Novel bacillus amyloliquefaciens strain having activity of promoting plant resistance against cold stress and uses thereof

Abstract

The present application relates to a novel Bacillus amyloliquefaciens strain, which enhances the stress resistance of plants and promotes the growth of plants. The Bacillus amyloliquefaciens N484 CP02-1006 strain, the Bacillus amyloliquefaciens N485 CP02-1007 strain, or a culture thereof of the present application can effectively mitigate the reductions in fresh weight, leaf length, and leaf width of plants caused by cold stress.

Description

Novel Bacillus amyloliquefaciens strain having activity for enhancing cold stress resistance of plants and use thereof

[0001] [Cross-reference with related applications]

[0002] This application claims the benefit of priority to Korean Patent Application No. 10-2024-0003016, filed January 8, 2024, the entire contents of which are incorporated herein by reference.

[0003] [Technical Field]

[0004] The present application relates to a novel Bacillus amyloliquefaciens strain having an activity of enhancing cold stress resistance of plants and its use.

[0005] Plants are significantly less able to avoid the various environmental stresses they face during their survival than animals. Therefore, environmental stresses such as low temperature, high temperature, salt, or drought significantly impact plant survival, growth, development, and yield. These environmental stresses are a significant factor that significantly reduces crop production in farms. Furthermore, biotic stresses, such as the onset of plant diseases, can significantly impact crop cultivation.

[0006] Among these, temperature is one of the most important factors that can affect the survival of all plants and serves as a primary driving force for plant growth and development. Low temperature stress is a major factor limiting plant growth and development. It can directly impact plant productivity by inhibiting normal plant growth through reduced photosynthetic rates, nutrient imbalances, secondary metabolic processes, and hormonal imbalances. Therefore, the development of products that minimize cold damage to plants caused by sudden low temperatures is necessary.

[0007] Meanwhile, with growing public concern over the adverse effects of pesticides, demand is growing for eco-friendly products that can replace chemical fertilizers and pesticides. Therefore, there is a pressing need to develop eco-friendly products that can enhance plant resistance to low-temperature stress and thereby promote plant production.

[0008] Korean Patent Publication No. 10-2023-0109295 discloses a Streptomyces anandii J6 strain, which has the property of conferring environmental stress tolerance to plants, and this strain is disclosed to confer tolerance to high temperature, drought, and low temperature stresses on grasses. Korean Patent Registration No. 10-2411304 discloses a Bacillus zanthoxyli HS1 strain, which is excellent in enhancing plant resistance to various stresses. Korean Patent Registration No. 10-2125456 discloses a Bacillus siamensis H30-3 strain, which enhances plant resistance to biotic and abiotic stresses.

[0009] The purpose of this application is to provide a novel strain that enhances stress resistance of plants and promotes plant growth.

[0010] In addition, the present application aims to provide a composition for enhancing stress resistance of plants.

[0011] In addition, the present application aims to provide a composition for promoting plant growth.

[0012] In addition, the present application aims to provide a method for enhancing stress resistance of plants.

[0013] In addition, the purpose of this application is to provide a method for promoting plant growth.

[0014] Each description and embodiment disclosed in this application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application is not limited by the specific descriptions set forth below. Furthermore, those skilled in the art will recognize or be able to ascertain, through routine experimentation alone, numerous equivalents to the specific embodiments of this application described in this application. Furthermore, such equivalents are intended to be encompassed by this application.

[0015] To achieve the above purpose, one aspect of the present application provides a Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the deposit number KCCM13395P.

[0016] In addition, another aspect of the present application provides Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under accession number KCCM13396P.

[0017] In addition, another aspect of the present application provides a composition for enhancing stress resistance of a plant, comprising as an active ingredient at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0018] In addition, another aspect of the present application provides a composition for promoting plant growth, comprising as an active ingredient at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0019] In addition, another aspect of the present application provides a method for enhancing stress resistance of a plant, comprising the step of treating a plant, an area around a plant, or both, with at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0020] In addition, another aspect of the present application provides a method for promoting plant growth, comprising the step of treating a plant, an area around a plant, or both, with at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with Accession No. KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with Accession No. KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0021]

[0022] The present application is described in detail below.

[0023]

[0024] One aspect of the present application provides Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under accession number KCCM13395P.

[0025] In addition, another aspect of the present application provides Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under accession number KCCM13396P.

[0026] The Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the above-mentioned accession number KCCM13395P was internationally deposited with the Korea Microbiological Conservation Center, an international depository under the Budapest Treaty, and was assigned the accession number KCCM13395P on September 21, 2023. In the present specification, the strain may also be used interchangeably as the Bacillus amyloliquefaciens N484 CP02-1006 strain, the N484 CP02-1006 strain, the N484 strain, or the CP02-1006 strain.

[0027] The Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the above-mentioned accession number KCCM13395P may include a nucleotide sequence of a 16S rRNA gene having a sequence identity of at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% with the nucleotide sequence of SEQ ID NO: 1. Specifically, the N484 CP02-1006 strain may include a nucleotide sequence of a 16S rRNA gene including the nucleotide sequence of SEQ ID NO: 1.

[0028] The Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the above-mentioned accession number KCCM13396P was internationally deposited with the Korea Microbiological Conservation Center, an international depository under the Budapest Treaty, and was assigned the accession number KCCM13396P on September 21, 2023. In the present specification, the strain may also be used interchangeably as the Bacillus amyloliquefaciens N485 CP02-1007 strain, the N485 CP02-1007 strain, the N485 strain, or the CP02-1007 strain.

[0029] The Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the above-mentioned accession number KCCM13396P may include a nucleotide sequence of a 16S rRNA gene having a sequence identity of at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% with the nucleotide sequence of SEQ ID NO: 2. Specifically, the N485 CP02-1007 strain may include a nucleotide sequence of a 16S rRNA gene including the nucleotide sequence of SEQ ID NO: 2.

[0030] In one embodiment, the N484 CP02-1006 strain and / or the N485 CP02-1007 strain can enhance stress resistance of a plant and / or promote plant growth.

[0031] In one embodiment, the stress may be low temperature stress. Low temperature stress can inhibit normal plant growth by causing a decrease in the plant's photosynthetic rate, nutrient imbalance, secondary metabolic processes, and hormonal imbalance.

[0032] In the present application, the low temperature stress may be a temperature condition of -20°C to 20°C. For example, the low temperature stress may be a temperature condition of -20°C to -12°C, -20°C to -4°C, -20°C to 8°C, -20°C to 12°C, -12°C to -4°C, -12°C to 4°C, -12°C to 12°C, -12°C to 20°C, -4°C to 4°C, -4°C to 12°C, -4°C to 20°C, 4°C to 12°C, 4°C to 20°C, or 12°C to 20°C, but is not limited thereto.

[0033] In this application, the term “plant” includes organisms belonging to the plant kingdom that perform photosynthesis, and is not taxonomically limited to a specific plant.

[0034] In the present application, the "plant" may be at least one selected from the group consisting of Cucurbitaceae, Brassicaceae, Amaranthaceae, Amaryllidaceae, Apiaceae, Asparagusaceae, Asteraceae, Pinaceae, Convolvulaceae, Leguminosae, Malvaceae, Muscariaceae, Poaceae, Polygonaceae, Rosaceae, Rubiaceae, Solanaceae, and Grapeaceae.

[0035] The above "Cucurbitaceae" plants may include, for example, cucumbers, melons, watermelons, cantaloupes, honeydews, melons, squashes, zucchinis, and pumpkins, and specifically, the above cucurbitaceae plants may be, but are not limited to, cucumbers, melons, watermelons, or melons.

[0036] The above "Brassicaceae" plants may include, for example, cabbage, arugula, broccoli, bok choy, Brussels sprouts, cabbage, cauliflower, canola, collard greens, white radish, Chinese cabbage, horseradish, kale, mustard, radish, rapeseed, rutabaga, turnip, wasabi, watercress, and Arabidopsis thaliana, and specifically, the above cruciferous plants may be, but are not limited to, cabbage, radish, mustard, rapeseed, or Arabidopsis thaliana.

[0037] The above "Amaranthaceae" plants may include, but are not limited to, for example, scallions, spinach, sugar beets, and quinoa.

[0038] The above "Amaryllidaceae" plants may include, but are not limited to, chives, onions, garlic, chives, leeks, and shallots.

[0039] The above "Apiaceae" plants may include, but are not limited to, anise, caraway, carrot, celery, chervil, coriander, cumin, dill, fennel, parsley, and parsnips.

[0040] The above "Asparagaceae" plants may include, but are not limited to, agave and asparagus.

[0041] The above "Asteraceae" plants may include, but are not limited to, artichokes, aster, chamomile, chicory, chrysanthemums, dahlias, daisies, echinacea, chrysanthemums, guajul, lettuce, marigolds, safflowers, sunflowers, and zinnias.

[0042] The above "Bromeliaceae" plants may include, but are not limited to, pineapples, etc.

[0043] The above "Convolvulaceae" plants may include, but are not limited to, morning glory, and sweet potato.

[0044] The above "Fabaceae" plants may include, but are not limited to, alfalfa, beans, carob, clover, guar, lentils, mesquite, peas, peanuts, soybeans, tamarind, tragacanth, and vetch.

[0045] The above "Malvaceae" plants may include, but are not limited to, cacao, cotton, durian, hibiscus, kenaf, cola, and okra.

[0046] The above "Musaceae" plants may include, but are not limited to, bananas and plantains.

[0047] The above "Poaceae" plants may include, but are not limited to, bamboo, barley, corn, fonio, millet, oats, ornamental grasses, rice, rye, sorghum, sugarcane, triticale, wheat, and grasses such as bahia grass, umbrella grass, bluegrass, buffalo grass, centipede grass, fescue, or zoysia.

[0048] The above "Polygonaceae" plants may include, but are not limited to, buckwheat, etc.

[0049] The above "Rosaceae" plants may include, but are not limited to, almonds, apples, apricots, blackberries, blueberries, cherries, peaches, plums, quinces, raspberries, roses, and strawberries.

[0050] The above "Rubiaceae" plants may include, but are not limited to, coffee, etc.

[0051] The above "Solanaceae" plants may include, but are not limited to, bell peppers, peppers, eggplants, petunias, potatoes, tobacco, and tomatoes.

[0052] The above "Vitaceae" plants may include, but are not limited to, grapes, for example.

[0053] In the present application, the term “plant growth” refers to the process in which a plant seed germinates and produces roots, stems, leaves, and flowers, or thereafter gradually increases in size, weight, and quantity, and the term “promotion of plant growth” refers to promoting the germination of a plant seed or an increase in the size and / or weight of a germinated plant.

[0054] According to one embodiment, the N484 CP02-1006 strain and / or the N485 CP02-1007 strain may have an activity of enhancing stress resistance of a plant and / or promoting growth of a plant. Specifically, when cucumber (Cucumis sativus) is grown under low temperature stress conditions, the fresh weight, leaf length, and leaf width of the cucumber are all reduced, but when the N484 CP02-1006 strain or the N485 CP02-1007 strain is treated to the cucumber, the reduced fresh weight, leaf length, and leaf width of the cucumber can all be recovered. For example, the N484 CP02-1006 strain or the N485 CP02-1007 strain can increase the fresh weight of cucumbers reduced by low temperature stress by 5 to 30%, increase leaf length by 5 to 20%, and increase leaf width by 2 to 10%. That is, the N484 CP02-1006 strain or the N485 CP02-1007 strain can enhance plant resistance to stress and promote plant growth, and thus can be usefully utilized for the purpose of enhancing plant stress resistance and promoting plant growth.

[0055] According to another aspect of the present application, a bacterial composition comprising the Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the above-mentioned deposit number KCCM13395P or the Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under the above-mentioned deposit number KCCM13396P is provided.

[0056] In one embodiment, the composition may further comprise a cryoprotectant.

[0057] The Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the deposit number KCCM13395P of the present application or the Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under the deposit number KCCM13396P can be prepared in the form of a lyophilized composition to preserve the activity of the microbial cells, and the composition can further include a cryoprotectant to prevent damage to the cells.

[0058] In one embodiment, the cryoprotectant may include one or more selected from the group consisting of glycerol, ethylene glycol, propylene glycol, trehalose, sucrose, glucose, maltodextrin, sorbitol, skimmed milk powder, and starch.

[0059] In one embodiment, the cryoprotectant may be present in an amount of 0.01 to 30 wt% relative to the total composition, specifically 0.01 to 30 wt%, 0.01 to 25 wt%, 0.01 to 20 wt%, 0.01 to 15 wt%, 0.1 to 30 wt%, 0.1 to 25 wt%, 0.1 to 20 wt%, 0.1 to 15 wt%, 1 to 30 wt%, 1 to 25 wt%, 1 to 20 wt%, or 1 to 15 wt% relative to the total composition.

[0060] In one embodiment, the strain comprises 10 5 May contain more than CFU / mL, specifically 1 x 10 5 CFU / mL to 1 x 10 6 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 11 CFU / mL, 10 5 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 6 CFU / mL to 1 x 1011 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 10 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 10 CFU / mL to 1 x 10 12 It can be included in the amount of CFU / mL.

[0061] In one embodiment, the bacterial composition may be in a lyophilized form.

[0062] In one embodiment, the bacterial composition may be in the form of a lyophilized powder.

[0063]

[0064] Another aspect of the present application provides a composition for enhancing stress resistance of plants.

[0065] The composition for enhancing stress resistance of the plant of the present application comprises, as an active ingredient, at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0066] In this application, the "culture solution" refers to a product produced by culturing the strain, and the culture solution may include the culture solution itself in which the strain is cultured, the culture filtrate obtained by filtering or centrifuging the culture solution to remove the strain, and the supernatant obtained by centrifuging the culture solution or culture filtrate.

[0067] In this application, the "concentrate" refers to a product obtained by concentrating the culture solution, and the "extract" refers to a product obtained by extracting the culture solution using an appropriate solvent according to a known method. The "dry product" refers to a product obtained by removing moisture from the culture solution or concentrate, and may be, for example, a dried cell form of the strain, but is not limited thereto.

[0068] In addition, the above "pulverized material" means the result of pulverizing the dried material from which the moisture of the culture medium or the cells of the strain has been removed, and may be, for example, a dried cell powder of the strain, but is not limited thereto.

[0069] The culture solution of the above strain can be prepared by inoculating the strain into a culture medium and according to a culture method known in the art, and the concentrate, extract, dried product or pulverized product can be prepared according to a treatment or drying method of the strain or its culture solution known in the art.

[0070] In one embodiment, the stress in the composition for enhancing stress resistance of the plant may be low temperature stress.

[0071] In one embodiment, in the composition for enhancing stress resistance of the plant, the plant may be at least one selected from the group consisting of Cucurbitaceae, Brassicaceae, Amaranthaceae, Amaryllidaceae, Apiaceae, Asparagusaceae, Asteraceae, Pinaceae, Convolvulaceae, Leguminosae, Malvaceae, Muscariaceae, Poaceae, Polygonaceae, Rosaceae, Rubiaceae, Solanaceae, and Grapeaceae.

[0072] According to one embodiment, at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the deposit number KCCM13395P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product can effectively increase the reduction in fresh weight, leaf length, and leaf width of a plant induced by low temperature stress, and thus can be usefully utilized for the purpose of enhancing stress resistance of a plant.

[0073] In addition, according to one embodiment, at least one selected from the group consisting of Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product can effectively alleviate a decrease in fresh weight, leaf length, and leaf width of a plant caused by low temperature stress, and thus can be usefully utilized for the purpose of enhancing stress resistance of a plant.

[0074] In one embodiment, the concentration of the Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P in the composition for enhancing stress resistance of the plant is 1 x 10 5 CFU / mL to 1 x 10 12 CFU / mL. For example, the concentration of the strain may be 1 x 10 5 CFU / mL to 1 x 10 6 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 6 CFU / mL to 1 x 109 CFU / mL, 1 x 10 6 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 6 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 8 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 9 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 9 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 9 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 9 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 9 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 10 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 10 CFU / mL 내지 1 x 10 12 CFU / mL, 또는 1 x 10 11CFU / mL to 1 x 10 12 It can be CFU / mL.

[0075] In addition, in one embodiment, the concentration of the Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P in the composition for enhancing stress resistance of the plant is 1 x 10 5 CFU / mL to 1 x 10 12 CFU / mL. For example, the concentration of the strain may be 1 x 10 5 CFU / mL to 1 x 10 6 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 8CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 10 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 10 CFU / mL to 1 x 10 12 CFU / mL, or 1 x 10 11 CFU / mL to 1 x 10 12 It can be CFU / mL.

[0076] In one embodiment, when the composition for enhancing stress resistance of a plant is in a solid state rather than a liquid state, the unit "CFU / mL" used to describe the concentration of the strain may be changed to "CFU / g".

[0077] In the present application, the composition for enhancing stress resistance of the plant may be prepared in the form of a directly sprayable solution, powder, and suspension, or as a highly concentrated aqueous, oily, or other suspension, dispersion, emulsion, oily dispersion, paste, dust, dustable material, or granule, but is not limited thereto.

[0078] The composition for enhancing stress resistance of plants in the present application can be formulated in various forms. For example, the formulation can be prepared by adding a solvent and / or carrier, and inert additives and surface-active substances, such as emulsifiers or dispersants, can be mixed into the formulation. Suitable surface-active substances are aromatic sulfonic acids (e.g. lignosulfonic acid, phenolsulfonic acid, naphthalene- and dibutylnaphthalenesulfonic acid), fatty acids, alkyl- and alkylarylsulfonates, alkyl lauryl ethers, alkali metal, alkaline earth metal and ammonium salts of fatty alcohol sulfates, sulfated hexa-, hepta- and octadecanol, salts of fatty alcohol glycol ethers, sulfonated naphthalene and derivatives thereof, condensates with formaldehyde, condensates of naphthalene or naphthalenesulfonic acid, phenol and formaldehyde, polyoxyethyleneoctyl phenol ethers, ethoxylated isooctyl-, octyl- or nonylphenols, alkylphenyl or tributylphenyl polyglycol ethers, alkylarylpolyether alcohols, isotridecyl alcohol, fatty alcohol / ethylene oxide condensates, ethoxylated castor oil, It may be, but is not limited to, polyoxyethylene alkyl ether or polyoxypropylene, lauryl alcohol polyglycol ether acetate, sorbitol ester, lignin-sulfite waste liquor or methylcellulose.

[0079] The carrier may be, but is not limited to, an agriculturally acceptable carrier, such as mineral earth (e.g., silica, silica gel, silicate, talc, kaolin, limestone, lime, chalk, boll, loess, clay, dolomite, diatomaceous earth, calcium sulfate, magnesium sulfate, magnesium oxide, ground synthetic material), fertilizer (e.g., ammonium sulfate, ammonium phosphate, ammonium nitrate, urea), plant products (e.g., grain flour, bark flour, wood meal, and nut shell flour), or cellulose powder, and the carrier may be used alone or in combination of two or more.

[0080] The composition for enhancing stress resistance in plants in the present application may additionally include any other ingredient used in the art to enhance stress resistance in plants. For example, the composition may additionally include at least one selected from the group consisting of plant growth regulators, plant growth stimulants, and plant nutrients.

[0081] The plant growth regulator, plant growth stimulant or plant nutrient may be another strain of a chemical or bacteria, and may be a herbicide, fungicide, insecticide, fertilizer or any combination thereof.

[0082] In the present application, the composition for enhancing stress resistance of the plant may additionally include at least one selected from the group consisting of other beneficial microorganisms, microbial activators, organic fertilizers, and chemical fertilizers.

[0083] Other beneficial microorganisms may include Bacillus spp., Azotobacter spp., Trichoderma spp., and Saccharomyces spp., which are known to enhance stress resistance in plants.

[0084] The above microbial activators may include enzyme precursors, microbial metabolites, organic acids, carbohydrates, enzymes, and trace elements. For example, the microbial activators may include processed enzyme products such as yeast autolysates, humic materials, seaweed extracts, starch, amino acids, and trace elements such as zinc, iron, copper, manganese, bromine, and molybdenum.

[0085] The above chemical fertilizers may include various chemicals that can provide nitrogen, phosphorus, and potassium nutrients to help increase plant stress resistance. For example, chemical fertilizers may include urea, calcium phosphate, potassium phosphate, and mixed nitrogen-phosphate-potassium (NPK) fertilizers, as well as other ingredients known in the art.

[0086] In the present application, the composition for enhancing stress resistance of the plant can be mixed with a dispersing agent, a penetrating agent, or a surfactant to increase absorption and effectiveness of the plant.

[0087] In the present application, the composition for enhancing stress resistance of the plant may additionally include a preservative, stabilizer, wetting agent, emulsifier, cryoprotectant, or excipient.

[0088] For example, the composition for enhancing stress resistance of the plant may include an effective amount of a preservative, stabilizer, or excipient sufficient to reduce quality deterioration of the composition, thereby allowing the N484 CP02-1006 strain and / or the N485 CP02-1007 strain to survive or remain active for a longer period of time in the composition, unlike the N484 CP02-1006 strain and / or the N485 CP02-1007 strain that exist in nature.

[0089] The preservative, stabilizer, or excipient may be any one commonly used in the art without limitation, as long as it can reduce the deterioration of the quality of the composition for enhancing stress resistance of the plant. For example, the stabilizer may be at least one selected from the group consisting of alginate, sodium alginate, casein, sodium caseinate, cellulose, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, pectin, gelatin, gum arabic, xanthan gum, dextran, cyclodextrin, and starch. The excipient may include a pharmaceutically acceptable solid carrier, liquid carrier, liquid diluent, liquefied gas diluent, solid diluent, or other auxiliary agent, for example, a surfactant such as an emulsifier, a dispersant, or a foaming agent.

[0090] The cryoprotectant may be included in the composition in an effective amount sufficient to reduce quality deterioration of the composition when the composition for enhancing stress resistance of the plant is in a freeze-dried state. For example, unlike the N484 CP02-1006 strain and / or the N485 CP02-1007 strain existing in a natural state, the composition may include an effective amount of the cryoprotectant sufficient to reduce quality deterioration of the composition in a freeze-dried state, thereby allowing the N484 CP02-1006 strain and / or the N485 CP02-1007 strain to survive or remain active for a longer period of time in the freeze-dried composition.

[0091] As the cryoprotectant, any agent commonly used in the art may be used without limitation, as long as it is intended to reduce quality deterioration of the composition in a lyophilized state. For example, the cryoprotectant may be at least one selected from the group consisting of glycerol, ethylene glycol, propylene glycol, dimethyl sulfoxide (DMSO), glucose, trehalose, maltodextrin, dextrin, skim milk, and starch.

[0092] In one embodiment, the composition for enhancing stress resistance of the plant may be in the form of a freeze-dried powder.

[0093] In one embodiment, the composition for enhancing stress resistance of a plant can be added to a plant fertilizer by immersion, spraying, or mixing.

[0094] The above plant fertilizer can be manufactured by a manufacturing method commonly used in the art. For example, the plant fertilizer can further include excipients or carriers commonly used in the art, and can be manufactured in a formulation of a conventional plant fertilizer known in the art, such as a wettable powder, granule, powder, emulsion, spray, mist, capsule, or gel.

[0095] In one embodiment, the plant fertilizer may be an eco-friendly fertilizer that can replace chemical fertilizers, and the plant fertilizer may be used to enhance stress resistance of plants.

[0096]

[0097] Another aspect of the present application provides a method for enhancing stress resistance of plants.

[0098] The method for enhancing stress resistance of the plant of the present application comprises a step of treating a plant, an area around the plant, or both of the plant, with at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with Accession No. KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with Accession No. KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0099] In the present application, the plant may include any plant part or type, including roots, stems, leaves, seeds, fruits, or any part or combination thereof. Specifically, the plant may be a plant capable of growing under stress conditions or may be a plant individual grown under stress conditions, but is not limited thereto.

[0100] In this application, the "area surrounding a plant" refers to the range within which the active ingredient of this application can exert its effect, and includes both above-ground and underground areas. For example, it may be an area within about 2 m, about 1 m, about 70 cm, about 50 cm, about 25 cm, about 10 cm, or about 5 cm around a plant, plant part, or fruit.

[0101] According to one embodiment, the method for enhancing stress resistance of a plant can effectively alleviate the decrease in fresh weight, leaf length and leaf width of a plant caused by low temperature stress, thereby effectively enhancing stress resistance of the plant.

[0102] At least one treatment selected from the group consisting of the strain, the culture solution of the strain, the concentrate of the culture solution, the extract of the culture solution, the dried product of the culture solution, and the pulverized product of the dried product may be applied to the plant by a method such as immersion, irrigation, spraying, injection, infusion, mixing, or mixing with soil, but is not limited thereto as long as it can directly or indirectly contact the inside or outside of the plant. In addition, the treatment may be performed simultaneously or at the same time as the plant grows under stress conditions, but is not limited thereto. For example, the treatment may be performed once or twice or more, but is not limited thereto. In addition, when the treatment is performed twice or more, the treatment may be performed at intervals of 3 to 15 days, 3 to 10 days, 3 to 7 days, 4 to 7 days, 5 to 7 days, or 6 days, but is not limited thereto.

[0103] Another aspect of the present application provides a use for enhancing stress resistance of a plant, comprising a composition comprising at least one selected from the group consisting of Bacillus amyloliquefaciens strain N484 CP02-1006 deposited under accession number KCCM13395P, or Bacillus amyloliquefaciens strain N485 CP02-1007 deposited under accession number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0104]

[0105] Another aspect of the present application provides a composition for promoting plant growth.

[0106] The composition for promoting plant growth of the present application comprises, as an active ingredient, at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0107] In one embodiment, in the composition for promoting plant growth, the plant may be at least one selected from the group consisting of Cucurbitaceae, Brassicaceae, Amaranthaceae, Amaryllidaceae, Apiaceae, Asparagusaceae, Asteraceae, Pinaceae, Convolvulaceae, Leguminosae, Malvaceae, Muscariaceae, Poaceae, Polygonaceae, Rosaceae, Rubiaceae, Solanaceae, and Grapeaceae.

[0108] According to one embodiment, at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the deposit number KCCM13395P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product can effectively increase a decrease in fresh weight, leaf length, and leaf width of a plant caused by low temperature stress, and thus can be usefully utilized for the purpose of promoting plant growth.

[0109] In addition, according to one embodiment, at least one selected from the group consisting of Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product can effectively increase the reduction in fresh weight, leaf length, and leaf width of a plant caused by low temperature stress, and thus can be usefully utilized for the purpose of promoting plant growth.

[0110] In one embodiment, the concentration of the Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P in the composition for promoting the growth of the plant is 1 x 10 5 CFU / mL to 1 x 10 12 CFU / mL. For example, the concentration of the strain may be 1 x 10 5 CFU / mL to 1 x 10 6 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 9CFU / mL, 1 x 10 6 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 6 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 8 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 9 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 7 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 9 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 8 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 9 CFU / mL 내지 1 x 10 10 CFU / mL, 1 x 10 9 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 9 CFU / mL 내지 1 x 10 12 CFU / mL, 1 x 10 10 CFU / mL 내지 1 x 10 11 CFU / mL, 1 x 10 10 CFU / mL 내지 1 x 10 12 CFU / mL, 또는 1 x 10 11 CFU / mL 내지 1 x 1012 It can be CFU / mL.

[0111] In addition, in one embodiment, the concentration of the Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P in the composition for promoting the growth of the plant is 1 x 10 5 CFU / mL to 1 x 10 12 CFU / mL. For example, the concentration of the strain may be 1 x 10 5 CFU / mL to 1 x 10 6 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 5 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 7 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 6 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 8 CFU / mL, 1 x 10 7CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 7 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 9 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 8 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 10 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 9 CFU / mL to 1 x 10 12 CFU / mL, 1 x 10 10 CFU / mL to 1 x 10 11 CFU / mL, 1 x 10 10 CFU / mL to 1 x 10 12 CFU / mL, or 1 x 10 11 CFU / mL to 1 x 10 12 It can be CFU / mL.

[0112] In one embodiment, when the composition for promoting plant growth is in a solid state rather than a liquid state, the unit "CFU / mL" used to describe the concentration of the strain may be changed to "CFU / g".

[0113] In the present application, the composition for promoting plant growth may be prepared in the form of a directly sprayable solution, powder, and suspension, or a highly concentrated aqueous, oily, or other suspension, dispersion, emulsion, oily dispersion, paste, dust, sprinkling material, or granule, but is not limited thereto, and may be formulated in various forms.

[0114] The plant growth promoting composition in the present application may additionally include any other ingredient used in the art to enhance stress resistance in plants. For example, the composition may additionally include at least one selected from the group consisting of plant growth regulators, plant growth stimulants, and plant nutrients.

[0115] In the present application, the composition for promoting plant growth may additionally include at least one selected from the group consisting of other beneficial microorganisms, microbial activators, organic fertilizers, and chemical fertilizers.

[0116] In the present application, the composition for promoting plant growth can be mixed with a dispersing agent, a penetrating agent, or a surfactant to increase absorption and effectiveness of the plant.

[0117] In the present application, the composition for promoting plant growth may additionally include a preservative, stabilizer, wetting agent, emulsifier, cryoprotectant, or excipient.

[0118] In one embodiment, the plant growth promoting composition can be added to plant fertilizer by immersion, spraying or mixing.

[0119] In one embodiment, the plant fertilizer may be an environmentally friendly fertilizer that can replace chemical fertilizers, and the plant fertilizer may be used to promote plant growth.

[0120]

[0121] Another aspect of the present application provides a method for promoting plant growth.

[0122] The method for promoting plant growth of the present application includes a step of treating a plant, an area around the plant, or both of the plant, with at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited with the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited with the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0123] In the present application, the plant may include any plant part or type, including roots, stems, leaves, seeds, fruits, or any part or combination thereof. Specifically, the plant may be a plant individual requiring growth promotion, but is not limited thereto.

[0124] According to one embodiment, the method for promoting plant growth can effectively increase the reduction in the fresh weight, leaf length and leaf width of the plant, thereby effectively promoting plant growth.

[0125] At least one treatment selected from the group consisting of the strain, the culture solution of the strain, the concentrate of the culture solution, the extract of the culture solution, the dried product of the culture solution, and the pulverized product of the dried product may be performed by immersing, irrigating, spraying, injecting, pouring, mixing, or mixing with soil, but is not limited thereto as long as it can directly or indirectly contact the inside or outside of the plant. For example, the treatment may be performed once or twice or more, but is not limited thereto. In addition, when the treatment is performed twice or more, the treatment may be performed at intervals of 3 to 15 days, 3 to 10 days, 3 to 7 days, 4 to 7 days, 5 to 7 days, or 6 days, but is not limited thereto.

[0126] Another aspect of the present application provides a use for promoting plant growth of a composition comprising at least one selected from the group consisting of Bacillus amyloliquefaciens strain N484 CP02-1006 deposited under accession number KCCM13395P, or Bacillus amyloliquefaciens strain N485 CP02-1007 deposited under accession number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

[0127] The Bacillus amyloliquefaciens N484 CP02-1006 strain, the Bacillus amyloliquefaciens N485 CP02-1007 strain, or a culture thereof of the present application can effectively restore the reduction in fresh weight, leaf length, and leaf width of plants caused by low temperature stress. Therefore, the strain of the present application or the culture thereof can be usefully utilized as an eco-friendly product that can promote plant production by enhancing plant resistance to low temperature stress.

[0128] However, the effects of the present application are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below.

[0129] Figure 1 shows the results of measuring the fresh weight of cucumbers after treating them with a culture solution of Bacillus amyloliquefaciens N484 CP02-1006 strain or a culture solution of Bacillus amyloliquefaciens N485 CP02-1007 strain under low temperature stress conditions.

[0130] Figure 2 shows the results of measuring the leaf length of cucumbers after treating them with a culture solution of Bacillus amyloliquefaciens N484 CP02-1006 strain or a culture solution of Bacillus amyloliquefaciens N485 CP02-1007 strain under low temperature stress conditions.

[0131] Figure 3 shows the results of measuring the leaf width of cucumbers after treating them with a culture solution of Bacillus amyloliquefaciens N484 CP02-1006 strain or a culture solution of Bacillus amyloliquefaciens N485 CP02-1007 strain under low temperature stress conditions.

[0132] Hereinafter, the present application will be described in detail by way of examples. However, the following examples are intended to specifically illustrate the present application, and the contents of the present application are not limited by the following examples.

[0133]

[0134] Example 1. Isolation and cultivation of Bacillus amyloliquefaciens strains

[0135] The strains used in this experiment were obtained from field soil. Specifically, approximately 50 g of soil was collected from a depth of 5 to 10 cm from the ground, and the collected soil was sieved through a 2 mm mesh sieve to remove large stones and foreign substances. Each 5 g of the sieved soil sample was cultured in a flask, and the cultured solution was diluted 1 / 10 in sterile distilled water to isolate the microorganisms.

[0136] Among the isolated microorganisms, six strains with high antagonism against representative plant pathogens, Rhizoctonia solani (Rs), Pythium ultimum (Pu), and Colletotrichum truncatum (Ct), were selected. To confirm the bactericidal effect against plant pathogens, four paper discs were first placed in an 8.5 cm diameter Petri dish containing PDA (potato dextrose agarose) medium, and three paper discs were inoculated with the standard strain, Bacillus amyloliquefaciens KACC 10116 strain, Serenade® (Bayer) (B. velezensis QST713 strain), and the strain to be evaluated, respectively. The remaining paper disc was used as an untreated control. The three plant pathogens (Rs, Pu, Ct) were cultured on potato dextrose agarose (PDA) medium at 28°C for 5 to 8 days, and then the pathogen colonies were spread on the center of a petri dish using a 5 mm diameter cork borer. After culturing in an incubator at 25°C for 7 to 20 days, the diameter of the zone of inhibition was examined to compare the antibacterial effects of the Bacillus strains. Finally, six strains with antibacterial effects equal to or greater than those of Serenade were selected (Table 1).

[0137] Antibacterial effect (unit: mm) Rhizoctonia solani Phythium ultium Colletotrichum trucatum N31 19.73.712.7 N47 410.33.311 N48 19.35.012.3 N48 48.71.011.7 N48 54.34.76.7 N49 09.74.010.7 serenade 9.74.011.7

[0138]

[0139] To identify the six strains selected as described above, base sequence analysis of the 16S rRNA gene was performed. Identification was performed by analyzing sequence identity with the previously reported Bacillus amyloliquefaciens standard strain KACC 10116 using the CLC genomics workbench 21 program (Qiagen). As a result of the analysis, each of the six strains was confirmed to have 99% sequence homology with the above standard strain, and were finally identified as Bacillus amyloliquefaciens strains: Bacillus amyloliquefaciens N474 CP02-1004 strain (accession number KCCM13393P), Bacillus amyloliquefaciens N481 CP02-1005 strain (accession number KCCM13394P), Bacillus amyloliquefaciens N484 CP02-1006 strain (accession number KCCM13395P), Bacillus amyloliquefaciens N485 CP02-1007 strain (accession number KCCM13396P), Bacillus amyloliquefaciens N490, and Bacillus amyloliquefaciens N311. Each identified strain was suspended in 20% glycerol and stored.

[0140] Among the six strains finally selected, strains N484 and N485 were found to have the best cold stress alleviation efficacy in follow-up experiments. The N484 strain was named Bacillus amyloliquefaciens N484 CP02-1006 and was internationally deposited with the Korea Microbiological Conservation Institute, an international depository under the Budapest Treaty, and was assigned the accession number KCCM13395P on September 21, 2023. In addition, the N485 strain was named Bacillus amyloliquefaciens N485 CP02-1007 and was internationally deposited with the Korea Microbiological Conservation Institute, an international depository under the Budapest Treaty, and was assigned the accession number KCCM13396P on September 21, 2023.

[0141] The nucleotide sequences of the 16s rRNA genes of the N484 and N485 strains are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

[0142] The six Bacillus amyloliquefaciens strains selected and identified as described above and the Bacillus amyloliquefaciens strain isolated from Serenade (Bayer) as a control were cultured. One colony of the Bacillus amyloliquefaciens strain was streaked on an LB (Luria-Bertani broth) medium and cultured for primary culture in an incubator at 28°C for 20 hours. A colony was picked from the primary culture medium and streaked on an LB medium for a second time, and cultured for secondary culture in an incubator at 28°C for 20 hours. Afterwards, 10 mL of sterilized distilled water was added to the secondary culture medium to perform cell resuspension, and the cells were recovered using a spreader. After measuring the OD (optical density) value of each strain, it was diluted to OD = 1.0 and used in the experiment.

[0143]

[0144] Example 2. Evaluation of the effect of Bacillus amyloliquefaciens strains on alleviating plant cold stress.

[0145] An experiment was conducted to confirm the effect of the six Bacillus amyloliquefaciens strains cultured in Example 1 above on alleviating plant low-temperature stress.

[0146]

[0147] 2-1. Experimental method

[0148] Cucumber (Cucumis sativus) was used as the plant, and cucumber seeds were sown in a 7:3 mixture of horticultural soil and vermiculite. The normal growth conditions for cucumbers were set as temperature 25℃, humidity 60-65%, and photoperiod 16h / 8h (day / night), and the low temperature stress growth conditions were set as temperature 10℃, humidity 60-65%, and photoperiod 16h / 8h (day / night). The control group was set as normal condition + DW treatment group, low temperature stress condition + DW treatment group, and low temperature stress condition + Bacillus amyloliquefaciens strain culture solution treatment group isolated from Serenade (Bayer Co., Ltd.), which is the product control group. The test group was set as low temperature stress condition + culture solution treatment group of six Bacillus amyloliquefaciens strains cultured in Example 1.

[0149] 10 days after sowing cucumbers, the Bacillus amyloliquefaciens strain culture was cultured to an OD = 1.0 value (OD = 1.0, 1Х10 8 CFU / ml) was applied twice at 5 mL / plant at 7-day intervals to the cucumber leaves. One day after the second application, the cucumbers were subjected to cold stress. After growing the cucumbers under cold stress conditions for 7 days, the fresh weight, leaf length, and leaf width of the cucumbers were measured and compared. Specifically, the fresh weight was measured as the total weight of the above-ground part of the cucumber, and the leaf length and leaf width were measured as the leaf length and leaf width of the largest leaf in the individual, respectively.

[0150]

[0151] 2-2. Live weight measurement results

[0152] As a result of measuring fresh weight, when cold stress was treated, the fresh weight of cucumbers was significantly reduced compared to normal conditions, but the fresh weight decrease caused by cold stress was recovered in the Bacillus amyloliquefaciens N484 CP02-1006 strain treatment group and the N485 CP02-1007 strain treatment group (Fig. 1). Specifically, the fresh weight of the N484 strain treatment group increased by approximately 20.4% and 17.7% compared to the cold stress + DW treatment group and the cold stress + Serenade treatment group, and the fresh weight of the N485 strain treatment group increased by approximately 9.8% and 7.3% compared to the cold stress + DW treatment group and the cold stress + Serenade treatment group, respectively. In addition, the fresh weights of the other strains N474, N481, N490, and N311 were lower than or equal to those of the untreated group or the Serenade treatment group.

[0153]

[0154] 2-3. Leaf length measurement results

[0155] As a result of measuring leaf length, when cold stress was applied, the leaf length of cucumbers was significantly reduced compared to normal conditions, but the reduction in leaf length caused by cold stress was recovered in the Bacillus amyloliquefaciens N484 strain treatment group and the N485 strain treatment group (Fig. 2). Specifically, the N484 strain treatment group showed an increase in leaf length of approximately 13.3% and 12.8% compared to the cold stress + DW treatment group and the cold stress + Serenade treatment group, and the N485 strain treatment group showed an increase in leaf length of approximately 6.9% and 6.4% compared to the cold stress + DW treatment group and the cold stress + Serenade treatment group, respectively. In addition, the N474, N481, N490, and N311 strain treatment groups showed leaf lengths that were lower or equivalent to those of the untreated group or the Serenade treatment group.

[0156]

[0157] 2-4. Leaf width measurement results

[0158] As a result of measuring leaf width, when cold stress was treated, the leaf width of cucumbers was significantly reduced compared to normal conditions, but the leaf width reduction caused by cold stress was recovered in the Bacillus amyloliquefaciens N484 strain treatment group and the N485 strain treatment group (Fig. 3). Specifically, the leaf width of the N484 strain treatment group increased by approximately 3.5% and 5.6% compared to the cold stress + DW treatment group and the cold stress + Serenade treatment group, and the leaf width of the N485 strain treatment group increased by approximately 3.0% and 5.1% compared to the cold stress + DW treatment group and the cold stress + Serenade treatment group, respectively. In addition, the leaf widths of the other strains N474, N481, N490, and N311 were lower than or equal to those of the untreated group or the Serenade treatment group.

[0159]

[0160] Through the above experiments, it was confirmed that Bacillus amyloliquefaciens strains N484 CP02-1006 and N485 CP02-1007 can effectively alleviate low-temperature stress by promoting plant growth when treated with low-temperature stress. Therefore, it is expected that the strains can be usefully utilized for enhancing stress resistance and promoting growth in plants.

[0161]

[0162] Although representative embodiments of the present application have been described above as examples, the scope of the present application is not limited to the specific embodiments described above, and a person with ordinary knowledge in the relevant field will be able to make appropriate changes within the scope described in the claims of the present application.

[0163]

[0164] [Accession number]

[0165] Name of depositor: Korea Microbiological Conservation Center

[0166] Accession number: KCCM13395P

[0167] Date of acceptance: 20230921

[0168]

[0169] Name of depositor: Korea Microbiological Conservation Center

[0170] Accession number: KCCM13396P

[0171] Date of acceptance: 20230921

[0172]

[0173] [Original text of the deposit certificate]

[0174] [Correction pursuant to Rule 91, February 19, 2025]

[0175] [Correction pursuant to Rule 91, February 19, 2025]

Claims

1. Bacillus amyloliquefaciens strain N484 CP02-1006 deposited under the deposit number KCCM13395P or Bacillus amyloliquefaciens strain N485 CP02-1007 deposited under the deposit number KCCM13396P.

2. In claim 1, The above strain is a strain that enhances stress resistance of plants.

3. In claim 2, The above stress is a low temperature stress, strain.

4. In claim 1, The above strain is a strain that promotes plant growth.

5. A composition for enhancing stress resistance of a plant, comprising as an effective ingredient at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

6. In claim 5, A composition for enhancing stress resistance of plants, wherein the above stress is low temperature stress.

7. In claim 5, A composition for enhancing stress resistance of a plant, wherein the plant is at least one selected from the group consisting of Cucurbitaceae, Brassicaceae, Amaranthaceae, Amaryllidaceae, Apiaceae, Asparagusaceae, Asteraceae, Pinaceae, Convolvulaceae, Legumeaceae, Malvaceae, Muscariaceae, Gramineae, Rosaceae, Rubiaceae, Solanaceae, and Grapeaceae.

8. A composition for promoting plant growth, comprising as an effective ingredient at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

9. In claim 8, A composition for promoting plant growth, wherein the plant is at least one selected from the group consisting of Cucurbitaceae, Brassicaceae, Amaranthaceae, Amaryllidaceae, Apiaceae, Asparagusaceae, Asteraceae, Pinaceae, Convolvulaceae, Legumeaceae, Malvaceae, Muscariaceae, Gramineae, Rosaceae, Rubiaceae, Solanaceae, and Grapeaceae.

10. A method for enhancing stress resistance in a plant, comprising a step of treating a plant, an area around a plant, or both, with at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

11. A method for promoting plant growth, comprising the step of treating a plant, an area around a plant, or both, with at least one selected from the group consisting of Bacillus amyloliquefaciens N484 CP02-1006 strain deposited under the deposit number KCCM13395P, or Bacillus amyloliquefaciens N485 CP02-1007 strain deposited under the deposit number KCCM13396P, a culture solution of the strain, a concentrate of the culture solution, an extract of the culture solution, a dried product of the culture solution, and a pulverized product of the dried product.

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