CA2107098A1 - Antifungal micro-organism - Google Patents
Antifungal micro-organismInfo
- Publication number
- CA2107098A1 CA2107098A1 CA002107098A CA2107098A CA2107098A1 CA 2107098 A1 CA2107098 A1 CA 2107098A1 CA 002107098 A CA002107098 A CA 002107098A CA 2107098 A CA2107098 A CA 2107098A CA 2107098 A1 CA2107098 A1 CA 2107098A1
- Authority
- CA
- Canada
- Prior art keywords
- antifungal agent
- plant
- antifungal
- ncib
- fungus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 229940121375 antifungal agent Drugs 0.000 title claims abstract description 13
- 244000005700 microbiome Species 0.000 title claims abstract description 9
- 230000000843 anti-fungal effect Effects 0.000 title claims abstract description 7
- 241000589540 Pseudomonas fluorescens Species 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 11
- 239000000203 mixture Substances 0.000 claims abstract description 11
- 241000233866 Fungi Species 0.000 claims abstract description 10
- 241000233639 Pythium Species 0.000 claims abstract description 7
- 241001361634 Rhizoctonia Species 0.000 claims abstract description 5
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 claims abstract description 4
- 241000223218 Fusarium Species 0.000 claims abstract description 4
- 239000003429 antifungal agent Substances 0.000 claims abstract 8
- 241000894006 Bacteria Species 0.000 claims description 6
- 230000002538 fungal effect Effects 0.000 claims description 4
- 238000009331 sowing Methods 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 241001149475 Gaeumannomyces graminis Species 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 10
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 10
- 241000918585 Pythium aphanidermatum Species 0.000 abstract description 5
- 230000000855 fungicidal effect Effects 0.000 abstract description 5
- 239000000417 fungicide Substances 0.000 abstract description 5
- 206010017533 Fungal infection Diseases 0.000 abstract description 3
- 208000031888 Mycoses Diseases 0.000 abstract description 3
- 244000038559 crop plants Species 0.000 abstract description 3
- 238000000576 coating method Methods 0.000 abstract description 2
- 238000012216 screening Methods 0.000 abstract description 2
- 239000011248 coating agent Substances 0.000 abstract 1
- 238000001228 spectrum Methods 0.000 abstract 1
- 238000011282 treatment Methods 0.000 description 23
- 241000196324 Embryophyta Species 0.000 description 17
- 230000000694 effects Effects 0.000 description 14
- 239000002689 soil Substances 0.000 description 14
- 235000010582 Pisum sativum Nutrition 0.000 description 13
- 241000219843 Pisum Species 0.000 description 12
- 244000052769 pathogen Species 0.000 description 12
- 230000001717 pathogenic effect Effects 0.000 description 12
- 240000004713 Pisum sativum Species 0.000 description 8
- 241001646398 Pseudomonas chlororaphis Species 0.000 description 8
- 239000000126 substance Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 239000008187 granular material Substances 0.000 description 6
- 239000002054 inoculum Substances 0.000 description 6
- 239000001965 potato dextrose agar Substances 0.000 description 6
- 241000918584 Pythium ultimum Species 0.000 description 5
- 241000209140 Triticum Species 0.000 description 5
- 235000021307 Triticum Nutrition 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 244000068988 Glycine max Species 0.000 description 4
- 235000010469 Glycine max Nutrition 0.000 description 4
- 239000005807 Metalaxyl Substances 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- ZQEIXNIJLIKNTD-UHFFFAOYSA-N methyl N-(2,6-dimethylphenyl)-N-(methoxyacetyl)alaninate Chemical compound COCC(=O)N(C(C)C(=O)OC)C1=C(C)C=CC=C1C ZQEIXNIJLIKNTD-UHFFFAOYSA-N 0.000 description 4
- OGYFATSSENRIKG-UHFFFAOYSA-N pencycuron Chemical compound C1=CC(Cl)=CC=C1CN(C(=O)NC=1C=CC=CC=1)C1CCCC1 OGYFATSSENRIKG-UHFFFAOYSA-N 0.000 description 4
- 230000008635 plant growth Effects 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 239000004576 sand Substances 0.000 description 3
- 230000003442 weekly effect Effects 0.000 description 3
- LDVVMCZRFWMZSG-OLQVQODUSA-N (3ar,7as)-2-(trichloromethylsulfanyl)-3a,4,7,7a-tetrahydroisoindole-1,3-dione Chemical compound C1C=CC[C@H]2C(=O)N(SC(Cl)(Cl)Cl)C(=O)[C@H]21 LDVVMCZRFWMZSG-OLQVQODUSA-N 0.000 description 2
- 239000005745 Captan Substances 0.000 description 2
- 241000427940 Fusarium solani Species 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 2
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 229940117949 captan Drugs 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- GPUADMRJQVPIAS-QCVDVZFFSA-M cerivastatin sodium Chemical compound [Na+].COCC1=C(C(C)C)N=C(C(C)C)C(\C=C\[C@@H](O)C[C@@H](O)CC([O-])=O)=C1C1=CC=C(F)C=C1 GPUADMRJQVPIAS-QCVDVZFFSA-M 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 235000009973 maize Nutrition 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000003415 peat Substances 0.000 description 2
- 230000000885 phytotoxic effect Effects 0.000 description 2
- YNCMLFHHXWETLD-UHFFFAOYSA-N pyocyanin Chemical compound CN1C2=CC=CC=C2N=C2C1=CC=CC2=O YNCMLFHHXWETLD-UHFFFAOYSA-N 0.000 description 2
- 229910052709 silver Inorganic materials 0.000 description 2
- 239000004332 silver Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- NWXMGUDVXFXRIG-WESIUVDSSA-N (4s,4as,5as,6s,12ar)-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O NWXMGUDVXFXRIG-WESIUVDSSA-N 0.000 description 1
- KUWPCJHYPSUOFW-YBXAARCKSA-N 2-nitrophenyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1[N+]([O-])=O KUWPCJHYPSUOFW-YBXAARCKSA-N 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 108010076119 Caseins Proteins 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 102000000634 Cytochrome c oxidase subunit IV Human genes 0.000 description 1
- 108050008072 Cytochrome c oxidase subunit IV Proteins 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 244000228957 Ferula foetida Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 239000005814 Pencycuron Substances 0.000 description 1
- 108010093965 Polymyxin B Proteins 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000022563 Rema Species 0.000 description 1
- 241000813090 Rhizoctonia solani Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- IPBVNPXQWQGGJP-UHFFFAOYSA-N acetic acid phenyl ester Natural products CC(=O)OC1=CC=CC=C1 IPBVNPXQWQGGJP-UHFFFAOYSA-N 0.000 description 1
- WNLRTRBMVRJNCN-UHFFFAOYSA-L adipate(2-) Chemical compound [O-]C(=O)CCCCC([O-])=O WNLRTRBMVRJNCN-UHFFFAOYSA-L 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 230000000853 biopesticidal effect Effects 0.000 description 1
- CKAAOCRYHRAMEC-UHFFFAOYSA-N butanoic acid;propane-1,2-diol Chemical compound CC(O)CO.CCCC(O)=O CKAAOCRYHRAMEC-UHFFFAOYSA-N 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- 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 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- 229940097572 chloromycetin Drugs 0.000 description 1
- 239000002361 compost Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- GHVNFZFCNZKVNT-UHFFFAOYSA-M decanoate Chemical compound CCCCCCCCCC([O-])=O GHVNFZFCNZKVNT-UHFFFAOYSA-M 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 229940049920 malate Drugs 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- YBVAXJOZZAJCLA-UHFFFAOYSA-N nitric acid nitrous acid Chemical compound ON=O.O[N+]([O-])=O YBVAXJOZZAJCLA-UHFFFAOYSA-N 0.000 description 1
- 229940049953 phenylacetate Drugs 0.000 description 1
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 239000004476 plant protection product Substances 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108010009004 proteose-peptone Proteins 0.000 description 1
- HEBKCHPVOIAQTA-ZXFHETKHSA-N ribitol Chemical compound OC[C@H](O)[C@H](O)[C@H](O)CO HEBKCHPVOIAQTA-ZXFHETKHSA-N 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- RVUXIPACAZKWHU-UHFFFAOYSA-N sulfuric acid;heptahydrate Chemical compound O.O.O.O.O.O.O.OS(O)(=O)=O RVUXIPACAZKWHU-UHFFFAOYSA-N 0.000 description 1
- 229910021653 sulphate ion Inorganic materials 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- PXSOHRWMIRDKMP-UHFFFAOYSA-N triaziquone Chemical compound O=C1C(N2CC2)=C(N2CC2)C(=O)C=C1N1CC1 PXSOHRWMIRDKMP-UHFFFAOYSA-N 0.000 description 1
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
- A01N63/27—Pseudomonas
-
- 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
- C12R2001/38—Pseudomonas
- C12R2001/39—Pseudomonas fluorescens
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Plant Pathology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Biochemistry (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
2107098 9218613 PCTABS00017 A new strain of Pseudomonas fluorescens which displays antifungal activity on a wide spectrum of disease fungi has been isolated. Screening, testing and field trial results are provided. The antifungal agent is particularly active against "damping-off"
disease caused by fungi of such genera as Rhizoctonia, Pythium, and Fusarium. Methods of using such micro-organisms and fungicidal compositions containing same to protect crop plants from fungal infection are described. The antifungal agent can be used as an agricultural fungicide, particularly as a seed coating.
disease caused by fungi of such genera as Rhizoctonia, Pythium, and Fusarium. Methods of using such micro-organisms and fungicidal compositions containing same to protect crop plants from fungal infection are described. The antifungal agent can be used as an agricultural fungicide, particularly as a seed coating.
Description
WO 92/18C13 1 PCI~/GB92/QOSg7 ~ 21~7~8 ANTIFUNGAr, MICRO--ORGANISM
This invention relates to an agricultural ; plant protection product. More specifically, the invention relate$ to an antifungal micro-organ$~m, in partlcular to a novel strain of Pseudomonts fluorescens, and the uses thereof for the protection of plants against fungal attack.
Certa~n 6trains of the micro-organism Pseudomonas fluorescens are known to be useful agricultural fungicides. The antifungal activity varies greatly amongst the innumerable strains of the micro-organism. Agriculturally useful strains are those which posse6s useful levels of activity against specific target fungi. One such target is the group of fungi which are associated with the disease known as ~damping-off~ which is partlcularly troublesome in certa$n crops and under particular climatic conditions. U80 of Biological Control Agents (BCAs) or biopesticides to control disease may be more effective or more preferable to the use of standard chemical control agents.
An ob~ect of the present invention i8 to provide an agriculturally useful fungicide.
According to the present invention there is provided a novel strain of Pseudomonas fluorescens Biovar I, deposited at The National Collection of Industrial and Marine Bacteria under the Accession Number NCIB 40189 on 1st September 1989.
~he invention also comprises an antifungal agricultural composition containing Pseudomonas fluQrescens Biovar r, strain NCIB 40189 as active ingredient in admixture with a carrier composit1on WO~lK13 2 21 0 7 ~ 9 8 PCT~GB9~g7 acceptable in agricultural practice. Exam~les of the types of agricultural formulation~ which may be employed are seed coatings compositions, liguid for root or soil drenching and granular or powder compositions. The base materials for these are well known in the art.
Additionally, the invention provides a method of protecting crop plants from fungal infection comprising applying to the plants, the roots or seedg thereof or the growing medium for the plant a fungicidally effective dosage of the said Pseudomonas fluorescens Biovar I, gtrain NCIB
40189. The treatment may be effected by application of a preparation containing organismg of the strain of this invention.
The antifungal micro-organism of this invention is particularly effective against fungal infections asgociated with ~damping-off~ disease, guch as fungi of the genera Rhizoctonia, Pythium, Fusarium. It is also active against .
Gaeumannomvces ~raminis which causes take-all digease .
The strain of this invention (NCIB 40189) was isolated from volunteer wheat roots collected from plants grown at Chiseldon, Swindon, Wiltshire, United Ringdom. It was deposited at The National Collections of Industrial and Marine Bacteria ~23 St Machar Drive, Aberdeen, Scotland, AB2 lRY) under the terms of the Budapest Treaty on 1st September 1989.
There now follows a description of the isolation, characterisation and screening of the bacterial isolate with examples of its activity against several fungi which infect crop plants.
WO g2/18613 3 PC~GB~2/00597 ISOLATION OF Tlll~ MICRO--ORGANISM
The bacterium was isolated from volunteer wheat root~ dug from a field at a farm known a~
Draycott Farm, Chiseldon, Swindon, Wiltshire, United Kingdom.
Th~ root6 were washed to remo~e all adhering soil and placed in sterile distilled water in a flask containing glass beads. The roots were shaken with the glas~ beads for 15 minutes and the wa~hing& then plated out on to Ring~ B agar ~Proteose Peptone No.3, 20g; glycerol lOml~
pota8sium sulphate, 1.5g; maqnesium sulphate heptahydrate, 1.5g; technical agar, 12gS and, one lltre of d~stilled water at p~ 7.2 and $ncludlng cycloheximide 75 micrograms/ml5 chloromycetin 12.5 micrograms/ml; and ampicillin 50 micrograms/ml) at 10:1 and 10:3 dilutions. The plates were incubated for three days at 20C.
The bacterium was taken at random from the agar plate and ~ub-cultured on to 1/10 trypt~c soya broth agar (TSA) (3g tryptic 80ya broth; 15g agar; one l~tre water). The bacterium was maintained an 1/10 TSA and 6tored at 4C until required.
C~ARACTERISATION OF TEIB MICRO--ORGANISM
The morphological characteri6tlcs of Pseudomonas fluorescens strain NCIB 40189 were determined by the National Collection of Industrial and Marine ~acteria and are #hown in Table 1 below.
, , ' - ' .
- ~ ~
21~ 7 ~ 9 8 PCT/GB~K97 Morphology of P iluore wens strain NCIB 40189 Cell Morphology: Gram negative rods Fluorescence: +ve - RAPID TEST (~P~) - Nitrate reduction Indole production Acid from glucose (~) Arginine dehydrolase +
ureage Aesculin hydrolysis Gelatin hydrolysis +
~-Galactos~dase Glucose ass~milation +
Arabinose assimilation +
Mannose assimilation +
Nannitol assimilation +
N-acetylglucosamine assimilation +
Naltose assimilation Gluconate assimilation +
Caprate assimilation +
Adipate assimilation Malate assimilation +
Citrate assimilation +
Phenylacetate assimilation Cytochrome oxidase +
Pyocyanin Fluorescence +
Gas Glucose Acid Glucose PNS +
ONPG
Nitrate to nitrite Nitrate to nitrogen : WO ~VIK13 5 PCT/GB~00597 ~`- 21~7~98 Residual nitrogen +
DNAa~e CSU:--Acetate +
Trehalose +
meso-Inositol Adonitol Butyrate Propylene glycol Gel Stab 20C
Gel Plate +
Casein (+) Starch Leclth egg ~ipase egg H2S ~TSI) Tween 80 +
Vrease +
Phenylalanine deaminase Growth factor required Peniclllln G
8treptomycln +
Chloramphenicol +
Tetracycl~ne +
Novoblocin Polymyxin B +
~evan +
~Tests are described in: R E Buchanan, N E Gibbons 30 (eds), 1974, Bergey's Manual of Determinatlve Bacteriology, Bth ed; S T Cowan, R J Steel, 1974, Manual for Identification of Medical 8acteria).
WO92/18613 6 210 7 0 9 8P~/GB92/00S97 PRII~ARY SOIL PEq'RI--DISEI SCRI~N
Pseudomonas fluorescens strain ~CIB 40189 was tested for activity against Pythium ultimum, using a petri-dish soil screen.
S Petri-dlshes (5cm) were filled with 10ml of potato dextrose agar (PDA). Agar plugs (5mm) from a 7 day old culture of Pythium ultimum grown at 20C were placed centrally on to the PDA. The pl~te~ were incubated for 5 days at 20C during which the P ultimum colonised the plate. Minster Mendip ~oam was autoclaved at 120C for 60 minutes. To this sterile wheatgerm was added (1%
w/w) and mixed well. Approximately 8g of ~oil was placed into each Petri dish to cover the Pythium lawn.
The following treatments were used (2 ml applied per petri-dish):
1) Pseudomonas fluorescens strain NCIB 40189 (culture grown in 1/10 tryptic soya broth (TS8J for 48 h at 12 C);
This invention relates to an agricultural ; plant protection product. More specifically, the invention relate$ to an antifungal micro-organ$~m, in partlcular to a novel strain of Pseudomonts fluorescens, and the uses thereof for the protection of plants against fungal attack.
Certa~n 6trains of the micro-organism Pseudomonas fluorescens are known to be useful agricultural fungicides. The antifungal activity varies greatly amongst the innumerable strains of the micro-organism. Agriculturally useful strains are those which posse6s useful levels of activity against specific target fungi. One such target is the group of fungi which are associated with the disease known as ~damping-off~ which is partlcularly troublesome in certa$n crops and under particular climatic conditions. U80 of Biological Control Agents (BCAs) or biopesticides to control disease may be more effective or more preferable to the use of standard chemical control agents.
An ob~ect of the present invention i8 to provide an agriculturally useful fungicide.
According to the present invention there is provided a novel strain of Pseudomonas fluorescens Biovar I, deposited at The National Collection of Industrial and Marine Bacteria under the Accession Number NCIB 40189 on 1st September 1989.
~he invention also comprises an antifungal agricultural composition containing Pseudomonas fluQrescens Biovar r, strain NCIB 40189 as active ingredient in admixture with a carrier composit1on WO~lK13 2 21 0 7 ~ 9 8 PCT~GB9~g7 acceptable in agricultural practice. Exam~les of the types of agricultural formulation~ which may be employed are seed coatings compositions, liguid for root or soil drenching and granular or powder compositions. The base materials for these are well known in the art.
Additionally, the invention provides a method of protecting crop plants from fungal infection comprising applying to the plants, the roots or seedg thereof or the growing medium for the plant a fungicidally effective dosage of the said Pseudomonas fluorescens Biovar I, gtrain NCIB
40189. The treatment may be effected by application of a preparation containing organismg of the strain of this invention.
The antifungal micro-organism of this invention is particularly effective against fungal infections asgociated with ~damping-off~ disease, guch as fungi of the genera Rhizoctonia, Pythium, Fusarium. It is also active against .
Gaeumannomvces ~raminis which causes take-all digease .
The strain of this invention (NCIB 40189) was isolated from volunteer wheat roots collected from plants grown at Chiseldon, Swindon, Wiltshire, United Ringdom. It was deposited at The National Collections of Industrial and Marine Bacteria ~23 St Machar Drive, Aberdeen, Scotland, AB2 lRY) under the terms of the Budapest Treaty on 1st September 1989.
There now follows a description of the isolation, characterisation and screening of the bacterial isolate with examples of its activity against several fungi which infect crop plants.
WO g2/18613 3 PC~GB~2/00597 ISOLATION OF Tlll~ MICRO--ORGANISM
The bacterium was isolated from volunteer wheat root~ dug from a field at a farm known a~
Draycott Farm, Chiseldon, Swindon, Wiltshire, United Kingdom.
Th~ root6 were washed to remo~e all adhering soil and placed in sterile distilled water in a flask containing glass beads. The roots were shaken with the glas~ beads for 15 minutes and the wa~hing& then plated out on to Ring~ B agar ~Proteose Peptone No.3, 20g; glycerol lOml~
pota8sium sulphate, 1.5g; maqnesium sulphate heptahydrate, 1.5g; technical agar, 12gS and, one lltre of d~stilled water at p~ 7.2 and $ncludlng cycloheximide 75 micrograms/ml5 chloromycetin 12.5 micrograms/ml; and ampicillin 50 micrograms/ml) at 10:1 and 10:3 dilutions. The plates were incubated for three days at 20C.
The bacterium was taken at random from the agar plate and ~ub-cultured on to 1/10 trypt~c soya broth agar (TSA) (3g tryptic 80ya broth; 15g agar; one l~tre water). The bacterium was maintained an 1/10 TSA and 6tored at 4C until required.
C~ARACTERISATION OF TEIB MICRO--ORGANISM
The morphological characteri6tlcs of Pseudomonas fluorescens strain NCIB 40189 were determined by the National Collection of Industrial and Marine ~acteria and are #hown in Table 1 below.
, , ' - ' .
- ~ ~
21~ 7 ~ 9 8 PCT/GB~K97 Morphology of P iluore wens strain NCIB 40189 Cell Morphology: Gram negative rods Fluorescence: +ve - RAPID TEST (~P~) - Nitrate reduction Indole production Acid from glucose (~) Arginine dehydrolase +
ureage Aesculin hydrolysis Gelatin hydrolysis +
~-Galactos~dase Glucose ass~milation +
Arabinose assimilation +
Mannose assimilation +
Nannitol assimilation +
N-acetylglucosamine assimilation +
Naltose assimilation Gluconate assimilation +
Caprate assimilation +
Adipate assimilation Malate assimilation +
Citrate assimilation +
Phenylacetate assimilation Cytochrome oxidase +
Pyocyanin Fluorescence +
Gas Glucose Acid Glucose PNS +
ONPG
Nitrate to nitrite Nitrate to nitrogen : WO ~VIK13 5 PCT/GB~00597 ~`- 21~7~98 Residual nitrogen +
DNAa~e CSU:--Acetate +
Trehalose +
meso-Inositol Adonitol Butyrate Propylene glycol Gel Stab 20C
Gel Plate +
Casein (+) Starch Leclth egg ~ipase egg H2S ~TSI) Tween 80 +
Vrease +
Phenylalanine deaminase Growth factor required Peniclllln G
8treptomycln +
Chloramphenicol +
Tetracycl~ne +
Novoblocin Polymyxin B +
~evan +
~Tests are described in: R E Buchanan, N E Gibbons 30 (eds), 1974, Bergey's Manual of Determinatlve Bacteriology, Bth ed; S T Cowan, R J Steel, 1974, Manual for Identification of Medical 8acteria).
WO92/18613 6 210 7 0 9 8P~/GB92/00S97 PRII~ARY SOIL PEq'RI--DISEI SCRI~N
Pseudomonas fluorescens strain ~CIB 40189 was tested for activity against Pythium ultimum, using a petri-dish soil screen.
S Petri-dlshes (5cm) were filled with 10ml of potato dextrose agar (PDA). Agar plugs (5mm) from a 7 day old culture of Pythium ultimum grown at 20C were placed centrally on to the PDA. The pl~te~ were incubated for 5 days at 20C during which the P ultimum colonised the plate. Minster Mendip ~oam was autoclaved at 120C for 60 minutes. To this sterile wheatgerm was added (1%
w/w) and mixed well. Approximately 8g of ~oil was placed into each Petri dish to cover the Pythium lawn.
The following treatments were used (2 ml applied per petri-dish):
1) Pseudomonas fluorescens strain NCIB 40189 (culture grown in 1/10 tryptic soya broth (TS8J for 48 h at 12 C);
2) M~TAIAXYL (100 ppm AI);
3) CONTRO~: 1/10 TSB alone.
Duplicate plates were prepared, and lncubated for 4 days at 10C.
Results are shown in Table 2 below. Actlvity was scored on a scale of zero to 3. A score of zero meant there was considerable growth of Pythium; a score of 3 meant there wa~ no mycelial growth in the dish.
WO~UI8613 7 ~CT/~B9~97 ~ 1 ~ 7 8 9 8 Activity of NCIB 40189 against P ulti~u~
TREATMENT ACTIVITY SCORE
NCIB 40189 . 3 Metalaxyl 3 Control O
The results show that strain NCIB 40189 inhibited fungal growth in the soil. Its activity is comparable to Metalaxyl, a standard chemical fungicide.
S~CONDARY POT SCR~FNING
.
~he effectiveness of Pseudomonas fluorescens strain NCIB 40189 was further tested against Pythium ultimum and Rhizoctonia solani. Results show that strain NCSB 40189 has activity against P
ultimum and R solan~ under glasshouse conditions.
ACTIVITY AGAINST P U~TIMUM:
Cultures of P ultimum were raised on PDA
plates for 7 days at 20C. A mixture of 200g of silver sand, Sg cornmeal, 4g Bemax wheatgerm and 40ml water wAs autoclaved at 120C for 20 minutes then placed in a flask and inoculated with 1/4 of a plate of Pythium. The flasks were incubated at 20C for 7 days, then the contents of each flask wa~ mixed with fine sand to a final weight of 300g. One flask of Pythium was mixed with 8 litres of Mendip Minster Loam to give a standard dilution (X). This was further diluted with clean :, :
, . .
~VO~VIK13 ~ 7~98 Mendip loam to give a dilut~on of X/16 used in the tests.
Tests were carried out in pots (3 inches diameter), filled to 3/4 with infested soil. Five peas were sown in each pot and covered with clean 80il.
The following treatments were used (35 ml added as a drench to each pot):
1) Pseudomonas fluorescens strain NCIB 40189 (culture grown $n 1/10 tryptic soya broth (TSB) for 4~ h at 20C);
2) METAhAXYL (0.1-5.0 ppm AI);
3) CONTROL: 1/10 TSB alone.
Five ~eplicates were prepared for each treatment.
Pea plants were grown for 14 days at 15C, and seedling emergence was monitored weekly. The plants were watered da$1y: particular care was taken to ensure that the soil was kept moist during the tests as the disease inc$tes damping-off under cool, damp conditions.
Results are shown in Table 3 below.
.' '' ' " ' ~- . '.:' - .
WO 9V18C13 9 PCl'~GB92/00597 ,-- 2107~98 Effect of strain NCI8 40189 on e~tablish~ent of pea~ ln the presence of P ultimum TREATMENT Percentage ~%) emergence of peas in pots wlth P ultimum present Metalaxyl 0.1 ppm 64 0.5 ppm 84 1.0 ppm 84 5.0 pp~ 96 Control 28 Result~ for strain NCIB 40189 and for the untreated control show that application of the Pseudomonas fluorescens strain allows slgnificantly more pea plants to emerge. Act~vlty of ~train NCIB 40189 is comparable wlth that of the chemical ~ungicide Metalaxyl at tho disoa~e levels expressed.
ACTIVITY AGAINST R SOLANI:
Cultures of R solani were raised on PDA plates for 7 days at 20C. A mixture of 200g of silver ~and, 4g Bemax wheatgerm and 40ml water was autoclaved at 120C for 20 minutes then placed in a fla~k and inoculated with l/4 of a plate of Rhizoctonla. ~he flasks were incubated at 20C
for 7 days, then the contents of each flask was . .
' WO9~18613 l0 2 1 ~ 7 ~ 9 ~CT/GB9~97 mixed with fine sand to a final weight o~ 300g.
Three quarters of a flask of Rhizoctonia inoculum was mixed with 6 litres of Mendip Minster ~oam to give a standard dilution (Y). This was further diluted with clean Mendip loam to give a dilution of Y/B used in the tests.
Tests were carried out in pots (3 inches diametsr), filled to 3/4 with infested soil. Five peas were sown in each pot and covered with clean soil.
The following treatments were used (35 ml added as a drench to each pot):
l) Pseudomonas fluorescens strain NcrB 401~9 (culture grown in l/10 tryptic soya broth (TSB) for 48 h at 20C);
2~ PENCYCURON (Monceren at l0 or l00 ppm AI);
3) DISEASED CONTRO~: l/l0 TSB alone;
Duplicate plates were prepared, and lncubated for 4 days at 10C.
Results are shown in Table 2 below. Actlvity was scored on a scale of zero to 3. A score of zero meant there was considerable growth of Pythium; a score of 3 meant there wa~ no mycelial growth in the dish.
WO~UI8613 7 ~CT/~B9~97 ~ 1 ~ 7 8 9 8 Activity of NCIB 40189 against P ulti~u~
TREATMENT ACTIVITY SCORE
NCIB 40189 . 3 Metalaxyl 3 Control O
The results show that strain NCIB 40189 inhibited fungal growth in the soil. Its activity is comparable to Metalaxyl, a standard chemical fungicide.
S~CONDARY POT SCR~FNING
.
~he effectiveness of Pseudomonas fluorescens strain NCIB 40189 was further tested against Pythium ultimum and Rhizoctonia solani. Results show that strain NCSB 40189 has activity against P
ultimum and R solan~ under glasshouse conditions.
ACTIVITY AGAINST P U~TIMUM:
Cultures of P ultimum were raised on PDA
plates for 7 days at 20C. A mixture of 200g of silver sand, Sg cornmeal, 4g Bemax wheatgerm and 40ml water wAs autoclaved at 120C for 20 minutes then placed in a flask and inoculated with 1/4 of a plate of Pythium. The flasks were incubated at 20C for 7 days, then the contents of each flask wa~ mixed with fine sand to a final weight of 300g. One flask of Pythium was mixed with 8 litres of Mendip Minster Loam to give a standard dilution (X). This was further diluted with clean :, :
, . .
~VO~VIK13 ~ 7~98 Mendip loam to give a dilut~on of X/16 used in the tests.
Tests were carried out in pots (3 inches diameter), filled to 3/4 with infested soil. Five peas were sown in each pot and covered with clean 80il.
The following treatments were used (35 ml added as a drench to each pot):
1) Pseudomonas fluorescens strain NCIB 40189 (culture grown $n 1/10 tryptic soya broth (TSB) for 4~ h at 20C);
2) METAhAXYL (0.1-5.0 ppm AI);
3) CONTROL: 1/10 TSB alone.
Five ~eplicates were prepared for each treatment.
Pea plants were grown for 14 days at 15C, and seedling emergence was monitored weekly. The plants were watered da$1y: particular care was taken to ensure that the soil was kept moist during the tests as the disease inc$tes damping-off under cool, damp conditions.
Results are shown in Table 3 below.
.' '' ' " ' ~- . '.:' - .
WO 9V18C13 9 PCl'~GB92/00597 ,-- 2107~98 Effect of strain NCI8 40189 on e~tablish~ent of pea~ ln the presence of P ultimum TREATMENT Percentage ~%) emergence of peas in pots wlth P ultimum present Metalaxyl 0.1 ppm 64 0.5 ppm 84 1.0 ppm 84 5.0 pp~ 96 Control 28 Result~ for strain NCIB 40189 and for the untreated control show that application of the Pseudomonas fluorescens strain allows slgnificantly more pea plants to emerge. Act~vlty of ~train NCIB 40189 is comparable wlth that of the chemical ~ungicide Metalaxyl at tho disoa~e levels expressed.
ACTIVITY AGAINST R SOLANI:
Cultures of R solani were raised on PDA plates for 7 days at 20C. A mixture of 200g of silver ~and, 4g Bemax wheatgerm and 40ml water was autoclaved at 120C for 20 minutes then placed in a fla~k and inoculated with l/4 of a plate of Rhizoctonla. ~he flasks were incubated at 20C
for 7 days, then the contents of each flask was . .
' WO9~18613 l0 2 1 ~ 7 ~ 9 ~CT/GB9~97 mixed with fine sand to a final weight o~ 300g.
Three quarters of a flask of Rhizoctonia inoculum was mixed with 6 litres of Mendip Minster ~oam to give a standard dilution (Y). This was further diluted with clean Mendip loam to give a dilution of Y/B used in the tests.
Tests were carried out in pots (3 inches diametsr), filled to 3/4 with infested soil. Five peas were sown in each pot and covered with clean soil.
The following treatments were used (35 ml added as a drench to each pot):
l) Pseudomonas fluorescens strain NcrB 401~9 (culture grown in l/10 tryptic soya broth (TSB) for 48 h at 20C);
2~ PENCYCURON (Monceren at l0 or l00 ppm AI);
3) DISEASED CONTRO~: l/l0 TSB alone;
4) HEALTHY CONTROL: l/l0 TSB alone on soil not infested with Rhizoctonia.
Five replicates were prepared for each treatment.
Pea plants were grown for 14 days at 24C, and seedling emergence was monitored weekly. The plants were watered daily.
Results are shown in Table 4 below.
' . ' :. . ' .,, .
.
~ ~ ' : - ' ' wo ~UI8613 11 210 7 0 9 8 PCT/GB9~97 Effect of ~train NCIB 40189 on establish~ent of peas in the presence of R solani TREATMENT Percentage.(%) emergence of peas Monceren 1010 ppm 40 100 ppm 60 Diseased control 20 Healthy control 92 Application of the Pseudomonas fluorescens strain NCIB 40189 allows significantly more pea plants to emerge when campared to the d~seas~d control. ~n fact, application of strain NCIB
40189 allows pea establishment at a level comparable to the hoalthy control ~treatment wlthout R solani). Activity of the BCA also appears to bo higher than that of the chomical fungicide Monceren.
PROTECTION AGAINST Pr,ANT D~SEASE: FVRT~IER TESTS
Pseudomonas fluorescens strain NCIB 40189 was further tested for its activity against Rhizactonia solani and its abillty to protect pea plants from disease.
Tho trial was carried out in rectangular trays ~165mm x 95mm), half-filled with infested soil (prepared according to the method described in the . ' . ! .
- - ' :- :
. .
WO ~VI86I3 12 2i~7~9~ PCT/GB~U~97 previous section). Twenty-five peas were sown in each tray and covered with clean soil.
The B~A (strain NCIB 40189) and Control treatments were as follows:
1) DRENC~ 100 ml bacterial suspension (109 cells/ml) added to each tray;
2) GRANU~ES lO g pre-formed granules added to each tray;
granules (types A, ~, C) carrying bacterial cells were inoculated at a maximum capacity of BCA
concentrate: 4.5x101 cells/g;
3) UNTREATED 100 ml water added as a drench to each tray as a control.
Three replicates were prepared for each treatment.
Pea plants were grown for 7 to 14 days at 24C. The plants were watered daily and seedling emergence was monitored weekly.
Test results were collated from all experiments and are shown in Table 5 below.
- .
;;
, ~ . .
WO~U18613 13 PCT/GB~OOSs7 210709~
T~LE 5 Activity of strain NCIB 40189 again~t R solanl TREATMENT Mean percentage emergence (%) of pea~
s from R solani infested compost Drench 73 Granule A 65 Granule ~ ~4 Granule C 66 Untreated 31 Application of stra$n NCI~ 40189 significantly $ncreased the percentage emergence of the plants when compared to the untreated control. ~here is no statistically significant difference between re~ults obtained with the drench or granule BCA treatments.
FIELD TRIA~S
Pseudomonas fluorescens straln NCIB 40189 Wa8 used ln field trlals as a biological control agent (BCA) again~t ~damping-off~ d~sease ~n peas and maize. Field trials of strain NC~B 40189 have boen conducted on its potential for inhibltlng Pythlum and Fusarlum spp. Results were compared to standard chemical treatments, and are reported below. ~n summary, these results show that disease caused by Pythium ultimum or Fusarium solani is reduced by application of strain NCIB
40189 to a level comparable with that obtained by standard chemlcal treatment.
' WO~Ut8613 14 2 1 0 7 ~ 9 ~CT/GB~K97 ~. . .
.
EFFICACY AGAINST PYTEII~
Trials were conducted in France using peas as the test crop. Pathogen was added at various rates as whaat grain inoculum, prepared as follows. satches of wheat seed (1 Kg) were soaked in water overnight, drained and autoclaved at 121~C. Fungal spores which rema~ned viable were allowed to germinate and grow for two days before the 6eed was re-autoclaved at 121~C. A 7 day old plate of PYthium ultimum (grown on potato dextrose agar at 20C) was added to the wheat seed and allowed to grow for 10 days, shaken to redistribute the mycelia, and left for a further 11 day growth period. The inoculum was then ready for use within the next 14 days.
Strain NCIB 40189 was applied as a gum/peat formulation (107 cell/seed). APRON (Trade Nark) was used as a chemical control, applied at a rate of 30g AI/lOORg seed. Two types of plant control were used: untreated pea seed, and blank pea seed (which had undergone seed treatment without addit~on of BCA or chemical).
Results are shown in Table 6 below. ~Number emerged~ refers to the mean number of plants which emerged 22 days after sowing from 2ao ceeds on a strip 5 metres long. The letters appearing after the numbers ~emerged~ indicate statistical significance: there is no significant difference at the 5% probability level between entries with a letter in common.
WO~U18613 15 2 1 0 7 G 9 8 PCT/GB9~K97 Field ~ctivity of NCIB 40189 again8t P ulti~u :
e~ergence of pea plants ;-TREATMENT NUMBE~ EMERGED WHEN P ULTIMUM
ADDED AT T~E FO~BOWING RATES(g inoculum per metre row):
NCIB 40189 180.2~ AB 153.00 CDG 134.50 FG~
APRON 171.50 AC 169.50 ACE 157.25 BCDF
Untreated 172.50 AC 130.25 GHI 111.25 I
alank 176.00 AC 144.34 DE~ 119.00 ~I
The fir6t column in Table 6 shows results for the control experiment where tests were conducted in normal field soil, without any added pathogen.
The absence of pathogen in the ~oil allowed normal plant growth. There i~ no real significant d~fference between treatments, indicatlng the addition of strain NCIB 40189 had no phytotoxic effect.
The other figures in Table 6 givo the results for two test treatments at different pathogen rates. The presence of pathogen in the 80il decreased the number of plants emerging from untreated or blank seed when compared to the control experiment. This shows disease was present and was adversely affectlng plant growth.
~owever, the presence of strain NCIB 40189 ~ignificantly increases the number of plants emerging in the presence of the pathogen.
.
WO~U18613 16 21 0 7 o ~/GBg~g7 EFFICACY AGAINST FUSARIUM:
Field trials were carried out in the USA
(Mississippi), using maize as the test crop.
Pathogen (F solani) was added at various rates as wheat grain inoculum, prepared as descrlbed above.
Stra~n NCIB 40189 was applied as a gum/peat formulation (107 cell/seed). The chemical control was CAPTAN (Trade Mark), used as a seed treatment at the recommended rate. Untreated seed was used as the plant control.
Results are shown in Table 7 below. Emergence is quoted as percentage (%) of plants l5 days after sowing. The letters appearing after the percentage emerged indicate statistical s~gnificance. There is no significant dif~erence at the 5% probability level between entries w~th a letter in common.
TA~LE 7 Field ~ctlYity of NCIB 40189 again~t F solani:
emergence of maize plants T~EATMENT PERCENTAGE ~%) EMERGED W~EN F SOhANT
ADDED AT THE FOL~OWING RATES
(g inoculum per metre row):
NCIB 40189 65.208 C 74.583 AC
CAPTAN 69.583 BC 69.375 BC
Vntreated 65.833 C 51.250 D
The first column of Table 7 shows results for the control tria}s where no test pathogen was added. The absence of pathogen ~n the so~l W0~18613 17 21 0 7 0 9 8 allowed normal plant growth. There ~8 no real significant difference between treatments, indicatlng the addit$on of strain NCIB 40189 had no phytotoxic effect.
S The presence of pathogen, Fusarium solan~, in the soil decreased the number of plants emerging from untreated seed when compared to the control experlment. This shows disease was present and wa~ adversely affect~ng plant growth. ~owever, the presence of ~tra~n NCIB 40189 significantly increases the percentage of plants emerging in the presence of the pathogen. In fact, treatment wlth strain NCIB 40189 improved plant performance 80 that the percentage emerging was not significantly different from the percentage emerging in the absence of pathogen.
Five replicates were prepared for each treatment.
Pea plants were grown for 14 days at 24C, and seedling emergence was monitored weekly. The plants were watered daily.
Results are shown in Table 4 below.
' . ' :. . ' .,, .
.
~ ~ ' : - ' ' wo ~UI8613 11 210 7 0 9 8 PCT/GB9~97 Effect of ~train NCIB 40189 on establish~ent of peas in the presence of R solani TREATMENT Percentage.(%) emergence of peas Monceren 1010 ppm 40 100 ppm 60 Diseased control 20 Healthy control 92 Application of the Pseudomonas fluorescens strain NCIB 40189 allows significantly more pea plants to emerge when campared to the d~seas~d control. ~n fact, application of strain NCIB
40189 allows pea establishment at a level comparable to the hoalthy control ~treatment wlthout R solani). Activity of the BCA also appears to bo higher than that of the chomical fungicide Monceren.
PROTECTION AGAINST Pr,ANT D~SEASE: FVRT~IER TESTS
Pseudomonas fluorescens strain NCIB 40189 was further tested for its activity against Rhizactonia solani and its abillty to protect pea plants from disease.
Tho trial was carried out in rectangular trays ~165mm x 95mm), half-filled with infested soil (prepared according to the method described in the . ' . ! .
- - ' :- :
. .
WO ~VI86I3 12 2i~7~9~ PCT/GB~U~97 previous section). Twenty-five peas were sown in each tray and covered with clean soil.
The B~A (strain NCIB 40189) and Control treatments were as follows:
1) DRENC~ 100 ml bacterial suspension (109 cells/ml) added to each tray;
2) GRANU~ES lO g pre-formed granules added to each tray;
granules (types A, ~, C) carrying bacterial cells were inoculated at a maximum capacity of BCA
concentrate: 4.5x101 cells/g;
3) UNTREATED 100 ml water added as a drench to each tray as a control.
Three replicates were prepared for each treatment.
Pea plants were grown for 7 to 14 days at 24C. The plants were watered daily and seedling emergence was monitored weekly.
Test results were collated from all experiments and are shown in Table 5 below.
- .
;;
, ~ . .
WO~U18613 13 PCT/GB~OOSs7 210709~
T~LE 5 Activity of strain NCIB 40189 again~t R solanl TREATMENT Mean percentage emergence (%) of pea~
s from R solani infested compost Drench 73 Granule A 65 Granule ~ ~4 Granule C 66 Untreated 31 Application of stra$n NCI~ 40189 significantly $ncreased the percentage emergence of the plants when compared to the untreated control. ~here is no statistically significant difference between re~ults obtained with the drench or granule BCA treatments.
FIELD TRIA~S
Pseudomonas fluorescens straln NCIB 40189 Wa8 used ln field trlals as a biological control agent (BCA) again~t ~damping-off~ d~sease ~n peas and maize. Field trials of strain NC~B 40189 have boen conducted on its potential for inhibltlng Pythlum and Fusarlum spp. Results were compared to standard chemical treatments, and are reported below. ~n summary, these results show that disease caused by Pythium ultimum or Fusarium solani is reduced by application of strain NCIB
40189 to a level comparable with that obtained by standard chemlcal treatment.
' WO~Ut8613 14 2 1 0 7 ~ 9 ~CT/GB~K97 ~. . .
.
EFFICACY AGAINST PYTEII~
Trials were conducted in France using peas as the test crop. Pathogen was added at various rates as whaat grain inoculum, prepared as follows. satches of wheat seed (1 Kg) were soaked in water overnight, drained and autoclaved at 121~C. Fungal spores which rema~ned viable were allowed to germinate and grow for two days before the 6eed was re-autoclaved at 121~C. A 7 day old plate of PYthium ultimum (grown on potato dextrose agar at 20C) was added to the wheat seed and allowed to grow for 10 days, shaken to redistribute the mycelia, and left for a further 11 day growth period. The inoculum was then ready for use within the next 14 days.
Strain NCIB 40189 was applied as a gum/peat formulation (107 cell/seed). APRON (Trade Nark) was used as a chemical control, applied at a rate of 30g AI/lOORg seed. Two types of plant control were used: untreated pea seed, and blank pea seed (which had undergone seed treatment without addit~on of BCA or chemical).
Results are shown in Table 6 below. ~Number emerged~ refers to the mean number of plants which emerged 22 days after sowing from 2ao ceeds on a strip 5 metres long. The letters appearing after the numbers ~emerged~ indicate statistical significance: there is no significant difference at the 5% probability level between entries with a letter in common.
WO~U18613 15 2 1 0 7 G 9 8 PCT/GB9~K97 Field ~ctivity of NCIB 40189 again8t P ulti~u :
e~ergence of pea plants ;-TREATMENT NUMBE~ EMERGED WHEN P ULTIMUM
ADDED AT T~E FO~BOWING RATES(g inoculum per metre row):
NCIB 40189 180.2~ AB 153.00 CDG 134.50 FG~
APRON 171.50 AC 169.50 ACE 157.25 BCDF
Untreated 172.50 AC 130.25 GHI 111.25 I
alank 176.00 AC 144.34 DE~ 119.00 ~I
The fir6t column in Table 6 shows results for the control experiment where tests were conducted in normal field soil, without any added pathogen.
The absence of pathogen in the ~oil allowed normal plant growth. There i~ no real significant d~fference between treatments, indicatlng the addition of strain NCIB 40189 had no phytotoxic effect.
The other figures in Table 6 givo the results for two test treatments at different pathogen rates. The presence of pathogen in the 80il decreased the number of plants emerging from untreated or blank seed when compared to the control experiment. This shows disease was present and was adversely affectlng plant growth.
~owever, the presence of strain NCIB 40189 ~ignificantly increases the number of plants emerging in the presence of the pathogen.
.
WO~U18613 16 21 0 7 o ~/GBg~g7 EFFICACY AGAINST FUSARIUM:
Field trials were carried out in the USA
(Mississippi), using maize as the test crop.
Pathogen (F solani) was added at various rates as wheat grain inoculum, prepared as descrlbed above.
Stra~n NCIB 40189 was applied as a gum/peat formulation (107 cell/seed). The chemical control was CAPTAN (Trade Mark), used as a seed treatment at the recommended rate. Untreated seed was used as the plant control.
Results are shown in Table 7 below. Emergence is quoted as percentage (%) of plants l5 days after sowing. The letters appearing after the percentage emerged indicate statistical s~gnificance. There is no significant dif~erence at the 5% probability level between entries w~th a letter in common.
TA~LE 7 Field ~ctlYity of NCIB 40189 again~t F solani:
emergence of maize plants T~EATMENT PERCENTAGE ~%) EMERGED W~EN F SOhANT
ADDED AT THE FOL~OWING RATES
(g inoculum per metre row):
NCIB 40189 65.208 C 74.583 AC
CAPTAN 69.583 BC 69.375 BC
Vntreated 65.833 C 51.250 D
The first column of Table 7 shows results for the control tria}s where no test pathogen was added. The absence of pathogen ~n the so~l W0~18613 17 21 0 7 0 9 8 allowed normal plant growth. There ~8 no real significant difference between treatments, indicatlng the addit$on of strain NCIB 40189 had no phytotoxic effect.
S The presence of pathogen, Fusarium solan~, in the soil decreased the number of plants emerging from untreated seed when compared to the control experlment. This shows disease was present and wa~ adversely affect~ng plant growth. ~owever, the presence of ~tra~n NCIB 40189 significantly increases the percentage of plants emerging in the presence of the pathogen. In fact, treatment wlth strain NCIB 40189 improved plant performance 80 that the percentage emerging was not significantly different from the percentage emerging in the absence of pathogen.
Claims (10)
1. An antifungal agent comprising the micro-organism Pseudomonas fluorescens strain NCIB 40189, a culture of which was deposited on 1st September 1989 under the terms of the Budapest Treaty with the National Collection of Industrial and Marine Bacteria Limited, Aberdeen, United Kingdom, under the Accession Number 40189.
2. An antifungal agricultural composition containing as active ingredient an antifungal agent claimed in claim 1, in admixture with a carrier composition acceptable in agricultural practice.
3. A method of inhibiting fungal attack on a plant, comprising applying an effective dose of an antifungal agent as claimed in claim 1 to the locus of a plant or seed.
4. A method as claimed in claim 3 where the antifungal agent is applied to a plant.
5. A method as claimed in claim 3 where the antifungal agent is applied to a seed prior to sowing.
6. A method as claimed in claim 3 where the antifungal agent is applied to a growing medium in which a plant is growing or is to be grown.
7. A method as claimed in claim 3 where the fungus is from the genus Rhizoctonia.
8. A method as claimed in claim 3 where the fungus is from the genus Pythium.
9. A method as claimed in claim 3 where the fungus is from the genus Fusarium.
10. A method as claimed in claim 3 where the fungus is Gaeumannomyces graminis.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB919107678A GB9107678D0 (en) | 1991-04-11 | 1991-04-11 | Antifungal micro-organism |
GB9107678.6 | 1991-04-11 |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2107098A1 true CA2107098A1 (en) | 1992-10-12 |
Family
ID=10693067
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002107098A Abandoned CA2107098A1 (en) | 1991-04-11 | 1992-04-03 | Antifungal micro-organism |
Country Status (13)
Country | Link |
---|---|
EP (1) | EP0579635A1 (en) |
JP (1) | JPH06506345A (en) |
AU (1) | AU656369B2 (en) |
BG (1) | BG61654B1 (en) |
CA (1) | CA2107098A1 (en) |
CZ (1) | CZ280653B6 (en) |
FI (1) | FI934441A (en) |
GB (1) | GB9107678D0 (en) |
HU (1) | HUT67770A (en) |
NO (1) | NO933623L (en) |
NZ (1) | NZ242291A (en) |
RU (1) | RU2104645C1 (en) |
WO (1) | WO1992018613A1 (en) |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
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GB9207352D0 (en) * | 1992-04-03 | 1992-05-13 | Ici Plc | Method to control fungal disease |
FI95598C (en) * | 1994-01-31 | 1996-02-26 | Kemira Agro Oy | Microorganism for biological control of plant diseases |
FI97148C (en) * | 1994-07-14 | 1996-10-25 | Lahden Polttimo Ab Oy | Process for the treatment of plants to improve the quality properties of seeds |
ES2186963T3 (en) * | 1998-12-24 | 2003-05-16 | Gabriele Dr Berg | ISOLATED OF RIZOBACTERIAS FOR ITS USE AGAINST FITOPATOGENOS FLOORS AND PROCEDURE FOR THE USE OF ISOLATED OF RIZOBACTERIAS. |
BR0009629A (en) | 2000-12-08 | 2002-08-20 | Embrapa - Empresa Brasileira De Pesquisa Agropecua | Biocontrol of plant diseases caused by fusarium species with new isolates of bacillus megaterium and pantoea agglomerans |
HU230555B1 (en) * | 2001-08-13 | 2016-12-28 | Biofil Kft. | Environment-friend micro-organism produce and producing thereof |
EP2236037A1 (en) * | 2009-03-30 | 2010-10-06 | Technische Universität Graz | Fungicide preparation |
MD508Z (en) * | 2012-01-06 | 2012-12-31 | Институт Генетики, Физиологии И Защиты Растений Академии Наук Молдовы | Process for presowing treatment of grape seeds |
US9877486B2 (en) | 2014-01-31 | 2018-01-30 | AgBiome, Inc. | Methods of growing plants using modified biological control agents |
KR102281836B1 (en) * | 2014-01-31 | 2021-07-26 | 아그바이오메, 인크. | Modified biological control agents and their uses |
Family Cites Families (4)
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US4456684A (en) * | 1982-09-08 | 1984-06-26 | The United States Of America As Represented By The Secretary Of Agriculture | Method for screening bacteria and application thereof for field control of diseases caused by Gaeumannomyces graminis |
US4647533A (en) * | 1984-09-14 | 1987-03-03 | The United States Of America As Represented By The Secretary Of Agriculture | Method for screening bacteria and application thereof for field control of Pythium spp. on small grain crops |
IL87322A0 (en) * | 1988-08-02 | 1989-01-31 | Univ Ramot | Method of controlling foliar diseases caused by fungal pathogens with fungicidal bacteria and novel pure cultures of fungicidal bacteria |
US4996049A (en) * | 1988-12-19 | 1991-02-26 | Pioneer Hi-Bred International, Inc. | Biological control of corn seed rot and seedling blight |
-
1991
- 1991-04-11 GB GB919107678A patent/GB9107678D0/en active Pending
-
1992
- 1992-04-03 EP EP92907473A patent/EP0579635A1/en not_active Withdrawn
- 1992-04-03 HU HU9302694A patent/HUT67770A/en unknown
- 1992-04-03 AU AU15315/92A patent/AU656369B2/en not_active Ceased
- 1992-04-03 RU RU93058396A patent/RU2104645C1/en active
- 1992-04-03 CA CA002107098A patent/CA2107098A1/en not_active Abandoned
- 1992-04-03 CZ CS932106A patent/CZ280653B6/en unknown
- 1992-04-03 JP JP4507168A patent/JPH06506345A/en active Pending
- 1992-04-03 WO PCT/GB1992/000597 patent/WO1992018613A1/en not_active Application Discontinuation
- 1992-04-09 NZ NZ242291A patent/NZ242291A/en unknown
-
1993
- 1993-10-07 BG BG98144A patent/BG61654B1/en unknown
- 1993-10-08 NO NO933623A patent/NO933623L/en unknown
- 1993-10-08 FI FI934441A patent/FI934441A/en unknown
Also Published As
Publication number | Publication date |
---|---|
HU9302694D0 (en) | 1994-01-28 |
WO1992018613A1 (en) | 1992-10-29 |
GB9107678D0 (en) | 1991-05-29 |
AU656369B2 (en) | 1995-02-02 |
BG98144A (en) | 1994-06-30 |
AU1531592A (en) | 1992-11-17 |
RU2104645C1 (en) | 1998-02-20 |
EP0579635A1 (en) | 1994-01-26 |
JPH06506345A (en) | 1994-07-21 |
CZ280653B6 (en) | 1996-03-13 |
NZ242291A (en) | 1993-08-26 |
BG61654B1 (en) | 1998-02-27 |
NO933623D0 (en) | 1993-10-08 |
FI934441A0 (en) | 1993-10-08 |
FI934441A (en) | 1993-10-08 |
CZ210693A3 (en) | 1994-04-13 |
NO933623L (en) | 1993-10-08 |
HUT67770A (en) | 1995-04-28 |
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