WO2012124863A1 - Préparation microbienne utilisant la souche ob-1 de trichoderma atroviride - Google Patents

Préparation microbienne utilisant la souche ob-1 de trichoderma atroviride Download PDF

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Publication number
WO2012124863A1
WO2012124863A1 PCT/KR2011/005003 KR2011005003W WO2012124863A1 WO 2012124863 A1 WO2012124863 A1 WO 2012124863A1 KR 2011005003 W KR2011005003 W KR 2011005003W WO 2012124863 A1 WO2012124863 A1 WO 2012124863A1
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WO
WIPO (PCT)
Prior art keywords
strain
plant
trichoderma atroviride
present
kccm
Prior art date
Application number
PCT/KR2011/005003
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English (en)
Korean (ko)
Inventor
이동현
이극래
이준석
박기병
문준혁
Original Assignee
(주)오비트
Priority date (The priority date 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 date listed.)
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Application filed by (주)오비트 filed Critical (주)오비트
Publication of WO2012124863A1 publication Critical patent/WO2012124863A1/fr

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Classifications

    • 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
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A01N63/38—Trichoderma
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal 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
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/885—Trichoderma

Definitions

  • the present invention relates to a novel microorganism and a microbial preparation using the same, and more particularly, to a novel Trichoderma atroviride strain derived from soil, and a plant growth promoter or plant disease control agent using the microorganism.
  • microorganisms in fertilizers can help to replenish soil fertility and maintain long-term by providing high quality soil microbial activity, help to suppress pathogenic soil organisms, and increase plant mass.
  • the present inventors have developed a microbial agent using soil microorganisms, Trichoderma atroviride strain (Accession Number: KCCM 11173P) isolated from domestic soil acts on the roots of crops to block harmful microorganisms and grow conditions It was confirmed that it improves the environment suitable for the growth, promotes growth by increasing the root adhesion of crops, shows excellent water efficiency, and shows resistance to some plant pathogens. By confirming this possibility, the present invention was completed by confirming that it can be usefully used as a microbial preparation for plant growth promotion or plant disease control.
  • Still another object of the present invention is to provide a microbial agent for promoting plant growth or controlling plant diseases using the novel Tricorderma atroviride strain.
  • Still another object of the present invention is to provide a method of promoting plant growth or controlling plant diseases using the microbial agent.
  • the present invention provides a Trichoderma atroviride OB-1 strain deposited with accession number KCCM 11173P.
  • the present invention provides a microbial agent comprising the strain or its culture as an active ingredient.
  • the present invention provides a method for promoting plant growth, comprising treating the strain, its culture solution or the microbial agent to soil, a plant or a seed of a plant.
  • the present invention provides a method for controlling plant diseases, comprising the step of treating the strain, its culture or the microbial agent to soil, plants or seeds of plants.
  • the present invention provides a Trichoderma atroviride OB—1 strain or a culture thereof deposited with Accession No. KCCM 11173P for use as a microbial agent.
  • KCCM 11173P for use as a microbial agent.
  • the present invention provides Trichoderma atroviride OB-1 strain deposited with accession number KCCM 11173P.
  • the strain according to the present invention is preferably derived from soil, but is not limited thereto.
  • the strain according to the present invention acts on the roots of crops to block harmful microorganisms, improve the environment suitable for growth conditions, increase the root adhesion of the crops to promote growth, and characterized by having excellent activity efficiency
  • the crop is preferably grass, cabbage, lettuce and tomato, but is not limited thereto.
  • the strain according to the present invention is characterized by having an antimicrobial activity against plant pathogens, wherein the plant pathogens are preferably any one selected from the group consisting of gray bear fungus, wearing disease, anthrax and wilt disease, but not limited thereto. .
  • the strain of the present invention is a novel strain, named Trichoderma atropudide 0B-1, and deposited on February 11, 2011 under the name Trichoderma atroviride OB-1 to the Korea Microbiological Conservation Center ( KCCM 11173P).
  • the present invention provides a microorganism preparation comprising Trichoderma atroviride 0B-1 strain deposited with Accession No. KCCM 11173P or a culture thereof as an active ingredient.
  • the present invention provides a method for promoting plant growth, comprising the step of treating the soil, plant or plant seed with a strain according to the present invention, its culture solution, or a microbial preparation comprising the same.
  • the present invention provides a plant disease control method comprising the step of treating the strain according to the present invention, its culture solution or microbial preparation comprising the same to soil, plants or seeds of plants.
  • the present invention provides a Trichoderma atroviride 0B-1 strain or a culture thereof deposited with Accession No. KCCM 11173P for use as a microbial agent.
  • the microbial agent according to the present invention is preferably for promoting plant growth or controlling plant diseases, but is not limited thereto.
  • the plant is preferably selected from the group consisting of grass, cabbage, lettuce, tomato, potato, rice crop and ginseng, but is not limited thereto, and includes all crops.
  • the plant disease is preferably selected from the group consisting of gray bear disease, wearing disease, anthrax and wilted disease, but is not limited thereto, and includes all plant pathogens.
  • the Trichoderma atrovirite 0B-1 strain (Accession No .: KCCM 11173P) isolated from the soil acts on the root of the crop.
  • Alternative Site Blocking harmful microorganisms, improving the environment suitable for growth conditions, increasing the root adhesion of crops to promote growth, show excellent water efficiency, and showed resistance to some plant pathogens.
  • the strain or its culture solution can be usefully used as a microbial preparation for plant growth promotion or plant disease control.
  • the microbial agent according to the present invention can be used not only Trichoderma atroviride 0B-1 strain or its culture solution itself, but additionally includes an agriculturally acceptable carrier.
  • Strains, cultures thereof, or microbial preparations according to the present invention may be used to spray the soil around the crop in a solid state, irrigation to plants in a liquid state, immersed or sprayed into the seeds of the crop, or coated on the seed, but is not limited thereto. .
  • Strains, cultures or microbial preparations thereof according to the invention can be poured into the soil around the plant (irrigation treatment) or the seeds can be immersed in the culture medium and preparation (immersion treatment), and sprayed (foliar treatment) in the art
  • the well-known technique can be sprayed to flow through the plant, but is not limited thereto.
  • the microbial agent according to the present invention is preferably used by diluting the strain according to the present invention or its culture solution about 10 to 1000 times, and more preferably about 100 to 500 times dilution, but is not limited thereto.
  • the microbial agent according to the present invention is preferably sprayed 3 to 10 times at intervals of 3 to 10 days during plant cultivation, and more preferably sprayed 5 times at 7 day intervals, but is not limited thereto.
  • Tricorderma atroviride 0B-1 strain deposited with Accession No. KCCM 11173P according to the present invention promotes growth by increasing the root adhesion of crops, and shows excellent growth efficiency, against various plant pathogens.
  • the strain or its culture may be usefully used as a microbial agent for promoting plant growth or controlling plant diseases.
  • FIG. 1 is a diagram showing the results of observing the thick film spores and conidia spores under a microscope before and after homogenization of the Trichoderma atroviride 0B-1 strain according to the present invention.
  • Figure 2 is a diagram showing the results of microscopic observation of the thick film spores and conidia from the Trichoderma atroviride 0B-1 strain according to the present invention.
  • Figure 3 is a diagram showing the results of the growth test for the grass of Trichoderma atroviride OB-1 strain according to the present invention.
  • Figure 4 is a graph showing the results of growth promotion experiments for tomatoes of the Trichoderma atroviride 0B-1 strain according to the present invention.
  • 5 is a diagram showing the results of experiments on the effect of increasing the potato for the Trichoderma atroviride 0B-1 strain according to the present invention.
  • Figure 6 is a diagram showing the results of experiments on the antimicrobial activity of the gray bear disease and wearing disease of the Trichoderma atroviride 0B-1 strain according to the present invention.
  • Figure 7 is a diagram showing the results of experiments on the antimicrobial activity of anthracnose and wilt disease of Trichoderma atropudide OB-1 strain according to the present invention.
  • the isolated strain was inoculated in a basal medium of ⁇ 7 ⁇ 0 and incubated at 25 ° C. for 72 hours. Preservation of the strain was carried out to contain glycerol at 20% final concentration.
  • EF elongation factor
  • PCR reactions were performed using 2 (5'-GGAGGTACCAGTGATCATGTT-3 ') (SEQ ID NO: 3). PCR reactions were performed with 20 ng of genomic DNA as compared to 30 ⁇ React ion mixtures including EF-Taq (Sol Gent, Korea). PCR was performed as follows. Activate Taq-Polymerase at 95 0 C for 2 minutes, perform 35 cycles for 1 minute at 95 ° C, perform 1 minute at 55 0 C and 72 ° C, and then 10 at 72 0 C. One cycle was performed for a minute. The amplified product was purified using Millipore Corp., Bedford, MA, USA, and the sequencing reaction was performed using a PRISM BigDye Terminator v3.1 Cycle sequencing Kit.
  • Hi-Di formamide (Applied Biosysterns, FosterCity, CA) was added to the DNA samples containing the amplified products, incubated at 95 0 C for 5 minutes, soaked in ice bucket for 5 minutes, and cooled down.
  • the sequence was analyzed by Prism 3730XL DNA analyzer (Appl ied Biosys terns, FosterCity, CA).
  • the nucleotide sequence thus obtained (SEQ ID NO: 1) was analyzed for homology using NCBI's BLAST program, and systematically using the CLUSTAL X program (Thompson et al., 1994) and the PHYLIP program (Felsenstein, 1993). The location was confirmed.
  • nucleotide sequence of the strain was 99% compared to Trichoderma atroviride compared to the microorganisms registered in NCBI GenBank.
  • Thick-cell spores and conidia are adhered to Trichoderma atheroviride cells cultured in a liquid incubator, and the cells, thick-film spores and conidia are first-order at low pressure of about 200 bar using a Homogenizer Separation and centrifuge at high speed at 8000 rpm to recover the thick film spores and conidia (FIGS. 1 and 2).
  • the dipping process is carried out by diluting 500 times diluted tomato seedlings in plastic seedlings (diameter X height: 30 X 25 cm) and seedling seedlings grown in a tray for growing seedlings for 1 month.
  • the irrigation treatment was once irrigated with 1 L of distillate solution per lm 2 after planting.
  • Immersion + irrigation is done once in the above two methods
  • the thick film spore concentration was 3.0 using a culture medium in which the strain was cultured in a liquid incubator containing a food source. After making the product to X 10 5 cfu / ml or more 3,000 times, it was immersed and compared with the untreated. After immersion in 100 L of distilled liquor per 100 kg of seed potatoes in Gangwon-do for 10-20 seconds, it was dried immediately and settled after 4 to 5 days.
  • the thick film spore concentration was 3.0 X using the culture medium in which the strain was cultured with a liquid incubator containing a food source.
  • the product was made to be 10 5 cfu / ml or more, diluted 500 times, and subjected to topsoil treatment (thick film spores) and immersion treatment (conidia spores and thick film spores), and compared with the untreated.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Mycology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Engineering & Computer Science (AREA)
  • Botany (AREA)
  • Biomedical Technology (AREA)
  • Agronomy & Crop Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne une souche de Trichoderma atroviride et une préparation microbienne l'utilisant. Plus particulièrement, l'invention concerne une nouvelle souche OB-1 de Trichoderma atroviride déposée sous le numéro d'enregistrement KCCM 11173P, permettant la croissance de plantes cultivées par l'augmentation de la capacité d'enracinement de la plante cultivée et présentant une efficacité de rendement élevé et une résistance élevée à divers pathogènes végétaux. Par conséquent une souche ou culture de la présente invention peut être utile en tant que préparation microbienne pour la promotion de la croissance végétale ou la lutte contre les pathogènes végétaux.
PCT/KR2011/005003 2011-03-14 2011-07-08 Préparation microbienne utilisant la souche ob-1 de trichoderma atroviride WO2012124863A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020110022547A KR20120104875A (ko) 2011-03-14 2011-03-14 트리코데르마 아트로비리데 ｏｂ?１ 균주를 이용한 미생물제제
KR10-2011-0022547 2011-03-14

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WO2012124863A1 true WO2012124863A1 (fr) 2012-09-20

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104630072A (zh) * 2014-12-19 2015-05-20 江苏省农业科学院 一株深绿木霉ta-9菌株及其在水稻病害防控中的应用
WO2020245154A1 (fr) 2019-06-07 2020-12-10 Bayer Cropscience Biologics Gmbh Procédés d'augmentation du taux de germination de spores fongiques
WO2021239777A2 (fr) 2020-05-28 2021-12-02 Bayer Cropscience Biologics Gmbh Nouveau substrat de fermentation pour fermentation à l'état solide
WO2021249972A1 (fr) 2020-06-08 2021-12-16 Bayer Cropscience Biologics Gmbh Nouvelles formulations pour augmenter le taux de germination de spores fongiques
WO2022040510A1 (fr) 2020-08-21 2022-02-24 Bayer Cropscience Lp Combinaisons de trichoderma et de bradyrhizobium
CN114644986A (zh) * 2022-03-29 2022-06-21 云南农业大学 一种深绿木霉及其在防治三七病害发生中的应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004346021A (ja) * 2003-05-23 2004-12-09 Nitto Denko Corp 植物病予防剤または除草剤、並びに植物病予防方法もしくは除草方法
US6890530B2 (en) * 2000-04-28 2005-05-10 Newbiotechnic, S.A. Composition comprising fungi of genus trichoderma used as biological control agent and the applications thereof
KR100665539B1 (ko) * 1999-04-08 2007-01-09 야스하루 사사키 식물의 활성부여제 및 그 제조방법과, 식물의 활성촉진제 및 그 사용방법
US20110020286A1 (en) * 2008-03-21 2011-01-27 Ilaria Pertot Trichoderma atroviride sc1 for biocontrol of fungal diseases in plants

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100665539B1 (ko) * 1999-04-08 2007-01-09 야스하루 사사키 식물의 활성부여제 및 그 제조방법과, 식물의 활성촉진제 및 그 사용방법
US6890530B2 (en) * 2000-04-28 2005-05-10 Newbiotechnic, S.A. Composition comprising fungi of genus trichoderma used as biological control agent and the applications thereof
JP2004346021A (ja) * 2003-05-23 2004-12-09 Nitto Denko Corp 植物病予防剤または除草剤、並びに植物病予防方法もしくは除草方法
US20110020286A1 (en) * 2008-03-21 2011-01-27 Ilaria Pertot Trichoderma atroviride sc1 for biocontrol of fungal diseases in plants

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104630072A (zh) * 2014-12-19 2015-05-20 江苏省农业科学院 一株深绿木霉ta-9菌株及其在水稻病害防控中的应用
WO2020245154A1 (fr) 2019-06-07 2020-12-10 Bayer Cropscience Biologics Gmbh Procédés d'augmentation du taux de germination de spores fongiques
WO2021239777A2 (fr) 2020-05-28 2021-12-02 Bayer Cropscience Biologics Gmbh Nouveau substrat de fermentation pour fermentation à l'état solide
WO2021249972A1 (fr) 2020-06-08 2021-12-16 Bayer Cropscience Biologics Gmbh Nouvelles formulations pour augmenter le taux de germination de spores fongiques
WO2022040510A1 (fr) 2020-08-21 2022-02-24 Bayer Cropscience Lp Combinaisons de trichoderma et de bradyrhizobium
CN114644986A (zh) * 2022-03-29 2022-06-21 云南农业大学 一种深绿木霉及其在防治三七病害发生中的应用

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