[go: up one dir, main page]

WO2024076222A1 - Composition pour réduire la contrainte de sécheresse comprenant une solution de culture de la souche aureobasidium pullulans ak-10 ou un extrait de la solution de culture de la souche, et procédé pour induire la réduction de la contrainte de sécheresse - Google Patents

Composition pour réduire la contrainte de sécheresse comprenant une solution de culture de la souche aureobasidium pullulans ak-10 ou un extrait de la solution de culture de la souche, et procédé pour induire la réduction de la contrainte de sécheresse Download PDF

Info

Publication number
WO2024076222A1
WO2024076222A1 PCT/KR2023/095051 KR2023095051W WO2024076222A1 WO 2024076222 A1 WO2024076222 A1 WO 2024076222A1 KR 2023095051 W KR2023095051 W KR 2023095051W WO 2024076222 A1 WO2024076222 A1 WO 2024076222A1
Authority
WO
WIPO (PCT)
Prior art keywords
drought stress
strain
aureobasidium pullulans
culture
treatment
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.)
Ceased
Application number
PCT/KR2023/095051
Other languages
English (en)
Korean (ko)
Inventor
박애란
김진철
전효성
서영수
한길
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University Industry Cooperation Foundation of Pusan National University
Industry Foundation of Chonnam National University
Original Assignee
University Industry Cooperation Foundation of Pusan National University
Industry Foundation of Chonnam National University
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.)
Filing date
Publication date
Application filed by University Industry Cooperation Foundation of Pusan National University, Industry Foundation of Chonnam National University filed Critical University Industry Cooperation Foundation of Pusan National University
Publication of WO2024076222A1 publication Critical patent/WO2024076222A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
    • 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
    • A01N63/32—Yeast
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • 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
    • C12N1/16—Yeasts; Culture media therefor
    • 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

Definitions

  • This invention was made under the support of the Korea Forest Service under task number 1405005918 and detailed task number 20211334B10-2323-CD02.
  • the research management agency for the project is the Korea Forestry Promotion Agency, and the research project name is "Resolution of pending issues of disasters and disasters in the forest field.”
  • Research and Development the title of the research project is "Development of a biological agent for aerial spraying to reduce drought damage to fir trees", the host organization is Chonnam National University Industry-Academic Cooperation Foundation, and the research period is 2023.01.01 to 2023.12.31.
  • this invention was made under the support of the Korea Forest Service under task number 1405005335 and detailed task number 20211334B10-2223-CD02.
  • the research management agency for the above project is the Korea Forestry Promotion Agency, and the research project name is "Resolving pending issues of disasters and disasters in the forest field.”
  • “Type research and development” the research project name is “Development of a biological agent for aerial spraying to reduce drought damage to fir trees”, the host organization is Chonnam National University Industry-Academic Cooperation Foundation, and the research period is 2022.01.01 to 2022.12.31.
  • this invention was made under the support of the Korea Forest Service under task number 1405004810 and detailed task number 2021334B10-2123-CD02.
  • the research management agency for the project was the Korea Forestry Promotion Agency, and the research project name was "Resolving pending issues of disasters and disasters in the forest field.”
  • “Type research and development” the research project name is "Development of a biological agent for aerial spraying to reduce drought damage to fir trees”
  • the host organization is Chonnam National University Industry-Academic Cooperation Foundation
  • the research period is 2021.04.01 to 2021.12.31.
  • this invention was made under the support of the Korea Forest Service under task number 1405004872 and detailed task number 2021334C10-2123-CD02.
  • the research management agency for the project was the Korea Forestry Promotion Agency, and the research project name was "Resolving pending issues of disasters and disasters in the forest field.”
  • “Type research and development” the research project name is “Transcriptome and microbial community analysis for reducing drought damage in fir trees”, the host organization is Pusan National University Industry-Academic Cooperation Foundation, and the research period is 2021.04.01 to 2021.12.31.
  • the present invention relates to a composition for reducing drought stress comprising Aureobasidium pullulans AK-10 strain, the strain, a culture thereof, or an extract thereof, a method for producing the composition, and drought stress using the composition. It concerns a method of inducing reduction.
  • the average temperature of the Earth's surface has increased by about 0.93 degrees compared to 1980, and the average temperature of the Earth by the end of the 21st century is expected to rise by about 3.7 degrees compared to the present.
  • Korea's fir tree is a species endemic to the Korean Peninsula that thrived during the Ice Age and survived by moving to higher mountains as temperatures rose after the Ice Age ended. It is also known as an internationally endangered species designated by the International Union for Conservation of Nature (IUCN) in 2011 as it is at high risk of extinction due to its small population. there is.
  • the Hallasan Research Institute analyzes that the decline in snow cover due to climate change and water shortage due to cold winds since the 2000s are the direct effects of the death of fir trees. As winter temperatures rise, snow cover decreases and evergreen trees, which must continue growing activities, are unable to photosynthesize. The cause of death is suggested to be that growth is greatly hindered by lack of necessary moisture.
  • the present inventors confirmed that the drought stress reduction activity was significantly superior when plants were treated with Aureobasidium pullulans AK-10 strain, the strain, its culture, or its extract.
  • the purpose of the present invention is to provide an Aureobasidium pullulans AK-10 strain deposited under the accession number KCTC 15106BP, which has drought stress reduction activity.
  • Another object of the present invention is to provide a composition for reducing drought stress comprising the Aureobasidium pullulans AK-10 strain deposited under the deposit number KCTC 15106BP, its culture, or its extract.
  • Another object of the present invention is to provide a method for producing a composition for reducing drought stress, including a culturing step of culturing the Aureobasidium pullulans AK-10 strain deposited under the deposit number KCTC 15106BP.
  • Another object of the present invention is to provide a method for inducing drought stress reduction, which includes a treatment step of treating the Aureobasidium pullulans AK-10 strain deposited under the accession number KCTC 15106BP, its culture, or its extract.
  • Another object of the present invention relates to the use of the Aureobasidium pullulans AK-10 strain, its culture, or its extract, deposited under deposit number KCTC 15106BP, for reducing drought stress.
  • the present invention relates to a composition for reducing drought stress comprising Aureobasidium pullulans AK-10 strain, a culture thereof, or an extract thereof, a method for producing the composition, and a method for inducing drought stress reduction using the composition. It's about.
  • One aspect of the present invention relates to the Aureobasidium pullulans AK-10 strain, which has drought stress reduction activity in plants.
  • the Aureobasidium pullulans AK-10 strain may be the Aureobasidium pullulans AK-10 strain deposited under the deposit number KCTC 15106BP.
  • the Aureobasidium pullulans AK-10 strain may contain 16S rRNA containing the base sequence of SEQ ID NO: 3.
  • the Aureobasidium pullulans AK-10 strain of the present invention was deposited with the Korea Research Institute of Bioscience and Biotechnology KCTC (Korean Collection for Type Cultures) under the accession number KCTC 15106BP on September 26, 2022.
  • the plant may be a coniferous tree, but is not limited thereto.
  • the coniferous tree may be a cypress tree, a yew tree, a pine tree, a metasequoia, a fir tree, a larch tree, a cypress tree, or a cypress tree.
  • it may be a cypress tree or a yew tree, but is not limited thereto. .
  • the plant may be a dicotyledonous plant, but is not limited thereto.
  • the dicotyledonous plant may be Arabidopsis thaliana, cucumber, tomato, soybean, mung bean, red bean, radish, Chinese cabbage, lettuce, pigweed, plantain, carrot, clover, peach tree, apple tree, persimmon tree, potato, or sweet potato. , for example, it may be Arabidopsis thaliana, cucumber or tomato, but is not limited thereto.
  • Another aspect of the present invention relates to a composition for reducing drought stress comprising Aureobasidium pullulans AK-10 strain, a culture thereof, or an extract thereof, which has the activity of reducing drought stress in plants.
  • the Aureobasidium pullulans AK-10 strain may be the Aureobasidium pullulans AK-10 strain deposited under the deposit number KCTC 15106BP.
  • the Aureobasidium pullulans AK-10 strain may contain 16S rRNA containing the base sequence of SEQ ID NO: 3.
  • the plant may be a coniferous tree, but is not limited thereto.
  • the coniferous tree may be a cypress tree, a yew tree, a pine tree, a metasequoia, a fir tree, a larch tree, a cypress tree, or a cypress tree.
  • it may be a cypress tree or a yew tree, but is not limited thereto. .
  • the plant may be a dicotyledonous plant, but is not limited thereto.
  • the dicotyledonous plant may be Arabidopsis thaliana, cucumber, tomato, soybean, mung bean, red bean, radish, Chinese cabbage, lettuce, pigweed, plantain, carrot, clover, peach tree, apple tree, persimmon tree, potato, or sweet potato. , for example, it may be Arabidopsis thaliana, cucumber or tomato, but is not limited thereto.
  • the composition can be formulated by conventional methods and can be prepared in the form of dry powder or liquid fertilizer, but is not limited thereto.
  • composition of the present invention can be prepared in liquid form, and can be used in the form of powder by adding an extender, or can be formulated and granulated, but the formulation is not particularly limited.
  • the composition may further include additives, extenders, nutrients, and/or disintegrants.
  • the additives are polycarboxylate, sodium lignosulfonate, calcium lignosulfonate, sodium dialkyl sulfosuccinate, sodium alkyl aryl sulfonate, polyoxyethylene alkyl phenyl ether, sodium tripolyphosphate, polyoxyethylene.
  • the extender and nutrient may be one or more selected from the group consisting of skim milk (medium), soybean flour, rice, wheat, red clay, diatomaceous earth, bentonite, dextrin, glucose, and starch, but is limited thereto. It doesn't work.
  • the disintegrant may be one or more selected from the group consisting of bentonite, talc, dialite, kaolin, and calcium carbonate, but is limited thereto. That is not the case.
  • the amount of composition used can be appropriately determined depending on its formulation, damage situation, application method, application location, etc.
  • culture in this specification means containing microorganisms after culturing them.
  • culture supernatant in this specification refers to the upper liquid obtained by removing most microorganisms from the culture medium through centrifugation, and is also referred to as “supernatant liquid.”
  • culture filtrate in this specification refers to the liquid remaining after filtering and removing bacterial cells from the culture medium by performing centrifugation and filtration. Culture filtrate contains substances formed and excreted during the growth of microorganisms, so the substances can be purified or extracted.
  • extract in this specification refers to a culture containing bacterial cells, extracts of bacterial cells, concentrates, concentrates, dried products, or dilutions thereof, etc., in addition to concentrates of cultures, dried products of cultures, and/or culture supernatants of the strains. It may include, for example, all states obtained by processing a culture medium or culture.
  • Another aspect of the present invention relates to a method for producing a composition for reducing plant drought stress, which includes a culturing step of culturing Aureobasidium pullulans AK-10 strain having drought stress reducing activity.
  • the Aureobasidium pullulans AK-10 strain may be the Aureobasidium pullulans AK-10 strain deposited under the deposit number KCTC 15106BP.
  • the Aureobasidium pullulans AK-10 strain may contain 16S rRNA containing the base sequence of SEQ ID NO: 3.
  • the plant may be a coniferous tree, but is not limited thereto.
  • the coniferous tree may be a cypress tree, a yew tree, a pine tree, a metasequoia, a fir tree, a larch tree, a cypress tree, or a cypress tree.
  • it may be a cypress tree or a yew tree, but is not limited thereto. .
  • the plant may be a dicotyledonous plant, but is not limited thereto.
  • the dicotyledonous plant may be Arabidopsis thaliana, cucumber, tomato, soybean, mung bean, red bean, radish, Chinese cabbage, lettuce, pigweed, plantain, carrot, clover, peach tree, apple tree, persimmon tree, potato, or sweet potato. , for example, it may be Arabidopsis thaliana, cucumber or tomato, but is not limited thereto.
  • the medium for cultivating the strain may typically contain milk proteins such as skim milk, whey, and casein, sugars, yeast extract, etc., but is not limited thereto.
  • various general aerobic or anaerobic methods can be used as appropriate for culturing strains.
  • the production method may further include the steps of concentrating, drying, and/or diluting the culture or its supernatant.
  • Another aspect of the present invention is to treat plants or soil with a composition for reducing drought stress, including Aureobasidium pullulans AK-10 strain, a culture thereof, or an extract thereof, which has drought stress reduction activity. It relates to a method for inducing plant drought stress reduction, including the treatment step of:
  • the Aureobasidium pullulans AK-10 strain may be the Aureobasidium pullulans AK-10 strain deposited under the deposit number KCTC 15106BP.
  • the Aureobasidium pullulans AK-10 strain may contain 16S rRNA containing the base sequence of SEQ ID NO: 3.
  • the plant may be a coniferous tree, but is not limited thereto.
  • the coniferous tree may be a cypress tree, a yew tree, a pine tree, a metasequoia, a fir tree, a larch tree, a cypress tree, or a cypress tree.
  • it may be a cypress tree or a yew tree, but is not limited thereto. .
  • the plant may be a dicotyledonous plant, but is not limited thereto.
  • the dicotyledonous plant may be Arabidopsis thaliana, cucumber, tomato, soybean, mung bean, red bean, radish, Chinese cabbage, lettuce, pigweed, plantain, carrot, clover, peach tree, apple tree, persimmon tree, potato, or sweet potato. , for example, it may be Arabidopsis thaliana, cucumber or tomato, but is not limited thereto.
  • the treatment steps include spraying (e.g., spraying, misting, atomizing, powder spraying, granule spraying, surface application, permanent use, etc.), soil irrigation (e.g., mixing, irrigation, etc.), and surface use. It may be performed by one or more methods selected from the group consisting of (e.g., application, smearing, coating, etc.), immersion, poisoning, smoke application, and seed treatment, but is not limited thereto.
  • the present invention relates to a composition for reducing drought stress comprising Aureobasidium pullulans AK-10 strain, the strain, a culture thereof, or an extract thereof, a method for producing the composition, and drought stress using the composition.
  • the reduction induction method it was experimentally confirmed that the drought stress reduction activity was excellent when treated with plants under a drought stress environment. Therefore, it can be usefully used as an active plant biological activator to reduce drought stress in various plants, and it can be sprayed over a wide area through foliar spraying, so it can be used to reduce drought stress damage to conifers and various crops, including fir trees, which are rare and endangered species in alpine areas, at low cost. It is expected that can be reduced.
  • Figure 1 shows the drought stress reduction effect of the AK-10 strain selected through secondary in vivo screening for drought stress reduction using fir tree seedlings according to an embodiment of the present invention.
  • Figure 2 shows a phylogenetic analysis based on the ITS gene sequence of the Aureobasidium pullulans AK-10 strain according to an embodiment of the present invention.
  • Figure 3 shows the drought stress reduction activity effect of the Aureobasidium pullulans AK-10 strain according to the type of culture medium in Arabidopsis thaliana according to an embodiment of the present invention.
  • Figure 4 shows the effect of drought stress reduction activity on yew leaves according to treatment with AK-10 strain culture medium, culture filtrate, and cell suspension in a drought stress environment according to an embodiment of the present invention.
  • Figure 5 shows the effect of drought stress reduction activity on the leaves of fir tree seedlings by treatment with various concentrations of AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention in terms of proline content.
  • Figure 6 shows the moisture content of the leaves of fir tree seedlings according to treatment with various concentrations of AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • Figure 7 is an observation of the effect of reducing drought stress on fir tree seedlings by treating various concentrations of AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • Figure 8 shows the MDA content of fir tree seedling leaves according to treatment with various concentrations of AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • Figure 9 shows the moisture content of cucumber leaves according to treatment with AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • Figure 10 is a photograph showing the effect of reducing drought stress on cucumbers according to treatment of AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • Figure 11 shows the moisture content of tomato leaves according to treatment with AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • Figure 12 is a photograph showing the effect of reducing drought stress on tomatoes according to treatment of AK-10 strain culture medium in a drought stress environment according to an embodiment of the present invention.
  • the present invention relates to the Aureobasidium pullulans AK-10 strain deposited under the accession number KCTC 15106BP, which has the activity of reducing plant drought stress.
  • % used to indicate the concentration of a specific substance is (weight/weight)% for solid/solid, (weight/volume)% for solid/liquid, unless otherwise specified. /Liquid is (volume/volume)%.
  • PEG 6000 was added to create an environment similar to drought treatment in Arabidopsis thaliana, and the proline content in the Arabidopsis thaliana was measured after treatment. The degree of stress relief was tested.
  • the Arabidopsis thaliana required for strain selection was first prepared. Arabidopsis seeds were surface sterilized with 70% ethanol for 3 minutes and then subjected to additional surface sterilization for 3 minutes using a bleach solution (2% NaOCl + 0.05% tween-20). After surface sterilization twice, it was washed three times with sterilized water, immersed in sterilized water, stored at 4°C for 2 days, and treated at low temperature.
  • a bleach solution 2% NaOCl + 0.05% tween-20
  • plant-derived microbial strain culture filtrate To prepare plant-derived microbial strain culture filtrate, plant-derived microbial strains were cultured in LB liquid medium (Tryptone 10 g, Yeast extract 5 g, Sodium chloride 10 g/L, Difco) at 30°C and 150 rpm for 2 days, and after cultivation, The OD 600 value of the culture medium was measured. The culture solution whose concentration was confirmed was filtered using a 0.2 ⁇ m membrane filter, 250 ppm of tween-20 was added, and the sample was diluted with distilled water so that the final microbial treatment concentration was 0.8 (OD 600 value) to prepare a culture filtrate sample.
  • LB liquid medium Teryptone 10 g, Yeast extract 5 g, Sodium chloride 10 g/L, Difco
  • Samples were treated with foliar spray on Arabidopsis thaliana twice: 4 days before the drought stress treatment (same as 12 days after planting Arabidopsis seeds) and 1 day before.
  • the sample processing volume was approximately 300 ⁇ L when culturing 10 Arabidopsis seeds on a plate with a diameter of 9 mm. After sample treatment, the plates were cultured in a plant growth incubator at 25°C (16 hours/8 hours light/dark cycle, 80% relative humidity).
  • the sulphosalicylic acid suspension was centrifuged at 13,000 rpm and 4°C for 5 minutes, and 200 ⁇ L of the supernatant was added to 200 ⁇ L acetic acid (Sigma-Aldrich, Saint Louis, MO, USA) and 200 ⁇ L ninhydrin reagent (Sigma-Aldrich, Saint Louis, MO, USA) was added, heat treated at 100°C for 45 minutes, and cooled on ice for 30 minutes. An equal amount of toluene (Sigma-Aldrich, Saint Louis, MO, USA) was added to the cooled mixture (600 ⁇ L), mixed for 1 minute, and centrifuged at 13,000 rpm at 4°C for 5 minutes.
  • the toluene layer of each sample was separated and the proline content was measured by measuring the absorbance at 532 nm using a Nano Photometer (Implen, Westlake Village, USA). Toluene was measured as a blank, and instead of the sample, proline was added at 1 ⁇ M, 10 ⁇ M, 50 ⁇ M, 100 ⁇ M, 150 ⁇ M, 200 ⁇ M, and 300 ⁇ M, and the absorbance was measured after going through the same processing as the sample and creating a standard curve. did.
  • the in vivo activity of 14 types of microorganisms selected based on their proline content in Arabidopsis thaliana leaves cultured under dry stress conditions was measured in reducing drought stress using Abies koreana .
  • 3-year-old fir trees were purchased from an international horticultural nursery and treated by foliar spraying of strain culture and culture filtrate. After watering was cut off and drought stress was applied, growth status was visually observed to conduct secondary in vivo selection of drought stress-reducing strains. did.
  • the microbial strains were cultured in LB liquid medium at 25°C and 150 rpm for 24 hours, and the OD 600 value of the culture medium was measured after cultivation. Samples were prepared by diluting the culture medium to a microbial concentration of 0.8 (OD 600 value) and adding tween-20 to a final concentration of 250 ppm. In the case of the culture filtrate, the culture liquid whose concentration was confirmed was filtered using a 0.2 ⁇ m membrane filter. Then, based on the concentration of microorganisms measured before filtration, the sample was diluted in the same ratio as the culture solution to 0.8 (OD 600 value).
  • the sample was diluted with distilled water and tween-20 was added to adjust the final concentration to 250 ppm to obtain a culture filtrate sample. was manufactured.
  • the samples were foliar sprayed on the fir trees three times: 7 days before (1st) and 4 days before (2nd) the drought stress treatment date, and 4 days after the drought stress treatment (3rd time). Drought stress of Korean fir trees was carried out by cutting off water during the stress period. The results are shown in Figure 1.
  • the AK-10 strain culture medium and culture filtrate treatment group showed an excellent drought stress reduction effect compared to the untreated group. According to the results of this experiment, among the 14 strains initially selected, the AK-10 strain was selected as the final drought stress reduction active strain.
  • Strain AK-10 isolated from pine leaves was identified molecularly through nucleotide sequence analysis of the ITS gene.
  • the strain was inoculated into YPD liquid medium (Yeast Extract 10 g, Peptone 20 g, Dextrose 20 g/L) and then cultured with shaking at 150 rpm at 25°C for 24 hours.
  • the harvested strain was iNtRON's I-genomic BYF DNA Extraction Mini. Genomic DNA (gDNA) of the strain was extracted using the kit according to the protocol.
  • the extracted gDNA of the strain the PCR-premix (Polymerase chain reaction-premix) from iNtRON Biotechnology, and the primer set (SEQ ID NO: 1 and SEQ ID NO: 2) of Table 2 below that can amplify the ITS of the strain. After mixing, the genes were amplified through PCR.
  • ITS gene PCR started at 95°C for 5 minutes, repeated 30 times at 95°C for 30 seconds, 50°C for 30 seconds, and 72°C for 90 seconds, and then amplified at 72°C for 10 minutes and ended at 4°C.
  • the amplified ITS gene PCR product was submitted to Genotech (Daejeon, Korea) for base sequence analysis, and a total base sequence of 551 bp (SEQ ID NO. 3) was obtained as the ITS coding base sequence of the isolated strain, AK-10 strain.
  • the AK-10 strain was identified as Aureobasidium pullulans.
  • the ITS base sequence of the strain was uploaded to the GenBank database as OP295075, and the strain selected through the present invention was named Aureobasidium pullulans AK-10 strain, and as of September 26, 2022, Korean It was deposited in the Collection for Type Cultures (KCTC) and given the accession number KCTC 15106BP.
  • Example 4 Aureobasidium pullulans ( Aureobasidium pullulans ) Activity test according to the type of culture medium of AK-10 strain
  • Aureobasidium pullulans Aureobasidium pullulans AK-10 strain was used in LB medium during the selection process, but as a result of molecular biological analysis, it was found to be yeast.
  • the activity of the culture filtrate cultured in YPD medium used for yeast culture and cultured in LB medium The activity of the culture filtrate was compared in Arabidopsis thaliana.
  • drought stress reduction activity of the strain was artificially promoted by treating PEG 6000 in the nutrient medium, so culture medium with possible contamination was not used and microorganisms were removed. Only the filtrate was used.
  • LB culture filtrate of Aureobasidium pullulans AK-10 strain culture was cultured in LB liquid medium (Tryptone 10 g, Yeast extract 5 g, Sodium chloride 10 g/L, Difco) at 30°C and 150 rpm for 2 days. After culturing, the OD 600 value of the culture medium was measured. The culture solution whose concentration was confirmed was filtered using a 0.2 ⁇ m membrane filter, 250 ppm of tween-20 was added, and the sample was diluted with distilled water so that the final microbial treatment concentration was 0.8 (OD 600 value) to prepare a culture filtrate sample.
  • LB liquid medium Teryptone 10 g, Yeast extract 5 g, Sodium chloride 10 g/L, Difco
  • AK-10 strain was inoculated into YPD liquid medium (Yeast Extract 10 g, Peptone 20 g, Dextrose 20 g/L) and incubated at 25°C. The culture was shaken at 150 rpm for 24 hours, and the OD 600 value of the culture medium was measured after culture.
  • the YPD culture solution whose concentration was confirmed was filtered using a 0.2 ⁇ m membrane filter, 250 ppm of tween-20 was added, and the sample was diluted with distilled water so that the final microbial concentration was 0.8 (OD 600 value) to prepare a culture filtrate sample.
  • the sulphosalicylic acid suspension was centrifuged at 13,000 rpm and 4°C for 5 minutes, and 200 ⁇ L of the supernatant was added to 200 ⁇ L acetic acid (Sigma-Aldrich, Saint Louis, MO, USA) and 200 ⁇ L ninhydrin reagent (Sigma-Aldrich, Saint Louis, MO, USA) was added, heat treated at 100°C for 45 minutes, and cooled on ice for 30 minutes.
  • An equal amount of toluene (Sigma-Aldrich, Saint Louis, MO, USA) was added to the cooled mixture (600 ⁇ L), mixed for 1 minute, and centrifuged at 13,000 rpm at 4°C for 5 minutes. After centrifugation, the toluene layer of each sample was separated and the proline content was measured by measuring the absorbance at 532 nm using a Nano Photometer (Implen, Westlake Village, USA). The results are shown in Table 3.
  • Toluene was measured as a blank, and instead of the sample, proline was added at 1 ⁇ M, 10 ⁇ M, 50 ⁇ M, 100 ⁇ M, 150 ⁇ M, 200 ⁇ M, and 300 ⁇ M, and the absorbance was measured after going through the same processing process as the sample and creating a standard curve. did.
  • both the LB culture filtrate of the AK-10 strain and the YPD culture filtrate treatment showed drought stress reduction activity in Arabidopsis.
  • the proline content was slightly lower when the YPD culture filtrate of the AK-10 strain was treated, making it superior to the other treatments. It was confirmed that it has drought stress reduction activity. Therefore, in subsequent examples, the Aureobasidium pullulans AK-10 strain was cultured in YPD liquid medium.
  • Example 5 Aureobasidium pullulans ( Aureobasidium pullulans ) Drought stress reduction activity assay in yew trees of culture medium, culture filtrate, and cell suspension of AK-10 strain
  • Example 2 The same experimental method described in Example 2 was performed to test the drought stress reduction activity in yew trees according to the treatment of the YPD culture medium, culture filtrate, and cell fraction of the Aureobasidium pullulans AK-10 strain.
  • YPD culture medium as the AK-10 sample to be processed, inoculate the YPD liquid medium with Aureobasidium pullulans AK-10 strain, culture with shaking at 150 rpm for 24 hours at 25°C, and measure the OD 600 value of the culture medium.
  • a culture sample was prepared by titrating the tween-20 to 250 ppm and diluting the sample with distilled water to obtain a final microbial concentration of 0.8 (OD 600 value).
  • the concentration of the YPD culture filtrate and cell suspension was confirmed by measuring the OD 600 value of the culture in the same manner as the YPD culture described above, and centrifuged at 10,000 rpm at 4°C for 10 minutes to separate into supernatant and cell fraction.
  • the supernatant obtained after centrifugation was filtered using a 0.2 ⁇ m membrane filter, then 250 ppm of tween-20 was added, and the concentration before centrifugation was applied to obtain a final microbial concentration of 0.8 (OD 600 value) at the same dilution ratio as the culture medium.
  • a culture filtrate sample was prepared by diluting the sample with distilled water, and the cell fraction obtained after centrifugation was suspended in the same volume of PBS before centrifugation and then diluted at the same dilution ratio as the culture solution so that the final microbial concentration was 0.8 (OD 600 value). was diluted with distilled water and 250 ppm of tween-20 was added to prepare a cell suspension to be used as a sample.
  • yew tree leaves were collected, weighed, and pulverized using liquid nitrogen.
  • the frozen yew tree leaves were pulverized using a Tissue Lyser (Qiagen, CA, USA), and after pulverization, the proline content was tested in the same manner as the proline content analysis method described at the top of Example 4.
  • the drought stress reduction activity of the YPD culture, culture filtrate, and cell suspension of the Aureobasidium pullulans AK-10 strain was measured by measuring the moisture content in the yew trees after drought treatment and visually observing the drought stress of each treatment. The reduction effect was measured.
  • leaves were collected from untreated and AK-10 strain treated yew trees 14 days after drought stress, the weight of the fresh leaves of the sample before drying was measured, and then stored at 65°C. The weight of the dried yew sample was measured after drying in an oven for 3 days. The moisture content of the treatment was measured using the following calculation formula.
  • Moisture content (%) (fresh weight of sample - dry weight of sample)/fresh weight of sample ⁇ 100
  • AK-10 YPD culture medium the proline content is 231.8 ⁇ M, which is under stress compared to the normal irrigation treatment group in which 115.7 ⁇ M proline was produced, but the moisture content is 37.2%, similar to the moisture content of the normal irrigation treatment group (37.8%). It was confirmed that it showed excellent drought stress tolerance. Therefore, AK-10 YPD culture medium was used in future experiments to test the effect of reducing drought stress in various plants.
  • Example 6 Reduction of drought stress at various concentrations of AK-10 strain using Korean fir tree in vivo Activity Assay - Proline Content
  • the AK-10 strain was inoculated into YPD liquid medium (10 g of Yeast Extract, 20 g of Peptone, 20 g/L of Dextrose) and then cultured with shaking at 150 rpm for 24 hours at 25°C. After culturing, the OD 600 value of the culture medium was measured.
  • a culture sample was prepared by adding 250 ppm of tween-20 to the culture medium whose concentration was confirmed and diluting the sample with distilled water so that the final microbial treatment concentration was 0.8, 0.08, and 0.008 (OD 600 value).
  • the samples were foliar sprayed on the fir trees three times: 7 days before (1st) and 4 days before (2nd) the drought stress treatment date, and 4 days after the drought stress treatment (3rd time).
  • Example 7 Reduction of drought stress at various concentrations of AK-10 strain using Korean fir tree in vivo Activity assay - moisture content determination and visual assay
  • Drought stress reduction in vivo activity of Aureobasidium pullulans AK-10 strain at various concentrations was measured by measuring the moisture content of Abies koreana leaves in a drought stress environment and measuring drought in Abies koreana seedlings. The stress reduction effect was observed with the naked eye. It is interpreted that the higher the moisture content, the lower the water loss rate within the fir tree and the less drought stress.
  • the fir tree plant for measuring moisture content was prepared in the same manner as in Example 6.
  • the culture medium of the AK-10 strain was inoculated into YPD liquid medium (10 g of Yeast Extract, 20 g of Peptone, 20 g/L of Dextrose) and then shaken at 150 rpm for 24 hours at 25°C.
  • the culture was performed, and the OD 600 value of the culture medium was measured after culture.
  • a culture sample was prepared by adding 250 ppm of tween-20 to the culture medium whose concentration was confirmed and diluting the sample with distilled water so that the final microbial treatment concentration was 0.8, 0.08, and 0.008 (OD 600 value).
  • the samples were foliar sprayed on the fir trees three times: 7 days before (1st) and 4 days before (2nd) the drought stress treatment date, and 4 days after the drought stress treatment (3rd time). Drought stress of Korean fir trees was carried out by cutting off water during the stress period.
  • the AK-10 strain sample was treated and the leaves of the fir tree under drought stress were collected 1, 2, and 3 weeks after the drought stress treatment, the weight of the fresh leaves of the sample was measured before drying, and then dried in an oven at 65°C for 3 days. I ordered it. The weight of the dried fir tree leaves was measured and the moisture content was measured using the following formula.
  • Moisture content (%) (fresh weight of sample - dry weight of sample)/fresh weight of sample ⁇ 100
  • the drought stress reduction activity according to the various concentration treatments of the YPD culture medium of the Aureobasidium pullulans AK-10 strain was analyzed in terms of the moisture content of the leaves of the fir tree seedlings, and the results showed that drought stress treatment 2 After a week, it was confirmed that the moisture content in the untreated group decreased rapidly, unlike the normal irrigation treated group.
  • the Aureobasidium pullulans AK-10 strain treatment group with OD 600 values of 0.8 and 0.08 showed high moisture content even after 3 weeks of drought stress treatment and showed a moisture content similar to that of the normal irrigation treatment group.
  • the moisture content of the Aureobasidium pullulans AK-10 strain treated group with an OD 600 value of 0.008 decreased over time similar to the untreated group, but as can be seen in Figure 7, the results were observed with the naked eye after 2 weeks of drought stress. , it was confirmed that the OD 600 value of 0.008 treatment group also showed healthy vitality compared to the untreated group, similar to the 0.8 and 0.08 treatments.
  • Example 8 Reduction of drought stress at various concentrations of AK-10 strain using Korean fir tree in vivo Activity assay - MDA content determination
  • the in vivo activity of Aureobasidium pullulans AK-10 strain in reducing drought stress at various concentrations was measured by the malondialdehyde (MDA) content of Abies koreana leaves in a drought stress environment.
  • MDA malondialdehyde
  • the preparation of the fir tree plant for measuring the MDA content of the fir tree was prepared in the same manner as in Example 6.
  • the culture medium of the AK-10 strain was inoculated into YPD liquid medium (10 g of Yeast Extract, 20 g of Peptone, 20 g/L of Dextrose) and then cultured with shaking at 150 rpm for 24 hours at 25°C. After culturing, the OD 600 value of the culture medium was measured.
  • a culture sample was prepared by adding 250 ppm of tween-20 to the culture medium whose concentration was confirmed and diluting the sample with distilled water so that the final microbial treatment concentration was 0.8, 0.08, and 0.008 (OD 600 value).
  • the samples were foliar sprayed on the fir trees three times: 7 days before (1st) and 4 days before (2nd) the drought stress treatment date, and 4 days after the drought stress treatment (3rd time). Drought stress of Korean fir trees was carried out by cutting off water during the stress period.
  • fir tree leaf samples were collected, weighed, and frozen in liquid nitrogen. Frozen fir leaves were pulverized using a Tissue Lyser (QIAGEN, Mettmann, Germany). 1 mL of 0.1% trichloroacetic acid (TCA) (Sigma-Aldrich, Saint Louis, MO, USA) was added to 0.1 g of the ground frozen sample and then homogenized. The homogenized sample was centrifuged at 10,000 rpm and 4°C for 10 minutes, and 200 ⁇ L of the supernatant was separated into 800 ⁇ L of 0.5% thiobarbituric acid (TBA) (Sigma-Aldrich, Saint Louis, MO, USA) containing 20% TCA.
  • TCA trichloroacetic acid
  • the drought stress reduction activity according to various concentration treatments of the YPD culture medium of the Aureobasidium pullulans AK-10 strain was analyzed based on the MDA content in the leaves of fir tree seedlings, and the drought stress treatment After two weeks, it was confirmed that the MDA content in the untreated group increased rapidly, unlike the normal irrigation treated group.
  • treatment with Aureobasidium pullulans AK-10 strain with OD 600 values of 0.8, 0.08, and 0.08 showed low MDA content even after 2 weeks of drought stress treatment. It was confirmed that the treatment with Aureobasidium pullulans AK-10 strain with OD 600 values of 0.08 and 0.008 showed a somewhat low MDA content even after 3 weeks of drought stress, although the MDA content increased over time.
  • Example 9 Reduction of drought stress in AK-10 strain using cucumber in vivo Activity assay - moisture content determination and visual assay
  • the in vivo activity of Aureobasidium pullulans strain AK-10 in reducing drought stress in cucumber seedlings was measured by water content, and the drought stress reduction effect was observed with the naked eye. It is interpreted that the higher the moisture content, the lower the moisture loss rate in cucumber seedlings and the less drought stress.
  • Samples and cucumber plants for measuring moisture content according to treatment with the AK-10 strain were prepared by treating them in the following manner. To prepare the culture medium of the AK-10 strain, the AK-10 strain was inoculated into YPD liquid medium (10 g of Yeast Extract, 20 g of Peptone, 20 g/L of Dextrose) and then cultured with shaking at 150 rpm for 24 hours at 25°C.
  • OD 600 value of the culture medium was measured.
  • a culture sample of which the concentration was confirmed was prepared by adding 250 ppm of tween-20 and diluting the sample with distilled water so that the final microbial treatment concentration was 0.8 (OD 600 value).
  • Samples were treated with foliar spray on cucumber seedlings twice: 7 days before the drought stress treatment date (1st, 17 days after sowing cucumber seeds, 4th leaf stage of cucumber seedlings) and 4 days before (2nd). Drought stress of cucumber seedlings was carried out by cutting off water during the stress period.
  • Moisture content (%) (fresh weight of sample - dry weight of sample)/fresh weight of sample ⁇ 100
  • the drought stress reduction activity according to the culture medium treatment of the Aureobasidium pullulans AK-10 strain was analyzed based on the water content of cucumber seedling leaves, and as a result, after 4 days of drought stress treatment, the untreated group (43.0 %), unlike the AK-10 treatment group (51.7%) and the normal irrigation treatment group (55.7%), the moisture content decreased significantly.
  • the untreated group showed symptoms of wilting.
  • the AK-10 treatment group showed healthy vitality similar to the normal irrigation treatment group. Therefore, it was confirmed that the Aureobasidium pullulans AK-10 strain treatment showed excellent drought stress reduction activity in cucumbers.
  • Example 10 Reduction of drought stress in AK-10 strain using tomatoes in vivo Activity assay - moisture content determination and visual assay
  • the in vivo activity of Aureobasidium pullulans strain AK-10 in reducing drought stress in tomato seedlings was measured by moisture content, and the drought stress reduction effect was observed with the naked eye. It is interpreted that the higher the moisture content, the lower the water loss rate in tomato seedlings and the less drought stress.
  • Samples and tomato plants for measuring moisture content according to treatment with the AK-10 strain were prepared by treating them in the following manner. To prepare the culture medium of the AK-10 strain, the AK-10 strain was inoculated into YPD liquid medium (10 g of Yeast Extract, 20 g of Peptone, 20 g/L of Dextrose) and then cultured with shaking at 150 rpm for 24 hours at 25°C.
  • OD 600 value of the culture medium was measured.
  • a culture sample of which the concentration was confirmed was prepared by adding 250 ppm of tween-20 and diluting the sample with distilled water so that the final microbial treatment concentration was 0.8 (OD 600 value).
  • Samples were treated with foliar spray on tomato seedlings twice, 7 days before the drought stress treatment date (1st, 26 days after sowing tomato seeds, 4th leaf stage of tomato seedlings) and 4 days before (2nd). Drought stress of tomato seedlings was carried out by cutting off water during the stress period.
  • Moisture content (%) (fresh weight of sample - dry weight of sample)/fresh weight of sample ⁇ 100
  • the drought stress reduction activity according to the culture medium treatment of the Aureobasidium pullulans AK-10 strain was analyzed based on the moisture content of tomato seedling leaves, and as a result, after 4 days of drought stress treatment, the untreated group (79.4 %), unlike the AK-10 treatment group (86.9%) and the normal irrigation treatment group (87.0%), the moisture content decreased significantly. Also, as can be seen in Figure 12, it was confirmed with the naked eye that the untreated group showed symptoms of wilting. On the other hand, it was confirmed that the AK-10 treatment group showed healthy vitality similar to the normal irrigation treatment group. Therefore, it was confirmed that the Aureobasidium pullulans AK-10 strain treatment showed excellent drought stress reduction activity in tomatoes.
  • the present invention relates to a composition for reducing drought stress comprising Aureobasidium pullulans AK-10 strain, the strain, a culture thereof, or an extract thereof, a method for producing the composition, and drought stress using the composition. It concerns a method of inducing reduction.

Landscapes

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

Abstract

La présente invention concerne une souche Aureobasidium pullulans AK-10, une composition pour réduire la contrainte de sécheresse comprenant la souche, une culture de celle-ci, ou un extrait de celle-ci, un procédé de préparation de la composition, et un procédé pour induire la réduction de la contrainte de sécheresse à l'aide de la composition.
PCT/KR2023/095051 2022-10-07 2023-08-24 Composition pour réduire la contrainte de sécheresse comprenant une solution de culture de la souche aureobasidium pullulans ak-10 ou un extrait de la solution de culture de la souche, et procédé pour induire la réduction de la contrainte de sécheresse Ceased WO2024076222A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2022-0129025 2022-10-07
KR1020220129025A KR102516279B1 (ko) 2022-10-07 2022-10-07 아우레오바시디움 플루란스 ak-10 균주의 배양액 또는 균주 배양액의 추출물을 포함하는 가뭄 스트레스 저감용 조성물, 및 가뭄 스트레스 저감 유도 방법

Publications (1)

Publication Number Publication Date
WO2024076222A1 true WO2024076222A1 (fr) 2024-04-11

Family

ID=85936697

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2023/095051 Ceased WO2024076222A1 (fr) 2022-10-07 2023-08-24 Composition pour réduire la contrainte de sécheresse comprenant une solution de culture de la souche aureobasidium pullulans ak-10 ou un extrait de la solution de culture de la souche, et procédé pour induire la réduction de la contrainte de sécheresse

Country Status (2)

Country Link
KR (1) KR102516279B1 (fr)
WO (1) WO2024076222A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102516279B1 (ko) * 2022-10-07 2023-04-03 전남대학교 산학협력단 아우레오바시디움 플루란스 ak-10 균주의 배양액 또는 균주 배양액의 추출물을 포함하는 가뭄 스트레스 저감용 조성물, 및 가뭄 스트레스 저감 유도 방법

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150119022A (ko) * 2013-02-11 2015-10-23 바이엘 크롭사이언스 엘피 고제로틴 및 생물학적 방제제를 포함하는 조성물
KR20150144779A (ko) * 2013-04-19 2015-12-28 바이엘 크롭사이언스 악티엔게젤샤프트 살충성 또는 농약성 2성분 혼합물
KR20200072886A (ko) * 2018-12-13 2020-06-23 주식회사 글루칸 흑효모 유래 바이오폴리머 생산을 위한 배지 조성물 및 그를 이용한 흑효모 유래 바이오폴리머의 제조방법
KR102516279B1 (ko) * 2022-10-07 2023-04-03 전남대학교 산학협력단 아우레오바시디움 플루란스 ak-10 균주의 배양액 또는 균주 배양액의 추출물을 포함하는 가뭄 스트레스 저감용 조성물, 및 가뭄 스트레스 저감 유도 방법

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150119022A (ko) * 2013-02-11 2015-10-23 바이엘 크롭사이언스 엘피 고제로틴 및 생물학적 방제제를 포함하는 조성물
KR20150144779A (ko) * 2013-04-19 2015-12-28 바이엘 크롭사이언스 악티엔게젤샤프트 살충성 또는 농약성 2성분 혼합물
KR20200072886A (ko) * 2018-12-13 2020-06-23 주식회사 글루칸 흑효모 유래 바이오폴리머 생산을 위한 배지 조성물 및 그를 이용한 흑효모 유래 바이오폴리머의 제조방법
KR102516279B1 (ko) * 2022-10-07 2023-04-03 전남대학교 산학협력단 아우레오바시디움 플루란스 ak-10 균주의 배양액 또는 균주 배양액의 추출물을 포함하는 가뭄 스트레스 저감용 조성물, 및 가뭄 스트레스 저감 유도 방법

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CENE GOSTIN?AR;ROBIN A OHM;TINA KOGEJ;SILVA SONJAK;MARTINA TURK;JANJA ZAJC;POLONA ZALAR;MARTIN GRUBE;HUI SUN;JAMES HAN;ADITI SHARM: "Genome sequencing of four Aureobasidium pullulans varieties: biotechnological potential, stress tolerance, and description of new species", BMC GENOMICS, BIOMED CENTRAL LTD, LONDON, UK, vol. 15, no. 1, 1 July 2014 (2014-07-01), London, UK , pages 549, XP021192796, ISSN: 1471-2164, DOI: 10.1186/1471-2164-15-549 *

Also Published As

Publication number Publication date
KR102516279B1 (ko) 2023-04-03

Similar Documents

Publication Publication Date Title
WO2013042900A2 (fr) Nouvelle souche bacillus vallismortis bso7m capable de stimuler la croissance de plante et conférant une résistance au froid à une plante et formulation microbienne comprenant celle-ci
WO2018021797A1 (fr) Souche dr-08 de bacillus methylotrophicus produisant un composé volatil naturel et présentant une activité antibactérienne et utilisation correspondante
WO2020262910A1 (fr) Composition pour prévenir une maladie chez les plantes en utilisant une souche de brevibacillus brevis hk544
WO2024076222A1 (fr) Composition pour réduire la contrainte de sécheresse comprenant une solution de culture de la souche aureobasidium pullulans ak-10 ou un extrait de la solution de culture de la souche, et procédé pour induire la réduction de la contrainte de sécheresse
WO2022114417A1 (fr) Souche de penicillium paxilli à effet de prévention des dommages causés aux plantes par une maladie et des insectes nuisibles, et utilisation associée
Panetsos et al. First analysis on allozyme variation in cedar species (Cedrus sp.)
WO2021162226A1 (fr) Souche bacillus subtilis jck-1398 induisant une résistance chez diverses plantes, et composition et procédé de lutte contre le flétrissement du pin utilisant ladite souche
KR102874731B1 (ko) 스트렙토마이세스 에스피 jck-8055 균주의 배양액 또는 이의 추출물을 포함하는 식물병 방제용 조성물, 이의 제조 방법 및 이를 이용한 방제 방법
WO2018093199A9 (fr) Nouveau micro-organisme bacillus oryzicola yc7011 produisant un lipopeptide cyclique en série de bacillopeptine, et formulation microbienne le comprenant
KR102650962B1 (ko) 스트렙토마이세스 에스피 jck-6131 균주의 배양액 또는 이의 추출물을 포함하는 식물 병해충 방제용 조성물, 이의 제조 방법 및 식물 병해충 방제 방법
WO2023191365A1 (fr) Souche d'irpex lacteus am003, effet de lutte contre la maladie des cultures provoquée par celle-ci, et applications agro-industrielles
WO2024106740A1 (fr) Composition pour lutter contre les champignons phytopathogènes ou réduire les mycotoxines, comprenant une solution de culture de bacillus velezensis jck-7158 ou son extrait, procédé de préparation, et procédé pour lutter contre les champignons phytopathogènes ou réduire les mycotoxines
WO2019117427A1 (fr) Composition comprenant une bactérie flavobacteriaceae trm1-10 ou une souche étroitement liée pour lutter contre une maladie de plante et favoriser la croissance de plante
Zhu et al. Screening, identification, and production application of endophytic Streptomyces W71 from tobacco plants in sanmenxia
WO2023191490A1 (fr) Composition contenant une souche de bacillus amyloliquefaciens pour améliorer la résistance des plantes aux maladies et son utilisation
KR20230172817A (ko) 화상병 또는 가지검은마름병 방제용 박테리오파지 혼합물 FireFighter-A
KR20250035670A (ko) 다양한 식물의 가뭄 스트레스에 저감 활성을 갖는 바실러스 벨레젠시스 ak-408 균주 및 이를 이용한 구상나무와 주목의 가뭄 스트레스 저감용 조성물 및 가뭄 스트레스 저감 유도방법
WO2024106754A1 (fr) Composition pour lutter contre les maladies fongiques des plantes, les maladies bactériennes ou les maladies provoquées par les nématodes, comprenant le fluide de culture de la souche lysobacter enzymogenes jck-1421 ou son extrait, procédé de préparation, et procédé pour lutter contre les maladies fongiques des plantes, les maladies bactériennes ou les maladies provoquées par les nématodes
WO2022005143A1 (fr) Composition pour lutter contre une maladie bactérienne de plante, comprenant un milieu de culture de souche de paenibacillus elgii jck-5075 ou d'un extrait de celle-ci, procédé de production de composition et procédé de lutte contre une maladie bactérienne de plante
KR20230021811A (ko) 애시도보락스 씨트룰라이에 길항력을 가지는 신규한 판토에 키프로페디 균주 및 이의 이용
KR20220167710A (ko) 토마토 시들음병 및 궤양병을 유발하는 클라비박터 미시가넨시스 아종 미시가넨시스에 의한 피해를 감소시키는 바실러스 시아멘시스 k203 및 이의 용도
Mori et al. Bois noir epidemiology and management
WO2023182732A1 (fr) Composition pour lutter contre des virus végétaux comprenant de l'organo-iode ou de l'organo-iode et du soufre en tant que principe(s) actif(s)
CN116035007B (zh) 一种脂肽类化合物kurstakin的应用、生物制剂及其制备方法和应用
KR102675203B1 (ko) 시들음병 또는 궤양병 방제용 바실러스 아리아바타이(Bacillus aryabhattai) H8-1 균주 및 이의 용도

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23875283

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 23875283

Country of ref document: EP

Kind code of ref document: A1