930182692 Strobilurin Natural products 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N 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
- 241000223259 Trichoderma Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 239000008121 dextrose Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000006194 liquid suspension Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 244000000003 plant pathogen Species 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 235000012015 potatoes Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000005063 solubilization Methods 0.000 description 2
- 230000007928 solubilization Effects 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- 241000194103 Bacillus pumilus Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- AFWTZXXDGQBIKW-UHFFFAOYSA-N C14 surfactin Natural products CCCCCCCCCCCC1CC(=O)NC(CCC(O)=O)C(=O)NC(CC(C)C)C(=O)NC(CC(C)C)C(=O)NC(C(C)C)C(=O)NC(CC(O)=O)C(=O)NC(CC(C)C)C(=O)NC(CC(C)C)C(=O)O1 AFWTZXXDGQBIKW-UHFFFAOYSA-N 0.000 description 1
- 241000222199 Colletotrichum Species 0.000 description 1
- 241001133184 Colletotrichum agaves Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 241000588697 Enterobacter cloacae Species 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 241000961787 Josa Species 0.000 description 1
- 241000863031 Lysobacter Species 0.000 description 1
- 241000878006 Miscanthus sinensis Species 0.000 description 1
- 229910019093 NaOCl Inorganic materials 0.000 description 1
- 241001556089 Nilaparvata lugens Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 241001506910 Paenibacillus barcinonensis Species 0.000 description 1
- 241000605564 Paenibacillus brasilensis Species 0.000 description 1
- 241000611801 Paenibacillus illinoisensis Species 0.000 description 1
- 241000189604 Paenibacillus jamilae Species 0.000 description 1
- 241000233727 Paenibacillus koreensis Species 0.000 description 1
- 241000706216 Paenibacillus kribbensis Species 0.000 description 1
- 241000285107 Paenibacillus massiliensis Species 0.000 description 1
- 241001429255 Paenibacillus panacisoli Species 0.000 description 1
- 241001140060 Paenibacillus peoriae KCTC 3763 Species 0.000 description 1
- 241001016862 Paenibacillus polymyxa ATCC 842 Species 0.000 description 1
- 241001587294 Paenibacillus polysaccharolyticus Species 0.000 description 1
- 241001365836 Paenibacillus qingshengii Species 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 108010040201 Polymyxins Proteins 0.000 description 1
- 240000005809 Prunus persica Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 239000005869 Pyraclostrobin Substances 0.000 description 1
- 241000589157 Rhizobiales Species 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 244000000231 Sesamum indicum Species 0.000 description 1
- 235000003434 Sesamum indicum Nutrition 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 235000002597 Solanum melongena Nutrition 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 108020004566 Transfer RNA Proteins 0.000 description 1
- 241000223260 Trichoderma harzianum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 229910021536 Zeolite Inorganic materials 0.000 description 1
- 230000036579 abiotic stress Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000013272 agar well diffusion method Methods 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 230000032770 biofilm formation Effects 0.000 description 1
- 230000004790 biotic stress Effects 0.000 description 1
- 239000001511 capsicum annuum Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 241001576300 endophytic bacterium Species 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000008686 ergosterol biosynthesis Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 244000000004 fungal plant pathogen Species 0.000 description 1
- 229930193868 fusaricidin Natural products 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- -1 iron complex compound Chemical class 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 101150078797 luxS gene Proteins 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000004066 metabolic change Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 239000006877 oatmeal agar Substances 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000003415 peat Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- HZRSNVGNWUDEFX-UHFFFAOYSA-N pyraclostrobin Chemical compound COC(=O)N(OC)C1=CC=CC=C1COC1=NN(C=2C=CC(Cl)=CC=2)C=C1 HZRSNVGNWUDEFX-UHFFFAOYSA-N 0.000 description 1
- 108700022487 rRNA Genes Proteins 0.000 description 1
- 230000008261 resistance mechanism Effects 0.000 description 1
- 230000003938 response to stress Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000019654 spicy taste Nutrition 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000005082 stem growth Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- NJGWOFRZMQRKHT-UHFFFAOYSA-N surfactin Natural products CC(C)CCCCCCCCCC1CC(=O)NC(CCC(O)=O)C(=O)NC(CC(C)C)C(=O)NC(CC(C)C)C(=O)NC(C(C)C)C(=O)NC(CC(O)=O)C(=O)NC(CC(C)C)C(=O)NC(CC(C)C)C(=O)O1 NJGWOFRZMQRKHT-UHFFFAOYSA-N 0.000 description 1
- NJGWOFRZMQRKHT-WGVNQGGSSA-N surfactin C Chemical compound CC(C)CCCCCCCCC[C@@H]1CC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)O1 NJGWOFRZMQRKHT-WGVNQGGSSA-N 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 108010050327 trypticase-soy broth Proteins 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000010457 zeolite Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/25—Paenibacillus
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/12—Bacillus polymyxa ; Paenibacillus polymyxa
Abstract
본 발명은 고추의 탄저병, 세균성 풋마름병, 점무늬병 등 방제의 어려움을 효과적으로 해결하기 위한 방법으로, 이 병을 일으키는 곰팡이 및 세균 병원균의 생장을 직접 억제할 뿐만 아니라, 기주 식물인 고추의 병 면역성 증진 및 생육 촉진 효과를 모두 갖는 내생 세균을 국내 근권 토양에서 분리, 배양 제제화함으로써 종합적인 식물 강화(plant strengthener) 효능을 가진 새로운 형태의 천연식물보호제를 개발하여 제공한다.The present invention is a method for effectively solving the difficulties in controlling pepper anthracnose, bacterial green blight, spot disease, etc., not only directly inhibits the growth of fungal and bacterial pathogens that cause this disease, but also enhances disease immunity of pepper, a host plant, and We develop and provide a new type of natural plant protection agent with a comprehensive plant strengthener effect by isolating and culturing endogenous bacteria that have both growth-promoting effects from domestic rhizosphere soil and formulating them.
Description
본 발명은 고추 탄저병 및 세균병 동시 방제용 다기능 천연식물보호제 개발에 관한 것으로, 보다 상세하게는 공생 세균 페니바실러스 폴리믹사 (Paenibacillus polymyxa) JG90 균주를 함유한 미생물 제제에 관한 것이다.The present invention relates to the development of a multifunctional natural plant protection agent for simultaneous control of pepper anthracnose and bacterial disease, and more particularly, to a microbial preparation containing the symbiotic bacterium Penibacillus polymyxa JG90 strain.
본 발명은 농림수산식품부에서 지원하는 '작물 바이러스 및 병해충 대응 산업화 기술개발' 사업과 과학기술정보통신부에서 지원하는 '고용위기기업부설연구소R&D전문인력활용지원(R&D)' 사업의 일환으로 수행된 연구로부터 도출된 것이다.The present invention is a research conducted as part of the 'Crop virus and disease pest response industrialization technology development' project supported by the Ministry of Food, Agriculture, Forestry and Fisheries and the 'Employment Crisis Enterprise Affiliated Research Institute R&D Professional Manpower Utilization Support (R&D)' project supported by the Ministry of Science and ICT is derived from
[과제고유번호: 1545022768, 과제명: 고추 탄저병균의 약제 저항성 메커니즘 구명 및 방제제 기술개발(2020~2024)][Project identification number: 1545022768, task title: Drug resistance mechanism of pepper anthrax and development of control agent technology (2020-2024)]
[과제고유번호: 1711140886, 과제명: 과수 세균구멍병(천공병) 방제를 위한 마이크로바이옴 분석 및 작용기작 연구(2021~2022)][Project identification number: 1711140886, task title: Microbiome analysis and mechanism of action study for fruit tree bacterial hole disease (boring disease) control (2021-2022)]
우리나라의 농산물 중 고추는 생산량이 약 1조 18억원(2018년)으로 벼, 시설 채소와 함께 중요한 환금작물이다. 대부분의 고추 재배는 동일한 지역에서 수 년간 연작하고 또한 연간 재배 기간이 길어서 재배 중 곰팡이, 세균, 바이러스 등 병원균에 의하여 다양한 병이 발생되기 때문에 안정적인 생산에 큰 피해를 입고 있다. 이 중에서도 고추의 탄저병에 의한 피해액은 총 생산액의 약 10% 정도인 1,000억원 정도이며, 심한 경우에는 50~80%의 손실을 가져오기도 한다(비특허문헌 1 참조). 이러한 손실은 탄저병의 발생 지역과 병원균의 농약 저항성에 따라서 많이 다를 수가 있는데, 국내의 탄저병균은 1990년대에는 콜레토트리쿰 글로에오스포리오이데스(Colletotrichum gloeosporioides)로 보고되었지만, 최근에는 주된 병원균이 콜레토트리쿰 아쿠타툼(C. acutatum)으로 변화되었다고 보고되었다(비특허문헌 2 참조). 그 동안 고추 탄저병의 방제를 위하여 탄저병 저항성 품종이 보급되고 있으나 높은 단가, 일부 품종의 매운 맛 상실과 고추 바이러스에 대한 복합저항성의 부재 등으로 사용에 다소 한계를 노출하고 있다. 반면에 가장 쉽게 효과적으로 방제할 수 있는 방법은 화학 살균제를 사용하는 것이다. 고추 재배 기간 중 평균적으로 사용한 살균제의 처리 횟수는 평균 20회 이상으로 그 중 탄저병 방제를 위해서는 평균 11.2회(권장 탄저병 방제력에서는 살균제 처리 횟수를 6~7회)를 처리하는 것으로 조사되었다(충북고추협력단 교육 자료, 2018). 이와 같은 살균제의 남용으로 인하여 현재 널리 사용되고 있는 에르고스테롤(ergosterol) 생합성 저해 살균제 및 스트로빌루린(strobilurin)계 살균제에 대한 저항성 병원균이 생겨 방제 효과가 떨어지고 있는 실정이다(비특허문헌 3 내지 4 참조). 따라서 살균제 저항성 탄저병균의 효과적인 방제를 위하여 길항 또는 공생 미생물을 이용한 천연식물보호제(생물농약) 개발 등 종합적인 방제법이 필요하다.Among agricultural products in Korea, red pepper is an important cash crop with a production of about 1.18 trillion won (2018), along with rice and vegetable vegetables. Most pepper cultivation is continuously cultivated in the same area for several years, and the annual cultivation period is long, causing various diseases caused by pathogens such as fungi, bacteria, and viruses during cultivation, resulting in great damage to stable production. Among them, the amount of damage caused by anthrax of red pepper is about 100 billion won, which is about 10% of the total production amount, and in severe cases, it may result in a loss of 50 to 80% (see Non-Patent Document 1). This loss can vary greatly depending on the area where anthrax occurs and the pesticide resistance of the pathogen. The domestic anthrax was reported as Colletotrichum gloeosporioides in the 1990s, but recently the main pathogen is Colletotrichum gloeosporioides . It has been reported that it has been changed to Totricum acutatum ( C. acutatum ) (see Non-Patent Document 2). In the meantime, anthrax-resistant varieties have been disseminated for the control of pepper anthrax, but their use is somewhat limited due to high unit price, loss of spicy taste of some varieties, and lack of complex resistance to pepper viruses. On the other hand, the easiest and most effective way to control is to use chemical fungicides. During the pepper cultivation period, the average number of disinfectant treatments used was 20 times or more, and among them, an average of 11.2 times for anthrax control (in the recommended anthrax control capacity, the number of fungicide treatments was 6 to 7 times) was investigated (Chungbuk Pepper Cooperation Foundation) Educational Materials, 2018). Due to the abuse of such fungicides, pathogens resistant to ergosterol biosynthesis inhibitors and strobilurin-based fungicides, which are currently widely used, are generated, and the control effect is reduced (see Non-Patent
탄저병과 함께 고추 생산에 많은 피해를 주는 병으로는 세균에 의한 풋마름병(청고병), 세균성 점무늬병 및 무름병이 있다(비특허문헌 1 참조). 풋마름병은 랄스토니아 솔라나세아룸(Ralstonia solanacearum)에 의하여 발생되는 토양 전염성 세균병으로 주로 지온이 25℃ 이상일 때 많이 발생되며 저온기에는 발병되지 않는다. 이 병원균은 생육 적온이 35~37℃로 고추 이외에 토마토, 가지, 참깨, 감자 등 400여 종의 식물에도 병을 일으키는 다범성으로 알려지고 있다. 이 병은 주로 7~8월 여름철에 발생되고 일단 감염된 이병주는 서서히 말라 죽으며 피해 줄기를 절단해 보면 도관이 갈색으로 변해 있고, 그 부분에서 우유 빛의 세균액이 분비된다. 일단 한 포기가 감염되면 병원균이 물을 따라 퍼져나가 순식간에 이웃 포기까지 발병시켜 밭 전체 작물을 고사시킨다. 현재 이 병 방제를 위하여 저항성 품종을 심는 것과 항생제를 관주하는 방법이 있지만, 항생물질에 대한 내성 발생 등 방제 효과가 낮아 길항 미생물을 이용한 천연식물보호제 개발이 절실한 실정이다(비특허문헌 5 내지 7 참조). Along with anthrax, diseases that cause a lot of damage to pepper production include bacterial blight (chill blight), bacterial spot disease and soft rot (see Non-Patent Document 1). Foot blight is a soil contagious bacterial disease caused by Ralstonia solanacearum, which occurs mainly when the ground temperature is 25 ° C or higher and does not occur in low temperatures. This pathogen has a growth optimum temperature of 35 ~ 37 ℃, and is known to cause diseases in more than 400 species of plants such as tomatoes, eggplants, sesame seeds, and potatoes in addition to peppers. This disease occurs mainly in the summer of July-August, and once infected, the diseased byeongju gradually withers and dies. Once one plant is infected, the pathogen spreads along the water and instantly infects neighboring plants, causing the entire field to die. Currently, there are methods of planting resistant varieties and drenching antibiotics to control this disease, but the control effect such as resistance to antibiotics is low, and the development of natural plant protection agents using antagonistic microorganisms is urgently needed (see Non-Patent
고추의 세균성 점무늬병은 잔쏘모나스 유베시카토리아(Xanthomonas euvesicatoria)에 의하여 발생되며 병원균은 병든 잎, 잎줄기 잔재나 종자에서 월동한 후 이듬해 봄 1차 전염원으로 주로 비, 바람, 농기구 등에 의해 퍼져 나간다. 일단 잎의 기공과 상처를 통해 병원균이 침입하면 처음에는 병반이 회갈색의 작은 점무늬로 시작하여 발병이 진전되면 잎 전체가 갈색으로 말라 죽는다(비특허문헌 8 참조). 이 외에도 펙토박테리움 카로토보룸(Pectobacterium carotovorum)에 의한 무름병이 있으며, 이 병은 고추 외에도 배추, 양배추, 파프리카 등에 발생되는 토양 전염병이다. 고추 세균병 방제 약제는 구리제나 농용신수화제, 옥소리닉 산 등 몇 종류에 불과하다. 이처럼 고추의 병은 다양한 종류의 여러 병원균에 의해 발생되며, 병 발생으로 인하여 생산에 큰 피해를 입게 되므로 발생 전 잘 예방해야 한다. 그러나 화학 농약은 곰팡이나 세균성 병원균들을 동시에 방제하지 못하고 작용 기작에 따라 주성분이 다른 원제를 혼합하여 합제 형태로 제품이 개발되어 사용된다.Bacterial spot disease of pepper is caused by Xanthomonas euvesicatoria, and the pathogen overwinters in diseased leaves, leaf stem remnants or seeds, and then spreads mainly by rain, wind, agricultural tools, etc. as the primary source of infection the following spring. Once the pathogen invades through the stomata and wounds of the leaves, the lesions initially begin as small grayish brown dots, and as the disease progresses, the entire leaf turns brown and dies (see Non-Patent Document 8). In addition to this, there is soft rot caused by Pectobacterium carotovorum , which is a soil epidemic that occurs in cabbage, cabbage, paprika, etc. in addition to pepper. There are only a few types of pepper bacterial disease control agents, such as copper agents, agricultural hydrolyzing agents, and oxalic acid. As such, diseases of peppers are caused by various types of pathogens, and the disease causes great damage to production, so it is necessary to prevent it well before it occurs. However, chemical pesticides cannot simultaneously control fungal or bacterial pathogens, and products are developed and used in the form of mixtures by mixing original agents with different main components depending on the mechanism of action.
이들 화학 농약은 인축 독성, 환경 오염과 생태계 영향 및 병원균의 살균제 저항성 발생이 지속적으로 문제가 되고 있어 친환경 천연식물보호제에 대한 관심과 개발이 최근 국내외적으로 급격히 늘어나고 있다. 우리나라에서는 그 동안 고추를 비롯한 채소, 과수, 곡류 등 농작물의 병, 해충 및 잡초 방제를 위하여 많은 양의 농약을 사용해 왔는데 이에 따른 여러 문제들을 해결하기 위하여 화학 비료 및 농약의 사용량을 줄이는 친환경농업 육성 정책을 추진해오고 있다. 이 계획은 2001년부터 5년 단위로 진행되어 현재까지 제 4차 계획(2016~2020년)이 추진되었으며, 목표는 이 기간 중 친환경농산물(무농약 이상)의 재배면적 비율을 현재 4.5%(7만 5천 ha)에서 8%(13만 3천 ha)로 약 78% 확대하고, 매년 약 1.5% 이상의 화학 비료 및 농약 사용량을 감축하는 것이다. 화학 농약을 대체할 수 있는 병해충 방제용으로 국내에 등록 가능한 친환경농자재는 유기농업자재와 천연식물보호제가 있다. 사용 재료는 미생물 자체를 직접 이용하는 생균제 및 미생물 배양액 또는 식물체를 추출하여 사용하는 종류로 크게 나눌 수 있으며, 천연식물보호제의 경우 전자는 미생물 농약, 후자는 생화학 농약으로 구분한다. 미생물 기반 제품들은 토양, 식물근권, 해양 등 특정 자연환경의 전체 미생물 종(microbiome)을 분석하여 새로운 기능의 미생물을 선발하고 대량 배양 후 엽면 살포 또는 토양 처리용으로 제제화하여 병해충 방제제로 사용된다(비특허문헌 9 참조).These chemical pesticides continue to be a problem in terms of human toxicity, environmental pollution and ecological impact, and fungicide resistance of pathogens, and interest in and development of eco-friendly natural plant protection agents are rapidly increasing at home and abroad. In Korea, a large amount of pesticides have been used to control diseases, pests, and weeds in crops such as peppers, vegetables, fruit trees, and grains. has been promoting This plan has been carried out every 5 years since 2001, and the 4th plan (2016-2020) has been implemented so far. 5,000 ha) to 8% (133,000 ha), expanding by about 78%, and reducing the use of chemical fertilizers and pesticides by about 1.5% or more every year. Eco-friendly agricultural materials that can be registered in Korea for pest control that can replace chemical pesticides include organic agricultural materials and natural plant protection agents. Materials used can be largely divided into probiotics that directly use microorganisms themselves and types that extract and use microbial cultures or plants. Microbial-based products are used as pest control agents by analyzing the entire microbiome of a specific natural environment such as soil, plant rhizosphere, and marine to select microorganisms with new functions, mass-cultivate them, and formulate them for foliar spraying or soil treatment (B See Patent Document 9).
여기에 사용되는 미생물은 주로 식물 병원균을 억제할 수 있는 천적(길항) 미생물, 식물생육 촉진 근권 세균(plant growth promoting rhizobacteria, PGPR) 및 식물조직에서 공생하는 내생(endophytic) 미생물들이 있다. 이러한 미생물 중에서도 최근 10여년 간 식물 내생 세균에 대한 연구가 많이 되고 있는데, 특히, 종자나 모종에 처리하여 식물 백신 역할을 함으로써 병해충, 염류 및 고온건조 스트레스에 대한 저항성이 생기게 하는 기술이 국내에서 개발된 바 있다(비특허문헌 10 내지 12 참조). 식물 내생 세균은 기주에 어떠한 해도 주지 않고 공생하면서 유익한 작용을 하는 세균을 말하며 보통 세포 사이의 공간이나 세포 내부에 존재하기도 한다. 작용 기작으로는 질소 고정, 인산의 가용화, 철 착화합 물질(siderophore) 생산, 식물호르몬, 지질펩티드(lipopeptide) 등 항균물질을 생산하여 병해충 저항성을 유도하거나 식물의 생장을 촉진하는 것으로 알려졌다. 현재까지 많이 연구된 내생 세균은 바실러스(Bacillus), 페니바실러스(Paenibacillus), 슈도모나스(Pseudomonas), 리조박터(Lysobacter), 아그로박테리움(Agrobacterium) 등 130여 종이 알려져 있으며, 이들은 벼, 밀 등 곡류와 고추, 토마토, 감자 등 채소 작물에 작용하여 병 발생을 억제하며 작물의 생산을 증진시킨다(비특허문헌 13 내지 17 참조). Bacillus oryzicola YC7007은 벼의 병 방제 및 생장 촉진 효과를 가진 복합 기능 내생 세균으로 기주 식물에 따라 서로 다른 작용 기작을 나타냄이 밝혀졌다(비특허문헌 18 참조).Microorganisms used herein include natural enemy (antagonist) microorganisms capable of inhibiting plant pathogens, plant growth promoting rhizobacteria (PGPR), and endophytic microorganisms that coexist in plant tissues. Among these microorganisms, a lot of research has been done on plant endogenous bacteria for the past 10 years. There is a bar (see Non-Patent
현재 우리나라에 등록된 식물병 방제용 천연식물보호제는 총 18개(2019년 기준)로 주성분이 주로 세균 바실러스(Bacillus)와 곰팡이 트리코데르마(Trichoderma)이다. 이중 실제로 제품이 판매되고 있는 것은 바실러스 서브틸리스(Bacillus subtilis) 10개, 곰팡이 트리코데르마 하지아눔(Trichoderma harzianum) 1개뿐이다(비특허문헌 9 참조). 이 중에서 5개는 해외 수입 미생물을 이용하여 제품화한 것으로, 이외에도 외국에서는 바실러스 아밀로리퀘페이션스(B. amyloliquefaciens) 및 바실러스 푸밀러스(B. pumilus) 등이 사용되고 있다. 미생물을 제품으로 이용하기 위해서는 좋지 않은 환경이나 유통 중에 오랜 기간 동안 밀도를 유지할 수 있고 방제 효과도 좋아야 한다. 그런 점에서 볼 때 포자를 형성하는 바실러스와 트리코데르마는 천연식물보호제 개발에 적절한 미생물로 많은 연구자들이 병해충의 생물학적 방제제로 연구하였다. 바실러스 균주 대부분은 지질펩티드 항균물질인 펜지엔(fengyein), 이투린(iturin), 서팩틴(surfactin) 등을 분비하여 곰팡이 병원균을 직접 억제하거나 기주의 병 저항성을 유도하는 것으로 알려져 있다(비특허문헌 19 참조). 앞에서 언급한 바실러스 외에 최근 페니바실러스 폴리믹사(Paenibacillus polymyxa)의 식물병 방제 효과에 대한 연구 결과가 발표되었다. 이 균은 처음에는 바실러스 속(genus)으로 분류되었으나, 1993년부터 새로운 페니바실러스 속으로 재분류되었다. 이들 균주 중 어떤 종은 식물병원 곰팡이 억제 효과뿐만 아니라 세균병인 풋마름병도 효과적으로 억제하는 것으로 밝혀졌다. 그리고 질소고정이나 인산용해, 식물생장호르몬 분비 등을 통하여 여러 작물의 생장 촉진도 하는 것으로 확인되었다. 특히 푸자리시딘(fusaricidin), 박테리오신(bacteriocin), 지질펩티드 같은 여러 종류의 항생물질을 분비하는데, 그 중 폴리믹신(polymyxin)은 오래전부터 인체 병원 세균을 억제하는 의약용 제품으로 사용되고 있다(비특허문헌 20 내지 23 참조). Currently, there are a total of 18 natural plant protection agents for controlling plant diseases registered in Korea (as of 2019), the main components of which are the bacterium Bacillus and the fungus Trichoderma . Of these, only 10 products of Bacillus subtilis and 1 of the fungus Trichoderma harzianum are actually sold (see Non-Patent Document 9). Five of these were commercialized using foreign imported microorganisms, and in addition, Bacillus amyloliquefaciens and Bacillus pumilus are used in foreign countries. In order to use microorganisms as products, they must be able to maintain density for a long period of time during unfavorable environments or distribution, and have good control effects. In this respect, Bacillus and Trichoderma, which form spores, are suitable microorganisms for the development of natural plant protection agents, and many researchers have studied them as biological control agents for pests. Most of the Bacillus strains are known to secrete lipopeptide antibacterial substances such as fengyein, iturin, and surfactin to directly suppress fungal pathogens or induce disease resistance in the host (non-patent literature). 19). In addition to the above-mentioned bacilli, recent studies on the plant disease control effect of Penibacillus polymyxa have been published. This fungus was initially classified as a Bacillus genus, but since 1993 it has been reclassified as a new Penenicillus genus. Some of these strains have been found to effectively inhibit plant pathogenic fungi as well as bacterial blight. In addition, it has been confirmed that it promotes the growth of various crops through nitrogen fixation, phosphate dissolution, and plant growth hormone secretion. In particular, it secretes several types of antibiotics such as fusaricidin, bacteriocin, and lipopeptide, among which polymyxin has long been used as a pharmaceutical product to inhibit pathogenic bacteria in the human body. See Patent Documents 20 to 23).
현재 국내외적으로 바실러스 속 몇 종이 상품화되어 식물병 방제에 사용되고 있으나 방제 효과가 화학 농약에 비해 상대적으로 낮아 시장에서의 확산이 더디게 진행되고 있다. 보다 효과적인 제품 개발을 위해 기술적인 면에서 방제 대상 병원균을 잘 억제하는 우수한 미생물을 선발해야 되고 동시에 작용 기작을 고려한 개발 전략이 필요하다.Currently, several species of Bacillus have been commercialized domestically and internationally and are used for plant disease control, but their control effect is relatively low compared to chemical pesticides, and their spread in the market is slow. In order to develop more effective products, it is necessary to select excellent microorganisms that suppress the pathogens to be controlled from a technical point of view, and at the same time, a development strategy that considers the mechanism of action is required.
[비특허문헌][Non-patent literature]
비특허문헌 1: 고영진, 김상수, 박서기, 박종대 (2004). 조미 채소 병해충 발생 생태 및 방제. 순천대학교 농업생명과학대학 조미채소기술센터.Non-Patent Document 1: Youngjin Ko, Sangsu Kim, Seogi Park, Jongdae Park (2004). Ecology and Control of Pests and Diseases of Seasoned Vegetables. Seasoned Vegetable Technology Center, College of Agriculture and Life Sciences, Sunchon National University.
비특허문헌 2: Kim, J. T., Par, S. Y., Choi, W. B., Lee, Y. H., Kim, H. T. (2008). Characterization of Colletotrichum isolates causing anthracnose of pepper in Korea. Plant Pathol. J. 24:17-23.Non-Patent Document 2: Kim, JT, Par, SY, Choi, WB, Lee, YH, Kim, HT (2008). Characterization of Colletotrichum isolates causing anthracnose of pepper in Korea. Plant Pathol. J. 24:17-23.
비특허문헌 3: 강범관, 민지영, 김윤식, 박성우, 응웬 반바흐, 김흥태 (2005). 고추 탄저병균 Colletotrichum acutatum의 포장 밀도 조사를 위한 반선택 배지의 확립 및 활용. 식물병연구 11:21-27. Non-Patent Document 3: Kang Beom-Kwan, Min Ji-Young, Kim Yun-Sik, Park Sung-Woo, Nguyen Van Bach, Kim Heung-Tae (2005). Establishment and utilization of a semi-selective medium for the investigation of packing density of pepper anthracnose Colletotrichum acutatum . Plant disease research 11:21-27.
비특허문헌 4: 김신화, 민지영, 김흥태 (2019). Strobilurin계 살균제 Pyraclostrobin에 대한 고추탄저병균 Colletotrichum acutatum의 저항성 발현 및 기작. 농약과학회지 23:202-211.Non-Patent Document 4: Kim Shin-Hwa, Min Ji-Young, Kim Heung-Tae (2019). Expression and mechanism of resistance of the pepper anthracnose Colletotrichum acutatum to the strobilurin fungicide Pyraclostrobin. Journal of Pesticide Science 23:202-211.
비특허문헌 5: 이형주, 조은정, 김남희, 채영, 이선우 (2011). 한국에서 분리한 Ralstonia solanacearum에 대한 순계 토마토의 병 반응과 고온에서의 발병. 식물병연구 17:326-333. Non-Patent Document 5: Hyungjoo Lee, Eunjung Cho, Namhee Kim, Young Chae, Sunwoo Lee (2011). Disease response of purebred tomato to Ralstonia solanacearum isolated in Korea and outbreak at high temperature. Plant Disease Research 17:326-333.
비특허문헌 6: 이영기, 강희안 (2013). 고추에서 분리된 Ralstonia solanacearum 계통의 생리, 생화학 및 유전적 특성. 식물병연구 19:265-272. Non-Patent Document 6: Youngki Lee, Heian Kang (2013). Physiological, biochemical and genetic characteristics of Ralstonia solanacearum strains isolated from pepper. Plant Disease Research 19:265-272.
비특허문헌 7: 이수민, 곽연수, 이경희, 김흥태 (2015). 고추 풋마름병에 대한 살균제의 방제 효과. 농약과학회지 19:323-328. Non-Patent Document 7: Sumin Lee, Yeonsu Kwak, Kyunghee Lee, Heungtae Kim (2015). Control effect of fungicides against green pepper blight. Journal of Pesticide Science 19:323-328.
비특허문헌 8: Kyeon, M. S., Son, S. H., Noh, Y. H., Kim, Y. E., Lee, H. I., Cha, J. S. (2016). Xanthomonas euvesicatoria causes bacterial spot disease on pepper plant in Korea. Plant Pathol. J. 32:431-440.Non-Patent Document 8: Kyeon, MS, Son, SH, Noh, YH, Kim, YE, Lee, HI, Cha, JS (2016). Xanthomonas euvesicatoria causes bacterial spot disease on pepper plant in Korea. Plant Pathol. J. 32:431-440.
비특허문헌 9: 성재훈, 김태훈, 추성민 (2020). 친환경농자재 산업 실태와 정책과제. 한국농촌경제연구원. Non-Patent Document 9: Sung Jae-Hoon, Kim Tae-Hoon, Choo Seong-Min (2020). Environment-friendly agricultural materials industry and policy tasks. Korea Rural Economic Institute.
비특허문헌 10: Hossain, M. T, Khan, A., Chung, E. J., Harun, R. M., Chung, Y. R. (2016). Biological control of rice bakane by an endophytic Bacillus oryzicola YC7007. Plant Pathol. J. 32:228-241.Non-Patent Document 10: Hossain, M. T, Khan, A., Chung, EJ, Harun, RM, Chung, YR (2016). Biological control of rice bakane by an endophytic Bacillus oryzicola YC7007. Plant Pathol. J. 32:228-241.
비특허문헌 11: Harun, M. R., Kim, H. J., Yeom, S. I., Yu, H. A., Moon, S. S., Kang, Y. J., Chung, Y. R. (2018). Bacillus velezensis YC7010 enhances plant defenses against brown planthopper through transcriptomic and metabolic changes in rice. Front. Plant Sci. 9:1904. doi: 10.3389/fpls.2018.01904.Non-Patent Document 11: Harun, MR, Kim, HJ, Yeom, SI, Yu, HA, Moon, SS, Kang, YJ, Chung, YR (2018). Bacillus velezensis YC7010 enhances plant defenses against brown planthopper through transcriptomic and metabolic changes in rice. Front. Plant Sci. 9:1904. doi: 10.3389/fpls.2018.01904.
비특허문헌 12: Baek, D., Rokibuzzaman, M., Khan, A., Kim, M. C., Par, H. J., Yun, D. J., Chung, Y. R. (2020). Plant-growth promoting Bacillus oryzicola YC7007 modulates stress-response gene expression and provides protection from salt stress. Front. Plant Sci., 9 Jan. doi.org/10.3389/ fpls.2019.01646.Non-Patent Document 12: Baek, D., Rokibuzzaman, M., Khan, A., Kim, MC, Par, HJ, Yun, DJ, Chung, YR (2020). Plant-growth promoting Bacillus oryzicola YC7007 modulates stress-response gene expression and provides protection from salt stress. Front. Plant Sci., 9 Jan. doi.org/10.3389/ fpls.2019.01646.
비특허문헌 13: Costa, J. M., & Loper, J. E. (1994). Characterization of siderophore production by the biological control agent Enterobacter cloacae. Mol. Plant-Microbe Interact.7: 440-448.Non-Patent Document 13: Costa, JM, & Loper, JE (1994). Characterization of siderophore production by the biological control agent Enterobacter cloacae . Mol. Plant-Microbe Interact.7: 440-448.
비특허문헌 14: Timmusk, S., Wagner, E. G. H. (1999). The plant-growth promoting rhizobacterium Paenibacillus polymyxa induces changes in Arabidopsis thaliana gene expression: a possible connection between biotic and abiotic stress responses. Mol. Plant-Microbe Interact.12:951-959.Non-Patent Document 14: Timmusk, S., Wagner, EGH (1999). The plant-growth promoting rhizobacterium Paenibacillus polymyxa induces changes in Arabidopsis thaliana gene expression: a possible connection between biotic and abiotic stress responses. Mol. Plant-Microbe Interact. 12:951-959.
비특허문헌 15: Wakelin, S. A., Warren, R. A., Harvey, P. R., Ryder, M. H. (2004). Phosphate solubilization by Penicillium spp. closely associated with wheat roots. Biol. & Fertil. Soils 40:36-43.Non-Patent Document 15: Wakelin, SA, Warren, RA, Harvey, PR, Ryder, MH (2004). Phosphate solubilization by Penicillium spp. closely associated with wheat roots. Biol. & Fertil. Soils 40:36-43.
비특허문헌 16: Rosenblueth, M., & Martinez-Romero, E. (2006). Bacterial endophytes and their interactions with hosts. Mol. Plant-Microbe Interact. 19: 827-837.Non-Patent Document 16: Rosenblueth, M., & Martinez-Romero, E. (2006). Bacterial endophytes and their interactions with hosts. Mol. Plant-Microbe Interact. 19: 827-837.
비특허문헌 17: Bibi, F., Yasir, M., Song, G. C., Lee, S. Y., and Chung, Y. R. (2012). Diversity and characterization of endophytic bacteria associated with tidal flat plants and their antagonistic effects on oomycetous plant pathogens. Plant Pathol. J. 28:20-31.Non-Patent Document 17: Bibi, F., Yasir, M., Song, GC, Lee, SY, and Chung, YR (2012). Diversity and characterization of endophytic bacteria associated with tidal flat plants and their antagonistic effects on oomycetous plant pathogens. Plant Pathol. J. 28:20-31.
비특허문헌 18: Chung, E. J., Hossain, M. T, Khan, A., Kim, K. H., Jeon, C. O., Chung, Y. R. (2015). Bacillus oryzicola sp. nov., an endophytic bacterium isolated from the roots of rice with antimicrobial, plant growth promoting and systemic resistance inducing activities in rice. Plant Pathol. J. 31:152-164.Non-Patent Document 18: Chung, EJ, Hossain, M. T, Khan, A., Kim, KH, Jeon, CO, Chung, YR (2015). Bacillus oryzicola sp. nov., an endophytic bacterium isolated from the roots of rice with antimicrobial, plant growth promoting and systemic resistance inducing activities in rice. Plant Pathol. J. 31:152-164.
비특허문헌 19: Islam, M. T., Rahman, M. M., Pandey, P., Boehme, M. M., Haesaert, G. (2020). Bacilli and Agrobiotechnology: Phytostimulation and Biocontrol (Vol 2). Springer. 335p.Non-Patent Document 19: Islam, M. T., Rahman, M. M., Pandey, P., Boehme, M. M., Haesaert, G. (2020). Bacilli and Agrobiotechnology: Phytostimulation and Biocontrol (Vol 2). Springer. 335p.
비특허문헌 20: Chung, Y. R., Kim, C. H., Hwang, I., Chun, J. (2000). Paenibacillus koreensis sp. nov., a new species that produces an iturin-like antifungal compound. Inter. J. Syst. & Evol. Microb. 50:1495-1500. Non-Patent Document 20: Chung, YR, Kim, CH, Hwang, I., Chun, J. (2000). Paenibacillus koreensis sp. nov., a new species that produces an iturin-like antifungal compound. Inter. J. Syst. & Evol. Microb. 50:1495-1500.
비특허문헌 21: Jeong, H. Y., Choi, S. K., Ryu, C. M., Park, S. H. (2019). Chronicle of a soil bacterium: Paenibacillus polymyxa E681 as a tiny guardian of plant and human health. Front Microbiol. 15 March 2019 doi.org/10.3389/fmicb.2019.00467 Non-Patent Document 21: Jeong, HY, Choi, SK, Ryu, CM, Park, SH (2019). Chronicle of a soil bacterium: Paenibacillus polymyxa E681 as a tiny guardian of plant and human health. Front Microbiol. 15 March 2019 doi.org/10.3389/fmicb.2019.00467
비특허문헌 22: Yi, J., Zhang, D., Cheng, Y., Tan, J., Luo, Y. (2019). The impact of Paenibacillus polymyxa HY96-2 luxS on biofilm formation and control of bacterial wilt. App. Microb. & Cell Physiol. 103:9643-9657. Non-Patent Document 22: Yi, J., Zhang, D., Cheng, Y., Tan, J., Luo, Y. (2019). The impact of Paenibacillus polymyxa HY96-2 luxS on biofilm formation and control of bacterial wilt. App. Microb. & Cell Physiol. 103:9643-9657.
비특허문헌 23: Grady, E. N., MacDonald, J., Liu, L., Richman, A., Yuan, Z. (2016). Current knowledge and perspectives of Paenibacillus: a review. Microb. Cell Fact. 15:203-211.Non-Patent Document 23: Grady, EN, MacDonald, J., Liu, L., Richman, A., Yuan, Z. (2016). Current knowledge and perspectives of Paenibacillus : a review. Microb. Cell Fact. 15:203-211.
비특허문헌 24: Yoon, S. H., Ha, S. M., Lim, J. M., Kwon, S.J. & Chun, J. (2017). A large-scale evaluation of algorithms to calculate average nucleotide identity. Anton van Leeuwen. 110:1281-1286.Non-Patent Document 24: Yoon, SH, Ha, SM, Lim, JM, Kwon, SJ & Chun, J. (2017). A large-scale evaluation of algorithms to calculate average nucleotide identity. Anton van Leeuwen . 110:1281-1286.
이에, 본 발명은 고추의 탄저병, 세균성 풋마름병, 점무늬병 등 방제의 어려움을 효과적으로 해결하기 위한 방법으로, 이 병을 일으키는 곰팡이 및 세균 병원균의 생장을 직접 억제할 뿐만 아니라, 기주 식물인 고추의 병 면역성 증진 및 생육 촉진 효과를 모두 갖는 내생 세균을 국내 근권 토양에서 분리, 배양 제제화함으로써 종합적인 식물 강화(plant strengthener) 효능을 가진 새로운 형태의 천연식물보호제를 개발하여 제공하고자 한다.Therefore, the present invention is a method for effectively solving the difficulties of control of pepper anthracnose, bacterial green blight, spot disease, etc., not only directly inhibiting the growth of fungal and bacterial pathogens that cause this disease, but also the disease immunity of pepper, a host plant It is intended to develop and provide a new type of natural plant protection agent with a comprehensive plant strengthener effect by isolating and culturing endogenous bacteria having both growth and growth promoting effects from domestic rhizosphere soil and formulating them.
상기 과제를 해결하기 위하여 본 발명은, 신규 미생물 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공한다.In order to solve the above problems, the present invention provides a novel microorganism Phenibacillus polymyxa ( Paenibacillus polymyxa ) JG90 (accession number: KCTC14388BP) strain.
또한 상기 균주는 서열번호 1로 기재되는 염기서열을 갖는 16S rRNA 유전자를 포함하는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공한다.In addition, the strain provides a Penibacillus polymyxa JG90 (accession number: KCTC14388BP) strain, characterized in that it contains a 16S rRNA gene having the nucleotide sequence represented by SEQ ID NO: 1.
또한 상기 균주는 고추 탄저병균, 고추 풋마름병균 및 사과 탄저병균으로 이루어진 군에서 선택되는 1종 이상의 병원균에 대한 길항력을 갖는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공한다.In addition, the strain is Penibacillus polymyxa JG90 (Accession Number: KCTC14388BP ) strain.
또한 상기 고추 탄저병 및 상기 고추 풋마름병의 병원균은 각각 콜레토트리쿰 아쿠타툼(Colletotrichum acutatum) 및 랄스토니아 솔라나세아룸(Ralstonia solanacearum)인 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공한다.In addition, the pathogens of the pepper anthracnose and the pepper green blight are Colletotrichum acutatum and Ralstonia solanacearum, respectively . Paenibacillus polymyxa JG90 ( Accession number: KCTC14388BP) strain is provided.
또한 상기 균주는 과수 검은가지마름병균, 복숭아 구멍병균, 고추 무름병균 및 고추 점무늬병균으로 이루어진 군에서 선택되는 1종 이상의 병원균에 대한 길항력을 갖는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공하다.In addition, the strain has antagonistic activity against one or more pathogens selected from the group consisting of fruit tree blight, peach pit blight, pepper soft rot and pepper spot bacillus Penibacillus polymyxa JG90 (Accession number: KCTC14388BP) provides a strain.
또한 상기 고추 무름병균 및 상기 고추 점무늬병의 병원균은 각각 펙토박테리움 카로토보룸(Pectobacterium carotovorum) 및 잔쏘모나스 유베시카토리아(Xanthomonas euvesicatoria)인 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공한다.In addition, the pathogens of the pepper soft rot and the pepper spot disease are Pectobacterium carotovorum and Xanthomonas euvesicatoria, respectively . Paenibacillus polymyxa JG90 (acquisition Number: KCTC14388BP) strain.
또한 상기 균주는 고추에 대한 식물생육 촉진 및 면역성 유도 효능을 갖는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 제공한다.In addition, the strain provides a strain of Penibacillus polymyxa ( Paenibacillus polymyxa ) JG90 (accession number: KCTC14388BP), characterized in that it has plant growth promoting and immunity-inducing effects on pepper.
또한 본 발명은 상기 균주 또는 이의 배양액을 유효성분으로 함유하는 미생물 제제를 제공한다.In addition, the present invention provides a microbial preparation containing the strain or its culture medium as an active ingredient.
또한 상기 미생물 제제는 비료용 또는 천연식물보호제용인 것을 특징으로 하는 미생물 제제를 제공한다.In addition, the microbial preparation provides a microbial preparation, characterized in that for fertilizer or natural plant protection agent.
본 발명에 따르면, 국내 토착 세균으로서 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90은 항생물질 분비와 식물 면역성 유도 작용으로 고추의 중요한 식물병인 탄저병 및 세균성 풋마름병, 무름병, 점무늬병 발생을 동시에 억제할 수 있고, 또한 과수의 중요한 세균성 병원균 생장 억제 능력을 모두 지닌 다기능 세균으로서, 천연식물보호제 및 미생물 비료로서의 역할을 동시에 수행할 수 있는 우수한 미생물 제제를 제공할 수 있다.According to the present invention, Paenibacillus polymyxa JG90, as a domestic native bacterium, secretes antibiotics and induces plant immunity, thereby inhibiting the occurrence of anthrax, bacterial blight, soft rot, and spot disease, which are important plant diseases of pepper, at the same time, In addition, as a multifunctional bacterium possessing both the ability to inhibit the growth of important bacterial pathogens of fruit trees, it is possible to provide an excellent microbial preparation capable of simultaneously serving as a natural plant protection agent and microbial fertilizer.
또한 현재 전국적인 확산에도 불구하고 방제가 어려운 과수 가지검은마름병 (화상병) 및 궤양 병원균에 대한 우수한 항균 활성을 갖고 있으며, 동시에 기주 식물의 병 면역성 유도 및 생육 촉진 효과도 보이는 새로운 다기능 미생물 및 이를 이용한 미생물 제제를 제공할 수 있다.In addition, it has excellent antibacterial activity against fruit tree black blight (burn) and ulcer pathogens, which are difficult to control despite its nationwide spread, and at the same time, a new multifunctional microorganism that induces disease immunity and promotes growth of host plants, and using it Microbial preparations may be provided.
도 1은 본 발명의 실시예 1에서 고추 탄저병 병원균 억제 시험 결과를 나타낸 사진이다. 도 1에서 A는 분리 미생물과 병원균과의 대치 배양 시험 결과를 나타내고, B는 억제 활성이 좋은 균주의 억제 효과 시험 결과를 나타내고 있다.
도 2는 JG90 균주의 16S rRNA 유전자의 염기서열을 나타낸 도면이다.
도 3은 JG90 균주의 16S rRNA 유전자 서열 분석으로 만든 계통수를 나타낸 도면이다.
도 4는 JG90 균주의 유전자 분석(BOX-PCR)을 이용한 표준 균주 대비 유전자 비교 분석 결과를 나타낸 사진이다.
도 5는 JG90 균주의 고추 탄저병 방제효과를 설명하는 처리구별 비교 결과를 나타낸 사진이다.
도 6은 JG90 균주의 사과 탄저병 방제효과를 설명하는 처리구별 비교 결과를 나타낸 사진이다.
도 7은 JG90 균주의 고추 풋마름병 방제효과를 설명하는 처리구별 비교 결과를 나타낸 사진이다.
도 8은 JG90 균주의 주요 식물 병원 세균 억제 효과를 나타낸 사진이다.
도 9는 JG90 균주의 복숭아 세균구멍병 방제효과를 설명하는 처리구별 비교 결과를 나타낸 사진이다.
도 10은 JG90 균주의 고추 탄저병에 대한 면역성 유도 효과를 설명하는 처리구별 비교 결과를 나타낸 사진이다.
도 11은 JG90 균주의 고추 생육 및 열매 개수 증가 효과를 나타낸 사진이다. 도 11에서 (a)는 4주 후, (b)는 8주 후를 나타내고 있다.
도 12는 JG90 균주의 고추 생육 촉진 효과를 나타낸 그래프이다.
도 13은 JG90 균주의 배지별, 배양 기간별 생장을 나타낸 그래프이다.1 is a photograph showing the results of an anthrax pathogen inhibition test in pepper in Example 1 of the present invention. In Figure 1, A shows the results of the replacement culture test of the isolated microorganism and the pathogen, and B shows the test results of the inhibitory effect of strains with good inhibitory activity.
2 is a diagram showing the nucleotide sequence of the 16S rRNA gene of strain JG90.
Figure 3 is a diagram showing a phylogenetic tree created by 16S rRNA gene sequence analysis of strain JG90.
Figure 4 is a photograph showing the results of gene comparison analysis compared to the standard strain using genetic analysis (BOX-PCR) of the JG90 strain.
Figure 5 is a photograph showing the comparison results for each treatment group explaining the pepper anthrax control effect of the JG90 strain.
Figure 6 is a photograph showing the comparison results for each treatment group explaining the apple anthracnose control effect of the JG90 strain.
Figure 7 is a photograph showing the comparison results for each treatment group explaining the pepper green blight control effect of the JG90 strain.
Figure 8 is a photograph showing the inhibitory effect of major plant pathogenic bacteria of strain JG90.
Figure 9 is a photograph showing the comparison results of each treatment group explaining the peach bacterial hole disease control effect of the JG90 strain.
Figure 10 is a photograph showing the comparison results for each treatment group explaining the immunity inducing effect of the JG90 strain against pepper anthracnose.
Figure 11 is a photograph showing the effect of increasing the number of fruits and pepper growth of the JG90 strain. In Fig. 11, (a) shows 4 weeks later and (b) shows 8 weeks later.
12 is a graph showing the growth promoting effect of the JG90 strain on pepper.
13 is a graph showing the growth of the JG90 strain by culture medium and culture period.
이하, 실시예를 통하여 본 발명을 상세히 설명하기로 한다. 이에 앞서, 본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 아니 되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여, 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다. 따라서, 본 명세서에 기재된 실시예의 구성은 본 발명의 가장 바람직한 일실시예에 불과할 뿐이고 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원 시점에 있어서 이들을 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다.Hereinafter, the present invention will be described in detail through examples. Prior to this, the terms or words used in this specification and claims should not be construed as being limited to the usual or dictionary meaning, and the inventor appropriately uses the concept of the term in order to explain his/her invention in the best way. Based on the principle that it can be defined, it should be interpreted as meaning and concept consistent with the technical spirit of the present invention. Therefore, since the configurations of the embodiments described in this specification are merely the most preferred embodiments of the present invention and do not represent all of the technical spirit of the present invention, various equivalents and modifications that can replace them at the time of this application It should be understood that there may be
본 발명자들은 항생물질 분비와 식물 면역성 유도 작용으로 고추의 중요한 식물병인 탄저병 및 세균성 풋마름병, 무름병, 점무늬병 발생을 동시에 억제할 수 있고, 또한 과수의 중요한 세균성 병원균 생장 억제 능력을 모두 지닌 동시에 기주 식물의 병 면역성 유도 및 생육 촉진 효과도 보이는 새로운 다기능 미생물을 발견하였다.The inventors of the present invention can simultaneously inhibit the occurrence of anthracnose, bacterial blight, soft rot, and spotted disease, which are important plant diseases of pepper, by secreting antibiotics and inducing plant immunity, and also have both the ability to inhibit the growth of important bacterial pathogens of fruit trees, and at the same time, A new multifunctional microorganism that also induces disease immunity and promotes growth was discovered.
따라서, 본 발명은 신규 미생물 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주를 개시한다.Accordingly, the present invention discloses a novel microorganism Phenibacillus polymyxa JG90 (accession number: KCTC14388BP) strain.
이하, 실시예를 들어 본 발명을 상세히 설명하기로 한다.Hereinafter, the present invention will be described in detail with reference to examples.
실시예 1 : JG46 균주의 분리 및 동정Example 1: Isolation and identification of strain JG46
(1) JG90 균주의 분리(1) Isolation of strain JG90
JG90 균주는 진주시 문산읍 ㈜제일그린산업의 토양에서 자라는 잡초 억새(Miscanthus sinensis)의 뿌리 내부조직에서 분리되었다. 먼저, 식물 내생 세균을 분리하기 위하여 뿌리 시료를 수돗물에 잘 세척한 후 작은 조각으로 절단하여 1% NaOCl 용액에 10분간 담궈 표면 살균하였다. 이 조각들을 멸균수로 다시 세척한 후 표면 살균이 제대로 되었는지 확인하기 위하여 살균된 조각을 1/10 TSA 배지(Tryptic Soy Broth 3 g, 한천 16 g / 증류수 1 ℓ)에 올려 놓고, 2~3일 정도 배양하면서 표면에서 세균 생장 여부를 관찰하였다. 세균이 전혀 생장하지 않는 뿌리 부분의 조각 1 g을 취하여 고압멸균기에서 살균된 증류수 9 ㎖를 넣어 살균된 주발과 절구로 마쇄 한 후 이 액 1 ㎖를 증류수 9 ㎖에 넣어 순차적으로 10배씩 희석(10-3, 10-4, 10-5)하였다. 희석액 0.1 ㎖를 1/10 TSA 배지에 분주하여 고르게 도말 한 후 28℃ 배양기에서 1~2주 동안 배양하였다. 생장된 세균 350여 종을 순수 분리 하였고, 이 균주들을 고추 탄저병 병원균과 대치 배양한 결과, 하기 표 1과 같이 억제 효과가 좋은 균주 15종을 선발하였다(도 1 참조). 선발 균주 중 생장 속도 등 제품화에 가장 적합한 1종(JG No. 90)을 최종 선발하여 이를 JG90 균주로 명명하였다. 하기 표 1에서 세균의 길항력은 28℃에서 4~6일간 배양한 후 균사 생장 억제 정도로 측정하였고(+, < 3 mm; ++, 4-5 mm; +++, 6-7 mm), R은 살균제 저항성 균주, S는 살균제 감수성 균주이다.The JG90 strain was isolated from the root tissue of Miscanthus sinensis growing in the soil of Jeil Green Industry Co., Ltd., Munsan-eup, Jinju-si. First, in order to isolate plant endogenous bacteria, root samples were washed well with tap water, cut into small pieces, and surface sterilized by soaking in 1% NaOCl solution for 10 minutes. After washing these pieces with sterilized water again, in order to check whether the surface sterilization was properly performed, the sterilized pieces were placed on a 1/10 TSA medium (Tryptic Soy Broth 3 g, agar 16 g / distilled water 1 ℓ) and left for 2 to 3 days. While culturing, the growth of bacteria on the surface was observed. Take 1 g of a piece of the root part where bacteria do not grow at all, add 9 ml of distilled water sterilized in a high-pressure sterilizer, grind it with a sterilized bowl and mortar, and then add 1 ml of this solution to 9 ml of distilled water and sequentially dilute 10 times (10 -3 , 10 -4 , 10 -5 ). 0.1 ml of the diluted solution was dispensed into 1/10 TSA medium, spread evenly, and then cultured in an incubator at 28 ° C for 1 to 2 weeks. About 350 kinds of grown bacteria were purely isolated, and as a result of replacing these strains with the pepper anthracnose pathogen, 15 strains with good inhibitory effects were selected as shown in Table 1 below (see FIG. 1). Among the selected strains, one (JG No. 90) most suitable for commercialization, such as growth rate, was finally selected and named as JG90 strain. In Table 1 below, the antagonism of bacteria was measured by the degree of mycelial growth inhibition after culturing at 28 ° C for 4 to 6 days (+, < 3 mm; ++, 4-5 mm; +++, 6-7 mm), R is a fungicide resistant strain, S is a fungicide sensitive strain.
(2) JG90 균주의 동정(2) Identification of strain JG90
상기 분리된 JG90 균주의 동정을 위하여 16S rRNA 유전자의 염기서열(1499 bp, 도 2 참조)을 결정하고, GenBank/EMBL/DDBJ의 데이터베이스와 상동성 검색을 수행하여 계통학적 위치를 검토하였다. 도 3에서는 JG90 균주의 16S rRNA 유전자 서열 분석으로 만든 계통수를 나타내고 있다. 도 3에서 교점 숫자는 1,000 반복에서 나온 부트스트랩 값(bootstrap value)을 나타낸다.In order to identify the isolated JG90 strain, the base sequence of the 16S rRNA gene (1499 bp, see FIG. 2) was determined, and the phylogenetic location was reviewed by performing a homology search with the GenBank/EMBL/DDBJ database. Figure 3 shows a phylogenetic tree created by 16S rRNA gene sequence analysis of strain JG90. In FIG. 3, the number of intersection points represents the bootstrap value from 1,000 iterations.
한편, 하기와 같이 유사한 표준 균주(type species)를 대상으로 유전체 염기 서열의 평균 핵산 동일성(ANI, average nucleotide identity)을 비교한 결과를 하기 표 2에 나타내었다(비특허문헌 24 참조).On the other hand, the results of comparing average nucleotide identity (ANI) of genomic nucleotide sequences to similar standard strains (type species) are shown in Table 2 below (see Non-Patent Document 24).
표 2를 참조하면, 분리된 JG90 균주는 보다 정확히 유전학적 특성 중 평균 핵산 동일성 분석을 통해 16s rRNA 유사성을 분석한 결과 페니바실러스 폴리믹사(Paenibacillus polymyxa)로 확인되었다.Referring to Table 2, the isolated JG90 strain was identified as Paenibacillus polymyxa as a result of analyzing 16s rRNA similarity through average nucleic acid identity analysis among genetic characteristics more accurately.
도 4에서는 JG90 균주의 유전자 분석(BOX-PCR)을 이용한 표준 균주 대비 유전자 비교 분석 결과를 나타내고 있다. 유전자 분석(BOX-PCR)을 위한 폴리머라제(polymerase)는 Platinum Taq DNA polymerase High Fidelity(Invitrogen), 프라이머(primer)는 BOXAR1(5'-CATCGGCAAGGCGACGCTGACG-3')이 사용되었다. PCR 조건은 초기 변성(initial denaturation)은 95℃에서 7분, 35회 반복되는 과정인 변성(denaturation), 어닐링(annealing), 연장(extension)은 각각 90℃에서 30초, 40℃에서 1분, 72℃에서 3분, 마지막으로 final extension은 72℃에서 10분간 반응시켜 증폭시켰다. 이러한 과정을 거쳐 얻은 PCR 산물에 대해 1% LE agarose gel(Seakem)을 이용하여 증폭 유무를 확인하였다. 도 4에서 M은 1kb 마커(marker), 1은 JG22 균주, 2는 JG90 균주, 3은 JG91 균주, 4는 JG93 균주, 5는 페니바실러스 자밀리에(Paenibacillus jamiliae) KCTC13919, 6은 페니바실러스 폴리믹사(Paenibacillus polymyxa) KACC10098, 7은 페니바실러스 폴리믹사(Paenibacillus polymyxa) E681 표준 균주를 나타낸다.Figure 4 shows the results of gene comparison analysis compared to the standard strain using genetic analysis (BOX-PCR) of the JG90 strain. Platinum Taq DNA polymerase High Fidelity (Invitrogen) was used as a polymerase for gene analysis (BOX-PCR), and BOXAR1 (5'-CATCGGCAAGGCGACGCTGACG-3') was used as a primer. PCR conditions include initial denaturation at 95 ° C for 7 minutes, denaturation, annealing, and extension, which are repeated 35 times, at 90 ° C for 30 seconds, 40 ° C for 1 minute, 3 minutes at 72 ℃, finally the final extension was amplified by reacting at 72
도 4에 나타낸 바와 같이, JG90 균주와 학명이 같거나 유사한 표준 균주와 보다 세부적인 비교를 위해 유전자 분석(BOX-PCR)을 수행한 결과, JG90 균주는 표준 균주들과는 다른 것으로 판명되었다.As shown in FIG. 4, as a result of genetic analysis (BOX-PCR) for more detailed comparison with standard strains having the same or similar scientific name as JG90 strain, the JG90 strain was found to be different from the standard strains.
따라서, 본 발명의 미생물은 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90 균주로 명명하고 이를 기탁기관에 기탁하여, 2020년 11월 24일자로 한국생명공학연구원 생물자원센터(KCTC: Korean Collection for Type Culture)로부터 수탁번호 KCTC14388BP를 부여받았다.Therefore, the microorganism of the present invention was named as the Penibacillus polymyxa JG90 strain and deposited it in a depository institution, and as of November 24, 2020, the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center (KCTC: Korean Collection for Type Culture) was granted accession number KCTC14388BP from
(3) JG90 균주의 전유전체(whole genome) 분석(3) Whole genome analysis of strain JG90
상기 분리된 JG90 균주의 전유전체 분석을 위해 ㈜천랩의 Trueback ID-Genome 시스템과 ExBioCloud을 이용하여 동정 확인 및 전유전체를 분석하였고, 그 결과를 하기 표 3에 나타내었다.For the whole genome analysis of the isolated JG90 strain, identification was confirmed and whole genome was analyzed using the Trueback ID-Genome system and ExBioCloud of Cheonlab Co., Ltd., and the results are shown in Table 3 below.
실시예 2: JG90 균주 제제의 고추 탄저병 방제 효과 검정 Example 2: Examination of pepper anthracnose control effect of JG90 strain preparation
JG90 균주의 고추 탄저병 방제 효과를 검정하기 위하여, 분말 제제화한 시료를 하기 표 4에 나타낸 바와 같이 지역이 다른 2 포장에서 병 방제 효능을 조사하였다. 방제 효능 시험은 난괴법에 따라 무처리, JG90 미생물제 3반복으로 조사하였으며, 미생물제는 1,000배 및 2,000배 희석하여 발병 초 10일 간격으로 4회 경엽 처리하였다. 방제가는 최종 미생물제 처리 10일 후 처리구 당 200과 이상에 대한 발병과 수를 조사하여 발병과 율(%)로 나타내었다. 약해 조사는 JG90 미생물제 처리 후 7, 14, 21일에 외관상 약해 유무를 조사하였으나 모두 약해는 없었다(도 5 참조).In order to test the pepper anthracnose control effect of the JG90 strain, the powdered samples were examined for disease control efficacy in two fields with different regions, as shown in Table 4 below. The control efficacy test was investigated with no treatment and three repetitions of JG90 microorganisms according to the egg mass method, and the microorganisms were diluted 1,000-fold and 2,000-fold and treated with foliage four times at intervals of 10 days at the beginning of the outbreak. The control value was expressed as incidence and rate (%) by examining the incidence and number of more than 200 families per
하기 표 4에 JG90 미생물제의 고추 탄저병에 대한 방제 효과 조사 결과를 나타내었다. 이러한 결과로부터 본 미생물제는 고추 탄저병 방제용으로 사용 가능한 것을 확인할 수 있다.Table 4 below shows the results of investigation of the control effect of JG90 microorganisms against pepper anthracnose. From these results, it can be confirmed that this microbial agent can be used for controlling pepper anthracnose.
(DMRT)significant difference
(DMRT)
(%)control
(%)
실시예 3: JG90 균주 제제의 사과 탄저병 방제 효과 검정 Example 3: Test of apple anthracnose control effect of JG90 strain preparation
JG90 균주의 사과 탄저병 방제 효과를 검정하기 위하여, 분말 제제화한 시료를 하기 표 5와 같이 사과 노지 포장에서 병 방제 효능을 조사하였다. 방제 효능 시험은 완전임의배치법에 따라 무처리, JG90 미생물제 3반복으로 조사하였으며, 미생물제는 1,000배 및 2,000배 희석하여 발병 초 10일 간격으로 7회 경엽 처리하였다. 방제가는 최종 미생물제 처리 10일 후 처리구 당 전과에 대한 발병과수를 조사하여 발병과율(%)로 나타내었다. 약해 조사는 JG90 미생물제 처리 후 3, 5, 7일에 외관상 약해 유무를 조사하였으나 모두 약해는 없었다(도 6 참조).In order to test the apple anthracnose control effect of the JG90 strain, the powdered samples were examined for disease control efficacy in apple open field packaging as shown in Table 5 below. The control efficacy test was investigated with no treatment and 3 repetitions of JG90 microorganisms according to the completely randomized batch method, and the microorganisms were diluted 1,000-fold and 2,000-fold and treated with foliage seven times at intervals of 10 days at the beginning of the onset. The control value was expressed as an outbreak rate (%) by examining the number of outbreaks per
하기 표 5에 JG90 미생물제의 사과 탄저병에 대한 방제 효과 조사 결과를 나타내었다. 이러한 결과로부터 본 미생물제는 사과 탄저병 방제용으로 사용 가능한 것을 확인할 수 있다.Table 5 below shows the results of investigation of the control effect of JG90 microorganisms against apple anthracnose. From these results, it can be confirmed that this microbial agent can be used for controlling apple anthracnose.
(DMRT)significant difference
(DMRT)
(%)control
(%)
실시예 4: JG90 균주의 고추 풋마름병 방제 효과 검정Example 4: Test of pepper green blight control effect of strain JG90
JG90 균주의 고추 풋마름 방제 효과를 검정하기 위하여, 분말 제제화한 시료를 하기와 같이 지역이 다른 2 포장에서 병 방제 효능을 조사하였다. 방제 효능 시험은 난괴법에 따라 무처리, JG90 미생물제 처리구 3반복으로 조사하였으며 미생물제는 1,000배 및 2,000배 희석하여 발병 초 7일 간격으로 4회 관주 처리하였다. 방제가는 최종 미생물제 처리 7일 후 처리구 당 전주에 대한 발병주 수를 조사하여 발병주 율(%)로 나타내었다. 약해 조사는 JG90 미생물제 처리 후 7, 14, 21일에 외관상 약해 유무를 조사하였으나 모두 약해는 없었다(도 7 참조).In order to test the effect of the JG90 strain on green pepper blight control, the powdered samples were examined for disease control efficacy in two fields with different regions as follows. The control efficacy test was investigated with no treatment and JG90
하기 표 6에 JG90 미생물제의 고추 풋마름병에 대한 방제 효과 조사 결과를 나타내었다. 이러한 결과로부터 본 미생물제는 고추 풋마름병 방제용으로 사용 가능한 것을 확인할 수 있다.Table 6 shows the results of investigation of the control effect of JG90 microorganisms against green pepper blight. From these results, it can be confirmed that this microbial agent can be used for controlling green pepper blight.
(DMRT)significant difference
(DMRT)
(%)control
(%)
실시예 5: JG90 균주의 주요 식물 병원 세균 억제 효과 Example 5: Inhibitory effect of major plant pathogenic bacteria of strain JG90
JG90 균주의 고추 및 과수의 중요 식물 병원 세균인 점무늬병균, 무름병균, 과수 검은가지마름병균 및 복숭아 구멍병균의 억제 효능을 검정하기 위하여, 하기와 같이 병균 생장 억제 효과를 조사하였다.In order to test the inhibitory effect of JG90 strain on important plant pathogenic bacteria of peppers and fruit trees, namely spot blight, soft rot, fruit tree blight and peach pit bacillus, the germ growth inhibitory effect was investigated as follows.
JG90 균주를 1/10 TSB 액체배지에서 28℃로 72시간 동안 진탕(180 rpm) 배양하여 한천구멍확산법(agar well diffusion method)으로 배양액의 억제 정도를 조사하였다. 배지 위에 병원 세균을 깔고 직경 10 mm의 구멍을 뚫은 후 그 구멍 안에 JG90 균주 배양액 100 ㎕를 넣은 후 구멍 주위에 형성된 병원균 생장 억제 정도를 확인하고, 그 결과를 도 8에 나타내었다. 도 8에서 (a)는 과수 검은가지마름병균, (b)는 고추 무름병균 및 (c)는 복숭아 세균구멍병균에 대한 항균 효과를 나타내고 있다.The JG90 strain was cultured in 1/10 TSB liquid medium at 28° C. for 72 hours with shaking (180 rpm), and the degree of inhibition of the culture medium was examined by the agar well diffusion method. After spreading pathogenic bacteria on the culture medium and drilling a hole with a diameter of 10 mm, 100 μl of JG90 strain culture medium was put into the hole, and then the degree of inhibition of pathogen growth formed around the hole was confirmed, and the results are shown in FIG. 8 . In FIG. 8, (a) shows the antibacterial effect against fruit tree blight, (b) pepper soft rot, and (c) peach bacterium.
도 8을 참조하면, JG90 배양액은 고추의 탄저병뿐만 아니라 병원 세균 외에도 과수의 병원 세균 억제 효과도 좋은 것을 알 수 있다.Referring to FIG. 8 , it can be seen that the JG90 culture medium has a good inhibitory effect on not only anthrax of pepper but also pathogenic bacteria of fruit trees.
실시예 6: JG90 균주 제제의 복숭아 세균구멍병 방제 효과 검정 Example 6: Examination of peach bacterial hole disease control effect of JG90 strain preparation
JG90 균주의 복숭아 세균구멍병 방제 효과를 검정하기 위하여, 분말 제제화한 시료의 병 방제 효능을 조사하였다. 방제 효능 시험은 완전임의배치법에 따라 무처리, JG90 미생물제 3반복으로 조사하였으며, 미생물제는 1,000배 및 2,000배 희석하여 발병 직전 10일 간격으로 3회 경엽 처리하였다. 방제가는 최종 미생물제 처리 11일 후 처리구 당 200엽 이상에 대한 발병엽 수를 조사하여 발병엽율(%)로 나타내었다. 약해 조사는 JG90 미생물제 처리 후 3, 5, 7일에 외관상 약해 유무를 조사하였으나 모두 약해는 없었다(도 9 참조).In order to test the effect of the JG90 strain on peach bacterial hole disease control, the disease control efficacy of powdered samples was investigated. The control efficacy test was investigated with no treatment and three repetitions of JG90 microorganisms according to the completely randomized batch method, and the microorganisms were diluted 1,000-fold and 2,000-fold and treated with foliage three times at 10-day intervals immediately before the onset. The control value was expressed as an onset leaf ratio (%) by examining the number of onset leaves for more than 200 leaves per treatment group 11 days after the final microbial agent treatment. In terms of weakness, the presence or absence of weakness was examined on the 3rd, 5th, and 7th days after treatment with JG90 microorganisms, but there was no weakness in all of them (see FIG. 9).
하기 표 7에 JG90 미생물제의 복숭아 세균구멍병에 대한 포장 방제 효과 조사 결과를 나타내었다. 이러한 결과로부터 본 미생물제는 복숭아 세균구멍병 방제용으로 사용 가능한 것을 확인할 수 있다.Table 7 below shows the results of investigation of the field control effect of JG90 microorganisms against bacterial hole disease in peaches. From these results, it can be confirmed that the present microbial agent can be used for controlling peach bacterial hole disease.
(DMRT)significant difference
(DMRT)
(%)control
(%)
실시예 7: JG90 균주의 고추 면역성 유도 효능 검정Example 7: Test of JG90 strain's efficacy in inducing immunity in pepper
JG90 균주의 고추 면역성 유도 효과를 검정하기 위하여, 하기와 같이 방제효과를 조사하였다.In order to test the JG90 strain's immunity induction effect of red pepper, the control effect was investigated as follows.
고추 종자(칼라)를 발아시킨 후 원예용 상토를 넣은 폿트(직경 10 cm)에 파종하여 식물 생육상(28~30℃, 상대습도 80% 이상, 암 16시간, 광 8시간)에서 3~4엽기 될 때까지 육묘한 후 JG90 균주 배양액(0.5 TSB, 28℃, 48 h, 160 rpm, O.D ≥ 1) 10 ㎖를 폿트의 고추 근권에 분주 처리하였다. 균주 처리 3일 후에 오트밀 한천(oatmeal agar) 배지에서 약 3주 동안 배양한 고추 탄저병균 콜레토트리쿰 코코데스(Colletotrichum coccodes)의 균사 디스크를 고추 묘 잎에 올려 놓았다. 각 주당 잎 2개에 병균 디스크 1개를 올려 5반복, 총 10개 잎에 접종하였다. 병균이 접종 처리된 폿트를 비닐로 덮어 6일간 생육상에 둔 후 발생된 병반 괴사 정도를 조사하였다. 발병도는 0~3(0: 병징 없이 건전, 1: 균사 디스크 주위 잎의 10% 정도 병반, 2: 균사 디스크 주위 잎의 30~50% 정도 병반, 3: 균사 디스크 주위 잎의 50% 이상 괴사)으로 구분하였고, 이병률은 병 조사로부터 얻은 발병 정도에 따라 하기 수학식 1에 따라 계산하여 산출하였다.After germination of red pepper seeds (color), seeds are sown in pots (10 cm in diameter) containing horticultural soil, and in the growth phase (28-30℃, relative humidity 80% or more, 16 hours dark, 8 hours light), the 3rd to 4th leaf stage After seedlings were grown until seedlings were grown, 10 ml of JG90 strain culture medium (0.5 TSB, 28 ° C, 48 h, 160 rpm, OD ≥ 1) was dispensed into the root zone of the pepper in the pot. After 3 days of strain treatment, mycelial disks of pepper anthracnose Colletotrichum coccodes cultured for about 3 weeks in an oatmeal agar medium were placed on pepper seedling leaves. 1 disease disc was placed on 2 leaves for each stock and inoculated in 5 repetitions, for a total of 10 leaves. The pots inoculated with the pathogen were covered with vinyl and left in the growth phase for 6 days, and then the degree of necrosis of the lesions was examined. The degree of disease is 0 to 3 (0: healthy without symptoms, 1: diseased about 10% of the leaves around the mycelial disc, 2: diseased about 30-50% of the leaves around the mycelial disc, 3: necrosis of more than 50% of the leaves around the mycelial disc) It was classified into, and the morbidity was calculated by calculating according to
[수학식 1][Equation 1]
이병률(%) = {[(이병엽 수 × 1) + (이병엽 수 × 2) + (이병엽 수 × 3)]/조사잎 수 × 3} × 100Disease rate (%) = {[(Number of Byeongyeop Lee × 1) + (Number of Byungyeop Lee × 2) + (Number of Byungyeop Lee × 3)]/Number of Josa leaves × 3} × 100
하기 표 8에 JG90 균주의 토양 근권 처리에 의한 고추 탄저병 방제 효과 조사 결과를 나타내었으며, 도 10에 고추 탄저병에 대한 면역성 유도 효과를 설명하는 처리구 별 비교 사진을 나타내었다.Table 8 shows the results of investigation of the effect of controlling pepper anthracnose by the soil rhizosphere treatment of strain JG90, and FIG.
(이병률, %)average
(Prevalence, %)
C. coccodes pathogen
C. coccodes
표 8 및 도 10을 참조하면, JG90 균주 현탁액을 토양에 처리한 3일 후에 병균을 접종하여 방제 효과를 조사한 결과, 병원균만 처리한 잎의 발병도가 2.4±0.8 인 것에 반해, JG90 균주 처리구는 0.9±0.7로 62.5%의 방제가를 나타내어, 기주 탄저병 면역성 유도 효과가 우수한 것을 확인할 수 있다.Referring to Table 8 and FIG. 10, as a result of examining the control effect by inoculating the
실시예 8: JG90 균주의 고추 생육 촉진 효능 검정Example 8: Test of pepper growth promoting efficacy of JG90 strain
JG90 균주 처리에 의한 고추의 생육 촉진 효능을 검정하기 위하여, 하기와 같이 식물 생육 촉진 효능을 조사하였다.In order to test the growth promoting effect of red pepper by treatment with the JG90 strain, the plant growth promoting effect was investigated as follows.
고추 종자(칼라)를 발아시킨 후 원예용 상토를 넣은 폿트(직경 10 cm)에 파종하여 식물 생육상(28~30℃, 상대습도 80% 이상, 암 16시간, 광 8시간)에서 3~4엽기 될 때까지 육묘한 후 JG90 균주 배양액(0.5 TSB, 28℃, 48 h, 160 rpm, O.D ≥ 1) 10 ㎖를 폿트의 고추 근권에 분주 처리하였다. 균주 처리 후에 식물 생육상에서 2주간 재배 후에 각각 줄기와 뿌리 생장 정도 및 생중량을 조사하였다. 대조구는 같은 양의 배지(0.5 TSB)만을 분주하였다. 각 처리는 3반복으로 반복 당 5주의 식물을 사용하였다. 하기 표 9에 JG90균주의 고추 생육 촉진 효능 조사 결과를 나타내었다.After germination of red pepper seeds (color), seeds are sown in pots (10 cm in diameter) containing horticultural soil, and in the growth phase (28-30℃, relative humidity 80% or more, 16 hours dark, 8 hours light), the 3rd to 4th leaf stage After seedlings were grown until seedlings were grown, 10 ml of JG90 strain culture medium (0.5 TSB, 28 ° C, 48 h, 160 rpm, O.D ≥ 1) was dispensed into the root zone of the pepper in the pot. After the strain treatment, the stem and root growth and fresh weight were examined after 2 weeks of cultivation on the plant growth bed. For the control group, only the same amount of medium (0.5 TSB) was dispensed. Each treatment was repeated in triplicate, using 5 plants per repetition. Table 9 below shows the results of investigation of the growth promoting efficacy of JG90 strains of pepper.
표 9를 참조하면, JG90 균주 현탁액을 토양에 처리한 2주 후에 고추 생중량과 뿌리 및 줄기 길이를 조사한 결과, JG90 균주 처리에 의하여 생중량이 무처리 11.4 g에 비하여 12.0 g으로 다소 증가하였으나 유의성 있는 차이는 없었다. 그러나 뿌리의 길이는 무처리구의 14.0 ㎝에 비하여 18.0 ㎝로 훨씬 길었고, 줄기 길이는 차이가 없었다. 상기와 같이 생육상에서 키운 모종을 다시 큰 폿트(직경 18 cm)에 옮겨 심어 노지에서 2개월간 재배하면서 생육 상태를 조사하였고, 그 결과를 도 11 및 12에 나타내었다.Referring to Table 9, as a result of examining the fresh weight and root and stem lengths of pepper two weeks after the treatment of the JG90 strain suspension in the soil, the fresh weight was slightly increased to 12.0 g compared to the untreated 11.4 g by the JG90 strain treatment, but the significance there was no difference However, the length of the root was much longer at 18.0 cm compared to 14.0 cm of the untreated group, and there was no difference in stem length. The seedlings grown in the growth phase as described above were transferred to a large pot (18 cm in diameter) and grown in the open field for 2 months to investigate the growth condition, and the results are shown in FIGS. 11 and 12.
도 11 및 12를 참조하면, JG90균주를 처리한 고추는 무처리에 비하여 식물 높이 등 생육이 현저히 향상된 것을 알 수 있다. 이 결과로 볼 때 유묘기에 JG90 균주 처리에 의한 고추 뿌리의 생육 및 줄기 촉진 효능이 확인되므로, 성체로 생장 후 고추 수확량이 증가될 것으로 예상할 수 있다.Referring to Figures 11 and 12, it can be seen that the growth of the JG90 strain was significantly improved compared to the untreated pepper. From this result, since the growth and stem promotion effects of the JG90 strain in the seedling period were confirmed, it can be expected that the yield of red pepper will increase after growing into an adult.
실시예 9: JG90 균주의 배지별, 배양 기간별 생장 비교 Example 9: Comparison of growth of JG90 strain by medium and culture period
JG90 균주의 제제화를 위하여 대량 배양 및 적정 생장 조건을 찾고자 5가지 종류의 배지(GM, SYM, NB, TSB 및 0.5 TSB)에서 각각 12, 24, 36, 48, 60 및 72시간 별로 흡광도(600 nm)를 측정하고, 그 결과를 도 13에 나타내었다.Absorbance (600 nm) at 12, 24, 36, 48, 60 and 72 hours in 5 types of media (GM, SYM, NB, TSB and 0.5 TSB) to find mass culture and appropriate growth conditions for the formulation of JG90 strain ) was measured, and the results are shown in FIG. 13.
도 13을 참조하면, 배양 48시간째 JG90 균주는 3가지 SYM, TSB 및 0.5TSB 배지에서 가장 높은 흡광도를 보여 배양에 적합한 것으로 확인되었다. 사용한 배지는 판매중인 딥코(Difco) 배지를 사용하였으며 주성분은 다음과 같다. GM; Soypepton 2g, Yeast Extract 1g, Sucrose 0.5g, MgSO4 4g, MgCl2 2g, NaCl 10g, SYM: Yeast Extract 5g, Sucrose 10g, NB: Beef Extract 3g, Peptone 5g, TSB: Pancreatic digest of Casein 17g, Papaic digest of soybean 3g, Dextrose 2.5g, Sodium Chloride 5g, Dipotassium Phosphate 2.5g, 0.5TSB: Pancreatic digest of Casein 8.5g, Papaic digest of Soybean 1.5g, Dextrose 1.25g, Sodium Chloride 5g, Dipotassium Phosphate 1.25g.Referring to FIG. 13, at 48 hours of culture, strain JG90 showed the highest absorbance in three types of SYM, TSB and 0.5TSB media, and was confirmed to be suitable for culture. The medium used was a commercially available Difco medium, and the main components were as follows. GM; Soypepton 2g, Yeast Extract 1g, Sucrose 0.5g, MgSO 4 4g, MgCl 2 2g, NaCl 10g, SYM: Yeast Extract 5g, Sucrose 10g, NB: Beef Extract 3g, Peptone 5g, TSB: Pancreatic digest of Casein 17g, Papaic digest of soybean 3g, Dextrose 2.5g, Sodium Chloride 5g, Dipotassium Phosphate 2.5g, 0.5TSB: Pancreatic digest of Casein 8.5g, Papaic digest of Soybean 1.5g, Dextrose 1.25g, Sodium Chloride 5g, Dipotassium Phosphate 1.25g.
실시예 10: JG90 균주의 제제화Example 10: Formulation of JG90 strain
JG90 균주의 제품화를 위하여 활성과 보관 기간 중 밀도를 유지할 수 있도록, 하기와 같이 분말과 액제 형태로 제제화하였다.For commercialization of the JG90 strain, it was formulated in powder and liquid form as follows to maintain activity and density during storage period.
균주를 대형 발효기(1톤 이상)에서 배양하여 얻은 세균 배양액 또는 원심분리 후 얻은 균체를 지오라이트(zeolite), 카올린(kaolin), 벤토나이트(bentonite), 이탄(peat) 등의 점토 광물과 1:100의 중량비로 혼합하여 저온에서 말린 후 균일하게 분쇄하여 분말 형태의 고상제로 제조하였다. 동시에 균 배양체를 상기 점토 광물과 1:100의 중량비로 혼합하여 액제 현탁액 형태로도 제조하였다.Bacterial culture obtained by culturing the strain in a large fermentor (more than 1 ton) or the cell obtained after centrifugation is mixed with clay minerals such as zeolite, kaolin, bentonite, and peat at a ratio of 1:100. It was mixed at a weight ratio of, dried at low temperature, and uniformly pulverized to prepare a solid in powder form. At the same time, the bacterial culture was mixed with the clay mineral at a weight ratio of 1:100 to prepare a liquid suspension form.
상기와 같이 제제화한 시료의 밀도를 조사한 결과, 대부분의 제제에서 109 cfu/g 이상을 포함하고 있었으며, 점토 광물과 혼합하여 제제한 액제 현탁액도 109 cfu/㎖ 이상의 밀도를 함유한 것으로 확인되었다.As a result of examining the density of the samples formulated as described above, most of the formulations contained 10 9 cfu / g or more, and it was confirmed that the liquid suspension prepared by mixing with clay minerals also contained a density of 10 9 cfu / ml or more. .
이상에서 설명한 본 발명의 바람직한 실시예들은 기술적 과제를 해결하기 위해 개시된 것으로, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자라면 본 발명의 사상 및 범위 안에서 다양한 수정, 변경, 부가 등이 가능할 것이며, 이러한 수정 변경 등은 이하의 특허청구범위에 속하는 것으로 보아야 할 것이다.The preferred embodiments of the present invention described above have been disclosed to solve the technical problems, and those skilled in the art will be able to make various modifications, changes, additions, etc. within the spirit and scope of the present invention. , such modifications and changes should be regarded as belonging to the scope of the following claims.
[수탁기관][Entrusted institution]
기탁기관명 : 한국생명공학연구원Name of Depositary Institution: Korea Research Institute of Bioscience and Biotechnology
수탁번호 : KCTC14388BPAccession number: KCTC14388BP
수탁일자 : 20201124Entrusted date: 20201124
<110> Jgreen Inc. <120> Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects <130> NP21-10105 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1499 <212> DNA <213> Paenibacillus polymyxa <400> 1 gtttgatcct ggctcaggac gaacgctggc ggcgtgccta atacatgcaa gtcgagcggg 60 gttatttaga agcttgcttc taaataacct agcggcggac gggtgagtaa cacgtaggca 120 acctgcccac aagacaggga taactaccgg aaacggtagc taatacccga tacatccttt 180 tcctgcatgg gagaaggagg aaaggcggag caatctgtca cttgtggatg ggcctgcggc 240 gcattagcta gttggtgggg taatggccta ccaaggcgac gatgcgtagc cgacctgaga 300 gggtgatcgg ccacactggg actgagacac ggcccagact cctacgggag gcagcagtag 360 ggaatcttcc gcaatgggcg aaagcctgac ggagcaacgc cgcgtgagtg atgaaggttt 420 tcggatcgta aagctctgtt gccagggaag aacgtcttat agagtaactg ctataagagt 480 gacggtacct gagaagaaag ccccggctaa ctacgtgcca gcagccgcgg taatacgtag 540 ggggcaagcg ttgtccggaa ttattgggcg taaagcgcgc gcaggcggct ctttaagtct 600 ggtgtttaat cccgaggctc aacttcgggt cgcactggaa actggggagc ttgagtgcag 660 aagaggagag tggaattcca cgtgtagcgg tgaaatgcgt agagatgtgg aggaacacca 720 gtggcgaagg cgactctctg ggctgtaact gacgctgagg cgcgaaagcg tggggagcaa 780 acaggattag ataccctggt agtccacgcc gtaaacgatg aatgctaggt gttaggggtt 840 tcgataccct tggtgccgaa gttaacacat taagcattcc gcctggggag tacggtcgca 900 agactgaaac tcaaaggaat tgacggggac ccgcacaagc agtggagtat gtggtttaat 960 tcgaagcaac gcgaagaacc ttaccaggtc ttgacatccc tctgatcgct gtagagatat 1020 ggctttcctt cgggacagag gagacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga 1080 gatgttgggt taagtcccgc aacgagcgca acccttatgc ttagttgcca gcaggtcaag 1140 ctgggcactc taagcagact gccggtgaca aaccggagga aggtggggat gacgtcaaat 1200 catcatgccc cttatgacct gggctacaca cgtactacaa tggccggtac aacgggaagc 1260 gaagccgcga ggtggagcca atcctagaaa agccggtctc agttcggatt gcaggctgca 1320 actcgcctgc atgaagtcgg aattgctagt aatcgcggat cagcatgccg cggtgaatac 1380 gttcccgggt cttgtacaca ccgcccgtca caccacgaga gtttacaaca cccgaagtcg 1440 gtgaggtaac cgcaaggggc cagccgccga aggtggggta gatgattggg gtgaagtcg 1499 <110> Jgreen Inc. <120> Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects <130> NP21-10105 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1499 <212> DNA <213> Paenibacillus polymyxa <400> 1 gtttgatcct ggctcaggac gaacgctggc ggcgtgccta atacatgcaa gtcgagcggg 60 gttattaga agcttgcttc taaataacct agcggcggac gggtgagtaa cacgtaggca 120 acctgcccac aagacaggga taactaccgg aaacggtagc taatacccga tacatccttt 180 tcctgcatgg gagaaggagg aaaggcggag caatctgtca cttgtggatg ggcctgcggc 240 gcattagcta gttggtgggg taatggccta ccaaggcgac gatgcgtagc cgacctgaga 300 gggtgatcgg ccacactggg actgagacac ggcccagact cctacggggag gcagcagtag 360 ggaatcttcc gcaatgggcg aaagcctgac ggagcaacgc cgcgtgagtg atgaaggttt 420 tcggatcgta aagctctgtt gccagggaag aacgtcttat agagtaactg ctataagagt 480 gacggtacct gagaagaaag ccccggctaa ctacgtgcca gcagccgcgg taatacgtag 540 ggggcaagcg ttgtccggaa ttaattgggcg taaagcgcgc gcaggcggct ctttaagtct 600 ggtgtttaat cccgaggctc aacttcgggt cgcactggaa actggggagc ttgagtgcag 660 aagaggagag tggaattcca cgtgtagcgg tgaaatgcgt agagatgtgg aggaacacca 720 gtggcgaagg cgactctctg ggctgtaact gacgctgagg cgcgaaagcg tggggagcaa 780 acaggattag ataccctggt agtccacgcc gtaaacgatg aatgctaggt gttaggggtt 840 tcgataccct tggtgccgaa gttaacacat taagcattcc gcctggggag tacggtcgca 900 agactgaaac tcaaaggaat tgacggggac ccgcacaagc agtggagtat gtggtttaat 960 tcgaagcaac gcgaagaacc ttaccaggtc ttgacatccc tctgatcgct gtagagatat 1020 ggctttcctt cgggacagag gagacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga 1080 gatgttgggt taagtcccgc aacgagcgca acccttatgc ttagttgcca gcaggtcaag 1140 ctgggcactc taagcagact gccggtgaca aaccggagga aggtggggat gacgtcaaat 1200 catcatgccc cttatgacct gggctacaca cgtactacaa tggccggtac aacgggaagc 1260 gaagccgcga ggtggagcca atcctagaaa agccggtctc agttcggatt gcaggctgca 1320 actcgcctgc atgaagtcgg aattgctagt aatcgcggat cagcatgccg cggtgaatac 1380 gttcccgggt cttgtacaca ccgcccgtca caccacgaga gtttacaaca cccgaagtcg 1440 gtgaggtaac cgcaaggggc cagccgccga aggtggggta gatgattggg gtgaagtcg 1499
Claims (9)
상기 균주는 서열번호 1로 기재되는 염기서열을 갖는 16S rRNA 유전자를 포함하는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주.According to claim 1,
The strain is characterized in that it comprises a 16S rRNA gene having a nucleotide sequence represented by SEQ ID NO: 1 Penibacillus polymyxa ( Paenibacillus polymyxa ) JG90 (accession number: KCTC14388BP) strain.
상기 균주는 고추 탄저병균, 고추 풋마름병균 및 사과 탄저병균으로 이루어진 군에서 선택되는 1종 이상의 병원균에 대한 길항력을 갖는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주.According to claim 1,
The strain is characterized in that it has an antagonistic force against one or more pathogens selected from the group consisting of pepper anthrax, pepper green blight and apple anthracnose, Paenibacillus polymyxa JG90 (accession number: KCTC14388BP) strain.
상기 고추 탄저병 및 상기 고추 풋마름병의 병원균은 각각 콜레토트리쿰 아쿠타툼(Colletotrichum acutatum) 및 랄스토니아 솔라나세아룸(Ralstonia solanacearum)인 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주.According to claim 3,
The pathogens of the pepper anthracnose and the pepper green blight are Colletotrichum acutatum and Ralstonia solanacearum, respectively. Paenibacillus polymyxa JG90 (acquisition) Number: KCTC14388BP) strain.
상기 균주는 과수 검은가지마름병균, 복숭아 구멍병균, 고추 무름병균 및 고추 점무늬병균으로 이루어진 군에서 선택되는 1종 이상의 병원균에 대한 길항력을 갖는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주.According to claim 1,
The strain has antagonistic activity against one or more pathogens selected from the group consisting of fruit tree black spot blight, peach pit blight, pepper soft rot and pepper spot bacillus Penibacillus polymyxa ( Paenibacillus polymyxa ) JG90 ( Accession number: KCTC14388BP) strain.
상기 고추 무름병균 및 상기 고추 점무늬병의 병원균은 각각 펙토박테리움 카로토보룸(Pectobacterium carotovorum) 및 잔쏘모나스 유베시카토리아(Xanthomonas euvesicatoria)인 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주.According to claim 5,
The pepper soft rot and the pathogen of the pepper spot disease are Pectobacterium carotovorum and Xanthomonas euvesicatoria, respectively. Paenibacillus polymyxa JG90 (accession number : KCTC14388BP) strain.
상기 균주는 고추에 대한 식물생육 촉진 및 면역성 유도 효능을 갖는 것을 특징으로 하는 페니바실러스 폴리믹사(Paenibacillus polymyxa) JG90(수탁번호: KCTC14388BP) 균주.According to claim 1,
The strain is a Penibacillus polymyxa JG90 (accession number: KCTC14388BP) strain, characterized in that it has plant growth promoting and immunity-inducing efficacy for pepper.
상기 미생물 제제는 비료용 또는 천연식물보호제용인 것을 특징으로 하는 미생물 제제.According to claim 8,
The microbial agent is a microbial agent, characterized in that for fertilizer or natural plant protection agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020220000878A KR20230105465A (en) | 2022-01-04 | 2022-01-04 | Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020220000878A KR20230105465A (en) | 2022-01-04 | 2022-01-04 | Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20230105465A true KR20230105465A (en) | 2023-07-11 |
Family
ID=87159547
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020220000878A KR20230105465A (en) | 2022-01-04 | 2022-01-04 | Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20230105465A (en) |
-
2022
- 2022-01-04 KR KR1020220000878A patent/KR20230105465A/en not_active Application Discontinuation
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2736382C1 (en) | Compositions and methods for fusarium disease control | |
| Liu et al. | Biological control of take-all in wheat by endophytic Bacillus subtilis E1R-j and potential mode of action | |
| Wu et al. | Effects of bio-organic fertilizer on pepper growth and Fusarium wilt biocontrol | |
| KR100587447B1 (en) | 120 bacillus subtilis eb120 strain microorganism formulation for controlling plant diseases containing same and method for controlling plant diseases using same | |
| KR101569737B1 (en) | Novel endophytic bacteria Bacillus oryzicola isolated from rice rhizosphere and development of a natural biopesticide and plant strengthener using same | |
| Ganesan et al. | Integrated management of stem rot disease (Sclerotium rolfsii) of groundnut (Arachis hypogaea L.) using Rhizobium and Trichoderma harzianum (ITCC-4572) | |
| Karimi et al. | Bacillus amyloliquefaciens SB14 from rhizosphere alleviates Rhizoctonia damping-off disease on sugar beet | |
| EP3251512A1 (en) | Compositions comprising bacillus strains and methods of use to suppress the activities and growth of fungal plant pathogens | |
| KR101624628B1 (en) | Novel bacillus vallismortis bs07m with promoting effect of plant growth and improving effect of cold-tolerance, and microbial agent containing the same | |
| US20030228679A1 (en) | Compositions and methods for increasing plant growth by inoculation with bacillus strains | |
| WO2010109436A1 (en) | Microbial formulation for widespread uesd in agricultural practices | |
| US20170238552A1 (en) | Bacterial endophytes from cucurbit species | |
| KR20140127670A (en) | Endophytic bacteria Bacillus Methylotrophicus YC7007 and development of a multifunctional biopesticide and microbial fertilizer using same | |
| CN115867640A (en) | Novel microorganism belonging to the genus Lactobacillus, and agent and method for controlling plant diseases caused by Ralstonia solanacearum or Ralstonia pseudosolanacearum | |
| KR100735572B1 (en) | 32 A novel Streptomyces padanus VSP-32 KCTC BP active against plant fungal pathogens and preparation of microbial pesticide | |
| Gao et al. | Characterization of an endophytic antagonistic bacterial strain bacillus halotolerans LBG-1-13 with multiple plant growth-promoting traits, stress tolerance, and its effects on lily growth | |
| KR101922410B1 (en) | Novel compounds produced by Bacillus oryzicola YC7011 with activities of induced resistance against plant pathogens and insect and plant growth promotion | |
| KR101005484B1 (en) | Streptomyces sporoclivatus CJS-49 KCTC 11109BP active against plant fungal pathogens | |
| CN109609403B (en) | Biocontrol bacterium and application thereof in prevention and control of downy mildew of crops | |
| Türkölmez et al. | Clonostachys rosea Strain ST1140: An Endophytic Plant-Growth-Promoting Fungus, and Its Potential Use in Seedbeds with Wheat-Grain Substrate | |
| KR20110120748A (en) | Bacillus amyloliquefaciens cp1 and contro method of strawberry anthracnose using the same | |
| KR101027082B1 (en) | Streptomyces misonensis CJS-70 with antibacterial activity to plant pathogens and microbial agent for controlling plant pathogens using the same | |
| KR100705777B1 (en) | Streptomyces costaricanus vsc-67 kctc 10910bp active against plant fungal pathogens | |
| JP7468842B2 (en) | Pesticide composition for controlling plant diseases and method for controlling plant diseases using the same | |
| KR20230105465A (en) | Development of a multifunctional biopesticide controlling anthracnose and bacterial diseases with plant growth stimulating effects |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| E902 | Notification of reason for refusal |