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WO2013034940A2 - Compositions synergiques de lutte biologique utilisées contre les infections à xanthomonas - Google Patents

Compositions synergiques de lutte biologique utilisées contre les infections à xanthomonas Download PDF

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Publication number
WO2013034940A2
WO2013034940A2 PCT/HU2012/000086 HU2012000086W WO2013034940A2 WO 2013034940 A2 WO2013034940 A2 WO 2013034940A2 HU 2012000086 W HU2012000086 W HU 2012000086W WO 2013034940 A2 WO2013034940 A2 WO 2013034940A2
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Prior art keywords
plant
strain
bacterium
pathogens
bacillus
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PCT/HU2012/000086
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English (en)
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WO2013034940A3 (fr
Inventor
László MANCZINGER
Csaba VÁGVÖLGYI
Enikö SAJBEN
Árpád NAGY
Zoltán SZÖKE-KIS
Adrienn NAGY
György TURÓCZI
András Kovács
Original Assignee
Szegedi Tudományegyetem
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Publication of WO2013034940A2 publication Critical patent/WO2013034940A2/fr
Publication of WO2013034940A3 publication Critical patent/WO2013034940A3/fr

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    • 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
    • A01N63/22—Bacillus
    • 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
    • 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
    • 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
    • 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
    • C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus

Definitions

  • the present patent application relates to a synergistic biological material combination useful to antagonize vegetable pathogens, which comprises a Bacillus strain, preferably a Bacillus subtilis strain, more preferably a Bacillus subtilis B23 deposited under # NCAIM (P) BOO 1390 according to the Budapest Treaty or a Bacillus amyloliquefaciens strain, more preferably a Bacillus amyloliquefaciens B83 strain deposited under # NCAIM (P) B 001391 according to the Budapest Treaty, and a bacterium-predator bacterium strain, preferably a Peredihacter starrii X13 strain deposited under # NCAIM (P) B 001388 according to the Budapest Treaty, and optionally an excipient.
  • a Bacillus strain preferably a Bacillus subtilis strain, more preferably a Bacillus subtilis B23 deposited under # NCAIM (P) BOO 1390 according to the Budapest Treaty or a Bac
  • the biological material according to the invention is an antagonist of the pathogens of vegetables, preferably preferably tomato, pepper, lettuce and/or cabbage, particularly preferably Xanthomonas vesicatoria pathovars.
  • the invention further relates to a process for controlling plant pathogens, and the use of the composition according to the invention for the protection of vegetables, preferably tomato, pepper, lettuce and/or cabbage.
  • the Bacillus strain has long been used for vegetable pest control purposes.
  • the compositions sold are especially effective against vegetable pathogen fungi.
  • the compositions already been commercialized and patented comprise mainly the strains of Bacillus subtilis and Bacillus amiloliquefaciens species. Their effectiveness is explained by different peptide antibiotics, secreted outside of the cell walls, which are effective mainly against fungi: surfactin, iturin, fengycin, bacillomycin, mycosubtilin.
  • compositions comprising alginate-metal-salts, which are capable of controlling pests.
  • the alginate-metal-salts are useful to substitute for different pesticides.
  • the compositions are in particular useful for the treatment of tomato, furthermore, for inhibiting the spreading of Xanthomonas campestris pv. vesicatoria.
  • the disclosed invention is not a biological control of pathogens.
  • US patent publication document No. 2010281585 relates to a composition that increases the resistance of plants against plant pathogens, wherein the composition comprises as actice ingredient a bacterial genetic material. Furthermore, the invention relates to a plant produced by such a method, possessing increased resistance. The method is an effective remedy against a number of pathogen bacteria and fungi, such as, among others Xanthomonas campestris pv. vesicatoria.
  • the patent application discloses a composition containing biological material, as well as combination active compound different from those of our invention.
  • WO9802545 discloses a resistance gene useful against phytopathogens, which is the Prf gene of tomato. When using this gene in transgenic plants, it renders the plant resistant to different phytopathogens, such as Xanthomonas campestris pv. vesicatoria, among others.
  • US patent publication document No. US2003162731 relates to methods and compositions for the protection of plants and their seeds against different diseases.
  • the main components of the compositions are saponines, which are extracted from quinoa and quillaja.
  • the saponines as extracted are applied as pesticides to the seeds, tubers of the plant, or to the plant itself.
  • the method is especially useful to treat tomato and potato, and in particular useful against Xanthomonas campestris pv. vesicatoria.
  • European publication document discloses a peptide or peptidomimetics with a specific sequencethat possesses antimicrobial or antiviral effect.
  • the patent application contains furthermore a nucleic acid molecule, which encodes the peptide according to the invention, as well as a vector and a host cell for the material according to the invention.
  • the method disclosed in the reference is capable of treating infectious diseases. The method may be used against Xanthomonas vesicatoria as well.
  • US patent publication document No. 2010143316 discloses an isolated Bacillus amyloliquefaciens strain, which produces a number of enzymes and antibiotics, including fengycin. Said bacterium strain is capable of inhibiting a number of bacterial and fungal infections, among them Xanthomonas.
  • the application fields of the bacterium strain include e.g. waste water management, chemical industry and agriculture.
  • J ⁇ acillus subtilis denotes the strains of the Bacillis subtilis bacterium species belonging to the Bacillus genus.
  • Bacillus subtilis strain for the practice according to the invention the Bacillus subtilis B23 strain deposited under # NCAIM (P) BOO 1390 according to the Budapest Treaty is especially useful, but is not limited thereto.
  • the Bacillus subtilis strain, which may be used according to the invention, may be selected by the skilled person without undue experimentation.
  • Bacillus amyloliquefaciens denotes the strains of the Bacillis amyloliquefaciens bacterium species belonging to the Bacillus genus.
  • Bacillus amyloliquefaciens B83 strain deposited under # NCAIM (P) B 001391 according to the Budapest Treaty is especially useful, but is not limited thereto.
  • the Bacillus amyloliquefaciens strain, which may be used according to the invention may be selected by the skilled person without undue experimentation.
  • the term facedbacterium-predator bacterium denotes the strains of the Peredibacter starrii bacterium-predator bacterium species belonging to the Peredibacter genus.
  • the Peredibacter starrii X13 strain deposited under # NCAIM (P) B 001388 according to the Budapest Treaty is especially useful, but is not limited thereto.
  • the Peredibacter starrii strain which may be used according to the invention, may be selected by the skilled person without undue experimentation.
  • pathovar denotes a bacterium strain or a group of strains that possess identical or similar characteristics.
  • the pathovars have one or more host plant-specific pathogenity and this distincts them from other strains or sub-species of the same species.
  • infectious disease means a virus, bacterium or fungus, which lives parasitically in different living organisms, and settling and reproducing in the host or its body (e.g. in a human) causes a disease.
  • the measure of the ability to infect, the pathogenity may be variable even within one species.
  • anti-agonist effect means a relationship between microorganisms, in which the members of one species are killed, deterred, or their reproduction is inhibited by the representatives of another species, e.g. through the chemicals (e.g. antibiotics, extracellular enzymes) produced by them.
  • chemicals e.g. antibiotics, extracellular enzymes
  • excipient as used in the present description is not particularly limited, and the excipients may be selected from the group as follows:
  • a liquid formulation e.g. water or an organic solvent (e.g. xilene, methanol, ethylene-glycol or mineral oil), a dispersion stabilizator, a surfactant (e.g. calcium-dodecyl-benzene-sulphonate, polyglycol- ether, etoxylated alkyl-phenol or alkyl-aryl-sulphonates), optionally waxes,
  • a surfactant e.g. calcium-dodecyl-benzene-sulphonate, polyglycol- ether, etoxylated alkyl-phenol or alkyl-aryl-sulphonates
  • optionally waxes optionally waxes
  • a granular formulation montmorillonite, bentonite, wood flour, starch, cellulose and a binder, such as e.g. a mineral oil, polyvinyl-alcohol or saccharose,
  • excipient according to the invention may be selected by the skilled person without undue experimentation.
  • dose formulation characterises the type of formulation according to the composition, which is determined according to the Pesticide formulation types and international coding system catalogue (Crop-Life International Technical Monography, No. 2; 5 ,h Edition, 2002). This may be e.g. aqueous suspension, suspension concentrate, capsulated concentrate, emulsion forming liquid spray, granule, granule dispersible in water, microgranule, water soluble powder, but is not limited thereto.
  • the dose formulation which may be used according to the invention may be selected by the skilled person without undue experimentation.
  • compositions comprising only one component As the effectiveness of the compositions comprising only one component is generally lower than it is expected, our objection was to develop such multi-component compositionsthat in their effect surpass the commercially available compositions, furthermore, they are safer and more effective to use in combating the plant pathogen bacteria and fungi.
  • the invention relates to a composition comprising a synergistic biological material combination useful to antagonize vegetable pathogens, which comprises a Bacillus strain, preferably a Bacillus subtilis strain, more preferably a Bacillus subtilis B23 deposited under # NCAIM (P) B001390 according to the Budapest Treaty or a Bacillus amyloliqiiefaciens strain, more preferably a Bacillus amyloliqiiefaciens B83 strain deposited under # NCAIM (P) B 001391 according to the Budapest Treaty, and a bacterium-predator bacterium strain, preferably a Peretlibacter starrii X 13 strain deposited under # NCAIM (P) B 001388 according to the Budapest Treaty, and optionally an excipient.
  • a Bacillus strain preferably a Bacillus subtilis strain, more preferably a Bacillus subtilis B23 deposited under # NCAIM (P) B001390 according
  • the biological material according to the invention is an antagonist of the pathogens of vegetables, preferably tomato, pepper, lettuce and/or cabbage, such as Xanlhomonas vesicatoria, Pseitdomonas syringae and Clavibacter michiganensis plant pathogen bacteria es Pythium debaryaniim, Phytophthora infestans, Alternaria a!ternata and Fusarium oxysporum plant pathogen fungi, preferably Xanlhomonas vesicatoria pathovars.
  • the Bacillus subtilis strain according to the invention is not in particular limited, in condition that it does not decrease the effectiveness of the biological material combination of the invention.
  • a Bacillus subtilis strain which is preferred for the present invention is without limitation the Bacillus subtilis B23 deposited under # NCAIM (P) B001390 according to the Budapest Treaty.
  • the Bacillus subtilis strain which may be used according to the invention, may be selected by the skilled person without undue experimentation.
  • Bacillus amyloliquefaciens strain according to the invention is not in particular limited, in condition that it does not decrease the effectiveness of the biological material combination of the invention.
  • a Bacillus amyloliquefaciens strain, which is preferred for the present invention is without limitation the Bacillus amyloliquefaciens B83 deposited under # NCAIM (P) B 0013 1 according to the Budapest Treaty.
  • the Bacillus amyloliquefaciens strain, which may be used according to the invention, may be selected by the skilled person without undue experimentation.
  • the bacterium-predator bacterium including the Peredibacter starrii strain according to the invention is not in particular limited, in condition that it does not decrease the effectiveness of the biological material combination of the invention.
  • a Peredibacter starrii strain which is preferred for the present invention is without limitation the Peredibacter starrii X13 deposited under # NCAIM (P) B 001388 according to the Budapest Treaty.
  • the Peredibacter starrii strain which may be used according to the invention, may be selected by the skilled person without undue experimentation.
  • the unique effect of the biological material-combination according to the invention against the pathogens is on one hand inhibits the operation of pathogens, especially Xanthomonas vesicatoria patovars in the rhysosphere and in the buds of the plants and on the other hand, at the same time it induces resistance in the protected plant against the inhibited pathogens.
  • the excipient according to the present invention is not particularly limited, in condition that it does not decrease the effectiveness of the biological material according to the present invention as an active agent, or a biological and chemical active agent combination
  • the applicable excipients include without limitation the following: a) in case of a liquid formulation e.g. water or an organic solvent (e.g. xilene, methanol, ethylene-glycol or mineral oil), a dispersion stabilizator, a surfactant (e.g. calcium-dodecyl-benzene-sulphonate, polyglycol-ether, etoxylated alkyl-phenol or alkyl-aryl-sulphonates), optionally waxes,
  • a liquid formulation e.g. water or an organic solvent (e.g. xilene, methanol, ethylene-glycol or mineral oil), a dispersion stabilizator, a surfactant (e.g. calcium-dodecyl-benzene-sulphonate
  • a granular formulation montmorillonite, bentonite, wood flour, starch, cellulose and a binder, such as e.g. a mineral oil, polyvinyl-alcohol or saccharose,
  • the dose form according to the present invention is not particularly limited, provided that it is suitable for the application of the biological material according to the invention as active agent, or the composition containing said biological material according to the invention to the protected plant or any part thereof.
  • Such applicable dose forms include without limitation the following: aqueous suspension, suspension concentrate, capsulated concentrate, emulsion forming liquid spray, granule, granule dispersible in water, microgranule, water soluble powder.
  • the dose formulation which may be used according to the invention may be selected by the skilled person without undue experimentation.
  • the invention in its third aspect relates to a process for controlling vegetable pathogens according to which the biological material or composition according to the invention is applied to a plant, preferably to a vegetable, more preferably to tomato, pepper, lettuce and/or cabbage.
  • the biological material according to the invention is preferably applied to the seeds of the protected plant, roots of the protected plant, stem of the protected plant, leaves of the protected plant, blooms of the protected plant, the foliage of the protected plant or fruits of the protected plant, is mixed to the irrigation water of the plant, sprayed to the protected plant and/or with any combination of the above.
  • the invention relates to the use of the composition according to the invention for the control of pests, preferably for the control of the pests of vegetables, more preferably for the control of the pests of tomato, pepper, lettuce and/or cabbage, furthermore, for inducing resistance in said plants against the pathogens according to the present invention.
  • Figure I illustrates the test of the bacterium-predator bacteria against Xcinthomonas vesicatoria 01533.
  • the third inoculation strip is the Peredibacter starrii X13 strain.
  • the isolations were made from the root surface and rhizomes of different tomato and pepper species on bacterium selective culturing medium. 10-10 strains were isolated from the dominant colony types in case of each tested sample. The best antagonists were selected against Pseudomonas syringae, Xanthomonas vesicatoria, Erwinia carotovora, illetve Phytophthora infestans, Sclerotinia sclerotiorum, Alternaria solani and Botrytis cinerea with preliminary antagonism tests on culturing plates. These were identified on species level by partial sequencing their 16S RNS gene, in order to exclude the plant pathogens from the further examinations. The efficacy of the non pathogenic strains was tested against another plant pathogenic bacteria and fungi. 40 of the best antagonists were selected for the in vivo plant treatment examinations.
  • endophyte bacterium strains were isolated from the roots and seeds of different vegetables (parsley, carrot, tomato, pepper, white cabbage, lettuce, spring onion, cucumber). Their ability to antagonize the plant pathogen Fusarium oxysporum, Rhizoctonia solani; Xanthomonas campestris pv vesicatoria, Erwinia carotovora and Pseudomonas syringae strains was tested by in vitro antagonism tests. The antagonist potential of the best 10 strains was tested against the plant pathogen fungi Phytophtora infestans, Botrytis cinerea, Sclerotinia sclerotiorum and Alternaria tenuis as well.
  • Bacillus subtilis (B ⁇ 2, B 19)
  • the 16S RNS gene was amplified by PCR (primers: Eub8F and Eub534R), sequenced, then compared with the databases to exclude the plant or human pathogen strains.
  • the tests aimed at clarifying the rhysosphere tolerance of the anatagonist strains in tomato and pepper culture (culturing methods using soil or using no soil), and in other vegetable cultures (cabbage, lettuce), and especially the very important question of how the plant tolerates the treatment with the bacterium.
  • Using quantitative culturing from the rhizomes of the treated plants it was clarified if the bacterium strain is incorporated in the plant, if it makes colonies endogenously without adversely affecting the development of the plant.
  • the tests were run with 20 strains possessing excellent in vitro antagonism spectrum.
  • the tests were made in two periods, using 2x 10 bacterium strains.
  • the strains to be tested members of the Bacillus, Pseudomonas, Pantoea genus
  • the further tests were made with young plants already having leaves.
  • the plant samples were collected at two dates, where the leaves of the selected plants were counted, and the plants were collected without their roots and their fresh green mass was weighed.
  • the tests were made with pepper and tomato species, in soil and rock wood production system, considering the irrigation with three different bacterium concentrations. In cases of cabbage and lettuce the tests were set only to the soil production method. All tests were made in four sets, in random block arrangements.
  • the antagonist bacterium component it is an important requirement that it should provide protection against the bacterial and fungal pathogens not only at the rhizosphere, but also at the organs of the plants above the soil through the activation of the inducible resistance mechanisms of the plant.
  • the tests were made with tomato and pepper.
  • the roots of the young plants were treated with the suspension of the antagonist bacterium strains, and after planting, when different intervals in time have passed, artificial infections were provoked on the leaves with a cell and conidium suspension of the plant pathogen bacteria and fungi.
  • B23 and B83 Both excellent antagonist strains (B23 and B83) were capable of inducing the protection mechanism of the plant (SAR; Systemic Acquired, Resistance).
  • SAR Systemic Acquired, Resistance
  • the best results in controlling Xanthomonas vesicatoria were achieved with the B23 Bacillus subtilis and B83 Bacillus amyloliquefaciens strains used in combination with Peredibacter starrii strains.
  • bacterium parasite strains were isolated following differential centrifugal supplementation using Pseudomonas syringae and Xanthomonas vesicatoria as host bacterium.
  • the bacterium parasite bacterium strains were identified by molecular methods: by the partial sequencing of the 16S RNS gene and the restriction analysis thereof. The host spectrum of the strains was determined using the pathovars of other plant pathogen Pseudomonas, Xanthomonas and Erwinia species.
  • the isolation is based on the method of Jurkevitch et al. (2000).
  • a soil sample originated from the rhyzosphere of 5 g tomato was suspended in 50 ml distilled water and it was shaken at 100 rpm for 30 minutes.
  • 30 ml soil suspension was centrifuged at 3800 rpm for 5 minutes to settle the particles of the soil.
  • the supernatant was repeatedly centrifuged (20500 rpm, 4 °C).
  • the bacteria were collected while the majority of the viruses remained in the supernatant. It is important for the reason that the bacteriophages form a Bdellovibiio- ke stain on the bacterial lawn.
  • the pellet was suspended in 3 ml DNB culturing solution, and the suspension was filtered in a 2 ⁇ pore size filter ( illipore) to get rid of the larger bacteria. Then a 5 member, 10 magnitude set was prepared in sterile distilled water. On glucose yeast culturing medium, a
  • 10 ⁇ cell/ml concentration suspension was prepared from a P. syringae 01810 culture cultivated for 1 day, at ambient temperature, in NaCl solution, then 2 ml of this was admixed to 15 ml DNA culturing media cooled to 40 °C, containing 0.6 % agar. To these 5-5 ml of the dilution set was added, and 5 ml of it was pored on the DNA base plates, containing 1.2 % agar prepared in advance. Half of the dishes were incubated at room temperature for 5-7 days, the remainder at 30 °C.
  • the plaques received were not unambiguous, therefore in the following, 20 g soil sample was suspended in 100 ml 1 % NaCl solution, then separately were inoculated with the X. vesicatoria 01533 and P. syringae 01810 host bacteria, and in round flasks they were shaken for 4 days (30 °C, 120 rpm), to concentrate the amount of the predator bacteria in the presence of the host bacterium. After the incubation, the isolation was done as described above. The plaques appeared were inoculated in several subsequent steps onto the DNA culturing medium containing the suitable host bacteria until no extraneous bacterium colony grew on the dishes.
  • plaques were found. These were cultured on the suitable host bacteria, and purified in several steps, and finally 4 predator strains were isolated on the X. vesicatoria 01533 strain, which were denominated using X I 1 , X I 3, X23 and X26 codes.
  • a 10 7 cell/ml concentration suspension was prepared on glucose-yeast extract culturing medium from the cultures of host bacteria grown for 1 day at room temperature, and 10 ml amounts of it were admixed to 100 ml Tris-YP and PPYE culturing media cooled to 40 °C, containing 0.6 % agar, then 5 ml of them was poured onto the base plates containing 1.2 % agar in the same culturing medium.
  • the host-independent Bdellovibrio bacteriovorus DSM 12732 and Bacteriovorax stolpii DSM 12778 strains used for the reason of comparison were inoculated from a suspension made in 2 ml of 1 % NaCl, onto the culturing medium containing the respective host-bacteria; while the host-dependent Bdellovibrio bacteriovorus DSM 50701 strain and the predator bacteria isolated by us were inoculated from the cultures of the respective host bacteria ⁇ Pseudomonas putida DSM 50906, and Xanthomonas vesicatoria 01533) in stripes onto the culturing medium containing the respective host- bacteria. The dishes were incubated at room temperature for 7 days.
  • the width of the stains formed after 6-7 days of incubation on the respective culturing media (Tris-YP es PPYE) was measured.
  • the host-independent Bdellovibrio strain proved to be ineffective with each of the host bacterium, thus it was unsuitable for the further tests.
  • our isolates were ineffective on other host bacteria, they proved to be effective against all tested Xanthomonas vesicatoria strains, thus, they were suitable for further examinations.
  • the host spectra of the 5 Bdellovibrio strains isolated from the soil samples according to Jurkevitch et al. (2000) were much wider than those of our isolates, since they could parasite as much as 5- 10 different bacterium species.
  • a 10 7 cell/ml concentration suspension was prepared on glucose-yeast extract culturing medium from the cultures of Pseudomonas putida DSM 50906 P. syringae 01810 and Xanthomonas vesicatoria 01 533 host bacteria grown for 1 day at room temperature, and 200 ⁇ amounts of it were measured into 20 ml Tris-YP and PPYE cuituring media.
  • bacteriovonis DSM 50701 strain and the potential BALOs isolated by us were grown on the suitable host bacteria ⁇ Pseudomonas pntida DSM 50906, illetve Xanthomonas vesicutoria 01533) as described above, then agar cubes made from the plaques were inoculated into the cuituring solutions. The cultures were shaken for 6 days (room temperature, 120 rpm), and 200 ⁇ / day was evaluated for measuring the optical density (620 nm, Jupiter HD Microplate Reader).
  • Table 1 The results of the provoked infection performed with Xanthomonas vesicutoria in soil-free production, with pepper test plant, Peredibacter starrii X 13 strain + protecting bacteria
  • the result of the infection at blossoming in soil-free production system shows that there was a treatment significantly better than the control in all three protecting bacteria.
  • the ai ; a_i; a 4 treatments were significantly better as compared to the control; in case of the B83 antagonist, the effect of the ai ; a 2 treatments was excellent.
  • Table 2 The results of the provoked infection performed with Xanthomonas vesicatoria in soil-system, with pepper test plant, using Peredibacter starrii XI 3 strain and three kinds of antagonist protecting bacteria, with treatments performed in different times, Kecskemet, 05. 26. 2009. (blossoming)
  • the combination of the isolated strains is capable of achieving an effective pest control against bacteria and fungi, in the bio-production of tomato, pepper, lettuce and cabbage both in soil and in soil-free systems, especially against Xanthomonas vesicatoria pathovars.

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Abstract

L'invention concerne une combinaison synergiques de matériaux biologiques utilisée pour exercer un antagonisme contre des pathogènes de legumes, laquelle combinaison comprend une souche ¨de Bacillus, de préférence une souche de Bacillus subtilis, plus avantageusement, une souche de Bacillus subtilis B23 déposé sous # NCAIM (P) B001390 selon le Traité de Budapest ou une souche de Bacillus amyloliquefaciens, plus avantageusement, une souche de Bacillus amyloliquefaciens B83 déposée sous # NCAIM (P) B 001391 selon le Traité de Budapest, et une souche bactérienne de bactérie prédatrice, de préférence, une souche Peredibacter starrii X13 déposé sous # NCAIM (P) B 001388 selon le Traité de Budapest, et éventuellement un excipient. Le matériau biologique selon l'invention est un antagoniste contre les pathogènes des légumes. L'invention concerne également un procédé pour réguler des pathogènes de légumes ainsi que l'utilisation de la composition selon l'invention pour protéger les légumes.
PCT/HU2012/000086 2011-09-08 2012-08-30 Compositions synergiques de lutte biologique utilisées contre les infections à xanthomonas WO2013034940A2 (fr)

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HUP1100495 2011-09-08
HU1100495A HUP1100495A2 (en) 2011-09-08 2011-09-08 Synergistic biocontrol compounds against xanthomonas infections

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WO2013034940A3 WO2013034940A3 (fr) 2014-05-08

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Cited By (4)

* Cited by examiner, † Cited by third party
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US9622484B2 (en) 2014-12-29 2017-04-18 Fmc Corporation Microbial compositions and methods of use for benefiting plant growth and treating plant disease
US11089785B2 (en) * 2015-05-22 2021-08-17 Institut National De U Recherche Scientifique Bacterial and fungal metabolites possessing anti-microbial activity against Xanthomonas species, compositions, methods, kits and uses relating to same
CN113913337A (zh) * 2021-10-28 2022-01-11 浙江丰瑜生态科技股份有限公司 一种农用微生物菌液及其应用

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US9622484B2 (en) 2014-12-29 2017-04-18 Fmc Corporation Microbial compositions and methods of use for benefiting plant growth and treating plant disease
US10375964B2 (en) 2014-12-29 2019-08-13 Fmc Corporation Microbial compositions and methods of use for benefiting plant growth and treating plant disease
CN114617136A (zh) * 2014-12-29 2022-06-14 Fmc有限公司 用于促进植物生长和治疗植物疾病的微生物组合物和使用方法
CN114617136B (zh) * 2014-12-29 2024-03-22 Fmc有限公司 用于促进植物生长和治疗植物疾病的微生物组合物和使用方法
ES2584538A1 (es) * 2015-03-27 2016-09-28 Industrias Químicas Del Vallés, S.A. Cepa de Bacillus amyloliquefaciens y uso en el control de enfermedades causadas por bacterias y hongos en las plantas
US10548325B2 (en) 2015-03-27 2020-02-04 Industrias Químicas Del Vallés, S.A. Strain of Bacillus amyloliquefaciens and its use in the control of diseases caused by bacteria and fungi in plants
US11089785B2 (en) * 2015-05-22 2021-08-17 Institut National De U Recherche Scientifique Bacterial and fungal metabolites possessing anti-microbial activity against Xanthomonas species, compositions, methods, kits and uses relating to same
US12161121B2 (en) 2015-05-22 2024-12-10 Instut National De La Recherche Scientifique Bacterial and fungal metabolites possessing anti-microbial activity against Xanthomonas species, compositions, methods, kits and uses relating to same
CN113913337A (zh) * 2021-10-28 2022-01-11 浙江丰瑜生态科技股份有限公司 一种农用微生物菌液及其应用
CN113913337B (zh) * 2021-10-28 2023-06-23 浙江丰瑜生态科技股份有限公司 一种农用微生物菌液及其应用

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