Trichoderma harzianum ivf1 for the control of fungal diseases and use thereof
By screening Trichoderma harzianum strain IVF1 and adding inactivated pathogen cells as an inducer to the fermentation medium, its secondary metabolic pathways were activated, solving the problem of unstable biocontrol efficacy of Trichoderma harzianum strain and significantly improving the control effect against various pathogenic fungi and vegetable diseases.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- INSTITUTE OF VEGETABLES & FLOWERS CHINESE ACADEMY OF AGRICULTURAL SCIENCES
- Filing Date
- 2026-02-26
- Publication Date
- 2026-06-02
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Figure CN122128106A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of agricultural biological control technology, specifically relating to Trichoderma harzianum IVF1 for the control of fungal diseases and its application. Background Technology
[0002] Trichoderma fungi, especially Trichoderma harzianum, are important biocontrol fungi with various biocontrol mechanisms, including nutrient competition, hyperparasitism, antibiotic activity, and induction of systemic resistance in plants. However, wild-type Trichoderma harzianum strains isolated from the natural environment generally suffer from unstable biocontrol effects and limited production of antimicrobial metabolites, which restricts their large-scale production and practical application.
[0003] Currently, improving the biocontrol performance of Trichoderma harzianum mainly relies on physical or chemical mutagenesis, culture medium optimization, and fermentation condition control. However, traditional mutagenesis methods have limitations such as being time-consuming and labor-intensive, and having uncontrollable directions; conventional fermentation processes mostly focus on increasing cell biomass, failing to effectively activate secondary metabolic pathways related to the synthesis of target antimicrobial substances, resulting in low efficiency in the synthesis of active ingredients and the failure to fully realize the biocontrol potential.
[0004] Therefore, there is an urgent need in this field to screen for Trichoderma strains with strong antifungal disease control effects and to further develop a method that can effectively activate its secondary metabolic pathways and improve the synthesis efficiency of antifungal products, so as to achieve efficient and green control of plant fungal diseases and promote the widespread application of microbial pesticides in agricultural production. Summary of the Invention
[0005] The purpose of this invention is to provide a Trichoderma harzianum strain.
[0006] Another object of the present invention is to provide the application of the above-mentioned Trichoderma harzianum in the prevention and control of fungal diseases in vegetables.
[0007] Another object of the present invention is to provide an inhibitor of pathogenic fungi.
[0008] According to the present invention, Trichoderma harzianum ( Trichoderma harzianum The preservation number of this bacterium is CGMCC No. 42324.
[0009] This invention provides Trichoderma harzianum ( Trichoderma harzianum Applications that inhibit pathogenic fungi.
[0010] According to the application of the present invention, the pathogenic fungus is *Botrytis cinerea* (…). Botrytis cinerea Fusarium oxysporum ( Fusarium graminearum ), Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum ) or Rhizoctonia solani ( Rhizoctonia solani ).
[0011] According to the application of the present invention, the Trichoderma harzianum ( Trichoderma harzianum Fermentation of *Trichoderma harzianum* (to obtain a fermentation broth) is carried out, and the fermentation broth is used to inhibit pathogenic fungi, wherein the *Trichoderma harzianum* (to obtain a fermentation broth) is fermented. Trichoderma harzianum Inactivated bacterial cells of the pathogen were added to the fermentation medium.
[0012] According to the present invention, the pathogenic fungal inhibitor includes the Trichoderma harzianum ( Trichoderma harzianum ) fermentation broth.
[0013] According to the pathogenic fungal inhibitor of the present invention, the pathogenic fungus is *Botrytis cinerea* (…). Botrytis cinerea Fusarium oxysporum ( Fusarium graminearum ), Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum ) or Rhizoctonia solani ( Rhizoctonia solani ).
[0014] According to the pathogenic fungal inhibitor of the present invention, in the fermentation preparation of the Trichoderma harzianum ( Trichoderma harzianum When fermenting the broth, inactivated bacterial cells of the pathogen are added to the fermentation medium.
[0015] Compared with the prior art, the present invention has the following advantages: The *Trichoderma harzianum* strain of this invention effectively inhibits gray mold in cucumbers and wilt disease in cowpeas. Compared with the registered *Trichoderma harzianum* strain LTR-2, the *Trichoderma harzianum* strain of this application exhibits better inhibitory effects against pathogenic fungi. Further investigation revealed that under inactivated gray mold induction conditions, differentially expressed genes were significantly enriched in pathways related to secondary metabolite biosynthesis, antibiotic synthesis, and amino acid synthesis. The *Trichoderma harzianum* strain of this invention produced increased antibacterial active substances, further enhancing its preventative efficacy. Metabolomics results showed a significant increase in the production of bioactive organic acids such as citric acid, benzene ring compound emodin, organic heterocyclic compound 6-methylnicotinic acid, and trichoderma diol.
[0016] The present invention (Trichoderma harzianum) Trichoderma harzianum IVF1, the preservation number of this bacterium is CGMCC No. 42324, and its classification name is... Trichoderma harzianum, The deposit date is November 21, 2025. The depositary institution is the China General Microbiological Culture Collection Center, and the deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. Attached Figure Description
[0017] Figure 1 Morphological characteristics of Trichoderma harzianum IVF1; Figure 2 A phylogenetic tree of Trichoderma harzianum IVF1 constructed based on the ITS gene sequence; Figure 3A phylogenetic tree of Trichoderma harzianum IVF1 constructed based on the TEF-1α gene sequence; Figure 4 The results of plate confrontation comparison of Trichoderma harzianum IVF1 against different pathogenic fungi are shown; Figure 5 The assay showed the antifungal effect of Trichoderma harzianum IVF1 metabolites on pathogenic fungi; Figure 6 The efficacy of Trichoderma harzianum IVF1 against gray mold in cucumber was demonstrated; Figure 7 The efficacy of Trichoderma harzianum IVF1 in preventing cowpea wilt was demonstrated; Figure 8 The assay showed the antibacterial effect of Trichoderma harzianum IVF1 metabolites on Botrytis cinerea induced by inactivated Botrytis cinerea; Figure 9 The efficacy of inactivated Botrytis cinerea IVF1 metabolites in the prevention of gray mold in cucumber; Figure 10 Transcriptome analysis of Trichoderma harzianum IVF1 induced by inactivated Botrytis cinerea. Detailed Implementation
[0018] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Example 1: Isolation and purification of Trichoderma harzianum IVF1 strain
[0019] Soil samples collected in September 2024 from Qingyuan County, Lishui City, Zhejiang Province were analyzed using the dilution plating method. The culture medium was serially diluted to 10⁻⁶ with sterile water. -4 ~10 -7 For each concentration, 200 μL of the diluted solution was spread onto PDA solid medium and then incubated upside down in a 30°C constant temperature incubator. The colonies growing on the medium were observed. Based on the size, color, and surface pattern of the colonies, larger single colonies were selected and repeatedly streaked on PDA solid medium for 2-3 generations of purification until single colonies appeared. The Trichoderma strain was isolated and purified and named IVF1. Example 2: Identification of Trichoderma harzianum IVF1 strain
[0020] The morphological characteristics of the IVF1 of this invention are as follows: Figure 1 As shown, the colonies are round, initially white, gradually turning green; the hyphae are filamentous, growing radially from the center; the conidiophores are flask-shaped to ampoule-shaped, swollen in the middle, and the apex is the thinnest and can produce spores; the flask cells are solitary or in whorls of 2 to 5; the conidia are spherical or ellipsoidal, with a smooth surface, light green, turning dark green when mature.
[0021] Genomic DNA was extracted from *Trichoderma harzianum* IVF1 using a fungal genomic DNA extraction kit. Using this DNA as a template, PCR amplification was performed using universal primers for fungal ITS and primers for TEF-1α gene amplification. After passing 1% agarose gel electrophoresis, the amplified products were purified and sequenced. The obtained ITS and TEF-1α sequences were compared for homology using the Basic Local Alignment Search (BLAST) tool in the National Center for Biotechnology Information (NCBI) database to preliminarily verify sequence accuracy. After confirming sequence accuracy, reference sequences of *Trichoderma harzianum* and its closely related species were downloaded from NCBI. Multiple alignments of all sequences were performed using BioEdit software to ensure sequence locus consistency. Finally, a phylogenetic tree was constructed using the maximum likelihood method with MEGA 11.0 software, and the confidence level of each branch was assessed using a bootstrap test with 1000 replicates. The phylogenetic analysis results are shown below. Figure 2 , Figure 3 As shown, the strain IVF1 identified in this application clusters within a high-support evolutionary branch with the Trichoderma harzianum standard strain and is clearly separated from other Trichoderma species. This result definitively identifies strain IVF1 as Trichoderma harzianum. Example 3: Comparison of plate confrontation between Trichoderma harzianum IVF1 and different pathogenic fungi
[0022] The plate confrontation method was used to evaluate the resistance of Trichoderma harzianum strains to Botrytis cinerea ( Botrytis cinerea Fusarium oxysporum ( Fusarium graminearum ), Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum ) and Rhizoctonia solani ( Rhizoctonia solani The *Trichoderma harzianum* strains described herein are *Trichoderma harzianum* strain IVF1 and existing *Trichoderma harzianum* strain LTR-2, with pesticide registration certificate number PD20243467. Four pathogenic fungi preserved in the laboratory were activated, and the activated pathogens and *Trichoderma harzianum* were cultured separately on PDA plates at 28°C for 5 days. Subsequently, 0.5 cm diameter mycelial discs were prepared at the edge of the colonies using a sterile punch, and inoculated onto opposite sides of a 9 cm diameter PDA plate, with the centers of the two mycelial discs approximately 4.5 cm apart and the line connecting them passing through the center of the plate. Plates inoculated only with pure cultures of the corresponding pathogens were set up as controls. All treatments were repeated 3 times and cultured in a 28°C incubator.
[0023] After 4-5 days of cultivation, the colony diameter of the pathogen in each treatment was measured using the cross-multiplication method, and the inhibition rate of *Trichoderma harzianum* on the mycelial growth of the pathogen was calculated. The calculation formula is as follows: Inhibition rate (%) = (Coronavirus colony diameter in control group – Coronavirus colony diameter in treatment group) / (Coronavirus colony diameter in control group – 0.5 cm) × 100% Plate confrontation culture revealed that the inhibition rates of the *Trichoderma harzianum* strain IVF1 proposed in this application against four pathogenic fungi—*Botrytis cinerea*, *Fusarium oxysporum*, *Sclerotinia sclerotiorum*, and *Rhizoctonia solani*—were 71.3%, 70.4%, 68.1%, and 57.4%, respectively. The existing *Trichoderma harzianum* strain LTR-2 showed inhibition rates of 65.5%, 61.4%, 59.5%, and 46.8% against the same four pathogenic fungi. Compared with *Trichoderma harzianum* LTR-2, the overall control efficacy of the *Trichoderma harzianum* strain IVF1 proposed in this application against the four pathogenic fungi was improved by approximately 10%. Figure 4 ). Example 4: Determination of the antifungal effect of Trichoderma harzianum IVF1 metabolites on pathogenic fungi
[0024] Fermentation filtrate preparation: The *Trichoderma harzianum* IVF1 strain of this application was inoculated onto PDA plates and cultured at 28±1℃ for 7 days. Spores were collected and a spore suspension was prepared with sterile water, and the concentration was adjusted to 1×10⁻⁶ using a hemocytometer. 7 CFU / mL. A spore suspension was inoculated into PDB liquid medium at an inoculum of 2% (v / v) and cultured at 28°C with shaking at 180 rpm for 7 days to obtain the fermentation broth. The fermentation broth was then filtered through quantitative filter paper and a 0.22 μm microporous membrane to obtain sterile fermentation filtrate.
[0025] Preparation of drug-containing plates: The above fermentation filtrate was mixed with sterilized PDA medium cooled to approximately 50°C at a ratio of 1:9, and poured into sterile petri dishes to prepare drug-containing plates. The control group consisted of PDA plates with an equal volume of sterile PDB added.
[0026] Antibacterial test: Fresh, activated *Botrytis cinerea* and *Fusarium oxysporum* were used to create 5 mm diameter mycelial discs, which were then inoculated into the center of drug-containing and control plates, respectively. Each treatment was repeated three times and incubated at 28°C for four days.
[0027] Inhibition rate calculation: The diameter of pathogenic bacterial colonies was measured using the cross-multiplication method, and the inhibition rate was calculated using the following formula: Inhibition rate (%) = [(Coronary diameter of control group – Coronary diameter of treatment group) / (Coronary diameter of control group – 5mm)] × 100% Plate confrontation culture revealed that the fermentation broth of Trichoderma harzianum IVF1 inhibited the growth of two pathogenic fungi, Botrytis cinerea and Fusarium oxysporum, by 65.2% and 59.8%, respectively. Figure 5 ). Example 5: Efficacy of Trichoderma harzianum IVF1 in controlling cucumber gray mold
[0028] 5.1 Preparation of suspensions of pathogens and Trichoderma Botrytis cinerea suspension: After activating Botrytis cinerea on a PDA plate, take 5 mycelial cakes with a diameter of 5 mm and inoculate them into 250 mL of PDB medium. Incubate at 25℃ for 7 days, and then break up the fermentation broth before use.
[0029] Trichoderma harzianum IVF1 suspension: Five Trichoderma mycelial discs with a diameter of 5 mm were inoculated into PDB medium and cultured with shaking at 28℃ and 180 rpm for 7 days. After the culture was completed, the mycelium was broken up using a tissue homogenizer to prepare a homogeneous mycelial fragment suspension for later use, which was then diluted 500 times.
[0030] 5.2 Plant Induction Treatment and Design Cucumber seeds were sown in flowerpots, with 12 seedlings of uniform growth per pot. When the seedlings reached the two-leaf-one-heart stage, inoculation was performed: a suspension of *Botrytis cinerea* was inoculated onto the cucumber leaves. The plants were then placed in a culture room at 20-25℃ and 80-90% humidity for 24 hours to promote disease development. After the humidity period, the *Trichoderma* fermentation broth was diluted and sprayed onto the leaves, with 50 mL of the suspension applied to every 12 cucumber seedlings. The experimental treatment groups are detailed in Table 1 below.
[0031] 5.3 Disease Investigation and Data Analysis Disease severity was assessed 6 days after inoculation. The severity of disease on each leaf was recorded according to the following grading criteria: Grade 0: No lesions; Grade 1: Lesions cover ≤ 5% of the leaf area; Grade 3: Lesions cover 6% to 15% of the leaf area; Level 5: Lesions cover 16% to 25% of the leaf area; Level 7: Lesions cover 26% to 50% of the leaf area; Level 9: The area of lesions accounts for ≥ 51% of the leaf area.
[0032] Based on the survey results, the disease index and prevention and control effectiveness were calculated using the following formula: Disease index = [∑(number of diseased leaves at each level × representative value of that level) / (total number of leaves surveyed × 9)] × 100; Prevention and control effect (%) = [(Disease index of blank control group - disease index of treatment group) / disease index of blank control group] × 100 Table 1. Results of the control efficacy of different Trichoderma species against gray mold in cucumber. .
[0033] Trichoderma harzianum IVF1 spores and fermentation broth are more effective against cucumber gray mold than the mainstream Trichoderma harzianum LTR-2 on the market. Figure 6 ). Example 6: Control efficacy of Trichoderma harzianum IVF1 spores against cowpea wilt.
[0034] 6.1 Preparation of suspensions of pathogens and Trichoderma Suspension of Fusarium oxysporum pathogen for cowpea wilt: After activating Fusarium oxysporum on PDA plates, five mycelial cakes with a diameter of 5 mm were inoculated into 250 mL of PDB medium and cultured at 25℃ for 7 days. The fermentation broth was then broken up before use.
[0035] Trichoderma suspension: The concentration of Trichoderma harzianum IVF1 spores was adjusted to 1×10⁻⁶ with sterile water. 7 CFU / mL available for use.
[0036] 6.2 Plant Induction Treatment and Design Sow cowpea seeds in pots containing substrate, keeping 12 seedlings of uniform growth per pot. When the seedlings have fully developed their first pair of true leaves, inoculate them using the root-drenching method, applying 10 mL of the prepared pathogen spore suspension to each seedling. After inoculation, place the plants in a greenhouse at 25-28℃ and 70%-85% humidity for normal management to promote disease development. Twenty-four hours after pathogen inoculation, treat with Trichoderma. Apply a diluted Trichoderma solution to the cowpea seedlings via root drenching, applying 10 mL of the fungal suspension to each seedling.
[0037] 6.3 Disease Investigation and Data Analysis Disease severity was assessed starting 14 days after inoculation. The severity of disease on each cowpea plant was recorded according to the following grading criteria: Grade 0: The plant shows no symptoms and is growing normally; Grade 1: Cotyledons or true leaves are slightly yellowed, but the plant does not wilt significantly; Grade 3: One or more true leaves are obviously yellowed and drooping, or necrosis appears on the leaf margins; Level 5: The plant is wilted overall, and the vascular bundles in the stem show slight browning; Level 7: The plant is severely wilted, growth is stunted, and the vascular bundles in the stem show obvious browning; Level 9: Plant dies.
[0038] Similarly, referring to the investigation method for the control effect of cucumber gray mold, the disease index and control effect of wilt were calculated. Specifically, the control effect of Trichoderma harzianum IVF1 spores on cowpea wilt reached 56.0% ( Figure 7 ). Example 7: Determination of the antifungal effect of Trichoderma harzianum IVF1 metabolites on Botrytis cinerea induced by inactivated gray mold.
[0039] After activation of *Botrytis cinerea* on PDA medium, it was transferred to PDB liquid medium and cultured at 25°C and 150 rpm for 7 days with shaking. After culture, the mycelium was collected by filtration through a sterile filter membrane and rinsed three times with sterile water. The washed mycelium was then freeze-dried under vacuum to obtain inactivated *Botrytis cinerea* mycelium, which was then sealed and stored at 4°C for later use.
[0040] 7.2 Induced fermentation and metabolite preparation Spore suspensions of Trichoderma harzianum strains IVF1 and LTR-2 (1×10⁻⁶) 7 CFU / mL) were inoculated into the following two fermentation media respectively: Experimental group (induction group): 1% (w / v) of the above-mentioned inactivated Botrytis cinerea mycelium was added to PDB medium; Control group (non-induction group): PDB medium without any added inducers was used.
[0041] Both groups were cultured at 28°C and 180 rpm with shaking for 7 days. After fermentation, the fermentation broth was centrifuged, and the supernatant was filtered through a 0.22 μm microporous membrane to obtain sterile "induced metabolite" and "non-induced metabolite" filtrates, respectively.
[0042] 7.3 Determination of antibacterial effect The antibacterial activity of metabolite filtrates against *Botrytis cinerea* was evaluated using the mycelial growth rate method. Specifically, filtrates containing induced and non-induced metabolites were mixed with PDA medium at a ratio of 1:9 (v / v) and poured into plates. Plates containing equal volumes of sterile PDB and PDA served as blank controls. Fresh *Botrytis cinerea* mycelial discs with a diameter of 5 mm were inoculated into the center of each plate and incubated at 25°C for 5 days. Colony diameters were measured using the cross-crossing method, and the inhibition rate was calculated using the following formula: Inhibition rate (%) = [(Coronavirus diameter of control group - Coronavirus diameter of experimental group) / (Coronavirus diameter of control group)] ×100%.
[0043] The results showed that the *Trichoderma harzianum* IVF1-induced metabolites prepared by the method of this invention exhibited significant antibacterial effects against *Botrytis cinerea*, with an inhibition rate as high as 82.1%, representing an 18.1% improvement compared to the non-induced metabolites (64.0% inhibition rate). However, *Trichoderma harzianum* LTR-2 showed no change in its antibacterial effect against *Botrytis cinerea* under inactivated *Botrytis cinerea* induction conditions. These results fully demonstrate that using inactivated *Botrytis cinerea* as an inducer can effectively stimulate *Trichoderma harzianum* IVF1 to produce metabolites with stronger antibacterial activity. Furthermore, different *Trichoderma harzianum* strains, under inactivated *Botrytis cinerea* induction conditions, did not all produce increased antibacterial activity substances. Figure 8 ). Example 8: Control efficacy of Trichoderma harzianum IVF1 metabolites induced by inactivated gray mold against cucumber gray mold.
[0044] This method, referring to Example 5, focuses on providing a highly efficient and environmentally friendly biological control scheme for cucumber gray mold. The key treatment steps in the experimental group are as follows: First, *Trichoderma* was cultured under the induction of inactivated *Botrytis cinerea* spores to prepare a *Trichoderma* fermentation broth; simultaneously, *Trichoderma* cultured on potato medium served as a control fermentation broth. Both fermentation broths were diluted 500 times and sprayed onto cucumber plants infected with gray mold. Experimental results showed that the *Trichoderma* fermentation broth induced by inactivated *Botrytis cinerea* exhibited a significant control effect on cucumber gray mold, with a control efficacy as high as 85.7%. Compared to the control group of uninduced *Trichoderma* fermentation broth on potato medium, its control efficacy increased by 10.3% (…). Figure 9 This method not only significantly improved the biocontrol activity of Trichoderma fermentation broth, but also demonstrated the technical advantage of stimulating the disease resistance potential of Trichoderma through pathogen induction, providing an important basis for the development of new microbial pesticides. Example 9: Transcriptome analysis of Trichoderma harzianum induced by inactivated gray mold
[0045] This study investigated the effect of inactivated *Botrytis cinerea* mycelium as an inducer on the expression of the *Trichoderma harzianum* IVF1 gene. First, inactivated *Botrytis cinerea* mycelium was co-cultured in *Trichoderma harzianum* liquid medium, with potato broth as a control. All treatments were performed in at least three biological replicates. Samples were collected at 24, 48, and 72 hours post-induction. Mycelium was immediately flash-frozen in liquid nitrogen and sent to Yuanzi Biotechnology for transcriptome sequencing. Sequencing data were quality-controlled to obtain Clean Reads, which were aligned to the *Trichoderma harzianum* reference genome (*Trichoderma harzianum* CBS 226.95) using HISAT2 or STAR. The raw Read Counts for each gene were then calculated using StringTie or featureCounts, and expression levels were normalized using FPKM or TPM. Differentially expressed genes were analyzed using DESeq2 or edgeR, with a selection threshold of |log2(Fold Change)| > 1 and a corrected P-value (padj) < 0.05. The results showed that, compared with the control group, 2252, 2376, and 2255 differentially expressed genes were identified in the experimental group at 24, 48, and 72 hours, respectively. GO functional enrichment analysis of the differentially expressed genes (padj < 0.05) screened for significantly enriched entries based on biological processes, molecular functions, and cellular components, revealing the key biological functions and related molecular mechanisms activated by *Trichoderma harzianum* under inactivated *Botrytis cinerea* induction. KEGG pathway enrichment analysis (padj < 0.05) indicated that the differentially expressed genes were significantly enriched in pathways related to secondary metabolite biosynthesis, antibiotic synthesis, and amino acid synthesis. Figure 10This suggests that Trichoderma harzianum may regulate the above-mentioned metabolic pathways to enhance its biocontrol activity in response to Botrytis cinerea induction. Example 10 Metabolomics analysis of Trichoderma harzianum induced by inactivated Botrytis cinerea
[0046] First, inactivated *Botrytis cinerea* mycelium was prepared as an inducer and then inoculated into *Trichoderma harzianum* liquid culture medium for co-culture to simulate the antagonistic environment of fungi in nature, thereby specifically activating the defense metabolic pathway of *Trichoderma harzianum*. After culture, the fermentation supernatant was separated, and crude metabolite extracts were obtained by multiple extractions with ethyl acetate and concentration under reduced pressure. The above samples were sent to Yuanzi Biotechnology for non-targeted metabolomics analysis using an ultra-high performance liquid chromatography-mass spectrometry platform, with data acquired simultaneously in both positive and negative ion modes. The metabolomics analysis results significantly showed that the metabolomic profile of *Trichoderma harzianum* underwent drastic reconstruction under inactivated *Botrytis cinerea* induction conditions. Compared with the uninduced control group, 105 and 122 significantly upregulated differential metabolites were identified in negative and positive ion modes, respectively. Further focused analysis revealed that the key upregulated metabolites were widely distributed in organic acids, organic heterocyclic compounds, and benzene ring compounds. Among them, the relative contents of representative molecules with important biological activities, such as citric acid (an organic acid), emodin (a benzene ring compound), 6-methylnicotinic acid (an organic heterocyclic compound), and trichoderma diol, increased by approximately 48.8, 26.0, 9.8, and 5.3 times, respectively, which fully verified the excellent effect of this induction strategy in activating defense metabolic pathways and enhancing the synthesis of antibacterial active products.
[0047] The above embodiments are only used to understand the technical solution of this application and do not limit the scope of protection of this application.
Claims
1. Trichoderma harzianum ( Trichoderma harzianum ), characterized in that, The Trichoderma harzianum ( Trichoderma harzianum The accession number for this work is CGMCC No. 42324.
2. The Trichoderma harzianum as described in claim 1 ( Trichoderma harzianum Applications that inhibit pathogenic fungi.
3. The application according to claim 2, characterized in that, The pathogenic fungus is *Botrytis cinerea* (…). Botrytis cinerea Fusarium oxysporum ( Fusarium graminearum ), Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum ) or Rhizoctonia solani ( Rhizoctonia solani ).
4. The application according to claim 2, characterized in that, The Trichoderma harzianum ( Trichoderma harzianum Fermentation is carried out to obtain a fermentation broth, which is used to inhibit pathogenic fungi, wherein the *Trichoderma harzianum* ( Trichoderma harzianum Inactivated bacterial cells of the pathogen were added to the fermentation medium.
5. A pathogenic fungal inhibitor, characterized in that, The pathogenic fungal inhibitor includes the *Trichoderma harzianum* as described in claim 1. Trichoderma harzianum ) fermentation broth.
6. The pathogenic fungal inhibitor according to claim 5, characterized in that, The pathogenic fungus is *Botrytis cinerea* (…). Botrytis cinerea Fusarium oxysporum ( Fusarium graminearum ), Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum ) or Rhizoctonia solani ( Rhizoctonia solani ).
7. The pathogenic fungal inhibitor according to claim 5, characterized in that, The *Trichoderma harzianum* was prepared by fermentation. Trichoderma harzianum When fermenting the broth, inactivated bacterial cells of the pathogen are added to the fermentation medium.