Trichoderma asperellum source natural product as well as preparation method and application thereof

Abstract

The invention belongs to the field of trichoderma secondary metabolites, and discloses a trichoderma asperellum source natural product as well as a preparation method and application thereof. The natural product has a structure as shown in a formula I. The trichoderma asperellum source natural product (namely ergosterol) extracted by the method disclosed by the invention can be applied to promoting plant growth, inducing plants to generate disease resistance and reducing plant morbidity. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of Trichoderma secondary metabolites, and more specifically, relates to a natural product derived from Trichoderma asperellum, a preparation method thereof, and an application thereof. Background Art

[0002] As an important biocontrol fungus, Trichoderma has a diverse range of novel-structured active metabolites and serves as an important resource pool for antibacterial active substances. Mining active substances with biocontrol functions from it has become a research hotspot. The active metabolites from Trichoderma harzianum include 6-pentyl-2H-pyran-2-one (6-PP), 1,2-dibromo-4-tert-butylbenzene (harzianopyridone), 2(5H)-furanone, and δ-decanolactone, which can significantly inhibit the growth of plant pathogens such as Macrophomina phaseolina, Rhizoctonia solani, Sclerotium rolfsii, and Fusarium oxysporum. The azaphilone from Trichoderma harzianum T22 and butenolide from T39 have obvious inhibitory effects on Leptosphaeria maculans, Phytophthora cinnamomi, and Botrytis cinerea. A ten-membered lactone, cremenolide, from Trichoderma cremeum effectively inhibits the radial mycelial growth of Fusarium oxysporum, Botrytis cinerea, and Rhizoctonia solani. The metabolites of Trichoderma can not only antagonize pathogens but also promote plant growth and induce disease resistance in plants. Trichokonins isolated from the metabolites of Trichoderma pseudokoningii SMF2 by Luo et al. can increase the activities of PAL and POD enzymes in tobacco under pathogen stress, upregulate the expression of multiple defense enzyme genes, and induce systemic resistance of tobacco to tobacco mosaic virus (TMV). Pascale et al. found that spraying the active substances harzianic acid (HA) and 6-pentyl-α-pyrone (6-PP) from Trichoderma harzianum M10 and Trichoderma atroviride P1 on grape leaves can inhibit the incidence of grape powdery mildew, increase grape yield, and enhance antioxidant activity.

[0003] In agricultural production, crop diseases have long been an important cause of yield reduction. Wheat scab caused by Fusarium graminearum is an important disease of wheat, generally resulting in a yield reduction of 10% to 30% or even a complete crop failure. To prevent and reduce the occurrence of such diseases, the extensive application of chemical pesticides is still the control strategy in most regions. However, promoting the reduction of chemical pesticides and increasing their efficiency has become a research hotspot. Trichoderma, as an important biocontrol fungus, can grow rapidly and form a biological barrier around the plant roots, promoting plant growth and enhancing plant disease resistance. Therefore, it is very necessary to apply Trichoderma more widely and comprehensively in agricultural production. Hundreds of secondary metabolites have been found in Trichoderma, but the research on these active substances mainly focuses on the preliminary exploration of biological activities such as cytotoxicity, anti-tumor activity, and antibacterial activity, and there are relatively few studies on plant growth regulation and disease resistance.

[0004] Ergosterol is the main sterol molecule in the fungal cell membrane, and it is also a precursor for the synthesis of various microorganisms, carrier compounds, and hormones. At the same time, it is an important biomarker for evaluating fungal biomass. At present, the main production methods of ergosterol at home and abroad are microbial fermentation. The strains used for large-scale production are yeasts, and some are also produced using Aspergillus. The research on ergosterol mainly focuses on its anti-inflammatory, lipid-lowering, anti-cancer, and immunomodulatory activities, and there are relatively few reports on its antibacterial activity. And there are no research reports on the application of ergosterol in plant growth regulation and disease resistance.

[0005] Therefore, there is an urgent need to propose a method for extracting and separating ergosterol from Trichoderma, so that the extracted ergosterol can be applied in improving plant disease resistance and plant growth regulation. Summary of the Invention

[0006] The purpose of the present invention is to address the deficiencies of the prior art and propose a natural product derived from Trichoderma asperellum, its preparation method, and application. The natural product derived from Trichoderma asperellum (i.e., ergosterol) extracted by the method of the present invention can be applied to promote plant growth, induce plant disease resistance, and reduce the plant incidence rate.

[0007] To achieve the above purpose, in the first aspect of the present invention, a natural product derived from Trichoderma asperellum is provided. This natural product has the structure shown in Formula I:

[0008]

[0009] In the second aspect of the present invention, a preparation method of a natural product derived from Trichoderma asperellum is provided. This preparation method includes fermenting and culturing Trichoderma asperellum in rice in sequence and then performing extraction, separation, and purification to obtain the above-mentioned natural product derived from Trichoderma asperellum.

[0010] According to the present invention, preferably, the temperature for the rice fermentation culture is 25 - 35 °C, and the time is 28 - 35 days.

[0011] According to the present invention, preferably, the extraction, separation and purification include ethyl acetate extraction, normal-phase silica gel column chromatography and recrystallization carried out in sequence.

[0012] According to the present invention, preferably, the number of times of ethyl acetate extraction is 2 - 4 times.

[0013] According to the present invention, preferably, the reduced-pressure gradient elution system used in the normal-phase silica gel column chromatography is a petroleum ether - dichloromethane - methanol system.

[0014] According to the present invention, preferably, the solvent used for recrystallization is at least one of methanol, petroleum ether and dichloromethane; the temperature for recrystallization is 15 - 35 °C.

[0015] In the present invention, as a preferred embodiment, the preparation method of the Trichoderma asperellum-derived natural product (i.e., ergosterol) includes the following steps:

[0016] 1) Inoculate the spore suspension of Trichoderma asperellum into a wide-mouth bottle containing a rice solid medium for fermentation culture to obtain a fermented product;

[0017] 2) Ultrasonically extract the fermented product with ethyl acetate and filter it. The filtrate is concentrated to dryness under reduced pressure to obtain a total ethyl acetate extract;

[0018] 3) Dissolve the total ethyl acetate extract in an organic mixed solvent (dichloromethane and methanol (1:1)) to obtain a dissolved mixture; perform reduced-pressure gradient elution chromatography on the dissolved mixture by using normal-phase silica gel column chromatography to obtain 22 groups of fractions; separate the ergosterol from at least one group of the fractions by recrystallization, and dry it at 25 - 30 °C.

[0019] In the present invention, the preparation method of the spore suspension of Trichoderma asperellum includes: On a PDA plate containing the colony of Trichoderma asperellum CBS 433.97, cut out a mycelial disc with a 6 mm punch and inoculate it into a conical flask containing 100 mL of PDB medium. Place the conical flask in a constant-temperature shaker at 25 °C and 180 r / min for 7 days. Filter the Trichoderma asperellum fermentation broth cultured for 7 days with four layers of sterile gauze, count the spores in the filtrate with a hemocytometer under an optical microscope, and then dilute the filtrate with sterile distilled water to a spore suspension with a spore content of 2×10 8 spores / mL, and store it in a refrigerator at 4 °C for later use.

[0020] The third aspect of the present invention provides the application of the Trichoderma asperellum-derived natural product in the following aspects:

[0021] 1) Promote plant growth;

[0022] 2) Induce disease resistance in plants.

[0023] According to the present invention, preferably, the plant is wheat and / or balsam pear.

[0024] According to the present invention, preferably, the promoting plant growth includes using the Trichoderma asperellum - sourced natural product to promote the growth of wheat and balsam pear.

[0025] According to the present invention, preferably, the promoting the growth of wheat includes promoting the increase of wheat root length, the increase of the number of roots and the increase of plant height.

[0026] According to the present invention, preferably, the promoting the growth of balsam pear includes promoting the increase of balsam pear root length, the increase of the number of roots, the increase of plant height and the increase of leaf area.

[0027] According to the present invention, preferably, the inducing disease resistance in plants includes using the Trichoderma asperellum - sourced natural product to induce wheat to produce disease resistance against Fusarium head blight and induce balsam pear to produce disease resistance against Fusarium wilt.

[0028] According to the present invention, preferably, the way of inducing wheat to produce disease resistance against Fusarium head blight includes increasing the activity of wheat antioxidant enzymes and regulating the expression of wheat disease - resistant genes.

[0029] According to the present invention, preferably, the way of inducing balsam pear to produce disease resistance against Fusarium wilt includes regulating osmotic adjustment substances, reducing membrane lipid peroxidation, increasing the activity of balsam pear antioxidant enzymes and reducing cell damage.

[0030] According to the present invention, preferably, the application of the Trichoderma asperellum - sourced natural product is carried out in the form of a solution, and the concentration of the Trichoderma asperellum - sourced natural product in the solution is 0.1 - 10 mg / L. Preferably, the concentration of the Trichoderma asperellum - sourced natural product in the solution is 0.1, 1 and 10 mg / L.

[0031] The beneficial effects of the technical solution of the present invention are as follows:

[0032] The Trichoderma asperellum - sourced natural product (i.e., ergosterol) extracted by the method of the present invention can be applied to promote plant growth, induce disease resistance in plants and reduce the incidence of plant diseases.

[0033] The present invention isolates and purifies the Trichoderma asperellum - sourced natural product ergosterol from the secondary metabolites of Trichoderma asperellum, which can be applied to promote the growth of wheat and balsam pear, induce and improve the disease resistance of wheat and balsam pear, and provide a certain theoretical basis for the development of biological pesticides.

[0034] The Trichoderma asperellum - sourced natural product of the present invention can be mass - produced and is natural and safe.

[0035] Other features and advantages of the present invention will be described in detail in the following detailed implementation section. Description of the Drawings

[0036] By describing the exemplary embodiments of the present invention in more detail in conjunction with the accompanying drawings, the above and other objects, features, and advantages of the present invention will become more apparent. Among them, in the exemplary embodiments of the present invention, the same reference numerals generally represent the same components.

[0037] Figure 1 Shows the 1 1H NMR spectrum of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention.

[0038] Figure 2 Shows the 13 13C NMR spectrum of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention.

[0039] Figure 3 A-3C shows the results of detecting the effect of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention on promoting wheat growth (where a, b, ab indicate significant differences at the p<0.05 level, CK is the control group; the values in the bar graph are the average ± SD of 3 replicates).

[0040] Figure 4 A-4C shows the results of detecting the effect of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention on the physiological indexes of wheat seedlings under the stress of Fusarium graminearum PH-1 (where a, b, c indicate significant differences at the p<0.05 level).

[0041] Figure 5 A-5F shows the results of detecting the effect of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention on the disease-resistant related genes of wheat seedlings under the stress of Fusarium graminearum PH-1.

[0042] Figure 6 A-6D shows the results of detecting the effect of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention on promoting the growth of bitter gourd.

[0043] Figure 7 A-7D shows the results of detecting the effect of ergosterol derived from Trichoderma asperellum obtained in Example 1 of the present invention on the physiological indexes of bitter gourd seedlings under the stress of Fusarium graminearum PH-1. Detailed Implementation

[0044] The preferred embodiments of the present invention will be described in more detail below. Although the preferred embodiments of the present invention are described below, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to fully convey the scope of the present invention to those skilled in the art.

[0045] In each of the following examples,

[0046] where the reagent or instrument used is not indicated by the manufacturer, it is a conventional product that can be obtained through market purchase.

[0047] The Trichoderma asperellum is Trichoderma asperellum CBS 433.97, with the deposit number ACCC30536, and is purchased from the Agricultural Culture Collection of China (ACCC).

[0048] The wheat is Zhoumai 36, selected from Zhoukou Academy of Agricultural Sciences;

[0049] The balsam pear is Saibilv, purchased from Hongliang Seed Co., Ltd. in Shouguang City, Shandong Province.

[0050] Example 1

[0051] This example provides a method for preparing a natural product from Trichoderma asperellum, which includes the following steps:

[0052] S1: Preparation of the spore suspension of Trichoderma asperellum

[0053] (1) Under sterile conditions, use an inoculation loop to pick a piece of Trichoderma asperellum CBS433.97 mycelium at the edge of the colony and place it in the middle of a PDA medium, and incubate it in an inverted position at 28 °C for 3 days to obtain an activated plate colony;

[0054] (2) Use a punch with a diameter of 6 mm to punch a piece of the colony at the edge of the activated plate colony and inoculate it into 100 mL of PDB medium, and shake it at 25 °C and 180 rpm for 7 days to obtain a Trichoderma asperellum fermentation broth;

[0055] (3) In a laminar flow hood, filter the Trichoderma asperellum fermentation broth with four layers of sterile gauze to obtain a filtrate;

[0056] (4) Under an optical microscope, count the spores in the filtrate with a hemocytometer, and then dilute the filtrate with sterile distilled water to a spore content of 2×10 8 spores / mL to obtain the spore suspension of Trichoderma asperellum.

[0057] S2: Add 1×10 spore counts to a wide-mouth bottle containing a rice medium 8A spore solution of Trichoderma asperellum at a concentration of [X] spores / mL was placed in an incubator at a constant temperature of 28°C for 34 days to obtain a fermentation product.

[0058] S3: The fermentation product and ethyl acetate were placed in a wide-mouth bottle in equal volumes and ultrasonicated for 30 minutes. This operation was repeated three times. Then, the ethyl acetate containing the fermentation product was subjected to rotary evaporation under reduced pressure at 45°C to obtain 62 g of the total ethyl acetate extract, which was placed in a refrigerator at 4°C for later use.

[0059] S4: 62 g of the total ethyl acetate extract was first dissolved in a mixed solution of dichloromethane and methanol (1:1), silica gel powder was added, and the sample was mixed. After evaporation to dryness under reduced pressure at 45°C, it was added to a pre-packed silica gel column, and a gradient elution under reduced pressure was carried out using a petroleum ether - dichloromethane - methanol system. 22 fractions were eluted, and ergosterol was separated from the 7th fraction by recrystallization. The spectroscopic data of the obtained ergosterol are as follows: white crystals, positive ion ESI-MS m / z: 397 [M+H] + , with the molecular formula C 28 H 44 O, and the degree of unsaturation is 7; 1 H NMR (500 MHz, CDCl3) δ: 5.56 (1H, dd, J = 5.5, 2.2 Hz, H-6), 5.39 - 5.36 (1H, m, H-5), 5.19 (1H, dd, J = 11.7, 7.4 Hz, H-22, 23), 3.63 (1H, tt, J = 11.2, 4.1 Hz, H-3), 1.03 (3H, d, J = 6.7 Hz, H-21), 0.94 (3H, s, H-19), 0.91 (5H, d, J = 6.9 Hz, H-28), 0.85 - 0.81 (11H, m, H-26, 27), 0.62 (2H, s, H-18); 13 C NMR (126 MHz, CDCl3) δ: 141.23 (C-8), 139.66 (C-5), 135.44 (C-22), 131.85 (C-23), 119.46 (C-6), 116.17 (C-7), 77.16, 70.32 (C-3), 55.60 (C-17), 54.43 (C-14), 46.12 (C-9), 40.65 (C-4), 40.31 (C-20), 39.65 (C-12), 38.96 (C-24), 38.25 (C-1), 37.13 (C-10), 32.96 (C-25), 31.51 (C-2), 28.13 (C-16), 24.18 (C-15), 20.98 (C-11, 21), 19.70 (C-27), 17.49 (C-28), 16.15 (C-19), 11.92 (C-18). The obtained ergosterol1 The \(^1H\) NMR spectrum is as Figure 1 shown, 13 and the \(^{13}C\) NMR spectrum is as Figure 2 shown.

[0060] Test Example 1

[0061] This test example is used to test the effect of ergosterol extracted in Example 1 on promoting wheat growth.

[0062] The ergosterol extracted in Example 1 was separately formulated into solutions of 0.1, 1, and 10 mg / L with distilled water. Select plump wheat seeds, soak them in warm water at 45 °C for 2 h, and then evenly place the wheat seeds into sterile petri dishes (90 mm). A layer of filter paper was laid at the bottom of the petri dish, and 50 wheat seeds were placed in each petri dish. Pipette 5 mL of the prepared ergosterol solution into each petri dish, and the control was an equal amount of distilled water, with 3 replicates for each treatment. After culturing at room temperature for 72 h, the seeds were transplanted into flower pots, with 200 g of soil and 15 seeds in each pot.

[0063] After treating wheat seeds with ergosterol, on the 8th day of wheat growth, the number of roots, root length, and plant height of wheat seedlings were measured to evaluate whether the ergosterol extracted in Example 1 had a growth-promoting effect on wheat. When the concentration of ergosterol extracted in Example 1 was 0.1 mg / L, the number of wheat roots and plant height increased by 6% and 10% respectively ( Figure 3 A, 3B), but there was no significant effect on wheat root length ( Figure 3 C); when the ergosterol concentration was 1 mg / L, the wheat root length decreased by 21.5% ( Figure 3 A), but there was no significant effect on the number of roots and plant height; when the ergosterol concentration was 10 mg / L, the number of wheat roots and plant height increased by 9% and 6% respectively ( Figure 3 A, 3B), and the root length decreased by 16.0% ( Figure 3 C).

[0064] Test Example 2

[0065] This test example is used to test the effect of ergosterol isolated in Example 1 on the physiological indexes of wheat seedlings under the stress of Fusarium graminearum PH-1.

[0066] This test example used the method of Test Example 1 to treat wheat seedlings. After the wheat seeds grew in pots for 3 d, the hypocotyls of the wheat seedlings were punctured in the middle with an inoculation needle, and 10 μL of PH-1 bacterial solution (the number of spores was 2×10 7 cells / mL) was added dropwise. The activities of PPO (polyphenol oxidase), POD (peroxidase), and the content of Pro (free proline) in wheat leaves were measured 7 days after inoculating the wheat with the pathogen.

[0067] The measurement results showed that the ergosterol extracted in Example 1 significantly affected the activities of PPO and POD enzymes in wheat leaves. When the concentration of ergosterol extracted in Example 1 was 0.1 mg / L, the PPO activity in wheat leaves increased significantly, increasing by 58.4% compared with the control; when the concentrations of ergosterol extracted in Example 1 were 1 mg / L and 10 mg / L, there was no significant effect on the PPO activity in wheat leaves( Figure 4 A). Ergosterol significantly increased the POD enzyme activity in wheat leaves at all test concentrations, with the most increase at 0.1 mg / L, increasing by 73.7% compared with the control. When the ergosterol concentration was 1 mg / L and 10 mg / L, although the POD activity decreased significantly compared with that at 0.1 mg / L, it still increased significantly compared with the control, increasing by 53% and 15.8% respectively( Figure 4 B). However, ergosterol did not affect the change in the free proline content in wheat leaves at all test concentrations( Figure 4 C).

[0068] Test Example 3

[0069] This test example was used to determine the effect of the ergosterol isolated in Example 1 on the disease-resistant related genes of wheat seedlings under the stress of Fusarium graminearum PH-1

[0070] In this test example, the methods of Test Example 1 and Test Example 2 were used to culture and inoculate wheat seedlings. Total RNA of wheat leaves was extracted by the Trizol method. After DNase treatment, cDNA was synthesized by reverse transcription. Real-time fluorescence quantitative PCR (RT-qPCR) used ChamQ SYBR qPCR Master Mix (Q311-02 / 03 kit) from Vazyme. The difference in gene expression levels was calculated by the method of relative quantification2 -△△Ct Calculate the difference in gene expression levels

[0071] The analysis results showed that

[0072] Under the treatment of ergosterol, compared with the control, the expression levels of the LTP-1 gene were all significantly up-regulated. When the ergosterol concentration was 0.1 mg / L, the LTP-1 expression level increased the most, by 2.07 times. When the ergosterol concentrations were 1 mg / L and 10 mg / L, their expression levels increased by 1.26 times and 1.98 times respectively( Figure 5 A).

[0073] Under the treatment of ergosterol, compared with the control, the expression levels of the P5CS gene were all significantly up-regulated. When the ergosterol concentration was 10 mg / L, its expression level increased the most, by 1.79 times. When the ergosterol concentrations were 0.1 mg / L and 1 mg / L, their expression levels increased by 1.33 times and 1.28 times respectively( Figure 5 B).

[0074] Under ergosterol treatment, compared with the control, the expression levels of Glu1 gene were all significantly up-regulated. When the ergosterol concentration was 0.1 mg / L, its expression level was up-regulated the most, by 1.89 times. When the ergosterol concentrations were 1 mg / L and 10 mg / L, its expression levels were up-regulated by 1.33 times and 1.20 times respectively( Figure 5 C).

[0075] Under ergosterol treatment, compared with the control, the expression levels of PR4 gene were all significantly down-regulated. When the ergosterol concentration was 1 mg / L, its expression level was down-regulated the most, by 0.29 times. When the ergosterol concentrations were 0.1 mg / L and 10 mg / L, its expression levels were down-regulated by 0.58 times and 0.66 times respectively( Figure 5 D).

[0076] Under ergosterol treatment, compared with the control, the expression levels of Chi1 gene were all significantly down-regulated. When the ergosterol concentration was 1 mg / L, its expression level was down-regulated the most, by 0.36 times. When the ergosterol concentrations were 0.1 mg / L and 10 mg / L, its expression levels were down-regulated by 0.52 times and 0.53 times respectively( Figure 5 E).

[0077] Under ergosterol treatment, compared with the control, the expression levels of PR1.1 gene were all significantly down-regulated. When the ergosterol concentration was 1 mg / L, its expression level was down-regulated the most, by 0.22 times. When the ergosterol concentrations were 0.1 mg / L and 10 mg / L, its expression levels were down-regulated by 0.27 times and 0.28 times respectively( Figure 5 F).

[0078] Test Example 4

[0079] This test example was used to test the effect of the ergosterol extracted in Example 1 on promoting the growth of balsam pear.

[0080] The ergosterol was respectively formulated into solutions with concentrations of 0.1, 1, and 10 mg / L using distilled water. Select plump balsam pear seeds, soak them in warm water at 45 °C for 5 hours, then lay the balsam pear seeds flat on wet gauze, and cover them with another layer of wet gauze. Incubate them in the dark at 28 °C until the radicles grow out, and then soak the balsam pear seeds in the above-prepared solutions for 12 h respectively. Transfer the balsam pear seeds to flower pots, with 200 g of soil and 1 seedling in each pot.

[0081] After treating the balsam pear seeds with ergosterol, on the 10th day after the balsam pear germinated, measure the number of roots, root length, and leaf area of the balsam pear seedlings to evaluate whether the ergosterol extracted in Example 1 has an effect on promoting the growth of balsam pear. When the ergosterol concentration was 1 mg / L, it promoted the growth of balsam pear most significantly, with the root length increasing by 3.5%( Figure 6 A), the number of roots increasing by 7.6%( Figure 6 B), and the plant height increasing by 13.9%(Figure 6 C), the leaf area increased by 32.3%( Figure 6 D).

[0082] Test Example 5

[0083] This test example was used to test the effect of ergosterol isolated in Example 1 on the physiological indexes of bitter gourd seedlings under the stress of Fusarium graminearum PH-1.

[0084] In this test example, the method of Test Example 4 was used to treat bitter gourd seedlings. The root-dipping method was used to inoculate the pathogenic bacteria on the bitter gourd. The roots of the bitter gourd were immersed in the pathogenic bacteria suspension for half an hour, and then the bitter gourd seedlings were transplanted into the soil, with 200 g of soil and 1 plant seedling per pot. 14 days after inoculating the pathogenic bacteria, the activities of PPO and POD, the content of free proline and the content of malondialdehyde in the bitter gourd leaves were measured.

[0085] On the 14th day after inoculating the bitter gourd plants with the pathogenic bacteria of bitter gourd wilt, the activities of PPO and POD in the bitter gourd leaves decreased with the decrease of the concentration compared with the control( Figure 7 A, 7B), and the contents of free proline and malondialdehyde both increased. When the concentration of Compound 7 was 1 mg / L, the content of free proline increased by 36.99%, and the content of malondialdehyde increased by 36.03%( Figure 7 C, 7D).

[0086] The embodiments of the present invention have been described above. The above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments.

Claims

1. A natural product derived from Trichoderma asperellum, characterized in that, This natural product has the structure shown in Formula I:

2. A method for preparing a natural product derived from Trichoderma asperellum, characterized in that, This preparation method includes successively performing rice fermentation culture and extraction, separation, and purification on Trichoderma asperellum to obtain the natural product derived from Trichoderma asperellum described in Claim 1.

3. The preparation method of the Trichoderma asperellum-derived natural product according to claim 2, wherein, The temperature of the rice fermentation culture is 25 - 35 °C, and the time is 28 - 35 days.

4. The preparation method of the Trichoderma asperellum-derived natural product according to claim 2, wherein, The extraction, separation, and purification include ethyl acetate extraction, normal-phase silica gel column chromatography, and recrystallization performed successively.

5. According to the preparation method of the natural product derived from Trichoderma asperellum described in Claim 4, wherein, The number of times of ethyl acetate extraction is 2 - 4 times; The reduced-pressure gradient elution system used in the normal-phase silica gel column chromatography is a petroleum ether - dichloromethane - methanol system; The solvent used for recrystallization is at least one of methanol, petroleum ether, and dichloromethane; the temperature of the recrystallization is 15 - 35 °C.

6. Application of the natural product derived from Trichoderma asperellum described in Claim 1 in the following aspects: 1) Promoting plant growth; 2) Inducing disease resistance in plants.

7. The application according to claim 6, wherein The plants are wheat and / or balsam pear.

8. The application according to claim 6, wherein, The promoting plant growth includes using the natural product derived from Trichoderma asperellum to promote the growth of wheat and balsam pear; Preferably, the promoting the growth of wheat includes promoting the increase in wheat root length, the increase in the number of roots, and the increase in plant height; Preferably, the promoting the growth of balsam pear includes promoting the increase in balsam pear root length, the increase in the number of roots, the increase in plant height, and the increase in leaf area.

9. The application according to claim 6, wherein The inducing disease resistance in plants includes using the natural product derived from Trichoderma asperellum to induce wheat to produce disease resistance to Fusarium head blight and induce balsam pear to produce disease resistance to Fusarium wilt; Preferably, the way of inducing wheat to produce disease resistance to Fusarium head blight includes increasing the activity of wheat antioxidant enzymes and regulating the expression of wheat disease-resistant genes; Preferably, the way of inducing balsam pear to produce disease resistance to Fusarium wilt includes regulating osmotic adjustment substances, reducing membrane lipid peroxidation, increasing the activity of balsam pear antioxidant enzymes, and reducing cell damage.

10. The application according to claim 6, wherein, The application of the natural product derived from Trichoderma asperellum is carried out in the form of a solution, and the concentration of the natural product derived from Trichoderma asperellum in the solution is 0.1 - 10 mg / L.

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