Composition for enhancing plant disease resistance

Abstract

This invention provides a composition for enhancing plant disease resistance. [Solution] A plant disease resistance enhancing composition comprising an amino acid, wherein the amino acid is at least one selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine, and the plant disease is at least one selected from the group consisting of leaf spot disease, soft rot, stem blight, bacterial leaf spot, and rice blast.

Description

Technical Field

[0001] This application relates to a composition for enhancing plant disease resistance containing amino acids, a method for enhancing plant disease resistance using the same, a method for preventing or treating plant diseases, use for enhancing plant resistance, use for preventing or treating plant diseases, a composition for controlling plants containing amino acids, a method for controlling plants using the same, and use for controlling plants.

Background Art

[0002] Plants have developed a signal transduction system to effectively respond to changes in the external environment. For example, when a plant is exposed to a pathogen, resistance-related genes are expressed by a resistance reaction, and the signal transduction system is activated. Also, as plant hormones, jasmonic acid, ethylene, salicylic acid, etc. are known, and they play a major role in inducing a disease resistance reaction (Induced resistance) from the infection site to the whole plant. Salicylic acid and jasmonic acid move in plants in forms derived from them or in their own forms, respectively, and induce a systemic resistance reaction. By inducing such a systemic reaction, a resistance reaction is induced not only in the leaves invaded by the pathogen but also in the whole plant body that has not been invaded. (ethylene), salicylic acid, etc. are known, and they play a major role in inducing a disease resistance reaction (Induced resistance) from the infection site to the whole plant. Salicylic acid and jasmonic acid move in plants in forms derived from them or in their own forms, respectively, and induce a systemic resistance reaction. By inducing such a systemic reaction, a resistance reaction is induced not only in the leaves invaded by the pathogen but also in the whole plant body that has not been invaded.

[0003] In this regard, derivatives that induce various resistance responses, not just those related to pathogens, have been reported. Among these, BTH (benzo(1,2,3)-thiadiazile-7-carbothioic acid S-methyl ester, benzothiadiazole) is known to exhibit the most potent effect. However, treatment with BTH presents a serious problem: it inhibits plant growth and drastically reduces yield. Subsequent research has suggested that this problem is caused by BTH inducing the expression of many resistance genes in plants even in the absence of pathogens, leading to excessive energy consumption and a shortage of energy available for growth. Therefore, there is an urgent need to develop a substance that exhibits a similar effect to BTH, which induces systemic resistance responses, but without causing plant growth inhibition.

[0004] Furthermore, while some fungicides and insecticides for agricultural and horticultural use containing certain compounds have been studied (Patent Document 1), the development of substances that induce systemic resistance reactions without inhibiting plant growth remains insufficient. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] U.S. Patent No. 10123537 [Non-patent literature]

[0006] [Non-Patent Document 1] Denoux et al., Molecular Plant, Volume 1, Number 3, pages 423-445, May 2008 [Overview of the project] [Problems that the invention aims to solve]

[0007] The problem that this application aims to solve is to provide a composition containing amino acids for enhancing plant disease resistance, a method for enhancing plant disease resistance using the same, a method for preventing or treating plant diseases, a use for enhancing plant resistance, a use for preventing or treating plant diseases, a composition containing amino acids for plant control, a method for plant control using the same, and a use for plant control. [Means for solving the problem]

[0008] This application aims to provide a composition containing amino acids for enhancing plant disease resistance.

[0009] Furthermore, this application aims to provide a method for enhancing plant disease resistance using the aforementioned plant disease resistance enhancing composition.

[0010] Furthermore, this application aims to provide a method for preventing or treating plant diseases using the aforementioned plant disease resistance-enhancing composition.

[0011] Furthermore, this application aims to provide a plant control composition containing amino acids.

[0012] Furthermore, this application aims to provide a method for controlling plants using the aforementioned plant control composition.

[0013] Furthermore, this application aims to provide the use of amino acids or the aforementioned plant disease resistance enhancing composition for enhancing plant disease resistance.

[0014] Furthermore, this application aims to provide the use of amino acids or the aforementioned plant control composition for plant control.

[0015] Furthermore, this application aims to provide the use of amino acids, the plant disease resistance enhancing composition, or the plant control composition for the prevention or treatment of plant diseases. [Effects of the Invention]

[0016] It has the effect of enhancing the disease resistance of plants using amino acids.

Brief Description of Drawings

[0017] [Figure 1] It is a diagram showing the verification results of the induced resistance of 10 kinds of amino acids in the model plant (Arabidopsis thaliana). [Figure 2] It is a diagram showing the result of obtaining the valine concentration that is most excellent in induced resistance in the model plant. [Figure 3] It is a diagram showing that the pathogenic bacteria decrease by treating the model plant with valine, proline and isoleucine. [Figure 4] It is a diagram observing the degree of expression of disease resistance genes by treatment with valine, proline and isoleucine on the model plant. [Figure 5] It is a diagram confirming the control effect by treatment with valine and isoleucine on soybean, cucumber, spinach, rice, tomato, wheat, barley and pepper. [Figure 6] It is a diagram confirming the control effect by treatment with valine, proline and isoleucine on soybean, cucumber and spinach. [Figure 7] It is a diagram showing the test results of induced resistance by a hyperspectral camera. [Figure 8] It is a diagram showing the test results of the growth promoting ability of valine.

Modes for Carrying Out the Invention

[0018] Hereinafter, the present application will be specifically described. Note that the description and embodiments of one aspect disclosed in the present application are also applicable to the description and embodiments of other aspects for common matters. In addition, all combinations of various elements disclosed in the present application are included in the present application. Furthermore, the present application is not limited to the following specific descriptions.

[0019] One aspect of the present application provides a composition for enhancing plant disease resistance containing amino acids.

[0020] The aforementioned amino acid is at least one selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine, and the aforementioned plant disease is Select from the group consisting of leaf spot disease, bacterial soft rot, Phytophthora stem and root rot, bacterial leaf blight, and rice blast. The present invention provides a composition for enhancing plant disease resistance, which is at least one of the following:

[0021] In this application, "branched-chain amino acid" means an amino acid having a branched alkyl group in its side chain, and includes valine, leucine, and isoleucine. Specifically, in this application, the branched-chain amino acid is an L-branched-chain amino acid, and the L-branched-chain amino acid is at least one selected from L-valine, L-leucine, and L-isoleucine, but is not limited to these.

[0022] The proline is L-proline, the glutamic acid is L-glutamic acid, the aspartic acid is L-glutamic acid, and the histidine is L-histidine, but is not limited to these.

[0023] As an example, the amino acids of this application are used to treat plants at concentrations of 0.0001-10%, 0.0001-5%, 0.0001-1%, 0.005-10%, 0.005-1%, 0.005-0.5%, 0.001-10%, 0.001-1%, 0.001-0.5%, 0.01-10%, 0.01-1%, 0.001-0.5%, 0.01-10%, 0.01-1%, 0.01-0.5%, 0.05-10%, 0.05-1%, 0.05-0.5%, 0.1-10%, 0.1-1%, or 0.1-0.5% (w / w) relative to the entire plant disease resistance enhancing composition, its dilutions, or concentrates, but are not limited thereto.

[0024] This application is technically significant because it clarifies that amino acids, specifically branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine, have applications in enhancing plant disease resistance or controlling plant diseases.

[0025] In this application, "plant disease" refers to a disease that occurs in plants, meaning an abnormality in their original form or physiological function. The causes of the disease include pathogenic diseases, nutrient deficiencies in the soil, weather conditions, and toxic gases. The pathogens are fungal or bacterial pathogens, but are not limited to these.

[0026] Specifically, the plant diseases covered by this application are leaf spot and bacterial soft rot. ), Phytophthora stem and root rot, Bacterial leaf blight It is at least one selected from the group consisting of ) and rice blast.

[0027] In this application, "leaf spot disease" refers to a disease that causes large and small spots (spots or specks) to appear on the stems, fruits, and leaves of plants. The spots are black, light brown, or brown. This leaf spot disease mainly affects fruit vegetables, but can affect any type of plant. This leaf spot disease is caused by various species of fungi or bacteria, including living parasitic pathogens, specifically microorganisms of the genus Pseudomonas, and more specifically Pseudomonas syringae pv. toma Pseudomonas syringae pv. tomato, more specifically Pseudomonas syringae This condition is induced by, but is not limited to, Pseudomonas syringae pv. tomato DC3000.

[0028] In this application, "bacterial soft rot" refers to a disease in which plant tissue is eroded, emitting a foul odor, rotting, and becoming ulcerated. It is particularly common in watery vegetables such as lettuce and Chinese cabbage, but it can also occur in tomatoes, sweet potatoes, potatoes, and other plants, so it can affect any type of plant. The aforementioned soft rot is induced by a dead-parasitic fungus, specifically Pectobacterium carotovorum subsp. carotovorum, but is not limited to this species.

[0029] In this application, "stem blight (Phytophthora stem and root rot)" refers to a type of fungal disease that affects potatoes, tomatoes, and other plants, with external symptoms varying somewhat depending on the degree of infection. During the vigorous growing season, the plant body wilts, the lower leaves turn reddish-brown, and the young leaves become chlorotic and take on various colors. In cases where the infection is not severe, external symptoms are often not clear, the fruits of infected plants are small, and the formation of stolons is poor. Although stem blight mainly affects fruit vegetables, it can affect any type of plant. Stem blight is caused by pathogens of the genus Phytophthora, which belongs to the family Pythiaceae of flagellated fungi, specifically Phytophthora sojae, but is not limited to these.

[0030] In this application, "bacterial leaf blight" refers to a disease where lesions appear in a spotted pattern. This refers to a disease that causes discoloration to brown. Lesions form on leaves, stems, pods, petioles, and cotyledons. It can affect any type of plant, including soybeans, mung beans, and fruit vegetables. The aforementioned bacterial spot disease is caused by microorganisms of the genus Pseudomonas, specifically Pseudomonas syringum. Triggered by Gae pv. Lachrymans (Pseudomonas syringae pv. lachrymans) However, this is not the only example.

[0031] In this application, "rice blast" refers to the most important disease of cereals, especially rice, and can occur in any part of the cereal plant. Lesions form on above-ground parts such as leaves, panicles, stalks, nodes, and grains, with blast being most common on leaves, panicles, and stalks. Infection occurs via conidia, and the typical symptoms are the spread of irregular red, gray, black, and / or brown spots on the leaves, followed by the entire leaf turning brown and wilting. The panicle neck and stalks wither and turn light brown, and if there is high humidity, gray mold develops on the surface. Dark brown lesions form on the nodes, making them brittle, and in humid environments, the surface becomes covered with gray mold. The aforementioned spot disease is caused by microorganisms of the genus Magnaporthe, specifically Magnaporthe It is induced by, but is not limited to, the oryzae (Magnaporthe oryzae).

[0032] As one example, the plant disease was selected from the group consisting of Pseudomonas syringae pv. tomato, Pectobacterium carotovorum subsp. carotovorum, Phytophthora sojae, Pseudomonas syringae pv. lachrymans, and Magnaporthe oryzae. It is induced by at least one bacterium, but is not limited to these.

[0033] As one example, the plant disease is a leaf spot disease caused by Pseudomonas syringae pv. tomato, and Pectobacterium carotovora Induced by the subspecies Pectobacterium carotovorum subsp. carotovorum Soft rot caused by Phytophthora sojae, soybean stem blight caused by Pseudomonas syringae pv. lachrymans, cucumber bacterial spot disease caused by Pectobacterium carotoborum subsequence Induced by the species Pectobacterium carotovorum subsp. carotovorum. At least one selected from the group consisting of lettuce soft rot and rice blast disease induced by Magnaporthe oryzae, but not limited to these.

[0034] The aforementioned plant disease may affect any type of plant, and in one example, the plant may be selected from the group consisting of legumes, fruit vegetables, leafy vegetables, and cereals.

[0035] In this application, "legumes" refers to leguminous plants (Fabaceae) used as food or animal feed. This means that, for example, soybeans, wild beans, kidney beans, black beans, peas, adzuki beans, peanuts, and cowpeas are examples. Specifically, the aforementioned legumes are soybeans, but are not limited to soybeans.

[0036] In this application, "fruit vegetables" refers to vegetables whose fruit is used when classified by the part of the vegetable that is utilized. Examples include chili peppers, cucumbers, tomatoes, watermelons, strawberries, and cantaloupes. Specifically, the aforementioned fruit vegetables are chili peppers, cucumbers, or tomatoes, but are not limited to these.

[0037] In this application, "leafy vegetables" refers to vegetables whose leaves are used when classified by the part of the vegetable that is utilized. Examples include Chinese cabbage, lettuce, perilla, and spinach. Specifically, the leafy vegetables are Chinese cabbage or lettuce, but are not limited to these.

[0038] In this application, "cereals" refers to cultivated grasses whose seeds are used for food. Examples include rice, wheat, barley, and corn. Specifically, the aforementioned cereals are rice, wheat, or barley, but are not limited to these.

[0039] In this application, "plant disease resistance" means the property of preventing the invasion of pathogens, the property of preventing plant diseases from developing, and / or the property of not becoming diseased even if pathogens invade.

[0040] As a specific example, the amino acids of this application may prevent or treat plant pathogenic infections, or they may induce systemic resistance in plants, but are not limited to these.

[0041] As a specific example, the amino acids of this application enhance plant disease resistance without inhibiting plant growth, but are not limited to this. Specifically, while derivatives that induce various resistance reactions in plants are known, not just those to pathogenic bacteria, some of them (for example, benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester, benzothiadiazole; BTH) suppress plant growth and drastically reduce yield. It causes serious side effects such as a decrease in plant growth. However, the amino acids in this application do not inhibit plant growth, but rather promote it.

[0042] The plant disease resistance enhancing composition of this application may be any formulation or product necessary for enhancing plant disease resistance, such as pesticides, fertilizers, or control agents.

[0043] In this application, "agricultural chemicals" refers to chemicals used for the protection and growth of crops. Specifically, this includes not only formulations used to control diseases, pests, weeds, pathogens, nematodes, mites, etc. that occur during the cultivation and storage of crops, but also growth regulators and adjuvants used to enhance or suppress the physiological functions of crops.

[0044] In this application, "fertilizer" also refers to a nutrient that enriches the soil and promotes the growth of plants. The fertilizer containing the composition of this application can be easily used by mixing it into the soil before sowing, and can be applied together with pesticides and other chemicals.

[0045] The plant disease resistance enhancing composition of this application may further contain substances commonly used for enhancing disease resistance, and in addition to the amino acids, it may further contain pharmaceutically acceptable solid carriers, liquid carriers, liquid diluents, liquefied gaseous diluents, solid diluents, or other auxiliary agents as excipients, such as surfactants such as emulsifiers, dispersants, and foaming agents.

[0046] The plant disease resistance enhancing composition of this application can be used by mixing the active ingredient with the excipient to formulate it into an agricultural composition, and the method of formulation into an agricultural composition may be any method commonly used in the art.

[0047] Furthermore, the plant disease resistance enhancing composition of this application may be provided in the form of wettable powders, granules, powders, emulsions, sprays, fumigants, capsules, and gels, or may be provided as a donut-shaped contact agent for buoyancy, but is not limited thereto.

[0048] As an example, the composition of this application is a composition for enhancing plant disease resistance or plant control that contains valine, wherein the plant disease is at least one selected from the group consisting of leaf spot, bacterial soft rot, Phytophthora root rot, bacterial leaf blight, and rice blast, but is not limited to these.

[0049] As an example, the composition of this application is a composition for enhancing plant disease resistance or plant control that contains isoleucine, wherein the plant disease is at least one selected from the group consisting of leaf spot, bacterial soft rot, Phytophthora root rot, bacterial leaf blight, and rice blast, but is not limited to these.

[0050] As one example, the composition of this application is a composition containing leucine for enhancing resistance to bacterial soft rot or for plant control, but is not limited to these.

[0051] As one example, the composition of this application is a composition containing proline for enhancing resistance to bacterial soft rot or for plant control, but is not limited to these.

[0052] As one example, the composition of this application is a composition for enhancing resistance to leaf spot disease or a composition for plant control, comprising at least one selected from the group consisting of glutamic acid, aspartic acid, and histidine, but is not limited to these.

[0053] Another aspect of this application provides a method for enhancing plant disease resistance, comprising the step of treating a plant with a plant disease resistance enhancing composition containing amino acids.

[0054] As a specific example, the aforementioned method for enhancing plant disease resistance also promotes plant growth, but it is not limited to this.

[0055] The aforementioned amino acids, plant diseases, plant disease resistance, and compositions for enhancing plant disease resistance are as described above.

[0056] The plant body may be any part of the plant, such as roots, stems, leaves, or combinations thereof, and may also be of any type of plant. Specifically, the plant body may be a plant individual that is at risk of being infected with a plant disease or is infected with a pathogen, but is not limited to these.

[0057] Treatment with the plant disease resistance-enhancing composition may be carried out by spraying, injecting, mixing, or mixing the composition into the soil of the plant, but is not limited to these methods as long as the inside or outside of the plant comes into direct or indirect contact with the composition. Furthermore, the treatment may be carried out simultaneously with or at a different time from the invasion of the plant by the pathogen, or before or after the invasion of the plant, but is not limited to these methods. do not have.

[0058] The aforementioned plant disease resistance enhancing composition is applied once or two or more times, but is not limited to these. When applied two or more times, the application intervals are 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 to these.

[0059] As one example, the plant disease resistance-enhancing composition is subjected to a primary treatment 7 days before pathogen invasion and a secondary treatment 1 day before, but is not limited to this.

[0060] As one embodiment, the plant is at least one selected from the group consisting of legumes, fruit vegetables, leafy vegetables, and cereals, but is not limited thereto.

[0061] The aforementioned legumes, fruit vegetables, leafy vegetables, grains, etc., are as described above.

[0062] A further aspect of this application provides a method for preventing or treating plant diseases, comprising the step of treating a plant with a composition for enhancing plant disease resistance that contains amino acids.

[0063] The aforementioned amino acids, plant diseases, plant disease resistance, compositions for enhancing plant disease resistance, treatment with plant disease resistance-enhancing compositions, and plant bodies are as described above.

[0064] In this application, "prevention" means any action that suppresses or delays the onset or progression of a plant disease.

[0065] In this application, “treatment” means any action that improves or favorably alters the symptoms of a plant disease. In this application, treatment includes any action that results in the prevention of infection by plant pathogens or recurrence of a plant disease, alleviation of symptoms, reduction of the rate of progression, reduction of the disease state, temporary or continuous relief of the disease state, recovery, or improvement of the prognosis.

[0066] Further embodiments of this application provide a plant control composition comprising an amino acid. The amino acid is at least one selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine, and the plant disease is at least one selected from the group consisting of leaf spot, bacterial soft rot, Phytophthora root rot, bacterial leaf blight, and rice blast.

[0067] The aforementioned amino acids, plant diseases, etc., are as described above.

[0068] In this application, "pest control" means preventing crops and other plants from being damaged by pests and diseases, removing pathogens that have emerged, and preventing the spread of pathogens. This pest control is a series of actions to reduce damage caused by pests and includes, but is not limited to, physical (mechanical) control methods, cultural control methods, chemical control methods, biological control methods, and environmentally friendly control methods. This pest control includes not only chemical control using fungicides or insecticides that are toxic to pests and diseases, but also methods to create an environment that pathogens do not like through cultivation of resistant plants, crop rotation, etc., methods to remove pathogens by heat treatment of plants or steam treatment of soil, and methods to attract and kill pests using attractants or sex pheromones.

[0069] The plant control composition of this application may be any formulation or product necessary for plant control, such as a pesticide, fertilizer, or control agent.

[0070] The plant control composition of this application may further contain substances commonly used for pest control, and in addition to the amino acids, it may further contain pharmaceutically acceptable solid carriers, liquid carriers, liquid diluents, liquefied gaseous diluents, solid diluents, or other auxiliary agents as excipients, such as surfactants such as emulsifiers, dispersants, and foaming agents.

[0071] The plant control composition of this application can be used by mixing the active ingredient with the excipient to formulate it into an agricultural composition, and the method of formulating it into an agricultural composition may be any method commonly used in the art.

[0072] Furthermore, the plant control compositions of this application may be provided in the form of wettable powders, granules, powders, emulsions, sprays, fumigants, capsules, and gels, or they may be provided as donut-shaped contact agents for buoyancy.

[0073] Another aspect of this application provides a method for controlling plants, comprising the step of treating plants with a plant control composition containing amino acids. Specifically, the plant control method may also promote plant growth.

[0074] The aforementioned amino acids, plants, plant bodies, plant diseases, pest control, and plant control compositions are as described above.

[0075] Treatment with the plant control composition may be carried out by spraying, injecting, mixing, or blending the composition into the plant body or soil, but is not limited to these methods as long as the inside or outside of the plant body comes into contact with the composition. Furthermore, the treatment may be carried out simultaneously with or at a different time from the invasion of the plant by the pathogen, or before or after the invasion of the plant, but is not limited to these methods.

[0076] The aforementioned plant control composition is applied once or two or more times, but is not limited to these. When applied two or more times, the application intervals are 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 to these.

[0077] As one example, the plant control composition is subjected to a primary treatment 7 days before pathogen invasion and a secondary treatment 1 day before, but it is not limited to this.

[0078] Further embodiments of this application provide uses of amino acids or compositions containing them for enhancing plant disease resistance, for plant control, and for the prevention or treatment of plant diseases.

[0079] The aforementioned amino acids, plant diseases, plant disease resistance, control, prevention, and treatment are as described above. [Examples]

[0080] The present application will be described in more detail below with reference to examples. However, these examples are merely preferred embodiments illustrating the present application, and the application is not limited thereto. Technical matters not described herein can be fully understood and readily implemented by a skilled technician in the art of this application or a similar art. [Examples]

[0081] Verification of amino acid-induced resistance to plant diseases To investigate the induced resistance of various amino acids to plant diseases, Arabidopsis thaliana, a model plant, was used, and the pathogen causing leaf spot disease was identified. We used the known Pseudomonas syringae pv. tomato DC3000 (hereinafter referred to as Pst) and Pectobacterium carotovorum subsp. carotovorum SCC1 (hereinafter referred to as Pcc), which is known to cause soft rot.

[0082] To grow the model plant, Arabidopsis thaliana seeds were surface-sterilized by immersion in 3% sodium hypochlorite (NaOCl) for 5 minutes. They were then rinsed five times with sterile water, followed by 1 / 2 Murashige and Skoog (MS) medium (0.6% agar, The seeds were sown in 1.5% sucrose (pH 5.8). After 3 days, once the seeds germinated, six germinated seeds were transplanted onto plates (Falcon, USA) and cultured in a plant incubator (under 12 hours of light and a temperature of 23°C).

[0083] Two weeks after transplanting, the roots of each plant were treated with 50 μl each of 10 different amino acids at a 0.1% concentration. Seven days after amino acid treatment, five leaves from each plant were examined in 1 × 10⁶ steps. 8 Each sample was treated with 2 μl each of Pst (live parasitic, Figure 1a) and Pcc (dead parasitic, Figure 1b) pathogens at cfu / ml.

[0084] Pst pathogens were incubated on King's B (KB) solid medium at 30°C for 48 hours before treatment. The plants were cultured for several days. In the case of Pst pathogens, the disease severity was observed 7 days after pathogen treatment, and the degree of disease on each leaf was measured on a scale of 0 to 5 according to the following criteria, and the average value was calculated. 0 = No symptoms, 1 = Mild whitening symptoms, 2 = Whitening symptoms, 3 = Whitening symptoms and small area of ​​necrosis, 4 = Necrosis, 5 = Extensive necrosis

[0085] PCC pathogens were cultured in Luria-Bertani broth (LB) before treatment. The plants were incubated at 30°C for 48 hours. In the plants treated with PCC pathogen, the disease severity of each leaf was observed 24 hours and 48 hours after treatment with the pathogen. The degree of disease severity was measured on a scale of 0 to 5 according to the following criteria, and the average value was calculated. 0 = No symptoms, 1 = Mild soft rot symptoms at the site of pathogen inoculation, 2 = Soft rot symptoms at the site of pathogen inoculation, 3 = Severe soft rot symptoms at the site of pathogen inoculation, 4 = Soft rot symptoms extending to the area around the site of pathogen inoculation, 5 = Soft rot symptoms throughout the entire leaf

[0086] We selected amino acids that exhibited a statistically significant effect in inducing disease resistance compared to the water-treated control group.

[0087] As a result, valine, proline, isoleucine, Leucine, glutamic acid, histidine, and Seven amino acids in aspartic acid induce resistance to Pst. It was confirmed that four amino acids—toto (Figure 1a), valine, proline, isoleucine, and leucine—independ to resistance to PCC (Figure 1b).

[0088] In particular, valine was confirmed to maintain disease resistance even 48 hours after treatment with Pcc pathogens. Furthermore, valine, proline, isoleucine, and leucine were confirmed to induce resistance to both Pst (a live parasitic fungus) and Pcc (a dead parasitic fungus) in plants. [Examples]

[0089] Further verification of amino acids that induce resistance to plant diseases Using valine, which showed the best induced resistance to Pcc and Pst in Example 1, the effect of inducing resistance was reconfirmed.

[0090] Valine was used at concentrations of 0.5%, 0.1%, 0.05%, 0.01%, 0.005%, and 0.001%, and Pst was used as the pathogen. All other aspects were the same as the experimental method in Example 1.

[0091] As a selection criterion, we evaluated whether there was a statistically significant difference (p<0.05) in the induced resistance to disease between the valine-treated groups at each concentration and the water-treated groups.

[0092] As a result, the disease severity decreased overall in the valine-treated groups. Furthermore, while the disease severity in the 0.5% and 0.1% (0.95 mM) valine-treated groups was 2.4 and 2.5, respectively, the disease severity in the water-treated group reached 4.6, confirming a significant decrease in disease severity in both the 0.5% and 0.1% valine-treated groups. The 1 mM BTH-treated group, known as an induced resistance substance, showed a disease severity of 3.7, indicating that the 0.5% Furthermore, the 0.1% valine-treated group was confirmed to have superior disease resistance compared to the BTH-treated group (Figure 2). [Examples]

[0093] Verification of induced resistance by valine and isoleucine treatment. Proline and branched-chain amino acids (BCAAs), including valine and We investigated whether induced resistance using biisoleucine reduces the number of pathogenic bacteria.

[0094] To this end, plants were prepared in the same manner as in Example 1 and treated with amino acids (0.1% valine, 0.1% isoleucine, and 0.1% proline, which were selected in Example 2 for their low-concentration induction resistance effect) and pathogenic bacteria.

[0095] The number of pathogens was measured by crushing the parts of Arabidopsis thaliana (excluding the roots) in an e-tube containing beads, and then examining the pathogens contained in the juice.

[0096] When Arabidopsis thaliana plants were treated with Pst pathogens, the number of pathogens was measured on the 3rd and 6th days, respectively. The results showed that in the 0.1% valine treatment group, the 0.1% isoleucine treatment group, and the 0.1% proline treatment group, the number of pathogens was statistically significantly reduced on the 6th day compared to the untreated control group (Figure 3a).

[0097] When Arabidopsis thaliana plants were treated with Pcc pathogens, the number of pathogens was measured on the third day. The results showed that, for Pcc as well, the number of pathogens decreased in the 0.1% valine treatment group, the 0.1% isoleucine treatment group, and the 0.1% proline treatment group compared to the untreated control group. The number of pathogens also decreased in the valine, isoleucine, and proline treatment groups compared to the BTH treatment group (Figure 3b). [Examples]

[0098] Confirmation of gene expression responses related to plant disease resistance To investigate whether valine, isoleucine, and proline induce the expression of disease resistance-related genes, Arabidopsis thaliana (Non-Patent Literature 1) containing the pathogenesis-related protein (PR) 1a promoter GUS fusion construct, a marker gene for disease resistance, was used. Arabidopsis thaliana plants were prepared in the same manner as in Example 1, and the roots of the Arabidopsis thaliana were treated with 50 μl of valine, 50 μl of isoleucine, and 50 μl of proline, respectively. One week after treatment, the presence or absence of expression of the resistance marker gene PR1a was observed by GUS (β-glucuronidase; GUS reporter) activity.

[0099] Observations were made from Day 1 after inoculation (Dai) to 4Dai after pathogen treatment. In addition to the BTH-treated group, GUS was also expressed in the valine, isoleucine, and proline-treated groups, and PR1a was also confirmed to be expressed (Figure 4). [Examples]

[0100] Testing of induced resistance to valine and isoleucine using pot experiments We re-examined disease resistance in crops grown in pots.

[0101] The seeds of eight crops were surface-sterilized by immersing them in 3% sodium hypochlorite (NaOCl) for 5 minutes. They were then rinsed five times with sterilized water, sown in pots filled with soil, and cultured in a plant incubator (12 hours of light and 23°C). The plants were cultivated in pots for 3 weeks in a plant incubator (12 hours of light, 26°C, Vision), after which they were sprayed with 5 ml of each amino acid at a 0.1% concentration. One day after amino acid treatment, pathogens were inoculated as follows, and the control efficacy was calculated.

[0102] The study targeted nine pathogens from eight crops (soybeans, cucumbers, lettuce, rice, tomatoes, wheat, barley, and chili peppers). The target pathogens are as follows: Soybean stem blight (Phytophthora sojae), cucumber bacterial spot (Pseudomonas syringae pv. lachrymans), lettuce soft rot (Pectobacterium carotovorum pv. carotovora), rice blast (Magnaporthe oryzae), tomato gray mold (Botrytis cinerea), tomato stem blight (Phytophthora infestans), wheat rust (Puccinia recondite), barley powdery mildew (Blumeria graminis f. sp. hordei), chili pepper anthracnose (Colletotrichum coccodes)

[0103] a) Soybean stem blight The Phytophthora sojae strain, the pathogen of soybean stem blight, was cultured in V8 medium. Spores were inoculated into V8 juice (1.8% agar), cultured in a 25°C incubator for 2 weeks, and harvested with sterilized distilled water to form a spore suspension (5.0 × 10⁻⁶). 5 A spore suspension (spores / ml) was prepared. Soybeans were inoculated with 5 ml of the prepared spore suspension per plant by spraying, and the plants were allowed to develop the disease for 3 days in a constant temperature and humidity chamber at 26°C. The disease area percentage (%) was then investigated. The disease area percentage was calculated as follows.

[0104]

number

[0105] b) Cucumber bacterial spot The pathogen of cucumber bacterial spot disease, Pseudomonas syringae pv. lachrymans, was cultured on KB (King's B) solid medium at 28°C for 48 hours. The pathogen had an OD=1(10 9 Each plant was treated with 5 ml of the CFU / ml concentration by spray application. The inoculated cucumbers were allowed to develop the disease for 3 days in a constant temperature and humidity chamber at 26°C, and the diseased area percentage was investigated.

[0106] c) Lettuce soft rot Pectobacterium carotov is the pathogen that causes bacterial soft rot in lettuce. The Orum pv. carotovora strain was cultured on LB (Luria-Bertani broth) solid medium at 28°C for 48 hours. The pathogenic bacteria were OD=1(10 9 Lettuce was treated by spraying 5 ml of CFU / ml per plant. The inoculated lettuce was allowed to develop the disease for 1 day in a constant temperature and humidity chamber at 26°C, and the diseased area percentage was investigated.

[0107] d) Rice blast disease In rice blast disease (RCB), the pathogenic fungus is Magnaporthe oryz The ae KI-1113a strain was inoculated onto rice bran agar and cultured in an incubator at 25°C for 2 weeks. The formed spores were harvested with sterilized distilled water, and a spore suspension (5.0 × 10⁻⁶) was prepared. 5 Rice plants were inoculated with 5 ml of spores / ml per plant by spray application. These plants were then left in a humid chamber in the dark for 24 hours, and subsequently allowed to develop disease for 4 days in a constant temperature and humidity chamber at 80% relative humidity and 25°C. The disease lesion area percentage was then investigated.

[0108] e) Tomato gray mold In tomato gray mold (TGM), the pathogenic fungus is Botrytis ciner The ea strain was inoculated onto potato agar medium and cultured in a 20°C incubator (dark). The resulting spores were used as the inoculum. To inoculate with the pathogen, spores were harvested and the spore concentration was measured using a hemocytometer at 5.0 × 10⁶. 5 The sample was diluted to spores / ml and then spray-inoculated onto tomato seedlings (2-3 leaf stage) treated with the sample. The inoculated tomato seedlings were left in a humid chamber at 20°C (relative humidity 95% or higher) for 3 days to allow the disease to develop, and the disease lesion area percentage was investigated.

[0109] f) Tomato stem blight In tomato stem blight (TLB), the pathogenic fungus is Phytophthora inf The *Estans* strain was inoculated into oatmeal medium and cultured in a 20°C incubator to form zoosporangia. Sterilized distilled water was added, and the formed zoosporangia were harvested, with a spore concentration of 2.0 × 10⁶. 4 A spore suspension containing sporangia / ml was prepared. This was then subjected to cold treatment in a refrigerator to release zoospores, which was used to prepare a zoospore suspension. This suspension was spray-inoculated onto tomato seedlings (2-3 leaf stage) treated with the sample. The tomato seedlings inoculated with the pathogen were subjected to humid chamber treatment at 20°C for 2 days, then allowed to develop disease in a constant temperature and humidity chamber for 2 days, and the disease lesion area percentage (%) was investigated.

[0110] g) Wheat rust In wheat rust (WLR), the pathogenic fungus is Puccinia recondus Since ita is a parasitic fungus, spores (urediniospores) formed on wheat leaves by direct subculturing were used as the inoculum. For the efficacy study, five wheat seeds ("Kumugan wheat") were sown in disposable pots (diameter: 4.5 cm), and seedlings at the one-leaf stage, cultivated in a greenhouse for eight days, were treated with the sample. One day later, the inoculum (spores 0.67 g / L) was spray-inoculated. The inoculated wheat seedlings were treated in a humid chamber at 20°C for one day, and then moved to a constant temperature and humidity chamber at 20°C with a relative humidity of 70% to induce disease development. The lesion area percentage (%) was investigated seven days after inoculation.

[0111] h) Barley powdery mildew In barley powdery mildew (BPM), the pathogenic bacterium is Blumeria gram Since inis f.sp.hordei is a parasitic fungus, we will subculture barley seedlings. Spores formed on barley leaves were used as the inoculation source. For the efficacy study, five barley seeds ("Hanyong barley") were sown in disposable pots (diameter: 4.5 cm), and seedlings at the one-leaf stage, cultivated in a greenhouse for eight days, were sprayed with the sample, air-dried in the greenhouse, and then powdery mildew spores were sprinkled onto the barley treated with the sample to inoculate it. The inoculated barley seedlings were left in a constant temperature and humidity chamber at approximately 20°C and 50% relative humidity for seven days to allow the disease to develop, and the disease lesion area percentage (%) was investigated.

[0112] i) Anthracnose of chili peppers In pepper anthracnose (PAN), the pathogenic fungus is Colletotrichum Coccodes were inoculated into oatmeal medium and cultured at 25°C for 10 days to form the coccodes. Spores were harvested, and the spore concentration was 5 × 10 5 A spore suspension was prepared by adjusting the concentration to spores / ml. Young pepper seedlings (3-4 leaf stage) treated with the sample were spray-inoculated with the prepared spore suspension, left in a humid chamber (25°C) for 2 days, and then allowed to develop the disease in a constant temperature and humidity chamber (25°C, 80%RH). Three days after inoculation, the percentage of lesion area formed on the leaves of the pepper plants was investigated.

[0113] The control efficacy was calculated from the disease lesion area percentage obtained by the methods a) to i) described above using the following formula.

[0114]

number

[0115] As a result, when we tested for induced resistance in nine pathogens, we confirmed that the treatment provided control compared to the untreated group in soybean stem blight (Figure 5a), cucumber bacterial spot (Figure 5b), lettuce soft rot (Figure 5c), and rice blast (Figure 5d).

[0116] Compared to the untreated control group, the control efficacy of the valine-treated group and the isoleucine-treated group was 15% and 10% for soybean stem blight, 17% and 12% for cucumber bacterial spot, 13% and 8% for lettuce soft rot, and 13% and 16% for rice blast.

[0117] In the BTH-treated group, which is an induced resistance-inducing substance, the control efficacy for soybean stem blight, cucumber bacterial spot, lettuce soft rot, and rice blast was 10%, 9%, 10%, and 9%, respectively, compared to the untreated group. Furthermore, the control efficacy for tomato gray mold in the BTH-treated group was 20% compared to the untreated group. To enhance the induced resistance effect against the pathogens being controlled, the following experiments were conducted regarding the timing and method of treatment. [Examples]

[0118] Testing the optimal conditions for inducing resistance to valine and isoleucine. To improve the disease control efficacy of valine and isoleucine, the induction period for resistance was altered. This study targeted three crops (soybean, cucumber, and lettuce) and three pathogens (soybean stem blight, cucumber bacterial leaf spot, and lettuce soft rot). The preparation of each plant was carried out in the same manner as in Example 5.

[0119] For the pathogen of soybean stem blight, the disease severity was observed 7 days after treatment with the pathogen, and the degree of disease severity was measured on a scale of 0 to 5 according to the following criteria. 0 = No symptoms, 1 = 1 true leaf wilts, 2 = 2 true leaves wilt, 3 = 3 true leaves wilt, 4 = 4 true leaves wilt, 5 = the entire plant wilts significantly.

[0120] For the pathogen of cucumber bacterial spot disease, the disease severity was observed 7 days after treatment with the pathogen, and the degree of disease severity was measured on a scale of 0 to 5 according to the following criteria. 0 = No symptoms, 1 = Mild symptoms, 2 = 30% symptoms, 3 = Symptoms and small area of ​​necrosis, 4 = Necrosis, 5 = Extensive necrosis and death symptoms

[0121] For the pathogen of lettuce soft rot, the degree of disease development was observed 48 hours after treatment with the pathogen, and the degree of disease development was measured on a scale of 0 to 5 according to the following criteria. 0 = No symptoms, 1 = Mild soft rot symptoms at the site of pathogen inoculation, 2 = Soft rot symptoms at the site of pathogen inoculation, 3 = Severe soft rot symptoms at the site of pathogen inoculation, 4 = Soft rot symptoms extending to the area around the site of pathogen inoculation, 5 = Soft rot symptoms throughout the entire leaf

[0122] Based on the disease severity investigated using this method, the control efficacy was calculated using the following formula.

[0123]

number

[0124] To obtain the optimal control efficacy for inducing induced resistance, various amino acid treatment periods and frequencies were tested. Ultimately, it was confirmed that performing a primary amino acid treatment 7 days before pathogen treatment and a secondary treatment 1 day before was the most effective in inducing resistance.

[0125] The results of performing a primary amino acid treatment 7 days before pathogen treatment and a secondary treatment 1 day before are as follows:

[0126] In soybean stem blight, the disease incidence in the valine-treated group and the isoleucine-treated group was 1.1 and 2.3, respectively, which was lower than the disease incidence in the water control group (4.1). The calculated control efficacy was 73% and 44%, respectively, compared to the untreated group. Valine demonstrated superior disease control efficacy compared to the BTH-treated group (46%), while isoleucine demonstrated disease control efficacy equivalent to that of the BTH-treated group (Figure 6a).

[0127] In cucumber bacterial spot disease, the disease severity in the valine-treated group and the isoleucine-treated group was 2.7 and 3.6, respectively, which was lower than the disease severity in the water control group (4.4). The calculated control efficacy was 39% and 18%, respectively, compared to the untreated group. On the other hand, in the BTH-treated group, it was confirmed that not only disease susceptibility but also growth was suppressed compared to the untreated group. Valine and isoleucine have a disease control effect by inducing resistance to cucumber bacterial spot disease. Unlike the BTH-treated group, which exhibits growth suppression when induced resistance is developed, it was confirmed that no growth suppression occurred in the valine-treated and isoleucine-treated groups (Figure 6b).

[0128] In the case of lettuce soft rot, the disease severity in the valine-treated group and the isoleucine-treated group was 2.2 and 3, respectively, which was lower than the disease severity in the water control group (4.2). The calculated control efficacy was 48% and 29%, respectively, compared to the untreated group. These results suggest that the valine-treated group and the isoleucine-treated group have superior disease control efficacy compared to the BTH-treated group, which had a control efficacy of 17% (Figure 6c). [Examples]

[0129] Hyperspectral camera analysis of inductive resistance Induced resistance to soybean stem blight was verified using a hyperspectral camera and a visible light camera. Plant preparation was carried out in the same manner as in Example 6, and seven days after pathogen treatment, leaves were collected from the same location in each treatment group and hyperspectral images were taken. The hyperspectral camera was used to capture images in the wavelength range of 400 to 1000 nm. The difference in wavelength range between normal leaves and diseased leaves was utilized, as this difference can be displayed by color. Even when it is difficult to distinguish the degree of disease with the naked eye, it can be easily distinguished using a hyperspectral camera.

[0130] The Normalized Difference Vegetation Index (NDVI) was used to quantify the extent of disease on the total leaf surface area, with a scale of 0.1 to 0.9, and the number of pixels used was calculated accordingly. Generally, a value of 0.7 or higher is considered indicative of a healthy leaf.

[0131] In the visible light band, the untreated group showed a lighter yellow-green color and faster disease progression compared to the valine-treated group (Figure 7a, RGB photograph).

[0132] When photographed with a hyperspectral camera, normal leaves appeared green, while dead leaves appeared brown, with different colors indicating the severity of the disease (Figure 7a, NDVI image). In the untreated group, 26% of the leaves were healthy, compared to 60% and 46% in the valine-treated group and isoleucine-treated group, respectively, indicating a higher proportion of healthy leaves compared to the untreated group. This was observed (Figure 7b). Hyperspectral imaging also reconfirmed that valine and isoleucine induce resistance and provide resistance to the disease. [Examples]

[0133] Testing the growth-promoting ability of valine Soybeans and cucumbers were treated with valine, and their growth was observed. One week after sowing in pots, each plant was drenched with 5 ml of valine at a concentration of 0.1%. Each plant was drenched with valine twice at one-week intervals. Two weeks after the final treatment, the stems of each treatment group were cut and weighed.

[0134] As a result, in soybeans, the valine-treated group averaged 11g, which was 2.4g heavier than the untreated group's 8.6g. In contrast, the BTH-treated group averaged 7.3g, indicating a growth inhibition phenomenon (Figure 8a).

[0135] In cucumbers, the valine-treated group averaged 34g, which was 3g heavier than the untreated group's 31g. In contrast, the BTH-treated group averaged 26g, showing a growth inhibition phenomenon compared to the untreated group (Figure 8b).

[0136] The above results suggest that valine also promotes the growth of soybeans and cucumbers.

[0137] From the above explanation, a person skilled in the art to which this application pertains will understand that this application can be implemented in other specific forms without altering its technical idea or essential features. It should be understood that the above embodiments are merely illustrative and not limiting. This application should be interpreted as including all modified or altered forms derived from the meaning and scope of the claims and their equivalent concepts, rather than the specification.

Claims

1. A composition for enhancing plant disease resistance, containing amino acids, The aforementioned amino acid is at least one selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine. The plant diseases include leaf spot, Bacterial soft rot, Phytophthora stem and root rot, Bacterial leaf blight and Rice blast. A composition for enhancing plant disease resistance, which is at least one selected from the following group.

2. The aforementioned plant disease is caused by Pseudomonas syringae pv. tomato. tomato), Pectobacterium carotovorum subsp. carotovorum, Phytophthora sojae, Pseudomonas syringae pv. lachrymans The plant disease resistance enhancing composition according to claim 1, which is induced by at least one fungus selected from the group consisting of and Magnaporthe oryzae.

3. The plant disease resistance enhancing composition according to claim 1, wherein the branched-chain amino acid is at least one selected from the group consisting of isoleucine, leucine, and valine.

4. The plant disease resistance enhancing composition according to claim 1, wherein the plant is at least one selected from the group consisting of legumes, fruit vegetables, leafy vegetables, and cereals.

5. The composition for enhancing plant disease resistance according to claim 1, wherein the amino acid prevents or treats infection of plants with pathogenic fungi.

6. The composition for enhancing plant disease resistance according to claim 1, wherein the amino acid induces systemic induced resistance in plants.

7. The composition for enhancing plant disease resistance according to claim 1, wherein the amino acid does not inhibit plant growth.

8. The composition is a plant disease resistance enhancing composition comprising valine, wherein the plant disease is at least one selected from the group consisting of leaf spot, bacterial soft rot, Phytophthora root rot, bacterial leaf blight, and rice blast, as described in claim 1.

9. The composition is a plant disease resistance enhancing composition containing isoleucine, and the plant diseases include bacterial leaf spot, bacterial soft rot, Phytophthora root rot, and bacterial leaf spot. A small number of diseases selected from the group consisting of bacterial leaf blight and rice blast. The plant disease resistance enhancing composition according to claim 1, wherein there is at least one of these components.

10. The composition for enhancing resistance to plant diseases according to claim 1, wherein the composition contains leucine and is for enhancing resistance to bacterial soft rot.

11. The composition for enhancing resistance to plant diseases according to claim 1, wherein the composition contains proline and is for enhancing resistance to bacterial soft rot.

12. The plant disease resistance enhancing composition according to claim 1, wherein the composition comprises at least one selected from the group consisting of glutamic acid, aspartic acid, and histidine, and is for enhancing resistance to leaf spot disease.

13. A method for enhancing resistance to plant diseases, comprising the step of treating a plant with the plant disease resistance enhancing composition described in claim 1.

14. The method for enhancing plant disease resistance according to claim 13, wherein the plant is at least one selected from the group consisting of legumes, fruit vegetables, leafy vegetables, and cereals.

15. The method for enhancing plant disease resistance according to claim 13, wherein the plant disease resistance enhancing composition is treated two or more times.

16. The method for enhancing plant disease resistance according to claim 13, wherein the plant disease resistance enhancing composition is applied at intervals of 3 to 15 days.

17. The method for enhancing plant disease resistance according to claim 13, wherein the method for enhancing plant disease resistance promotes plant growth.

18. A method for preventing or treating a plant disease, comprising the step of treating a plant with the plant disease resistance enhancing composition described in claim 1.

19. A plant control composition containing amino acids, The aforementioned amino acid is at least one selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine. The plant disease composition may be used to treat spot diseases, Bacterial soft rot, and Phytophthora blight. A plant control composition for controlling at least one plant disease selected from the group consisting of stem and root rot, bacterial leaf blight, and rice blast.

20. A method for controlling plants, comprising the step of treating a plant with the plant control composition described in claim 19.

21. The use of at least one amino acid selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine for enhancing plant disease resistance, The plant disease is at least one selected from the group consisting of leaf spot, soft rot, stem blight, bacterial leaf spot, and blast disease, and is used for enhancing plant disease resistance.

22. The use of at least one amino acid selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine for plant control, The plant disease is at least one selected from the group consisting of leaf spot disease, soft rot, stem blight, bacterial leaf spot, and rice blast, and is used for plant control.

23. The use of at least one amino acid selected from the group consisting of branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine for the prevention or treatment of plant diseases, The plant disease is at least one selected from the group consisting of leaf spot disease, soft rot, stem blight, bacterial leaf spot, and blast disease, and is used for the prevention or treatment of plant diseases.

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