Trichoderma occidentalis agent and its application in preparing bio-organic fertilizer

Abstract

The present invention relates to the field of microbial technology, in particular to a Trichoderma agent and its application in preparing biological organic fertilizers. The present invention separates and screens the Trichoderma strain SY392 ( Trichoderma erinaceum ), demonstrating a trinity of growth-promoting, disease-resisting, and quality-enhancing effects. It efficiently produces IAA, solubilizes phosphate, adapts to acidic and alkaline environments, and exhibits strong antagonism against a variety of pathogens, particularly with an inhibition rate exceeding 70% against Fusarium oxysporum in cowpea. Field trials have shown that SY392 significantly improves cowpea growth indicators, yield, and quality, and reduces morbidity. After being formulated using shallow-plate fermentation technology, it has performed exceptionally well in cowpea fields at the Batou Base in Yacheng Town, Sanya City, Hainan Province, providing an innovative microbial solution for the green, high-yield development of tropical legume crops and promising broad prospects for widespread application.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, in particular to a Trichoderma ergifer agent and application thereof in preparing biological organic fertilizer. Background Art

[0002] cowpea( Vigna unguiculata (L.) Walp. ) is an important winter leguminous vegetable economic crop in Hainan Province, but its growth is often affected by insufficient soil fertility, pest and disease invasion and other problems, resulting in unstable yield and quality. Although excessive reliance on chemical fertilizers and pesticides in traditional agriculture can increase yields in the short term, it causes soil degradation, environmental pollution and increased pathogen resistance. Continuous cropping problems are very likely to occur in production, affecting economic and social benefits.

[0003] As a green and environmentally friendly alternative, microbial agents have received widespread attention in recent years. In the existing technology, we have found that some microorganisms can produce plant-stimulating substances and have the ability to dissolve phosphorus, significantly improve soil fertility, promote plant root development, and antagonize a variety of pathogens, thereby enhancing plant disease resistance. However, as a type of biological agent, different microbial strains have different effects on crops due to their different physiological characteristics. At present, the strains reported on the market have limited application effects on cowpea cultivation and lack broad-spectrum resistance to specialized pathogens in specific tropical regions. Therefore, in order to solve the continuous cropping obstacles caused by the application of chemical fertilizers and pesticides in the process of cowpea cultivation, it is necessary for us to conduct research on cowpea, screen out highly specific biocontrol and quality-enhancing microorganisms for cowpea plants, and use these microorganisms to produce bio-organic fertilizers, providing an efficient microbial solution for the green and high-yield of tropical legume crops. Summary of the Invention

[0004] In view of the above, it is necessary to conduct research on cowpea, screen out highly specific biocontrol and quality-enhancing microorganisms for cowpea plants, and apply these microorganisms to produce bio-organic fertilizers, providing an efficient microbial solution for the green and high-yield of tropical legume crops.

[0005] In order to achieve the above purpose, the present invention screened out a new strain: Trichoderma Trichoderma erinaceum SY392, whose classification is named as: Trichoderma erinaceum SY392, Chinese classification name: Trichoderma SY392, preservation number: CCTCC NO: M20242615; the strain is deposited in the China Center for Type Culture Collection, address: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, and the preservation date is November 21, 2024.

[0006] The present invention also includes the Trichoderma Trichoderma erinaceum SY392 bacterial agent.

[0007] The present invention also includes the Trichoderma Trichoderma erinaceum Application of SY392 or the bacterial agent in the preparation of bio-organic fertilizer.

[0008] The present invention also includes the Trichoderma Trichoderma erinaceum Application of SY392 or the bacterial agent in promoting the growth of cowpea.

[0009] The present invention also includes the Trichoderma Trichoderma erinaceum Application of SY392 or the bacterial agent in producing IAA, solubilizing phosphate and / or dissolving phosphate.

[0010] The present invention also includes the Trichoderma Trichoderma erinaceum Application of SY392 or the bacterial agent in improving cowpea quality and / or promoting cowpea root nodulation.

[0011] Furthermore, the cowpea quality indicators are: vitamin C, soluble solid content and / or soluble sugar.

[0012] The present invention also includes the Trichoderma Trichoderma erinaceum Use of SY392 or the bacterial agent in preparing an antibacterial agent.

[0013] Furthermore, the pathogens inhibited by the antibacterial agent are: Fusarium oxysporum ( Fusarium oxysporum f. sp. Tracheiphilum ), Ascomycetes ( Magnapothe oryzae ), Fusarium oxysporum melon-specific FOM ( Fusarium oxysporum f.sp.melonis ), Fusarium oxysporum cucumber-specific FOC ( Fusarium oxysporum f.sp. Cucumerinum ), Bakanae oxysporum ( Fusarium fujikuroi ), Alternaria ( Alternaria ​ ), Fusarium oxysporum tomato-specific type FOL ( ​ ) and the Vanilla-specific FOV of Fusarium oxysporum ( ​ )

[0014] The present invention also includes a method for growing cowpea using the bacterial agent, which comprises: ​ ​ The SY392 bacterial suspension or fermentation liquid is inoculated into mushroom residues for fermentation to obtain the bacterial agent, and then the bacterial agent is applied from the seedling stage to the vine stage.

[0015] Further, the Trichoderma ​ SY392 solid bacterial agent is diluted with water at a mass percentage of 1.6%; the dosage per hole is 0.4L, and it is applied from the seedling stage to the vine stage; the hedgehog ​ ​ The number of Trichoderma spores in SY392 solid inoculum is 10 8 cfu / g.

[0016] The present invention has the following beneficial effects: SY392, isolated from a mango orchard by the research team, exhibits excellent IAA production, phosphate solubilization, and phosphate dissolution capabilities. Experimental results show that SY392 not only directly promotes plant growth but also demonstrates excellent antagonism against 10 pathogens (e.g., specialized Fusarium oxysporum strains of cucumber, tomato, melon, and banana). Its broad-spectrum resistance to specialized pathogens effectively enhances plant disease resistance. Furthermore, the strain is acid and alkali tolerant. Furthermore, it was prepared using shallow tray fermentation technology and applied to cowpeas. Sampling during the vine emergence, vine emergence-pod formation transition, and pod formation stages of cowpeas revealed that SY392 improved cowpea growth indicators (stem diameter, SPAD value, nodule number, nodule weight, and root fresh-dry weight), increased yield (pod length, pod width, single pod mass, pod number, and pod weight), and quality (vitamin C and soluble protein), while also reducing the incidence of cowpea wilt in the field. This strain breaks through the technical bottleneck of the single function of traditional Trichoderma agents through the three-in-one mechanism of "growth promotion, disease resistance and quality improvement", and provides an efficient microbial solution for the green and high yield of tropical leguminous crops. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] ​ The colony morphology of SY392 is shown in Figure 1. The left picture is the front view of the plate, and the right picture is the back view of the plate.

[0018] ​ This is the phylogenetic tree of SY392.

[0019] ​ This is the result diagram of the acid resistance of Trichoderma strain SY392.

[0020] ​ This is the result diagram of the salt tolerance of Trichoderma strain SY392.

[0021] ​ This is the antagonistic graph of Trichoderma strain SY392 against different pathogens; among them, the pathogens in the 1st to 6th rows from top to bottom are: Fusarium oxysporum 1 ( ​ ), Fusarium oxysporum f.cocereus 2 ( ​ ), the pathogen of rice blast: Ascomycetes ( ​ ), Fusarium oxysporum melon-specific FOM ( ​ ​ ), Fusarium oxysporum cucumber-specific FOC ( ​ ).

[0022] ​ The antagonistic graph of Trichoderma strain SY392 against different pathogens; the pathogens in the 1st to 4th rows from top to bottom are: Bakanae oryzae ( ​ ), Alternaria (​ ), Fusarium oxysporum tomato-specific type FOL ( ​ ) and the Vanilla-specific FOV of Fusarium oxysporum ( ​ ).

[0023] ​ These are shallow tray fermentation images of different Trichoderma strains; the left image is the shallow tray fermentation image of strain SY392, and the right image is the shallow tray fermentation image of strain SY2.

[0024] ​ The following are the effects of different treatments at different growth stages on the SPAD values ​​of cowpea leaves; Figure A is the cowpea vine-sprouting stage, Figure B is the vine-sprouting-pod-setting transition period, and Figure C is the pod-setting stage.

[0025] ​ These are diagrams showing the effects of different treatments at different growth stages on the stem diameter of cowpea. Figure A shows the vine-forming stage, Figure B shows the vine-forming-pod-setting transition period, and Figure C shows the pod-setting stage.

[0026] ​ Figure 3 shows the effects of different treatments at different growth stages on the fresh weight of cowpea roots. Figure A shows the vine-forming stage, Figure B shows the vine-forming-pod-setting transition period, and Figure C shows the pod-setting stage.

[0027] ​ Figure 3 is the effect of different treatments at different growth stages on the dry weight of cowpea roots; Figure A is the cowpea vine-sprouting stage, Figure B is the vine-sprouting-pod-setting transition period, and Figure C is the pod-setting stage.

[0028] ​ Figure 3 is the effect of different treatments at different growth stages on the weight of cowpea nodules; Figure A is the cowpea vine-sprouting stage, Figure B is the vine-sprouting-pod-setting transition period, and Figure C is the pod-setting stage.

[0029] ​ These are diagrams showing the effects of different treatments at different growth stages on the number of cowpea nodules; Figure A is the cowpea vine-sprouting stage, Figure B is the vine-sprouting-pod-setting transition period, and Figure C is the pod-setting stage.

[0030] ​ Figure 3 is a graph showing the effects of different treatments on cowpea pod length, pod weight, number of pod grains and single pod grain weight; Figure A shows the cowpea pod length, Figure B shows the single pod weight of cowpea, Figure C shows the single pod grain number of cowpea, and Figure D shows the single pod grain weight of cowpea.

[0031] ​ Effects of different treatments on cowpea yield.

[0032] ​ Effects of different treatments on the incidence of cowpea.

[0033] ​Figure 3 shows the effects of different treatments on cowpea quality. Figure A shows vitamin C in cowpea, Figure B shows soluble protein in cowpea, Figure C shows soluble solids, Figure D shows soluble sugar, and Figure E shows soluble dry matter.

[0034] Biomaterial deposit information

[0035] The deposit information of the SY392 strain of this application is: ​ SY392, whose classification is named as: ​ SY392, Chinese classification name: Trichoderma SY392, preservation number: CCTCCNO: M20242615; the strain is deposited in the China Center for Type Culture Collection, address: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, and the preservation date is November 21, 2024. DETAILED DESCRIPTION

[0036] All features disclosed in this specification, or all steps in the disclosed methods or processes, except mutually exclusive features and / or steps, can be combined in any manner.

[0037] Any feature disclosed in this specification (including any accompanying claims and abstract), unless otherwise stated, is merely an example of a series of equivalent or similar features. Example 1

[0038] This example is the isolation and identification of Trichoderma hymenii SY392.

[0039] Isolation of SY392: In the mango orchard at the Fruit Island Experimental Base in Sanya City, Hainan Province, rhizosphere soil samples were collected from three key growth stages of mangoes: the pre-flowering stage (W), the flowering-young fruit stage (X), and the fruit expansion stage (G) to ensure the comprehensiveness and representativeness of the research data. Subsequently, the rhizosphere soil samples collected from the three periods were thoroughly mixed. 10 g of soil sample was weighed and mixed with 90 mL of sterile water in a triangular flask and incubated at 28°C and 170 rpm. -1 Under the conditions of shaking for 30 minutes, the soil suspension was obtained. On the clean bench, 1 mL of soil suspension was drawn into a test tube containing 9 mL of sterile water, and gradient dilution was performed in sequence. Then, 10 mL of soil suspension was drawn into the test tube. -3 , 10 -4 and 10 -5 100 μL of soil suspension was evenly spread on Trichoderma selective medium with 10 replicates for each gradient. The plate was inverted and incubated in a 28°C incubator in the dark for 2 d. Colonies with different morphologies were selected and inoculated on new PDA plates.

[0040] Purification of Trichoderma strain SY392: Use a sterile inoculating loop to dip the isolated Trichoderma mycelium onto a PDA plate. When the colony grows to 2-4 cm in diameter, transfer it to a new PDA plate until a single pure fungal colony is obtained.

[0041] 2. Identification of SY392: ① Morphological identification ​ As shown: SY392 strain was inoculated into PDA medium and cultured in a constant temperature incubator for 120 hours. The colonies on the solid medium were observed to be dark green and covered with green short-haired hyphae. Its morphological characteristics were consistent with the common characteristics of Trichoderma. ② The purified Trichoderma was inoculated onto a new PDA plate and cultured at 28°C for 3-5 days. An appropriate amount of hyphae was collected and genomic DNA (gDNA) was extracted according to the operating instructions of the Fungal Genomic DNA Rapid Extraction Kit. The PCR products were sent to Wuhan Qingke Biological Co., Ltd. for sequencing and alignment was performed based on the tef1 sequence. The alignment results are shown as follows: ​ As shown, this strain ​ They are highly similar, hence the name ​ ​ SY392. Example 2

[0042] This example is the determination of the growth-promoting function and antagonistic pathogenic bacteria ability of Trichoderma strain SY392.

[0043] 1. Qualitative determination of IAA production capacity of strains: IAA production was determined by the Salkowski colorimetric method. Fresh Trichoderma spores were added to a 1% inoculum containing 200 mg·L -1 In the PDB medium containing tryptophan, under dark conditions, 28 °C, 170 rpm -1 Oscillate for 5 days. Then incubate the bacterial suspension at 4°C and 8000 r / min. -1 Centrifuge for 10 minutes. Take 100 μL of the fermentation supernatant and add Salkowski colorimetric solution at a 1:1 ratio. Incubate in the dark for 30 minutes. If the reaction solution turns red, it indicates that the fermentation solution contains IAA analogs, and the darker the color, the higher the IAA content. Measure the absorbance at a wavelength of 530 nm using a microplate reader. Calculate the amount of IAA produced by the strain based on a standard curve prepared using an IAA standard (Sigma). The concentrations of the prepared IAA solutions are 0, 10, 20, 30, 40, and 50 mg·L. -1 100 μL of each gradient solution was aspirated, 100 μL of colorimetric solution was added, and the reaction was carried out in the dark for 30 minutes. The absorbance at a wavelength of 530 nm was measured. A PDB blank culture medium with colorimetric solution added was used as a control. Three replicates were set for each assay to plot a standard curve.

[0044] 2. Phosphate Solubilization Capacity Determination: During the experiment, well-grown Trichoderma strains were first selected from the plates, and their spores were eluted and collected. Subsequently, the spores were inoculated into organic and inorganic phosphorus liquid culture media at a 1% inoculum rate. The culture media were shaken in a shaker at 28°C and 180 rpm for three days. After incubation, the culture broth was filtered using a funnel and filter paper to obtain the supernatant. The soluble phosphorus content in the supernatant was then determined using a molybdenum antimony colorimetric method. Three replicates were performed for each strain to ensure data accuracy and reliability. Culture media without the inoculated strain served as a blank control (CK). To quantify soluble phosphorus content, a 5 mg / L phosphorus stock solution was prepared, and a phosphorus standard curve was constructed based on this solution. The absorbance of the supernatant samples was measured colorimetrically, and the actual soluble phosphorus content in the samples was calculated using the standard curve.

[0045] 3. pH Tolerance Assay: Inoculate Trichoderma spores onto standard PDA plates and incubate at 28°C for 2 days. Once mycelium has fully grown on the plate, use a microporator to extract the outermost mycelium and inoculate it onto MM medium (pH 3) in five replicates. Incubate at 28°C in the dark, and measure the colony diameters on the first, second, third, and fourth days.

[0046] 4. Salt Tolerance Assay: Inoculate Trichoderma spores onto standard PDA plates and incubate at 28°C for 2 days. Once mycelium has fully grown on the plate, use a microporator to extract the outermost mycelium and inoculate it onto PDA plates containing 0 mM, 400 mM, 800 mM, and 1000 M NaCl, with five replicates. Incubate in a 28°C incubator in the dark. After 120 hours, measure the colony diameter.

[0047] 5. Antagonistic pathogenic bacteria ability test: The Trichoderma strains were screened from the rhizosphere of cowpea and two strains of Fusarium oxysporum: cowpea Fusarium oxysporum 1 ( ​ ), Fusarium oxysporum f.cocereus 2 ( ​ ) Among them, the cowpea oxysporum Fusarium 1 and 2 of this application were screened from cowpea roots. Molecular identification showed that they both belonged to Fusarium oxysporum, with slight morphological differences. Therefore, the two strains were named 1 and 2. ​ ), Fusarium oxysporum melon-specific FOM ( ​ oxysporum f. sp. melonis ), Fusarium oxysporum cucumber-specific FOC ( Fusarium oxysporum f. sp. Cucumerinum ), Bakanae oxysporum ( Fusarium fujikuroi ), Alternaria ( Alternaria alternata ), Fusarium oxysporum tomato-specific type FOL ( Fusarium oxysporum f. sp. lycopersici) and the Vanilla-specific FOV of Fusarium oxysporum ( Fusarium oxysporum f. sp. vanillae ).

[0048] Inoculate onto PDA plates and incubate at 28°C for 7 days. Aseptically, use a 5 mm diameter borer to punch a hole on a plate infested with Trichoderma and the pathogen. Using tweezers, carefully transfer the strains onto fresh PDA plates, each 2 cm from the edge of the dish. A control plate inoculated with the pathogen alone was used. These plates were then incubated inverted in the dark at 28°C. Each standoff experiment was replicated three times. After 6 days, the colony radius of the pathogen control group and the radius of the pathogen pointing toward the Trichoderma strain in the standoff culture were measured, and the inhibition rate was calculated.

[0049] Inhibition rate (%) = (colony radius of control group − colony radius of Trichoderma group) / colony radius of control group × 100%.

[0050] Test results: 1. The IAA production, phosphate solubilization and phosphate dissolution abilities of Trichoderma strain SY392 are shown in Table 1.

[0051]

[0052] As shown in Table 1, Trichoderma Trichoderma erinaceum ) strain SY392 had strong IAA production and phosphate solubilization abilities, indicating that it had the potential to produce plant hormones and promote nutrient absorption.

[0053] 2. pH tolerance of Trichoderma strain SY392: In the experiment to evaluate the acid tolerance of Trichoderma strain SY392, we paid special attention to its growth under extremely acidic conditions (pH=3), because this is related to the adaptability and application potential of Trichoderma strains in natural acidic soil environments. Figure 3 As shown: The Trichoderma strain SY392 exhibited a certain acid resistance. Specifically, in an acidic environment with a pH value adjusted to 3, the Trichoderma strain SY392 was able to grow and cover the entire culture plate within the fourth day, showing good adaptability to the acidic environment.

[0054] 3. Salt tolerance of Trichoderma strain SY392: The experimental results are as follows Figure 4 As shown in the figure, the colony morphology of SY392 shows good growth at different salt concentrations (0mM, 400mM, 800mM, and 1000mM). As the salt concentration increases, the size and morphology of the colonies change, but it can still maintain a certain growth ability at a high salt concentration of 1000mM, indicating that it has good salt tolerance.

[0055] 4. Antagonistic ability of Trichoderma strain SY392 against 10 pathogens Figure 5-Figure 6 As shown, Figure 5In the figure, the pathogens in the 1st to 6th rows from top to bottom are: Fusarium oxysporum 1 ( Fusarium oxysporum f. sp. Tracheiphilum ), Fusarium oxysporum f.cocereus 2 ( Fusarium oxysporum f. sp. Tracheiphilum ), Rice blast: Ascomycetes ( Magnapothe oryzae ), Fusarium oxysporum melon-specific FOM ( Fusarium oxysporum f. sp. melonis ), Fusarium oxysporum cucumber-specific FOC ( Fusarium oxysporum f. sp. Cucumerinum ); Figure 6 In the figure, the pathogens in the 1st to 4th rows from top to bottom are: Bakanae oryzae ( Fusarium fujikuroi ), Alternaria ( Alternaria alternata ), Fusarium oxysporum tomato-specific type FOL ( Fusarium oxysporum f. sp. lycopersici ) and the Vanilla-specific FOV of Fusarium oxysporum ( Fusarium oxysporum f. sp. vanillae Plate confrontation tests showed that the strain SY392 exhibited significant antagonism against ten pathogens (Fusarium oxysporum). Specifically, the strain exhibited 72.10% inhibition against Fusarium oxysporum 1 and Fusarium oxysporum 2, 75.00% inhibition against rice blast, 84.77% inhibition against banana-specific FOC, 70.87% inhibition against melon-specific FOC, 61.70% inhibition against cucumber-specific FOC, 67.97% inhibition against rice bakanae pathogen, 68.59% inhibition against Alternaria alternata, 68.59% inhibition against tomato-specific FOL, and 73.88% inhibition against vanilla orchid-specific FOV. This result indicates that the Trichoderma strain SY392 not only has a broad-spectrum antibacterial activity, but can also effectively inhibit the growth of specific plant pathogens, showing its application prospect as a potential biocontrol agent. Example 3

[0056] This example describes a fungicide prepared by Trichoderma harbingerii strain SY392 and its field efficacy evaluation.

[0057] Trichoderma strains: The strains used were Trichoderma harbingerii strain SY392 and Trichoderma harbingerii SY2 (wherein SY2 was the same strain screened out from the same batch and at the same location and identified as Trichoderma harbingerii through molecular identification).

[0058] Solid fermentation preparation of bacterial agent: The screened Trichoderma strains SY392 and SY2 were inoculated into PDB medium and shaken at 28°C and 170 rpm. -1, and carry out liquid fermentation for 3 to 4 days to obtain Trichoderma fermentation liquid. Dilute the hydrolyzed amino acids 20 times and add them to the turnover box containing mushroom residue (the main component is corn cob, 70%), and soak for 24 hours. Wring the soaked mushroom residue dry and sterilize it at 115°C for 1 hour. After cooling, add the prepared Trichoderma fermentation liquid to a shallow dish, adjust the water content to about 65%, and culture at a constant temperature of 28°C for 5 days. During the fermentation period, maintain the water content at 60% to obtain Trichoderma solid strains, which are stored in a refrigerator at 4°C. After microscopic examination, the number of Trichoderma spores in each g (dry weight) of Trichoderma solid inoculant is about 10 8 cfu / g; SY392 Trichoderma agent and SY2 Trichoderma agent were obtained, and the appearance of the agent was as follows Figure 7 shown.

[0059] Evaluation of the effect of field application of Trichoderma: A field experiment was conducted on cowpea at the Batou base in Yacheng Town, Sanya City, Hainan Province. The cowpea variety was 'Jiangxing 708'. The experiment was conducted with three treatments: no inoculation (CK), inoculation with Trichoderma SY392 (SY392), and inoculation with Trichoderma SY2 (SY2). Each treatment was repeated three times, and the plot area was 30 m 2 , set up two compartments, with a hole distance of 30~40cm, about 63 holes in each compartment, about 126 holes in each plot, and 2 seedlings left in each hole.

[0060] Treatments with Trichoderma SY392 and SY2 were conducted by diluting the solid Trichoderma inoculum at a 1.6% mass percentage in water. Approximately 0.4 L of the solid inoculum was applied per hole. Cowpea plant samples were collected at different growth stages (vine emergence, vine emergence-pod formation transition, and pod formation) to measure SPAD values, stem diameter, underground dry and fresh weight, and nodule weight and number. During the harvest period, samples were collected every other day for a total of 18 harvests, and yield was recorded each time. Cowpea quality (vitamin C, soluble sugars, soluble solids, soluble protein, and dry matter content) was measured during the peak harvest period. Disease status was recorded in 50 plants per treatment during the late harvest period.

[0061] Experimental results: 1. Effects of different treatments on SPAD values ​​of cowpea Figure 8 As shown, Figure 8 Figure A shows the cowpea vine emergence stage, Figure B shows the transition period from vine emergence to pod formation, and Figure C shows the pod formation stage. The figures show that the different treatments (CK, SY392, and SY2) had different effects on the SPAD values ​​of cowpea leaves. During the vine emergence stage, the SPAD value of the SY392 treatment was significantly higher than that of the control (CK). During the transition period from vine emergence to pod formation, the SY392 treatment maintained a higher SPAD value, maintaining its clear advantage. At the pod formation stage, the SPAD value of the SY392 treatment was significantly higher than that of both the CK and SY2 treatments. Overall, the SY392 treatment demonstrated the best chlorophyll-enhancing effect across all three growth stages.

[0062] 2. Effects of different treatments on cowpea stem diameter Figure 9 As shown, Figure 9 Figure A shows the cowpea vine emergence stage, Figure B shows the transition period from vine emergence to pod formation, and Figure C shows the pod formation stage. As can be seen from the figures, the stem diameter of the SY392 group was significantly greater than that of the CK group during both the vine emergence and pod formation stages, but not significantly different from that of the SY2 group. At the pod formation stage, the stem diameter differences among the three groups were not significant. Overall, the microbial treatment group significantly increased cowpea stem diameter in the early stages.

[0063] 3. Effects of different treatments on the fresh weight and dry weight of cowpea roots Figure 10 and Figure 11 As shown, Figure 10 This is the experimental result of fresh weight of cowpea root. Figure 11 The experimental results of root dry weight are as follows: Figure 10 and Figure 11 In the figure, A is the vine-forming stage, B is the vine-forming to pod-forming stage, and C is the pod-forming stage. Figure 10-11 As can be seen from the results, during the vine-forming stage, SY2 had the highest root fresh and dry weights, significantly exceeding those of the other two groups. During the vine-forming to pod-setting transition, SY392 had significantly higher root fresh and dry weights than the CK group. However, at the pod-setting stage, no significant differences were found among the three groups in terms of root fresh and dry weight. Overall, SY392 was more effective in increasing root fresh and dry weights as the cowpeas grew and developed.

[0064] 4. Effects of different treatments on the weight and number of cowpea nodules Figure 12 and Figure 13 As shown, Figure 12 is the experimental result of cowpea nodule weight, Figure 13 is the experimental result of the number of nodules, Figure 12 and Figure 13 Figure A shows the cowpea vine emergence stage, Figure B the vine emergence-pod formation transition stage, and Figure C the pod formation stage. As can be seen from the figures, during the vine emergence stage, the SY392 treatment significantly increased nodule weight, outperforming the other treatments. During the vine emergence-pod formation transition stage, the SY392 treatment performed best in both nodule weight and number. At the pod formation stage, the SY392 treatment maintained high nodule weight and number. Overall, the SY392 treatment performed exceptionally well in increasing cowpea nodule weight and number.

[0065] 5. Effects of different treatments on cowpea pod length, single pod weight, single pod grain number and single pod grain weight Figure 14Figure 1 shows cowpea pod length, B shows cowpea pod weight, C shows cowpea kernel number per pod, and D shows cowpea kernel weight per pod. The results show that the treatments had no significant effect on cowpea pod length, kernel weight, or kernel number per pod. Regarding kernel weight per pod, SY392 performed best, followed by SY2, while the CK treatment performed relatively poorly. Overall, SY392 was the most effective treatment in increasing kernel weight per pod.

[0066] 6. Effects of different treatments on cowpea yield Figure 15 The results in the figure show that the SY392 treatment had the highest cowpea yield, with an average yield of approximately 162 kg per plot, 9.6 kg higher than the SY2 treatment and 13.1% higher than the CK treatment. Overall, the SY392 treatment demonstrated a certain advantage in increasing cowpea yield.

[0067] 7. Effects of different treatments on the incidence of cowpea Figure 16 As shown in the figure, the SY392 treatment group reduced the disease incidence by 9.90% compared to the CK treatment, and the SY2 treatment group reduced the disease incidence by 3.65% compared to the CK treatment. The SY392 treatment had the lowest disease incidence, indicating that it has certain advantages in suppressing cowpea diseases.

[0068] 8. Effects of different treatments on cowpea quality Figure 17 Figure 1 shows the cowpea vitamin C content (Figure A), soluble protein (Figure B), soluble solids (Figure C), soluble sugars (Figure D), and soluble dry matter (Figure E). The results show that the SY392 treatment significantly increased the vitamin C and soluble protein content of cowpeas. Soluble solids and soluble sugar content were higher in the SY392 treatment group than in the CK and SY2 treatments. Dry matter content showed little difference among the three groups. Overall, the SY392 treatment performed well in improving the nutritional quality of cowpeas.

[0069] In summary, the Trichoderma strain SY392 ( Trichoderma erinaceum ), demonstrating a trinity of growth promotion, disease resistance, and quality improvement. It efficiently produces IAA, solubilizes phosphates, adapts to acidic and alkaline environments, and possesses strong antagonism against a variety of pathogens, particularly Fusarium oxysporum in cowpea, with an inhibition rate exceeding 70%. Field trials have shown that SY392 can significantly improve cowpea growth indicators, yield, and quality, and reduce morbidity. After being produced through shallow-plate fermentation technology, it performed exceptionally well in cowpea fields at the Batou Base in Yacheng Town, Sanya City, Hainan Province, providing an innovative microbial solution for the green and high-yield production of tropical legume crops and possessing broad prospects for promotion and application.

[0070] The above-described embodiments merely illustrate several embodiments of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make various modifications and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. Trichoderma Trichoderma erinaceum ) SY392, its deposit number is CCTCC NO: M20242615.

2. comprising the Trichoderma serrata described in claim 1 ( Trichoderma erinaceum ) SY392 bacterial agent.

3. Trichoderma as claimed in claim 1 ( Trichoderma erinaceum ) Use of SY392 or the bacterial agent as described in claim 2 in the preparation of biological organic fertilizer.

4. Trichoderma as claimed in claim 1 ( Trichoderma erinaceum ) Use of SY392 or the bacterial agent as described in claim 2 in promoting the growth of cowpea.

5. Trichoderma as claimed in claim 1 ( Trichoderma erinaceum ) Use of SY392 or the bacterial agent as described in claim 2 in producing IAA and / or solubilizing phosphate.

6. Trichoderma as claimed in claim 1 ( Trichoderma erinaceum ) Use of SY392 or the bacterial agent as described in claim 2 in improving the quality of cowpea and / or promoting the nodulation of cowpea root nodules; the cowpea quality indicators are: vitamin C, soluble protein content and / or soluble sugar.

7. Trichoderma as claimed in claim 1 ( Trichoderma erinaceum )SY392 or the use of the bacterial agent according to claim 2 in the preparation of an antibacterial agent; the pathogens inhibited by the antibacterial agent are: cowpea oxysporum f. sp. Tracheiphilum, ascomycetes (Magnapothe oryzae), Fusarium oxysporum f. sp. melonis, Fusarium oxysporum f. sp. Cucumerinum, rice seedling pathogen (Fusarium fujikuroi), Alternaria alternata, Fusarium oxysporum f. sp. lycopersici and Fusarium oxysporum f. sp. vanillae.

8. The method for growing cowpea using the microbial agent according to claim 2, characterized in that: The method comprises the following steps: Trichoderma erinaceum ) The SY392 bacterial suspension or fermentation liquid is inoculated into the mushroom residue and fermented to obtain the bacterial agent, which is then applied from the seedling stage to the vine stage.

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