WO2021018145A1 - Trichoderma virens strain for preventing and treating phytophthora capsici leonian, use thereof, and method for cultivating capsicum frutescens l. - Google Patents

Abstract

Provided is a Trichoderma virens strain, HZA14, for preventing and treating Phytophthora capsici leonian, with the deposit number being CCTCC M 2019484. The Trichoderma virens strain has an inhibitory effect on the growth of mycelia of Phytophthora capsici, is used for preventing and treating Phytophthora capsici leonian by means of reducing the morbidity and the disease severity of Phytophthora capsici, and can be used for biological control applications.

Description

Trichoderma viride for preventing and treating pepper phytophthora, application and pepper cultivation method

This application requires the priority of the Chinese patent application submitted to the Chinese Patent Office on July 29, 2019, the application number is 201910688710.0, and the invention title is "A Trichoderma viride for the prevention and control of pepper phytophthora, application and pepper cultivation method". Right, the entire contents of which are incorporated in this application by reference.

Technical field

The present invention relates to the technical field of biological control of plant diseases, in particular to a Trichoderma viride used for preventing and treating capsicum Phytophthora, application and capsicum cultivation method.

Background technique

Pepper (Capsicum frutescens L.) has been cultivated in my country for many years, and it is one of the vegetables with the largest cultivated area in my country, with an annual planting area of 1.42 million hectares. Chili is rich in vitamin C and more antioxidants in nutritional value. It has the effects of accelerating metabolism, protecting skin, controlling heart disease, lowering cholesterol, preventing cancer and other chronic diseases. From the point of view of production value, it has strong production adaptability, high yield potential and good commercial property, and it is suitable for long-season, early and late autumn high-yield cultivation.

Pepper blight, commonly known as "dead seedling disease", is a devastating soil-borne disease caused by Phytophthora capsici. It was first discovered in New Mexico (Leonian, L.H. 1992) and California (Tompkins, C.m., and Tucker, C.M. 1937) in the United States in 1918. It is now found in pepper growing areas all over the world. When the disease occurs, the main and lateral roots of the plant are light brown or black, the root hairs are reduced, and they rot in severe cases. Generally, the mortality of diseased plants is 15%-30%, and in severe cases, it can be as high as 80% or even no harvest. This brings serious losses to pepper production. my country first reported the occurrence of pepper blight in Jiangsu in 1940. Since the 1980s, with the rapid development of my country’s pepper industrialization, the continuous strengthening of cultivation measures and the continuous promotion of new pepper varieties, the current pepper blight has been It is common in my country and the incidence is increasing, mainly in Xinjiang, Qinghai, Heilongjiang, Beijing, Shanghai, Gansu, Guizhou, Yunnan, Shaanxi, Guangdong and the Yangtze River Basin.

Phytophthora capsici belongs to the phylum Oomycota, Oomycota, Pythium, Pythium, and Phytophthora. The main component of its cell wall is cellulose, not chitin. The vegetative hypha of Phytophthora is thick and has no septum. The sporangia stem produces asexual zoosporangia; sexual reproduction produces oospores. The oospores are spherical, thick-walled, and have an ovipositor outside. The oospores can survive the winter and survive the bad environment. The lowest temperature for pathogen growth is 10°C, and the optimum temperature is 24-28°C and the highest temperature is 35°C.

Combination of agricultural cultivation measures and chemical pesticide control has always been the main method to control pepper fusarium wilt. Agricultural cultivation measures include crop rotation, soil insolation, soil improvement, and cultivation techniques to prevent the spread of pathogens (drip irrigation, high ridges and film mulching), but these methods can only reduce the occurrence of diseases. Fumigating the soil with bromomethane is the only effective control method. However, bromomethane has a destructive effect on the ozone layer of the atmosphere, and many countries have expressly banned or reduced its use. The disease is still mainly applied with chemically synthesized fungicides. These conventional prevention and control technologies have been difficult to effectively control and easily cause ecological and environmental problems.

Biological control is currently recognized as a safe and effective biological control measure. Trichoderma mycelium has a variety of antagonistic mechanisms, including reparasitisation and the production of enzymes that degrade the cell walls of pathogens (chitinase, xylanase, glucanase, etc.) And produce antibiotics, etc. However, for different pathogens, the antagonistic mechanisms and activities of different Trichoderma species and strains are different. In particular, the ability to produce antibiotics varies between different isolates of the same bacteria and between different isolates of different species. There will be changes (Dennis, C., Webster, J., 1971. Antagonistic properties of species-groups of Trichoderma: II. production of volatile antibiotics. Trans. Br. Mycol. Soc. 57, 25–39.), so For different pathogens, screen isolates of Trichoderma. Having a high antagonistic activity and characterizing its antagonistic mechanism is an important work for its application as a biological control agent (Vinale, F., Sivasithamparam, K., Ghisalberti, EL, Marra, R., Barbetti, MJ, Li, H., Woo, SL,Lorito,M.,2008.A novel role for Trichoderma secondary metabolites in the interactions with plants.Physiol.Mol.Plant Pathol.72,80–86.).

Summary of the invention

Aiming at the deficiencies in the prior art, the present invention provides a Trichoderma viride for preventing and treating capsicum Phytophthora, application and capsicum cultivation method.

A Trichoderma virens used to prevent and treat Capsicum Phytophthora, classified and named as Trichoderma virens, strain number HZA14, and preservation number as CCTCC NO: M 2019484.

The present invention Trichoderma virens HZA14 is collected from the soil where pepper seeds have been grown for many years in Hangzhou. The biological characteristics of the strain are as follows: the colony grows rapidly, with a light dark blue-green color, and the back is colorless to light yellow; the conidia monospores are smooth, oval, green in clusters, with a size of 3.2～6.1×2.6～4.3μm. Conidiophores are usually whorled, pyramid-shaped, with short intervals between the wheels; sporozoites are ampoule-shaped to flask-shaped. Usually a large number of chlamydospores are produced. In the PDA medium, a yellow pigment is usually produced, but there is no obvious smell. The optimum growth temperature is 25～30℃.

The present invention also provides the application of the Trichoderma viride in inhibiting the growth of Phytophthora capsici. When the Trichoderma viride HZA14 of the present invention is cultured in a confrontation with Phytophthora capsici, the HZA14 grows rapidly, obviously causes the mycelium to collapse and degrade, and shows high antagonistic activity.

The present invention also provides the application of the said Trichoderma viride in preventing and treating capsicum phytophthora.

The invention also provides a pepper cultivation method, which includes the following steps:

(1) Preparation of the Trichoderma spore suspension of Trichoderma viride;

(2) Watering the Trichoderma spore suspension in step (1) on the roots of pepper seedlings.

The preparation method of the Trichoderma spore suspension includes the following steps: cultivating the Trichoderma viride to obtain Trichoderma spore powder, dispersing the Trichoderma spore powder in water to prepare a Trichoderma spore suspension, wherein 1 g of Trichoderma spores is added to every 10 ml of water powder.

Step (2) Watering 10ml of Trichoderma spore suspension around each pepper seedling during watering.

Step (2) The watering time is that the pepper grows to the stage of 6-8 leaves.

The present invention obtains a Trichoderma viride that can be used to prevent and treat Phytophthora capsici through screening. The classification is named Trichoderma virens, the strain number is HZA14, and the preservation number is CCTCC NO: M 2019484. Efficient inhibitory effect, used to control pepper blight can significantly reduce the incidence and severity of Phytophthora capsici, and has a good application prospect for biocontrol.

Biological preservation instructions

Trichoderma virens (HZA14) was deposited in the China Type Culture Collection on June 24, 2019, at the Wuhan University School, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province. The deposit number is: CCTCC NO: M 2019484.

Description of the drawings

Figure 1 shows the antagonistic effect of 15 isolates (HZA1-HZA15) (bottom) with an antagonistic grade of 1 against Phytophthora capsici hypha (upper) on PDA after 5 days of inoculation. Figures a～o are the isolates respectively. HZA1～15;

Figure 2 shows the collapse (*) and degradation results of the isolate HZA14 (bottom) caused by Phytophthora capsici (top) colony, where A and B are the results of two repeated experiments respectively;

Figure 3 shows the morphological observation results of Trichoderma pseudo-dorothée in the PDA or CMD or SNA medium of a 9cm diameter petri dish under 12h dark/12 light conditions for 4 days, where a～c: on PDA medium (a), on CMD medium (b), on SNA medium (c); d～m: on CMD medium; d～f: conidia piles, which can be seen in the conidia piles Single pinnate conidiophore (f); g～k: conidiophore and phialophore. At g (arrow) and j (arrow) you can see the apical hyperplasia of phialophore; l: conidia; m: Chlamydospores, scale bars: d and e=1mm, f=500μm, g～m=10μm;

Figure 4 is the full scan mass spectrum of active component C. The peak of gliotoxin that has lost two sulfur atoms is at m/z263;

Figure 5 shows the results of detection of the inhibitory activity of different concentrations of gliotoxin on the mycelial growth of Phytophthora capsici on the culture medium, where a: 0.5μg/ml gliotoxin is used; b: 1.0μg/ml gliotoxin is used; c: use 5.0μg/ml gliotoxin; d: use 10.0μg/ml gliotoxin; e: use 15.0μg/ml gliotoxin; f: control group, without gliotoxin;

Figure 6 shows the results of inoculation of the spore powder suspension of Trichoderma viride HZA14 isolate on the incidence and severity of pepper blight, in which a ~ b: pathogenic bacteria inoculated 14 days later; a: Trichoderma viride HZA14 spore powder suspension and pepper blight The lesions of the plants treated with the zoospore suspension of fungal infection spread upward (arrow); b: plants only inoculated with the zoospore suspension of Phytophthora capsici; c: the incidence and severity of disease after 15 days of inoculation; severity (DS%) )=Σ(disease grade×number of plants)/(maximum grade value×total number of plants)×100, the vertical line represents the standard deviation of the average (n=3), according to the least square method (LSD), when p<0.05, There are significant differences in the values of different letters with the bars.

Detailed ways

Example 1

Isolation, screening and identification of strains.

1. Isolation of Trichoderma

Forty soil samples were collected from 8 fields in Hangzhou City, Zhejiang Province. These fields were severely infected by the pathogen of pepper fusarium. After retrieval, the samples were stored in a refrigerator at 4°C. Dissolve 1g soil sample in 9ml sterile water to prepare a series of soil suspensions. Take 1ml soil suspension and evenly spread it on the Trichoderma screening medium (TSM: 0.2g MgSO ₄ 7H ₂ O; 0.9g K ₂ HPO ₄ ; 3.0g glucose; 0.25g chloramphenicol; 0.3g sodium p-dimethylaminobenzodiazepine; 0.2g pentachloronitrobenzene; 0.15g bengal red; 20g agar; add water to make the volume to 1L), at 27± Cultivate for 4 days at 1°C, then transfer the colonies grown on the medium to potato dextrose medium (PDA) for monospore isolation. For short-term preservation of strains, place in a refrigerator at 4°C. For long-term storage, place in the refrigerator at -40°C (spores + 17% skimmed milk powder + silica gel particles).

2. Screening of Trichoderma

The antagonistic ability of the isolated strains was tested by the confrontation culture method. After the Trichoderma strain isolated from the soil sample was cultured on PDA medium for three days, a 5mm diameter bacterial cake was taken from the edge of the colony and placed on the side of a 9cm diameter PDA petri dish. Take the Phytophthora capsici cake on the other side of the petri dish and incubate at 25±1℃. The test was repeated three times, and the antagonistic effect was observed and recorded 7 days later. The evaluation of the antagonistic effect uses the grading standard of 1 to 5:

①Trichoderma grows completely on Phytophthora, covering the entire surface of the medium; ②Trichoderma grows on at least two-thirds of the medium; ③Trichoderma and Phytophthora each account for half of the medium, neither of which is obvious Growth advantage; ④Each species of Trichoderma and pathogens have nearly half of the colonization in the medium, and no one kind of microorganism is dominant in the growth of the medium; ⑤The pathogens are completely grown on the Trichoderma and in the medium Colonization of the entire surface.

The screening results showed that based on the antagonistic activity, 15 strains with significant antagonism to Phytophthora capsici were finally obtained (Figure 1). They were named HZA1, HZA2, HZA3, HZA4, HZA5, HZA6, HZA7, HZA8, HZA9, HZA10, HZA11, HZA12, HZA13, HZA14, HZA15. Among them, the isolated line HZA14 caused the colony collapse and degradation of Phytophthora capsici (Figure 2).

3. Morphological observation of the separated lines

Under 20～25℃, 12h light and 12h dark cycle conditions (Jaklitsch, W.M., 2009. European species of Hypocrea part I. the green-spored species. Stud. Mycol. 63, 1–91.). The isolated Trichoderma virens strain HZA14 was cultured on potato dextrose agar medium (PDA), oat agar medium (CMD) and synthetic low nutrient medium (SNA). A Zeiss microscope (AxioVision Software Release 3.1., v.3-2002; Carl Zeiss Vision Imaging Systems) with Axiocam CCD camera and Axiovision imaging system was used to observe and measure the asexual structure of fungi, such as conidiophores, phial stems, and subspores. Spores and chlamydospores. Compare closely related taxa on the phylogenetic tree through morphological taxonomic features.

The optimum temperature for growth on PDA and SNA medium is 27-30°C. The colonies grown on PDA produced conidia within 96 hours, but a large number of aerial mycelium did not have concentric rings (a in Figure 3); on CMD medium, non-concentric aerial mycelia were rich in Contains yellow conidia (b in Figure 3); on the SNA medium, a pile of microspores began to form on the conidia, and there were significant concentric circles around the inoculated bacterial cake (c in Figure 3). No diffuse pigments or unique odors were detected on any medium. On SNA, the conidia piles are green to dark green. Conidia production is almost continuous, tending to form dense, flocculent spore piles, the size of the spore piles is 1-2mm (d in Figure 3 and e in 3). Long and fully fertile branches are usually seen in the spore pile (f in Figure 3). Conidiophores have a recognizable main axis, along with fertile branches, which appear more or less in pairs in longer or shorter internodes (g in Figure 3). The longer branches appear near the base of the stem, and the shorter ones appear at the top (h in Figure 3); the branches re-branching or directly produce the bottle body (i in Figure 3); sometimes, one location Several bottles are clustered together (k in Figure 3). The size of the bottle is (9.76-)9.69-11.43(-11.92)×(2.54-)3.06-3.89(-4.36)μm, slender, straight and slightly enlarged in the middle. In some cases, the bottle body tends to proliferate and form a new bottle body (i and j in Figure 3). Conidia size (3.19-)3.34-3.91(-4.18) (average length 3.63μm)×(2.86-)3.07×3.48(-3.56) (average width 3.28μm)μm, spherical to nearly spherical, occasionally elliptical, Smooth (l in Figure 3). A large number of chlamydospores are produced at the end or inside of the hyphae, spherical to nearly spherical (m in Figure 3).

Example 2

Detection of antibacterial activity of metabolites.

The 15 strains that were screened and purified were taken with a puncher to take a bacterial cake with a diameter of 5 mm into an Erlenmeyer flask containing 100 ml PDB medium, and cultured in a ZWY-211B shaker at 27±1°C and 150rpm for 4 days. First use sterile gauze to filter and centrifuge the hyphae to obtain a supernatant containing metabolites. Dilute the obtained supernatant to 50% and 20% concentrations, filter with a bacterial filter with a pore size of 0.22μm, pour 1ml of 50% and 25% metabolite solutions together with 10ml of melted PDA medium into a 9cm petri dish, and then connect Into the Phytophthora capsici cake with a diameter of 5mm, culture at 25°C. The treatment without metabolites was used as a control, and the measurement was performed when the growth of the Phytophthora hyphae of the control treatment approached the edge of the petri dish.

The inhibitory effect test of 15 Trichoderma metabolites on the growth of Phytophthora capsici showed that the metabolites from different isolates showed different levels of inhibitory activity (p<0.05) (Table 1). The metabolites produced by T. virens HZA14 completely inhibited mycelial growth after being diluted 20 or 40 times, showing the highest inhibition percentage (100%). Followed by T. afroharzianum HZA3, the inhibition rates were 44.85% (40 times) and 78.96% (20 times), respectively. T. citrinoviride HZA9, the inhibition rates were 42.44% (40 times) and 77.81% ( 20 times), and T. dorothopsis (T. dorothopsis) HZA5, HZA8, HZA15, the inhibition rates were 34.67% ~ 39.59% (40 times) and 70.92% ~ 71.33% (20 times), T. dorothopsis (T. koningiopsis) HZA6, the inhibition rates were 37.29% (40 times) and 72.18% (20 times), respectively. The strains with the lowest inhibition percentages were T.atroviride HZA1, HZA2 and HZA13, and Trichoderma acnes (T.asperellum) HZA10 and Trichoderma harzianum (T.harzianum) HZA11, the inhibition rates were 0.62%～1.59% (40 times) and 5.18%～6.29% (20 times).

Table 1 The inhibitory effect of metabolites produced by 15 Trichoderma isolates on the growth of Phytophthora capsici

^a Percentage inhibition rate (%) of the radial growth of Phytophthora capsici hyphae on the PDA medium containing the diluted culture solution 5 days after inoculation, ^b Dilute the culture solution 40 times or 20 times, after LSD test, the letter after the median Indicates that the difference is significant when p<0.05.

Example 3

Purification and identification of active metabolites.

1. Purification and activity test of active metabolites

Based on the results of the inhibitory activity, the possible active compounds produced by Trichoderma viride HZA14 were isolated, purified and identified. The selected Trichoderma viride HZA14 was inoculated into an Erlenmeyer flask containing PDB, and cultured in a shaker for 14 days, using the above method. Obtain 2L of culture broth, and extract metabolites with ethyl acetate. The metabolite residue was obtained by evaporation under reduced pressure. The residue was purified by silica gel column chromatography (particle size 200-300 mesh), and purified by preparative silica gel TLC (GF254) to obtain four components A, B, C, and D. A small amount of components A, B, C, and D are dissolved in DMSO (dimethyl sulfoxide) for biological activity determination. Add 1ml of each component (100μg/ml) solution to a petri dish of 10ml molten PDA. A cake (5mm) containing Phytophthora capsici hyphae was placed in the center of each PDA plate. Observe after 4 days of cultivation at 25°C, and determine the biological activity of the four components A, B, C, and D by the amount of mycelium growth on the plate. The test results show that component C has high inhibitory activity, while the other three components are inactive.

2. Identification of the chemical structure of active metabolites

In order to identify the chemical structure of the C component, the C component was further purified. The C component was further purified by Waters 600 high performance liquid chromatography (HPLC) equipped with Shim-pack Prep ODS column (20×250mm). The eluent was monitored at 254 nm with a Waters 2487 Dual lambda absorbance detector. By isocratic elution with a mixture of methanol and distilled water (1:1 V/V), a good semi-preparative separation of the active fraction peak was obtained at a flow rate of 6 mL/min. Component C was analyzed with VG Autospec-3000 mass spectrometer (VG, Manchester, UK) and API QSTAR Pulsar 1 (Applied Bio-systems, Foster City, USA). The purified component C was dissolved in methanol (5mg/ml) and injected into the mass spectrometer. The results of the full scan mass spectrum are shown in Figure 4. The ion peak (m/z349) corresponds to the molecular weight of glioblastin [M+Na] ⁺ sodium, and the main product ion peak (m/z263) corresponds to the desulfurized glioblastin [M-2S] ⁺ , just like the original results of electron bombardment mass spectrometry (Bose, AK, Das, KG, Funke, PT, Kugajevsky, I., Shukla, OP, Khanchandani, KS, Suhadolnik, RJ, 1968. Biosynthetic studies on gliotoxin using stable isotopes and mass spectral methods. J. Am. Chem. Soc. 90, 1038-1041.). The isotope distribution of the product ion also confirms that its composition lacks two sulfur atoms (Bose, AK, Das, KG, Funke, PT, Kugajevsky, I., Shukla, OP, Khanchandani, KS, Suhadolnik, RJ, 1968. Biosynthetic studies on gliotoxin using stable isotopes and mass spectral methods. J. Am. Chem. Soc. 90, 1038-1041.). The production of the product ion peak (m/z245) is related to the molecular weight [M-2S-H ₂ O] ⁺ (Grovel, O., Pouchus, YF, Robiou du Pont, T., Montagu, M., Amzil, Z. ,Verbist,JF,2002.Ion trap MSn for identification of gliotoxin as the cytotoxic factor of a marine strain of Aspergillus fumigatus Fresenius.J.Microbiol.Meth.48,171–179.), a product ion sheet attributable to ion peak m/z28 (Svahn, KS,

U.,El-Seedi,H.,Bohlin,L.,Larsson,DGJ,Olsen,B.,Chryssanthou,E.,2012.Antimicrobial activity of filamentous fungi isolated from highly antibiotic-contaminated river sediment.Infect.Ecol.Epidemiol .2,1–6.). The chemical structure analysis showed that the C component is the reported glioblastin.

3. Determination of glioblastin activity

For further determination of glioblastin, the purified compound C was dissolved in DMSO to obtain different mother liquors, and they were mixed with molten V8 juice medium to prepare final concentrations of 0.5, 1.0, 5.0, 10.0 and 15.0 μg/ml flat. A cake (5mm) containing Phytophthora capsici hyphae was placed in the center of each V8 medium plate. Each treatment was repeated five times. After culturing for 4 days at 25°C, when the control colony was close to the edge of the plate, the diameter of the colony was measured (Figure 5).

The results of the activity test showed that the 0.5μg/ml component C had 40% mycelial growth inhibition after 4d culture, and the 1.0μg/ml glioblastin had 66% mycelial growth inhibition after 4d culture. However, glioblastin at a concentration of 5.0 μg/ml or higher completely inhibited mycelial growth (100% inhibition).

Example 4

Application of screening strains in preventing and controlling pepper blight.

1. Preparation of Trichoderma spore powder and pathogen inoculum

The selected Trichoderma viride HZA14 was inoculated into a wheat kernel medium made of sterilized wheat kernels, and cultured for 15 days at a temperature of 25° C. under a 12-h light/dark cycle to obtain Trichoderma spore powder.

Put 5 colonies containing Phytophthora capsici hyphae into a petri dish filled with 10 mL sterile water, and place the petri dish at 25°C under light conditions for 3 days to produce Phytophthora zoosporangium ; Then, put the cultured zoosporangia into a refrigerator at 4°C to release the zoospores. The suspension was filtered with cotton gauze to remove hyphae and debris, and observed with a hemocytometer to prepare a zoospore suspension of Phytophthora zoospores with a concentration of 2000 zoospores/mL.

2. Plant cultivation and control test

The pepper seeds are disinfected with 2% sodium hypochlorite solution, and then rinsed with sterile water for 2 to 3 times. The seeds are placed on a flat plate containing moist sterilized filter paper and cultured at 25°C for 5 to 6 days. After the seeds germinate, sowing In a plug containing a nutrient soil substrate. Among them, the nutrient soil substrate is composed of peat: vermiculite: field nutrient soil ratio of 2:1:1, and the plug size is 11×11×11cm. The plugs are placed in a light growth incubator with a humidity of 80% to 90% and a temperature of 28 to 30°C. When the plant is growing at 6 to 8 leaf stages, water 10 mL of Trichoderma spore suspension (1g of Trichoderma spore powder dissolved in 10 mL of sterile water) around the root of each seedling. One week after the inoculation of the Trichoderma spore suspension, each plant was inoculated with 2 ml of the Phytophthora capsici zoospore suspension. Inoculated sterile water and pathogenic bacteria were used as controls, and each treatment was repeated 3 times.

3. Evaluation of control effect

The inoculation test showed that the selected strain HZA14 can delay the occurrence of pepper blight and significantly reduce the incidence and severity of the disease. After 5 days of inoculation, brown lesions were observed on the stem base of a few seedlings in the control treatment. After 10 days of inoculation, the symptoms of leaf wilting were observed in the control treatment, but no symptoms were found in the pepper plants co-treated with the isolate HZA14 and the zoospore suspension. However, 12 days after inoculation, typical symptoms appeared at the base of the stem of the pepper plants co-inoculated with the isolate HZA14 and zoospore suspension, and 14 days later, the lesions were significantly expanded (a in Figure 6). During this period, the control plants withered and fell severely (b in Figure 6). 15 days after inoculation, the incidence and severity of pepper plants co-inoculated with isolate HZA14 and zoospore suspension were 29.84±2.6% and 14.18±0.6%, while the incidence and severity of the control treatment were 92.48±, respectively. 2.1% and 88.38±2.9% (c in Figure 6). Obviously, the isolated line Trichoderma viride HZA14 significantly reduced the incidence (62.64%) and the incidence (64.2%) of Phytophthora capsici.

The Trichoderma virens HZA14 isolated in this application was named Trichoderma virens, strain number HZA14, and was deposited at the China Center for Type Culture Collection (CCTCC) at Wuhan University, Wuhan, China on June 24, 2019. The deposit number: CCTCC NO: M 2019484.

It can be understood that for those of ordinary skill in the art, equivalent substitutions or changes can be made according to the technical solution and concept of the present invention, and all these changes or substitutions should fall within the protection scope of the appended claims of the present invention.

Claims (8)

1. A Trichoderma virens for the prevention and treatment of Phytophthora capsici, characterized in that it is classified and named as Trichoderma virens, strain number is HZA14, and preservation number is CCTCC NO: M2019484.
2. The use of the Trichoderma viride of claim 1 in inhibiting the growth of Phytophthora capsici.
3. The application according to claim 2, characterized in that the Trichoderma viride produces glioblastin through metabolism to inhibit the growth of Phytophthora capsici.
4. The use of the Trichoderma viride of claim 1 in the prevention and treatment of capsicum phytophthora.
5. A method for cultivating pepper, which is characterized by comprising the following steps:

   (1) Preparation of a Trichoderma spore suspension of Trichoderma viride according to claim 1;

   (2) Watering the Trichoderma spore suspension in step (1) on the roots of pepper seedlings.
6. The pepper cultivation method according to claim 5, wherein the preparation method of the Trichoderma spore suspension comprises the following steps: cultivating the Trichoderma viride according to claim 1 to obtain Trichoderma spore powder; The mold spore powder is dispersed in water to prepare a Trichoderma spore suspension, wherein 1 g of the Trichoderma spore powder is added to every 10 ml of water.
7. The pepper cultivation method according to claim 5, wherein in step (2), 10 ml of Trichoderma spore suspension is watered around the root of each pepper seedling during watering.
8. The pepper cultivation method according to claim 5, characterized in that the watering time of step (2) is that the pepper grows to the stage of 6-8 leaves.

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