Trichoderma genus strain
The Trichoderma sp. GS1 strain, used as an additive in hydroponic cultivation, effectively addresses the weak disease suppression issue in hydroponic systems, providing a robust solution for controlling water-borne diseases and enhancing plant health.
Patent Information
- Application Number
- JP2023204221
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- Filing Date
- 2023-12-01
- Publication Date
- 2025-06-12
AI Technical Summary
Trichoderma spp. in hydroponic cultivation do not exhibit a strong disease-suppressing effect, and existing solutions are inadequate for effectively controlling water-borne diseases in crops like spinach.
Utilization of Trichoderma sp. GS1 strain (Deposit number at the Patent Organism Depositary: NITE P-04001) as an additive for hydroponic cultivation, which is incorporated into a solid medium such as rock wool, to enhance disease suppression in hydroponically grown plants.
The Trichoderma sp. GS1 strain demonstrates a strong disease suppression effect against Pythium and other pathogens in hydroponic cultivation, leading to improved plant health and yield.
Smart Images
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Abstract
Description
Technical Field
[0001] The present disclosure relates to fungi of the genus Trichoderma and the like.
Background Art
[0002] Most of the hydroponic cultivation of vegetables in plant factories is limited to the Asteraceae family (such as lettuce) and the Brassicaceae family (such as Komatsuna). Although some vegetables such as the Chenopodiaceae family (spinach) and the Aizoaceae family (ice plant) are expensive, they have not been widely used as hydroponic crops. This is because crops such as spinach, which are vulnerable to water-borne diseases, are shunned. In addition, the application of pesticides to nutrient solutions is prohibited, and the lack of effective solutions is also a major factor. Although plant factories are closed facilities and it seems that there is no room for plant pathogens to enter, in fact, nutrient solution-transmissible plant diseases occur constantly.
[0003] Fungi of the genus Trichoderma have been known as biocontrol microorganisms for a long time (Non-Patent Documents 1 and 2). However, the disease-suppressing effect of Trichoderma fungi on diseases of hydroponically cultivated crops has hardly been clarified.
Prior Art Documents
Non-Patent Documents
[0004]
Non-Patent Document 1
Non-Patent Document 2
Summary of the Invention
Problems to be Solved by the Invention
[0005] In the course of the research, the present inventors found that Trichoderma spp. in hydroponic cultivation such as soilless culture do not necessarily exhibit a disease-suppressing effect, and even if they do exhibit such an effect, the effect is weak.
[0006] An object of the present disclosure is to provide Trichoderma spp. having a stronger disease-suppressing effect.
Means for Solving the Problems
[0007] As a result of intensive research in view of the above problems, the present inventors found that the above problems can be solved by using Trichoderma sp. GS1 strain (Deposit number at the Patent Organism Depositary: NITE P-04001), which is a Trichoderma spp. That is, the present disclosure includes the following aspects.
[0008] Item 1. A Trichoderma spp. which is Trichoderma sp. GS1 strain (Deposit number at the Patent Organism Depositary: NITE P-04001).
[0009] Item 2. An additive for hydroponic cultivation containing the Trichoderma spp. according to Item 1.
[0010] Item 3. The additive for hydroponic cultivation according to Item 2, which is used by being held in a solid medium for hydroponic cultivation.
[0011] Item 4. The additive for hydroponic cultivation according to Item 3, wherein the material of the solid medium for hydroponic cultivation contains at least one selected from the group consisting of rock wool, urethane, sand, gravel, perlite, peat moss, coconut coir, vermiculite, leaf mold, bark, bark compost, rice straw, rice straw compost, sawdust, sawdust compost, wood pellets, charcoal and bamboo charcoal.
[0012] Item 5. The additive for hydroponic cultivation according to any one of Items 2 to 4, which is used for disease suppression in hydroponic cultivation.
[0013] Item 6. A solid medium for hydroponics that holds the fungus of the genus Trichoderma described in Item 1.
[0014] Item 7. The solid medium for hydroponics described in Item 6, which is used for disease suppression.
[0015] Item 8. A method for producing a plant, which includes hydroponically cultivating the plant in a state where it can come into contact with the fungus of the genus Trichoderma described in Item 1.
Advantages of the Invention
[0016] According to the present disclosure, it is possible to provide a fungus of the genus Trichoderma having a stronger disease suppression effect.
Brief Description of the Drawings
[0017]
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Mode for Carrying Out the Invention
[0018] In this specification, the expressions "contain" and "include" include the concepts of "contain", "include", "consist essentially of", and "consist only of".
[0019] 1. Fungus of the genus Trichoderma In one aspect of the present disclosure, it relates to a Trichoderma genus bacterium which is Trichoderma sp. strain GS1 (Deposit Number at the Patent Organism Depositary: NITE P-04001) (which may also be referred to as "the Trichoderma genus bacterium of the present disclosure" in this specification). This will be described below.
[0020] The Trichoderma genus bacterium of the present disclosure is a type of filamentous fungus. The Trichoderma genus bacterium of the present disclosure is not dead, and as long as it is in a viable state, it is not particularly limited. Also, the properties are not particularly limited, and it can be in a solid state, pestle-like, gel-like, sol-like, or liquid state, etc. Further, the Trichoderma genus bacterium of the present disclosure can be in a state of the bacterium alone, or can be in a state combined with other substances and / or organisms (for example, a mixed state, a complex). Also, the Trichoderma genus bacterium of the present disclosure can be in a wet state, or can be in a dried or freeze-dried state.
[0021] The Trichoderma fungus of the present disclosure has been deposited at the Patent Microorganisms Depositary Center (NPMD) of the National Institute of Technology and Evaluation (NITE), and can be obtained from the center. Its culturing method is not particularly limited, and examples thereof include a method of static culturing on a PDA medium (potato dextrose agar medium) at about 25°C under aerobic conditions. When cultured by this culturing method, for example, by culturing for about several days (for example, 3 to 7 days), a sufficient amount of the fungus can be ensured for use as an additive for hydroponics described below.
[0022] 2. Additive for hydroponic cultivation In one aspect, the present disclosure relates to an additive for hydroponics (which may also be referred to as "the additive of the present disclosure" in this specification) containing the Trichoderma fungus of the present disclosure. This will be described below.
[0023] The additive of the present disclosure can consist only of the Trichoderma spp. of the present disclosure, or in addition to the Trichoderma spp. of the present disclosure, it may contain various other components according to the dosage form, application mode, etc. described later. The content ratio of the Trichoderma spp. of the present disclosure in the additive of the present disclosure can be appropriately determined according to the properties, application mode, etc. described later, but for example, it can be 0.01 to 100% by mass, 0.1 to 100% by mass, 1 to 100% by mass, 5 to 100% by mass, 10 to 100% by mass, 20 to 100% by mass, 40 to 100% by mass, 60 to 100% by mass, 80 to 100% by mass, or 90 to 100% by mass, and it can also be 0.01 to 90% by mass, 0.01 to 70% by mass, 0.01 to 50% by mass, 0.01 to 40% by mass, 0.01 to 30% by mass, 0.01 to 20% by mass, 0.01 to 10% by mass, 0.01 to 5% by mass, or 0.01 to 1% by mass. Examples of other components include carriers, fixing agents, dispersants, auxiliary agents, culture medium components, plant materials, etc. Examples of carriers include solid carriers such as talc, bentonite, clay, kaolin, diatomaceous earth, white carbon, vermiculite, and silica sand; liquid carriers such as water-soluble polymer compounds (polysaccharides such as agarose, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylic acid, etc.), water, vegetable oils, and liquid animal oils. Examples of fixing agents include casein, gelatin, gum arabic, alginic acid, etc. Examples of dispersants include alcohol sulfates, polyoxyethylene glycol ethers, etc. Examples of auxiliary agents include carboxymethyl cellulose, starch, lactose, etc. The culture medium components are not particularly limited as long as they are components of a culture medium that can be used for culturing Trichoderma spp., and examples include nutrient components in PDA medium. The plant materials are not particularly limited as long as they are materials composed of all or part of a plant body that can be used for culturing Trichoderma spp., and examples include gramineous plants such as barley. The above carriers, fixing agents, dispersants, auxiliary agents, culture medium components, and plant materials can be used alone or in appropriate combinations according to their respective purposes.
[0024] The properties of the additive of the present disclosure are not particularly limited. The additive of the present disclosure can be, for example, solid, paste-like, gel-like, sol-like, or liquid.
[0025] In a preferred embodiment, the additive of the present disclosure includes a culture of the Trichoderma genus bacteria of the present disclosure (for example, a mycelium material obtained by culturing the Trichoderma genus bacteria of the present disclosure on a solid medium, or a mycelium material obtained by inoculating and culturing the Trichoderma genus bacteria of the present disclosure on a plant). The culture may be in a wet state or a dry state. Further, as other specific embodiments of the additive of the present disclosure, for example, liquid agents, solid agents, powders, granules, pellets, wettable powders, flowables, emulsions, pastes, dispersants, etc. can be mentioned.
[0026] The additive of the present disclosure is for use in hydroponics of plants.
[0027] The target plants of the additive of the present disclosure are not particularly limited as long as they are plants capable of hydroponics. For example, it can be widely applied to angiosperms (dicotyledons, monocotyledons, etc.). Specific examples include nightshades such as tomatoes, peppers, chili peppers, and eggplants; cucurbits such as cucumbers, pumpkins, melons, and watermelons; brassicas such as cabbages, broccoli, and Chinese cabbages; leafy greens or herbs such as celery, parsley, and lettuce; alliums such as green onions, onions, and garlic; legumes such as soybeans, peanuts, kidney beans, peas, and azuki beans; other fruit vegetables such as strawberries; root vegetables such as daikon radishes, turnips, carrots, and burdocks; tubers such as taros, cassavas, potatoes, sweet potatoes, and yacon; leafy vegetables such as asparagus, spinach, Komatsuna, mizuna, water dropwort, purslane, morning glory, perilla, etc.; flower crops such as Turkish carnations, stocks, carnations, and chrysanthemums; cereals such as rice, wheat, barley, oats, corn, and quinoa; turfgrasses such as bentgrass and Korean lawn grass; oil crops such as rapeseed and peanuts; sugar crops such as sugarcane and sugar beets; fiber crops such as cotton and rush; forage crops such as clover, sorghum, and dent corn; deciduous fruit trees such as apples, pears, grapes, and peaches; citrus fruits such as Satsuma mandarins, lemons, and grapefruits; root vegetables such as ginger, taro, and sweet potato; flowering shrubs such as roses and hydrangeas, etc. From the perspective of being vulnerable to water-borne diseases, the target plants preferably include spinach, ice plant, perilla, quinoa, strawberry, etc. Although it is known that a specific Trichoderma (Trichoderma viride) causes seedling blight in gramineous plants such as rice, the GS1 strain is likely to be a different species from the Trichoderma (for example, Trichoderma asperellum). Therefore, the additive of the present disclosure is considered to be effective also for gramineous plants. On the other hand, the additive of the present disclosure can also target plants other than gramineous plants.
[0028] Hydroponics is a cultivation method in which crops are cultivated using a liquid (culture solution) in which fertilizer is dissolved in water, without using soil. Hydroponics methods include "hydroponics" in which roots grow in or on the surface of the culture solution without using a medium, "solid medium cultivation" in which crops are planted in various solid media that replace soil, and "spray cultivation" in which the culture solution is sprayed onto the roots in a mist form. The additives of the present disclosure can be applied to any of these methods. The additives of the present disclosure can be used in an appropriate manner such that the plant (especially the root or stem) can come into contact with the Trichoderma genus bacteria of the present disclosure according to various methods.
[0029] The application mode of the additive of the present disclosure is not particularly limited as long as the plant and the Trichoderma genus bacteria of the present disclosure can come into contact during or after application. Examples include addition to the plant growth environment, spraying on the plant, dripping onto the plant, coating on the plant, etc.
[0030] From the viewpoint of more effectively exerting the disease control effect, the additive of the present disclosure is preferably used by being held in a solid medium for hydroponics. The solid medium for hydroponics itself maintains a stable shape, can hold a part (usually a part or all of the root, stem, etc.) so that the plant does not fall over, and can penetrate the supplied culture solution to come into contact with the root or stem of the plant and be absorbed by the plant (has water permeability), and as long as it meets this requirement, it is not particularly limited.
[0031] The material of the solid medium for hydroponics is not particularly limited, and examples include rock wool, urethane, sand, gravel, perlite, peat moss, coconut coir, vermiculite, leaf mold, bark, bark compost, rice straw, rice straw compost, sawdust, sawdust compost, wood pellets, charcoal, bamboo charcoal, etc. Among these, inorganic materials, especially rock wool, are preferred from the viewpoint of being able to stably germinate, etc. The shape of the material of the solid medium for hydroponics is preferably fibrous from the viewpoint of water permeability, etc.
[0032] The solid medium for hydroponics is particularly preferably an aggregate of fiber materials (preferably inorganic materials, particularly preferably rock wool) from the viewpoint of more effectively exerting the disease inhibitory effect. The shape of the solid medium for hydroponics is not particularly limited, and can be, for example, a cube or a rectangular parallelepiped. Further, the solid medium for hydroponics is preferably divided into portions for one individual plant or used after being divided into portions for one individual plant.
[0033] The mode of holding the additive of the present disclosure in the solid medium for hydroponics is not particularly limited as long as the additive of the present disclosure does not immediately flow down due to the penetration of the nutrient solution and the plant (particularly the root or stem) can come into contact with the Trichoderma fungus of the present disclosure. From the viewpoints of simplicity, cost, etc., it is preferable to arrange the additive of the present disclosure around the stem or root of the plant on the surface of the solid medium for hydroponics or at the planned sowing site. The nutrient solution supplied during hydroponics continuously penetrates into the solid medium for hydroponics and the Trichoderma fungus of the present disclosure held therein.
[0034] The additive of the present disclosure has a disease inhibitory effect in hydroponics. Therefore, the additive of the present disclosure can be used for disease inhibition in hydroponics. Examples of the disease include a decrease in yield due to infection with pathogenic bacteria. The pathogenic bacteria are not particularly limited, and examples thereof include Pythium aphanidermatum and other Pythium bacteria, Orpidium bacteria, Phytophthora bacteria, Pseudomonas bacteria, Aphanomyces bacteria, Fusarium bacteria, soft rot bacteria, and bacterial wilt bacteria. Among these, particularly preferably, Pythium bacteria such as Pythium aphanidermatum are mentioned.
[0035] 3. Solid medium for hydroponic cultivation In one aspect, the present disclosure relates to a solid medium for hydroponics (which may also be referred to as "the medium of the present disclosure" in this specification) that holds the Trichoderma fungus of the present disclosure.
[0036] The medium of the present disclosure is for use in hydroponic cultivation of plants. The medium of the present disclosure can preferably be used for disease control in hydroponic cultivation.
[0037] Regarding the Trichoderma spp. of the present disclosure, the holding mode, hydroponic cultivation, solid medium for hydroponic cultivation, target plants, disease control, etc. in the medium of the present disclosure are as described in the above "1. Trichoderma spp." and "2. Additive for hydroponic cultivation".
[0038] 4. Method for producing plants In one aspect, the present disclosure relates to a method for producing a plant (which may also be referred to as "the production method of the present disclosure" in this specification) including hydroponically cultivating a plant in a state where it can come into contact with the Trichoderma spp. of the present disclosure.
[0039] Regarding the Trichoderma spp., target plants, hydroponic cultivation, etc. of the present disclosure are as described in the above "1. Trichoderma spp." and "2. Additive for hydroponic cultivation".
[0040] "Contactable" is not particularly limited as long as the Trichoderma spp. of the present disclosure and the plant can come into contact with each other at the time of or after application by the application of the Trichoderma spp. of the present disclosure.
[0041] In the production method of the present disclosure, the timing of applying the Trichoderma spp. of the present disclosure is not particularly limited. Cultivation can be started by applying the Trichoderma spp. of the present disclosure at the time of sowing, or after sowing and cultivating for a certain period, the Trichoderma spp. of the present disclosure can be applied for cultivation. In a preferred aspect of the present invention, it is preferable to apply the Trichoderma spp. of the present disclosure and cultivate at the seedling stage.
[0042] The production method of the present disclosure preferably includes hydroponically cultivating a plant using the solid medium for hydroponic cultivation of the present disclosure.
[0043] The nutrient solution used for hydroponics is a solution in which nutrients (essential elements) necessary for plant growth are dissolved in water at a concentration suitable for absorption, and it is a solution continuously supplied to plants.
[0044] As nutrients, nitrogen (nitrate form; NO 3 -N and ammonium form; NH 4 -N, etc.), phosphorus (phosphate form; PO 4 -P, etc.), potassium, calcium, magnesium, sulfur (sulfate form; SO 4 -S, etc.), and other macro elements, as well as trace elements such as iron, manganese, boron, copper, zinc, molybdenum, silicon, chlorine, aluminum, sodium, nickel, etc. Note that carbon, oxygen, and hydrogen can be mainly supplied in the form of carbon dioxide in the air and water absorbed from the roots.
[0045] Hydroponics conditions such as the composition of the nutrient solution, nutrient solution management, air temperature, humidity, light conditions, etc. can be appropriately set according to the type of crop to be cultivated, the quality of the target harvest, etc. Hydroponics conditions can be set with reference to, for example, "New Manual of Hydroponics" (edited by the Japan Society for Horticultural Facilities, Seibundo Shinkosha, July 2002), etc.
[0046] By the production method of the present disclosure, plants can be produced while suppressing diseases. According to the production method of the present disclosure, even in the presence of pathogenic bacteria, a higher yield can be obtained.
Examples
[0047] Hereinafter, the present invention will be described in detail based on examples, but the present invention is not limited by these examples.
[0048] Example 1. Isolation of Trichoderma genus strains In 2022, three Trichoderma strains (Trichoderma sp. GW1 strain, Trichoderma sp. GM1 strain, Trichoderma sp. GS1 strain) were isolated from the roots of hydroponically cultivated strawberries with symptoms of seedling blight confirmed in Sakai City, Osaka Prefecture, Japan. Among them, the Trichoderma sp. GS1 strain was deposited with the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE) on October 27, 2023 (Trichoderma sp. GS1 strain, Patent Organism Deposit Center Accession Number: NITE P-04001).
[0049] Example 2. Evaluation of the effect of Trichoderma genus strains on Pythium in hydroponic cultivation A hydroponic nutrient solution (manufactured by Otsuka House, product name: Otsuka House Fertilizer No. 1, a mixed solution of S1 and No. 2) was put into a simple hydroponic device in a plastic case (22 cm long x 14 cm wide x 4 cm deep), and a rock wool cube (23 mm long x 23 mm wide x 28 mm high) (manufactured by Grodan, product name: Grodan Taco Plug) was placed therein. The amount of the hydroponic nutrient solution was set to an amount such that the lower part of the rock wool cube was immersed. Spinach (variety: Mirrage) was sown in the depression at the center of the upper surface of the rock wool cube and grown for 10 days in a plant growing device with a daytime temperature of 24°C and a nighttime temperature of 23°C (11-hour day length). After 10 days of growth (seedling stage), it was divided into five test plots numbered 1 to 5 below. The presence or absence and outline of the treatments for each test plot are as shown in Table 1.
[0050]
Table 1
[0051] Details of the treatment in the case of inoculation with the HSP1 strain (Pythium aphanidermatum HSP1 strain = strain derived from hydroponic spinach) are as follows. The HSP1 strain in a 100 mL flask was cultured on a PDA plate medium at 25°C for 5 days, and a mycelial disk with a diameter of 5 mm was cut out. The obtained mycelial disk was cultured (for 5 days, at 31°C) in a Japanese millet seed medium (about 1 g of Bentgrass Highland, 0.175 g of agar, and 100 ml of DW were added and autoclaved), and then inoculated at the base of the hydroponic spinach (10 days after sowing).
[0052] Details of the treatment in the case of inoculation with Trichoderma strains (GW1, GM1, GS1) are as follows. Trichoderma strains (Trichoderma sp. GW1 strain, Trichoderma sp. GM1 strain, or Trichoderma sp. GS1 strain) were cultured on a PDA plate medium at 25°C for 5 days, and a mycelial disk with a diameter of 3 mm was cut out. The obtained mycelial disk was inoculated at the base of the hydroponic spinach (10 days after sowing).
[0053] After inoculation with the above HSP1 strain and Trichoderma strains, the hydroponic cultivation of spinach was continued for another 10 days. Three plants were cultivated in each experimental plot, and the experiment was repeated 4 times.
[0054] After 20 days of cultivation, the condition of the spinach was observed, and the symptoms of damping-off were evaluated. Also, the root length and the shoot length were measured. Further, a 5-mm section from the base of the root was taken out with tweezers and observed under a microscope to confirm the presence or absence of oospores and zoosporangia of the Pythium HSP1 strain.
[0055] External photos of the spinach are shown in Figures 1 and 2, the evaluation results of the symptoms of damping-off are shown in Table 2, the measurement results of the root length are shown in Figure 3, and the measurement results of the shoot length are shown in Figure 4.
[0056]
Table 2
[0057] The Trichoderma sp. GW1 strain and the Trichoderma sp. GM1 strain showed no or weak disease control effects against damping-off caused by Pythium in hydroponic cultivation (Figs. 1 to 4 and Table 2). On the other hand, the Trichoderma sp. GS1 strain showed a strong disease control effect against damping-off caused by Pythium in hydroponic cultivation (Figs. 1 to 4 and Table 2).
[0058] Also, as a result of checking for the presence or absence of oospores and zoosporangia of Pythium, when the Trichoderma sp. GW1 strain was used (test plot 3), a large number of oospores and zoosporangia were confirmed (Fig. 5), and oospores were also confirmed when the Trichoderma sp. GM1 strain was used (test plot 4), whereas no oospores and zoosporangia were found when the Trichoderma sp. GS1 strain was used (test plot 5) (Fig. 6). This indicates that the Trichoderma sp. GS1 strain has a unique effect of preventing the infection of Pythium.
[0059] Example 3. Confrontation culture test Confrontation culture was performed between each of the Trichoderma strains (Trichoderma sp. GW1 strain, Trichoderma sp. GM1 strain, Trichoderma sp. GS1 strain) and the Pythium strain (HSP1 strain). Specifically, the Trichoderma sp. GW1 strain, Trichoderma sp. GM1 strain, or Trichoderma sp. GS1 strain was inoculated at one end of a circular Petri dish containing PDA medium, and the Pythium strain was inoculated at the opposite end, followed by culturing at 31°C for 5 days.
[0060] A photograph of the Petri dish after the cultivation is shown in Fig. 7. When the Trichoderma sp. GS1 strain was used, the formation of a clear inhibition zone was confirmed.
Claims
1. A fungus of the genus Trichoderma, which is Trichoderma sp. strain GS1 (Deposit number at the Patent Biological Depositary Center: NITE P-04001).
2. An additive for hydroponics, containing the fungus of the genus Trichoderma according to Claim 1.
3. The additive for hydroponics according to Claim 2, which is used by being held in a solid medium for hydroponics.
4. The additive for hydroponics according to Claim 3, wherein the material of the solid medium for hydroponics contains at least one selected from the group consisting of rock wool, urethane, sand, gravel, perlite, peat moss, coconut coir, vermiculite, leaf mold, bark, bark compost, rice straw, rice straw compost, sawdust, sawdust compost, wood pellets, charcoal, and bamboo charcoal.
5. The additive for hydroponics according to any one of Claims 2 to 4, which is used for disease control in hydroponics.
6. A solid medium for hydroponics, which holds the fungus of the genus Trichoderma according to Claim 1.
7. The solid medium for hydroponics according to Claim 6, which is used for disease control.
8. A method for producing a plant, which includes hydroponically cultivating the plant in a state where it can come into contact with the fungus of the genus Trichoderma according to Claim 1.