Biocontrol streptomyces ITBB10008 sourced from wheat plants and application of biocontrol streptomyces ITBB10008
Through the biocontrol of Streptococcus ITBB10008 and its spore liquid isolated from wheat plants, the biological control problem of wheat gibberellia was solved, and effective inhibition of wheat gibberellia and environmentally friendly crop disease prevention and control were achieved.
Patent Information
- Application Number
- CN202411363870.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-28
- Publication Date
- 2024-12-27
AI Technical Summary
The existing technology is difficult to effectively prevent and control wheat gibberellia. Chemical control has drug resistance and environmental pollution problems, and lacks safe and environmentally friendly biological control methods.
A streptomyces of biodrugated streptomyces ITBB10008 was isolated from the wheat plants. The strains that have inhibitory effects on the pathogens of wheat gibberellia were screened and identified, and their spore liquid was prepared as a biological control agent to inhibit the expansion of wheat gibberellia.
This biological control agent significantly inhibits the expansion of wheat gibberellia, has broad-spectrum antibacterial effect, is low toxicity, no residue, environmentally friendly, and shows efficient control effects in potted plant experiments.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of microorganisms, and particularly relates to a wheat plant-derived biocontrol Streptomyces ITBB10008 and applications thereof. Background Art
[0002] Wheat is an important food crop in my country and around the world, and pasta made from its flour is our primary source of energy and protein. Fusarium head blight is a major disease that threatens wheat production safety. It can occur throughout the wheat growth period, primarily affecting the wheat ear. This disease not only causes grain discoloration, shrinkage, and yield reduction, and in severe cases, even total crop failure, but also produces various mycotoxins, including deoxynivalenol and zearalenone, upon infection. These mycotoxins can seriously harm the health of humans and animals, and can even cause their death.
[0003] During the epidemic period, wheat fusarium head blight can reduce wheat yield by about 40%-60%. my country's wheat has also been harmed by fusarium head blight, and there is still a lack of effective cures for wheat fusarium head blight. Currently, prevention is mainly carried out by screening disease-resistant varieties before planting and strengthening field cultivation management. Chemical control is an important means of field management, but chemical control is faced with some irresistible factors, such as precipitation and strong winds, which have a great impact on the occurrence and spread of wheat fusarium head blight, bringing many difficulties to chemical control. In addition, the repeated use of large-scale chemical agents leads to drug resistance of pathogens, environmental pollution and food safety issues. Biological control has become the development trend of wheat fusarium head blight prevention and control because it can effectively reduce environmental pollution, reduce the use of pesticides, and is safer for humans and animals.
[0004] Actinomycetes are renowned for their antibiotic production; approximately 70% of natural antibiotics are derived from them. Actinomycetes are not only a major source of bioactive compounds but also serve as biocontrol agents (live bacteria and / or their metabolites) for agricultural applications. For example, avermectin and jinggangmycin play a significant role in controlling plant pests and diseases. Antagonistic strains of Fusarium graminearum, the pathogen of wheat head blight, have been screened from actinomycetes and applied in agriculture to control wheat head blight. These strains are highly specific and prevent drug resistance and environmental pollution. Summary of the Invention
[0005] The object of the present invention is to provide a wheat plant-derived biocontrol Streptomyces ITBB10008 and a biocontrol product for controlling crop pathogens comprising the biocontrol Streptomyces ITBB10008. The wheat plant-derived biocontrol Streptomyces ITBB10008 and the biocontrol product thereof have good control effects on various crop pathogens, especially wheat fusarium.
[0006] The present invention also aims to provide the use of the above-mentioned wheat plant-derived biocontrol Streptomyces ITBB10008 or a biocontrol product comprising the above-mentioned wheat plant-derived biocontrol Streptomyces ITBB10008 in the preparation of a medicament having the effect of preventing and controlling crop pathogens.
[0007] The above-mentioned first purpose of the present invention can be achieved by the following technical scheme: a biocontrol Streptomyces ITBB10008 derived from a wheat plant, whose taxonomic name is Streptomyces sp., the preservation number is GDMCCNo: 64824, the preservation date is July 18, 2024, and the preservation unit is Guangdong Provincial Microbial Culture Collection Center.
[0008] The inventors of the present application collected wheat plants in Xinyang City, Henan Province, isolated and obtained multiple actinomycetes, and screened out an antagonistic strain that can effectively inhibit the wheat fusarium graminearum pathogen (Fusarium graminearum) through the plate confrontation method. The spore liquid of this strain can significantly inhibit the spread of wheat fusarium graminearum under greenhouse conditions, and has good application value for biological control of wheat fusarium graminearum. In the examples of the present invention, it is named Streptomyces sp. ITBB10008.
[0009] The present invention also provides a biocontrol product for controlling crop pathogens, comprising the biocontrol Streptomyces ITBB10008 and / or the spore liquid of the biocontrol Streptomyces ITBB10008.
[0010] The test results show that the biocontrol Streptomyces sp. ITBB10008 in the present invention has an antibacterial effect on a variety of crop pathogens.
[0011] Optionally, the spore liquid of the biocontrol Streptomyces ITBB10008 is prepared by the following method:
[0012] (1) activating the biocontrol Streptomyces ITBB10008 on an ISP2 plate and then transferring it to an MS medium for culturing until a large number of spores are produced;
[0013] (2) Sterile water was poured into the above ISP2 plate, and the spores were scraped with a sterile cotton swab to collect the spore liquid.
[0014] Optionally, in step (1), the culture is statically cultured at 28° C. in the dark for 7 to 10 days until a large number of spores are produced.
[0015] Furthermore, the collected spore solution can be diluted, and a suitable concentration can be selected for plating. After the spores germinate and grow into colonies, the original spore solution concentration can be calculated after the colonies are counted, and then the spore solution concentration can be adjusted to 10 7 CPU / mL, and place in a 4℃ refrigerator until use.
[0016] The test results show that the spore liquid of the biocontrol Streptomyces ITBB10008 of the present invention has a biological control effect on wheat fusarium wilt in a pot experiment.
[0017] The second object of the present invention can be achieved by the following technical solution: use of the above-mentioned biocontrol Streptomyces ITBB10008 or the above-mentioned biocontrol product in the preparation of a drug having the effect of preventing and controlling crop pathogens.
[0018] Optionally, the crop pathogens include one or a combination of several of the pepper anthracnose pathogen Colletotrichum capsici, wheat fusarium graminearum Schw, banana wilt pathogen Fusarium oxysporumf.sp.cubense, rice blast pathogen Pyricularia oryzae, papaya anthracnose pathogen Colletotrichum gloeosporioides, rubber anthracnose pathogen Colletotrichum acutatum, cowpea stem base rot pathogen Pythium aphanidmatum, rice sheath blight pathogen Thanatephorus cucumeris, mango anthracnose pathogen Colletotrichum gloeosporioides and asparagus stem blight pathogen Phomopsis asparagi.
[0019] The present invention can use the developed drug for biological control, specifically comprising: using the biocontrol Streptomyces sp. ITBB10008 and / or the spore liquid of the biocontrol Streptomyces sp. ITBB10008 to carry out biological control on crops.
[0020] Optionally, the crops include one or a combination of pepper, wheat, banana, rice, papaya, rubber, cowpea, mango and asparagus.
[0021] Compared with the prior art, the present invention has the following advantages:
[0022] (1) The biocontrol Streptomyces sp. ITBB10008 obtained by the present invention not only has a good biological control effect on wheat fusarium wilt, but also has a broad antibacterial spectrum and good antibacterial effect, has good development prospects, and has great potential for biological control;
[0023] (2) The biocontrol strain Streptomyces sp. ITBB10008 and its biocontrol products in the present invention have the advantages of no residue, low toxicity, and environmental friendliness compared to chemical pesticides; and have better biological control effects than other reported active strains. BRIEF DESCRIPTION OF THE DRAWINGS
[0024] Figure 1 The inhibitory effect of Streptomyces sp. ITBB10008 on Fusarium graminearum on the plate in Example 1;
[0025] Figure 2 This is the morphology of Streptomyces sp. ITBB10008 grown on IPS2 for 7 days in Example 1;
[0026] Figure 3 This is the phylogenetic tree of Streptomyces sp. ITBB10008 and similar typical strains in Example 2;
[0027] Figure 4 This is the biological control effect of Streptomyces sp. ITBB10008 on wheat fusarium wilt under greenhouse conditions in Example 4. DETAILED DESCRIPTION
[0028] Example 1 Strain isolation and screening
[0029] Wheat plants were collected from wheat fields in Xinyang City, Henan Province. The sample grinding dilution solution was spread on the selected TWYE medium to isolate actinomycetes using the grinding gradient dilution spreading method. Strains that met the actinomycete colony morphology were observed and selected daily and transferred to ISP2 medium. After isolation and purification, the strains were stored in glycerol tubes at -20°C.
[0030] The TWYE medium is prepared according to the following components in mass volume ratio: yeast extract 0.25 g / L, K2HPO4 0.5 g / L, agar 15.0 g / L, pH 7.2.
[0031] ISP2 medium was prepared according to the following components in mass volume ratio: yeast extract 4.0 g / L, malt extract 10.0 g / L, glucose 4.0 g / L, agar 15.0 g / L, pH 7.2±0.2.
[0032] All isolated actinomycete strains were cultured on ISP2 medium. After vigorous growth and sporulation, the antibacterial activity of the actinomycetes against Fusarium graminearum, the wheat head blight pathogen, was determined using the plate standoff method, using the indicator bacteria. A 7-mm-diameter F. graminearum cake was inoculated in the center of a 90-mm-diameter PDA plate, with 7-day-old actinomycete cakes (7 mm in diameter) placed 3 cm to the left and right of the center. A F. graminearum plate without actinomycete inoculation served as a control. The plates were incubated at 28°C until the control plate had grown to the entire surface. The diameter of the pathogen colonies was measured, and the inhibitory rate against the pathogen was calculated. Each treatment was replicated three times.
[0033] Bacterial inhibition rate (%) = [(pathogen growth diameter of control group - pathogen block diameter) - (pathogen growth diameter of treatment group - pathogen block diameter)] / (pathogen growth diameter of control group - pathogen block diameter) × 100%.
[0034] According to the results of the activity screening against wheat fusarium scaber pathogen, strain ITBB10008 had the strongest antagonistic activity against it, with an inhibition rate of 76.71±0.37% ( Figure 1 The strain was used as the target strain for the antibacterial spectrum and greenhouse biocontrol experiments. Actinomycete strain ITBB10008 grew well on ISP2 medium. After culturing at 28°C for 3 days, colonies were visible to the naked eye. The colonies were convex and wrinkled. After 5-6 days, spores began to form. The color of the spore pile was off-white. The colony morphology (7 days of culture) is shown in the figure below. Figure 2 shown.
[0035] Example 2 16S rRNA gene sequence analysis of strain ITBB10008
[0036] The ITBB10008 mycelia cultured for 7 days in Example 1 were collected and total DNA was extracted using an enzymatic mini-quantity method. 16S rRNA gene universal primers 27F / 1492R were used for 16S rRNA amplification.
[0037] The total volume of the PCR amplification reaction system was 50 μL, including 25 μL of 2×mix DNA polymerase, 1 μL of primer 27F, 1 μL of primer 1492R, 1 μL of DNA template, 5 μL of DMSO, and 17 μL of sterile water.
[0038] The amplification conditions were pre-denaturation at 98°C for 2 min, denaturation at 98°C for 10 s, annealing at 56°C for 15 s, and extension at 72°C for 10 s, for 35 cycles; and final extension at 72°C for 5 min.
[0039] 0.8% agarose gel was used to detect the approximately 1.5 Kb fragment obtained after the PCR reaction. The electrophoresis results were observed using a gel imager. Positive results were sent to the Kunming branch of Beijing Qingke Biotechnology Co., Ltd. for sequencing.
[0040] The 16S rRNA gene sequence of this strain is:
[0041] tgcagtcgaacgatgaaccggtttcggccggggattagtggcgaacgggtgagtaacacgtgggcaatct
[0042] gccctgcactctgggacaagccctggaaacggggtctaataccggatatgactgccgaccgcatggtctg
[0043] gtggtggaaagctccggcggtgcaggatgagcccgcggcctatcagcttgttggtggggtgatggcctac
[0044] caaggcgacgacgggtagccggcctgagagggcgaccggccacactgggactgagacacggcccagactc
[0045] ctacgggaggcagcagtggggaatattgcacaatgggcgcaagcctgatgcagcgacgccgcgtgaggga
[0046] tgacggccttcgggttgtaaacctctttcagcagggaagaagcgcaagtgacggtacctgcagaagaagc
[0047] gccggctaactacgtgccagcagccgcggtaatacgtagggcgcaagcgttgtccggaattattgggcgt
[0048] aaagagctcgtaggcggcttgtcgcgtcggatgtgaaagcccggggcttaactccgggtctgcattcgat
[0049] acgggcaggctagagttcggtaggggagatcggaattcctggtgtagcggtgaaatgcgcagatatcagg
[0050] aggaacaccggtggcgaaggcggatctctgggccgatactgacgctgaggagcgaaagcgtggggagcga
[0051] acaggattagataccctggtagtccacgccgtaaacgttgggaactaggtgtgggcgacattccacgttg
[0052] tccgtgccgcagctaacgcattaagttccccgcctggggagtacggccgcaaggctaaaactcaaaggaa
[0053] ttgacgggggcccgcacaagcggcggagcatgtggcttaattcgacgcaacgcgaagaaccttaccaagg
[0054] cttgacatacatcggaaacctctggagacaggggcccccttgtggtcggtgtacaggtggtgcatggctg
[0055] tcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccttgtcctgtgttgccag
[0056] catgccctttggggtgatggggactcacaggagactgccggggtcaactcggaggaaggtggggacgacg
[0057] tcaagtcatcatgccccttatgtcttgggctgcacacgtgctacaatggccggtacaatgagctgcgaag
[0058] ccgtgaggtggagcgaatctcaaaaagccggtctcagttcggattggggtctgcaactcgaccccatgaa
[0059] gtcggagtcgctagtaatcgcagatcagcattgctgcggtgaatacgttcccgggccttgtacacaccgc
[0060] ccgtcacgtcacgaaagtcggtaacacccgaagccggtggcccaacccttgtgggggagccgtcga1396 (as shown in SEQ ID NO: 1 in the sequence listing).
[0061] The obtained sequences were compared by BLAST on the EZBioCloud website for similarity search. The 16S rRNA gene sequences of typical strains with a similarity greater than 98.5% were selected as reference objects. Multiple sequence alignment was performed in MEGA 11 software. Cluster analysis and phylogenetic tree construction were performed by the neighbor-joining method (Kimura two parameter model was selected, and the bootstrap value was set to 1000 times). The phylogenetic tree of Streptomyces sp. ITBB10008 and similar typical strains is shown in Figure 2. Figure 3 As shown, based on the results, the strain was identified as Streptomyces sp., with a deposit number of GDMCC No: 64824 and a deposit date of July 18, 2024. The depository unit is Guangdong Microbial Culture Collection Center, and the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Yuexiu District, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences.
[0062] Example 3 Evaluation of the biological control effect of strain ITBB10008 on wheat scab in a greenhouse
[0063] Cultivation of potted wheat experimental materials: After cleaning and disinfecting the wheat seeds, the grains were scattered in a prepared culture dish lined with germination paper. The germination paper was moistened with sterile water and then placed in a 4°C refrigerator. An appropriate amount of sterile water was added regularly to keep the germination paper moist. After 2 days, the seeds were taken out and placed in the light for about 1-2 days. After the seeds sprouted, they were sown in seedling pots and placed in a 9°C light vernalization chamber (16h / 8h light / dark) for 21 days. After the vernalization was completed, the wheat seedlings were transplanted into large flower pots (inner diameter 215mm × bottom diameter 13mm × height 165mm), with 7 seedlings transplanted per pot. The pots were then cultured in a greenhouse (greenhouse cultivation conditions: 16h light (6:00-22:00), 8h dark, 20-25°C).
[0064] Preparation of pathogenic fungus F. graminearum spore liquid: Inoculate Fusarium graminearum into a 15 mL tube containing an appropriate amount of mung bean culture medium (30 g of mung beans are boiled with deionized water for about 5 minutes, and the volume is adjusted to 500 mL) and culture in a shaker at a constant temperature (25°C, 180 rpm) for 5-7 days. Count the spores under a microscope using a hemocytometer, dilute to 200 spores / μL, and store in a 4°C refrigerator until use.
[0065] Preparation of spore solution of Streptomyces ITBB10008: The glycerol tube of the strain stored in a -20°C refrigerator in Example 1 was inoculated on ISP2 for activation by the plate streak method, and then transferred to MS medium. After culturing at 28°C for 7-10 days, the spores were washed with sterile water in a clean bench and filtered with sterile cotton balls. After gradient dilution of the spore solution, 200 μL of 10 -5 , 10 -6 and 10 -7 The dilution was spread on ISP2 medium, and each gradient was repeated three times. The culture was carried out in an incubator for about 7 days. The plate with 30-300 colonies was selected and the concentration of the spore solution was calculated after counting. The spore solution was diluted to 8×10 7 CFU / mL is reserved.
[0066] Potted wheat experiments were conducted in a greenhouse. Wheat seedlings were transplanted after 21 days of vernalization. Seven seedlings were planted per pot and watered regularly. Heading was observed daily after wheat reached head formation. Inoculation was performed when the anthers turned from green to yellow. Equal amounts of pathogen F. graminearum spores and biocontrol agent ITBB10008 spores were mixed to a concentration of 10 spores for each agent. 5 / mL and 10 7 The experimental group was composed of 100 CFU / mL of F. graminearum spores per liter. A positive control group (CK) was formed by mixing equal amounts of sterile water and F. graminearum spores. A group inoculated with sterile water alone served as the negative control. Gently break open the wheat husks, inject 10 μL of the prepared mixture or sterile water near the florets, and then release the bag to recover. Wheat ears were placed in a ziplock bag to retain moisture for 48 hours. Inoculation information was recorded on the ear stem. Total spikelet counts and diseased spikelet counts were counted 14 and 21 days after inoculation, and the disease index (PPS) was calculated. Twenty wheat ears were inoculated in each group, and the experiment was repeated three times.
[0067] PPS = ∑ (number of diseased spikelets / total number of spikelets) / n×100
[0068] Preventive and therapeutic effect = (control group disease index - treatment disease index) / control group disease index × 100
[0069] By calculating and plotting the incidence rate and disease index and performing multiple comparisons through variance analysis, it was found that the wheat ears in the sterile water negative control group without pathogen inoculation had no lesions, while the experimental group and positive control group inoculated with pathogen spores suffered from wheat fusarium wilt. Among them, the disease index of the positive control group CK inoculated only with pathogen F. graminearum spores was higher, while the disease index of the experimental group inoculated with both ITBB10008 spores and pathogen spores was lower (Table 1), indicating that the spores of the biocontrol fungus ITBB10008 play an important role in the biological control of wheat fusarium wilt.
[0070] Table 1 Statistics of disease index and biocontrol effect of strain ITBB10008 on wheat fusarium head blight in greenhouse
[0071]
[0072] Note: Different letters in the same column indicate significant differences among treatments at the P < 0.05 level.
[0073] From the lesion point of view ( Figure 4 ), the number of wheat ears withered and whitened in the experimental group was less than that in the positive control group. Statistical analysis showed that the disease index of scab in the experimental group was significantly lower than that in the positive control group. Fourteen days after inoculation, the disease index of the control group was 51.57±14.63, while the disease index of the experimental group inoculated with ITBB10008 spores was 2.44±0.31, significantly different from the control group (P<0.05). Twenty-one days after inoculation, the disease index of all wheat plants increased, with the disease index of the positive control group at 65.88±8.45 and the disease index of the experimental group inoculated with ITBB10008 spores at 5.39±0.93, both significantly different from the control group (P<0.05). Overall, inoculation with ITBB10008 spores was able to prevent and control the spread of wheat scab, with a control rate of 95.25% 14 days after inoculation and a control rate of 91.82% 21 days after inoculation.
[0074] Example 4 Determination of the antibacterial spectrum of Streptomyces sp. ITBB10008
[0075] The pathogens of pepper anthracnose, Colletotrichum capsici, papaya anthracnose, Colletotrichum gloeosporioides, banana wilt, Fusarium oxysporum f.sp.cubense, rice blast, Pyricularia oryzae, rubber anthracnose, Colletotrichum acutatum, cowpea stem rot, Pythium aphanidermatum, rice sheath blight, Thanatephorus cucumeris, mango anthracnose, Colletotrichum gloeosporioides and asparagus stem blight, Phomopsis asparagi, were used as indicator bacteria for antimicrobial spectrum determination.
[0076] The antibacterial spectrum of Streptomyces sp. ITBB10008 was determined by the plate standoff method: using the above-mentioned crop pathogens as indicator bacteria, a 7 mm diameter cake prepared from the pathogen strain pre-cultured on PDA was inoculated in the center of a new PDA plate (90 mm diameter). 3 cm to the left and right of the center was a Streptomyces sp. ITBB10008 cake (7 mm diameter) cultured for 7 days in advance. A plate inoculated with the pathogen only in the center served as the control group. The plates were incubated in the dark at 28°C until the control group grew to cover the entire plate. The diameters of the pathogen colonies were measured, and the antibacterial rates of the strains against each pathogenic fungus were calculated according to the method in Example 2.
[0077] The inhibition rates of ITBB10008 against various plant pathogenic fungi are shown in Table 2 below:
[0078] Table 2 Antimicrobial spectrum determination of Streptomyces sp.ITBB10008
[0079]
[0080] As can be seen from the data in Table 2, the live strain ITBB10008 had a certain antibacterial effect on the nine plant pathogenic fungi tested: among the pathogens tested, strain ITBB20662 had the weakest inhibitory effect on banana wilt pathogen, with an inhibition rate of 70.17±0.82%; it had a stronger inhibitory effect on pepper anthracnose pathogen, rubber anthracnose pathogen, cowpea stem rot pathogen, and rice sheath blight pathogen, with inhibition rates of 73.28±0.86%, 74.79±0.98%, 77.35±0.37%, and 79.91±0.37%, respectively; it had a strong inhibitory effect on the other three pathogens, with inhibition rates all exceeding 80%. The inhibitory rates against rice blast reached 85.90±0.64%, against papaya anthracnose pathogens reached 87.95±1.18%, and against asparagus stem blight reached 89.20±1.08%. The highest inhibitory rate was 94.44±0.74% against mango anthracnose pathogens. The live bacterial pellets of strain ITBB10008 exhibited excellent broad-spectrum activity against the tested agricultural pathogens, thus showing great potential for the development of key technologies for the preparation of novel microbial control agents (microbial pesticides) for combating crop diseases.
[0081] It should be pointed out that the above embodiments are only further explanations of the present invention, rather than limitations. Any adjustments or changes by those skilled in the art within the meaning and scope equivalent to the technical solutions of the present invention should be considered to be included in the scope of protection of the present invention.
Claims
1. A wheat plant-derived biocontrol Streptomyces ITBB10008, characterized in that: Its taxonomic name is Streptomyces sp., the preservation number is GDMCC No: 64824, the preservation date is July 18, 2024, and the preservation unit is Guangdong Provincial Microbiological Culture Collection Center.
2. A biocontrol product for controlling pathogens of crops, characterized in that: It comprises the biocontrol Streptomyces ITBB10008 described in claim 1 and / or the spore liquid of the biocontrol Streptomyces ITBB10008.
3. The biocontrol product for controlling crop pathogens according to claim 2, characterized in that: The spore liquid of the biocontrol Streptomyces ITBB10008 is prepared by the following method: (1) activating the biocontrol Streptomyces ITBB10008 on an ISP2 plate and transferring it to an MS medium for culturing until a large number of spores are produced; (2) Pour sterile water into the above-mentioned ISP2 plate, scrape the spores with a sterile cotton swab and collect the spore liquid.
4. Use of the biocontrol Streptomyces ITBB10008 according to claim 1 or the biocontrol product according to any one of claims 2 to 3 in the preparation of a medicament having the effect of preventing and controlling pathogens of crops.
5. The use according to claim 4, characterized in that: The crop pathogens include one or a combination of the pepper anthracnose pathogen Colletotrichum capsici, the wheat fusarium graminearum Schw, the banana wilt pathogen Fusarium oxysporum f.sp.cubense, the rice blast pathogen Pyricularia oryzae, the papaya anthracnose pathogen Colletotrichum gloeosporioides, the rubber anthracnose pathogen Colletotrichum acutatum, the cowpea stem base rot pathogen Pythium aphanidermatum, the rice sheath blight pathogen Thanatephoruscucumeris, the mango anthracnose pathogen Colletotrichum gloeosporioides and the asparagus stem blight pathogen Phomopsis asparagi.
6. The use according to claim 4, characterized in that: The crops include one or a combination of pepper, wheat, banana, rice, papaya, rubber, cowpea, mango and asparagus.