Biocontrol monascus from rainforest and application of biocontrol monascus

Abstract

The invention discloses biocontrol monascus from rainforests and application of the biocontrol monascus. The name of the monascus is Monascus sanguineus BNMZJ6-6, the monascus sanguineus is classified and named as Monascus sanguineus, and the preservation number of the monascus sanguineus in the Guangdong Microbial Culture Collection Center (GDMCC) is GDMCC No.65369. The monascus sanguineus is named as Monascus sanguineus. The monascus has a very strong antagonistic effect on corynespora vigna, fusarium graminearum, rhizoctonia solani, phomopsis asparagi, verticillium dahliae and pyricularia oryzae.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to a biocontrol Monascus purpureus originating from a rainforest and application thereof. Background Art

[0002] Monascus is a saprophytic filamentous fungus belonging to the genus Monascus, class Ascomycetes, subclass Euascomycetes, and fungi. It is found in trees, soil, and sediments. Its optimal pH is typically between 3.5 and 5.5, and its optimal growth temperature is generally between 25 and 35°C. It is widely used in traditional Chinese medicine, winemaking, and food coloring. Its sweet flavor and warm nature have benefits such as digestion and stomach conditioning, promoting blood circulation and relieving pain, and strengthening the spleen and stomach. Modern research indicates that Monascus and its secondary metabolites, such as statins, fatty acids, sterols, and pigments, have pharmacological effects such as regulating blood lipids, providing anti-inflammatory, anti-tumor, antioxidant, and cholesterol-lowering effects. Furthermore, Monascus has been used as a food colorant and preservative in my country for over 1,000 years, primarily in fermented foods. Monascus, as well as its citrinin and monascus pigments, possess antibacterial activity and are highly safe.

[0003] Research on Monascus and its secondary metabolites has primarily focused on the pharmaceutical field. Monascus possesses numerous biological activities, including antibacterial, lipid-regulating, anti-atherosclerotic, anti-tumor, and neuroprotective properties. Monascus metabolites also exhibit antibacterial activity against fungi and bacteria, and their pharmacological effects are widely used in the pharmaceutical field. Studies have shown that WMD2424, a strain of Monascus purpurogenum isolated from the mangrove wetlands of Chiayi County, possesses unique morphology and antibacterial activity. Cultures of Monascus exhibit significant antibacterial activity against bacteria, particularly the Gram-positive Bacillus subtilis. L. Martinkova et al. found that Monascus pigment extracts have inhibitory effects on bacteria, yeast, and filamentous fungi. Yu Jiyuan discovered that Monascus yellow pigment can inhibit both Gram-positive and Gram-negative bacteria. Ren Hao found that Monascus yellow pigment has weak antibacterial activity against Staphylococcus aureus and Bacillus subtilis, while Monascus red pigment has weak antibacterial activity against Bacillus subtilis. The antimicrobial properties and safety of Monascus and its secondary metabolites suggest that they are promising candidates for biocontrol of plant diseases. However, current research on Monascus primarily focuses on pharmaceuticals and food products, with limited research and application of Monascus and its secondary metabolites in plant disease biocontrol. Effective biocontrol agents or their secondary metabolites would be crucial for effective plant disease biocontrol. Exploring more effective and safer biocontrol agents would provide new insights and theoretical foundations for the application of Monascus and its secondary metabolites in related fields. Given the high safety of Monascus and its antibacterial activity against a wide range of plant pathogens, the question remains as to whether its application in plant disease biocontrol could minimize plant damage and effectively inhibit disease, while also potentially providing other beneficial benefits. Summary of the Invention

[0004] This study isolated strains from rainforest humus samples and identified five Monascus strains through morphological observation and multi-gene molecular identification. Antagonistic tests with Monascus itself, antagonistic activity tests of Monascus fermentation products in different culture media, and product richness assays were performed to screen strains with significant biocontrol efficacy and optimal culture media. Pot culture experiments were also conducted to further demonstrate the biocontrol effects of Monascus. One strain was significantly more effective than the others, exhibiting red or dark red colonies on PDA plates and capable of extensive fermentation on media such as rice, cornmeal, and wheat bran.

[0005] Based on this, the present invention provides a Monascus strain, characterized in that: the Monascus is named Monascus sanguineus BNMZJ6-6, classified as Monascus sanguineus, and has a deposit number of GDMCC No. 65369 in the Guangdong Provincial Microbiological Culture Collection Center (GDMCC).

[0006] The present invention also provides a biological preparation containing the Monascus strain, which is used for preventing and controlling plant pathogens, wherein the plant pathogens are one or more of Corynespora vulgaris, Gibberella zeae, Sheath blight of rice, Phomopsis officinalis, Verticillium dahliae and Rice blast pathogens.

[0007] The application of the Monascus strain in preventing and controlling plant pathogens also falls within the protection scope of the present invention.

[0008] In the application, the plant pathogen is one or more of Corynespora vulgaris, Gibberella fusca, Sheath blight of rice, Phomopsis officinalis, Verticillium dahliae and Rice blast pathogen.

[0009] The present invention also provides a method for culturing the Monascus strain, comprising fermenting and culturing the Monascus strain at room temperature using a rice culture medium or an industrial fermentation culture medium; the rice culture medium is a culture medium prepared by adding 50 mL of water to every 50 g of rice; and the industrial culture medium is a culture medium prepared by mixing 25 g of corn flour, 25 g of wheat bran, and 50 mL of water to every 25 g of corn flour.

[0010] Beneficial effects of the present invention:

[0011] The Monascus sanguineus BNMZJ6-6 provided by the present invention is a Monascus strain with a biocontrol effect. As a traditional food fungus, the Monascus strain has good safety and can provide an efficient and safe strain selection for the research and development of plant disease biological control products. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 The spore and hyphae morphology of the strain under a microscope.

[0013] Figure 2 This is the phylogenetic tree of ITS sequences.

[0014] Figure 3 This is the phylogenetic tree of LSU sequences.

[0015] Figure 4 This is the phylogenetic tree of pksKS sequences.

[0016] Figure 5 The antagonistic effect of 5 tested Monascus strains co-cultured with different pathogens for 4 days.

[0017] Figure 6 Antagonistic effects of metabolites obtained by fermentation in different culture media and co-cultured with different pathogens for 5 days

[0018] Figure 7These are the HPLC analysis results of different culture medium products; Note: A is the HPLC result of CYA culture medium product, B is the HPLC result of MEA culture medium product, C is the HPLC result of PDB culture medium product, D is the HPLC result of industrial culture medium product, and E is the HPLC result of rice culture medium product.

[0019] Figure 8 These are the NMR results of products from different culture media; Note: A is the NMR result of the product from CYA culture media, B is the NMR result of the product from MEA culture media, C is the NMR result of the product from PDB culture media, D is the NMR result of the product from industrial culture media, and E is the NMR result of the product from rice culture media.

[0020] Figure 9 Shows the growth status of plants under different treatments (17d).

[0021] Figure 10 The graph shows the antibacterial test results of strains 6-4 and 6-6.

[0022] Note: The photos of Corynespora vulgaris and Gibberella tritici were taken on the 3rd day, the photos of Rhizoctonia solani, Phomopsis officinalis and Magnaporthe grisea were taken on the 4th day, and the photos of Verticillium dahliae were taken on the 7th day.

[0023] Biomaterial Deposit

[0024] Deposit number: GDMCC No.65369

[0025] Name: Monascus sanguineus BNMZJ6-6

[0026] Taxonomic nomenclature: Monascus sanguineus

[0027] Deposit date: October 30, 2024

[0028] Depository: Guangdong Provincial Microbiological Culture Collection Center (GDMCC)

[0029] Address: Building 59, No. 100, Xianlie Middle Road, Guangzhou

[0030] Survival: Yes. DETAILED DESCRIPTION

[0031] The methods in the following examples are all conventional methods unless otherwise specified.

[0032] The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the reagents used are all commercially available products.

[0033] Example 1. Isolation and Identification of Monascus sanguineus BNMZJ6-6

[0034] 1. Test Materials

[0035] 1. The test pathogens, including banana wilt pathogen, cowpea wilt pathogen, pepper phytophthora, and papaya anthracnose, were provided by the Microbial Resources and Utilization Laboratory, Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences. Cowpea seeds, Cuilong No. 3, were purchased from Beijing Zhongnong Lvheng Technology Co., Ltd. Potting soil was collected from the garden of the Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences.

[0036] 2. Test culture medium

[0037] PDA medium: 200 g potato, 20 g glucose, 15.0 g agar, 1000 mL water;

[0038] Rice culture medium: 50 g rice, 50 mL water;

[0039] Wheat bran medium (industrial medium): 25 g corn flour, 25 g wheat bran, 50 mL water;

[0040] MEA medium: malt extract 15.0 g, peptone 1.0 g, water 1000 mL, pH 7.0;

[0041] PDB medium: 200 g potato, 20 g glucose, 1000 mL water;

[0042] CYA medium: yeast extract 5.0 g, sucrose 30.0 g, agar 15.0 g, water 1000 mL, NaNO3 3.0 g, KH2PO4 1.0 g, KCl 0.5 g, MgSO4·7H2O 0.5 g, FeSO4·7H2O 0.01 g;

[0043] 2. Methods

[0044] 1. Sample collection and strain isolation

[0045] Samples were collected from the Bawangling tropical rainforest in September 2023. The fungi were isolated using the dilution-spreading method. After isolation, colonies with distinct morphologies were purified using the streak method. This step was repeated until a single colony was obtained. The colonies were then transferred to PDA solid medium for culture, numbered, and incubated in a constant-temperature incubator at 28°C. A total of 50 fungal strains were obtained, numbered 1-1 to 1-15, 2-1 to 2-8, 3-1 to 3-8, 4-1 to 4-4, 5-1 to 5-6, 6-1 to 6-6, and 7-1 to 7-3.

[0046] The antimicrobial activity of the fungal strains against common plant pathogens was evaluated using the plate standoff method, and five Monascus strains with antimicrobial activity were screened. The strain with the best activity was named BNMZJ6-6.

[0047] A total of five target Monascus strains were screened, of which the one with the best activity was named BNMZJ6-6.

[0048] 2. Morphological observation of Monascus strains

[0049] The colonies were pretreated using the insert method, and the morphology of hyphae and spores of the strains were observed under an upright microscope.

[0050] The morphological observation results of Monascus are as follows Figure 1 As shown, the morphology of hyphae and spores obtained after treatment by the insert method was observed under an upright microscope. A single spore was grown on the strain BNMZJ6-6.

[0051] 3. Molecular identification of Monascus strains

[0052] 3.1 DNA extraction of test strains

[0053] Take the bacterial culture in the logarithmic growth phase and place it in a 5 mL centrifuge tube, and centrifuge at 12000 r / min for 2 min; add 1.7 mL TE buffer to the precipitate, then add 1.8 g lysozyme, 90 μL 10% SDS and 9 μL 20 mg / mL proteinase K in sequence, and keep the temperature in a 37°C water bath for 1 h; add 300 μL 6 mol / L NaCl and 240 μL CTAB / NaCl, and keep the temperature in a 65°C water bath for 10 min; add an equal volume of chloroform / isoamyl alcohol, mix well, and centrifuge at 12000 r / min for 5 min; transfer the supernatant to another centrifuge tube, add 0.6 volumes of isopropanol to precipitate the DNA, and centrifuge at 12000 r / min for 1 min; discard the supernatant, wash twice with 70% ethanol, dissolve in TE buffer, and store at -20°C until use.

[0054] 3.2 Sequence amplification and PCR

[0055] The universal primers V9G and LS266 were used to amplify the ITS region of the five target strains, the primers LR5 and LR0R were used to amplify the LSU region, and the primers pks1 and pks2 were designed to amplify the pksKS region of the target strain.

[0056] Primers V9G, LS266, LR0R, LR5, pks1, and pks2 are shown in Table 1.

[0057] Table 1. Target gene fragments and primer sequences

[0058]

[0059]

[0060] The PCR reaction system (50 μL) consisted of 25 μL of 2× Es Taq Master Mix (Dye), 17.5 μL of ddH2O, 4.5 μL of DNA template, and 1.5 μL of each upstream and downstream primer. PCR reaction conditions were: 94°C for 5 min, followed by 35 cycles of 94°C for 45 s, 53°C for 30 s, and 72°C for 1 min 45 s, and 72°C for 10 min.

[0061] 3.3 Construction of phylogenetic tree

[0062] The PCR product was sent to Sangon Biotech (Shanghai) Co., Ltd. for gene sequencing. Sequencing yielded the corresponding ITS, LSU, and pksKS base sequences. The ITS sequence of BNMZJ6-6 is shown as Sequence 1 in the sequence listing. The LSU sequence of BNMZJ6-6 is shown as Sequence 2 in the sequence listing, and the pksKS base sequence is shown as Sequence 3 in the sequence listing.

[0063] The sequences obtained after sequencing the PCR products were compared on the NCBI website. At the same time, the base sequences of the reference strains were downloaded to make a phylogenetic tree. Figure 2 ), LSU sequence phylogenetic tree ( Figure 3 ), pksKS sequence phylogenetic tree ( Figure 4 ). After phylogenetic analysis, it was concluded that the target strains were similar to CGMCC 5845 based on the conserved gene ITS sequence and LSU sequence, and BNMZJ6-6 belonged to Monascus sanguineus. Figure 2 , 3, and 4 are represented as 6-6. 6-1, 6-2, 6-3, 6-4, and 6-5 are other strains screened.

[0064] The aforementioned BNMZJ6-6, named Monascus sanguineus BNMZJ6-6 and taxonomically designated Monascus sanguineus, was deposited with Guangdong Provincial Microbiological Culture Collection (GDMCC, address: Building 59, No. 100, Xianlie Middle Road, Guangzhou) on October 30, 2024. Deposit number: GDMCC No. 65369.

[0065] Example 2: Identification of the antibacterial activity of Monascus sanguineus BNMZJ6-6

[0066] 1. Analysis of antibacterial activity of Monascus strains

[0067] The antifungal activity of target Monascus strains against important plant pathogens, including Fusarium oxysporum sp. cubensis, Fusarium oxysporum sp. Tracheiphlium, Phytophthora capsici Leonian, and Coletotrichum gloeosporioides, was tested using a plate standoff method. A 5 mm diameter cake of the target pathogens was punched out with a hole punch and placed in the center of a PDA plate. Different Monascus strains were then inoculated at equal distances and sizes onto the left and right sides of the target pathogen cake. A control (CK) containing only the pathogens and no Monascus was used. The plates were incubated at 28°C, with three replicates for each treatment. Starting two days after inoculation, measure the colony diameter (dB) of the target pathogen strain and the colony diameter (dCK) of the control target pathogen daily. Calculate the inhibition rate of the suspected Monascus target strain using the following formula: Inhibition rate / % = (dCK - dB) / dCK × 100. Select the Monascus strain with the strongest overall effect for subsequent experiments.

[0068] The results of the antibacterial activity of Monascus strains are shown in Table 2 and Figure 2 As shown, strain 6-1 had a weak antagonistic effect on cowpea wilt pathogen; the antagonistic rate of the five Monascus strains was generally above 20% for banana wilt pathogen; strains 6-4, 6-5, and 6-6 had high antagonistic rates against pepper phytophthora; and strains 6-5 and 6-6 had a more significant antagonistic effect on papaya anthracnose. According to the calculation of the total inhibition rate of the four pathogens, strains 6-4 and 6-6 had a better overall inhibition rate (Table 2). Antagonistic pictures on the fourth day ( Figure 5 ). Among them, Monascus sanguineus BNMZJ6-6 is listed in Table 2 and Figure 5 It is represented as 6-6. 6-1, 6-2, 6-3, 6-4, and 6-5 are other strains screened.

[0069] Table 2. Inhibition rate of each strain against different pathogens

[0070]

[0071] 2. Screening of the optimal culture medium for Monascus antibacterial

[0072] 1. Antibacterial experiment of Monascus secondary metabolites

[0073] Monascus purpureus was inoculated into PDB, MEA, CYA, industrial, and rice culture media, respectively, and fermented for 28 days. After fermentation, the extract was extracted with an equal volume of ethyl acetate three times. The ethyl acetate phase was filtered and dried using a fermentation rotary evaporator. The crude metabolite extract was dissolved in ethyl acetate to obtain the metabolite. The antibacterial activity of the crude metabolite extract against the above-mentioned important plant pathogens was determined by observing the growth of the pathogens using a plate standoff method.

[0074] Based on the results of morphological observation, molecular identification and antagonism experiments, the better strain 6-6 was selected as the target strain for subsequent experiments. The plate confrontation experiment investigated the antagonistic effect of the crude extract of Monascus secondary metabolites on cowpea wilt pathogen, papaya anthracnose and pepper phytophthora co-cultured for 5 days ( Figure 6 The fermentation products of PDB medium have almost no inhibitory effect on pathogens, while the fermentation products of industrial medium have the most obvious inhibitory effect on pathogens.

[0075] 2. Evaluation of the richness of Monascus secondary metabolites

[0076] Prepare a total of 11.3 kg of rice culture medium and inoculate each bottle with 5 mL of Monascus culture. Ferment at room temperature for 28 days. After fermentation, extract the culture three times with ethyl acetate. Combine the extracts and rotary evaporate to dryness to obtain the crude secondary metabolite extract.

[0077] Use a syringe to inject the sample into the instrument and mix it with the mobile phase through the valve connected to the sample loop. Install the chromatographic column on the liquid chromatograph, paying attention to the flow direction and sealing. Use a syringe to inject the fermentation product liquid of the sample Monascus into the instrument and mix it with the mobile phase through the valve connected to the sample loop. The injection volume is 10μL. Install the chromatographic column on the liquid chromatograph. Column model: C 18 -MS-II 4.6×250mm. Methanol and water were selected as the mobile phases at a flow rate of 3 ml / min. Full-wavelength detection was performed from 200 nm to 580 nm, with a mobile phase elution gradient of 20% to 100% and an elution time of 25 min. Filtration and degassing were performed. After analysis, the resulting chromatograms were collected using HPLC software, and the richness of Monascus products obtained from fermentation in different culture media was analyzed based on the chromatograms. Simultaneously, the fermentation products of the bacteria were dissolved in deuterated methanol and analyzed by nuclear magnetic resonance (NMR). The solvent peak of deuterated methanol was 3.31 ppm, and the water peak was 4.87 ppm. Using the results of analytical liquid chromatography and NMR, the richness of Monascus secondary metabolites under different conditions was preliminarily determined by the number of peaks. Combined with the results of the plate standoff experiment, the appropriate fermentation medium was determined.

[0078] From the chromatogram ( Figure 7), under the same elution conditions, the rice culture medium chromatogram has the largest number of peaks and the highest richness. Figure 8 Similar to the HPLC results, rice and industrial fermentation medium (25g corn flour, 25g wheat bran, 50ml water) showed high substance richness in fermentation products, while PDB medium showed the lowest substance richness. Combined with these results, industrial medium was determined to be the optimal medium for Monascus fermentation, providing a richer fermentation product and the best antibacterial effect.

[0079] 3. Potted plant prevention effect experiment

[0080] 1) Preparation of cowpea wilt pathogen spore suspension

[0081] The colonies grown on the PDA plate were inoculated into 200 mL of PDB medium and cultured in a shaker at 28°C for 5 days. The number of spores in the suspension was counted using a hemocytometer to obtain the concentration of the spore suspension, which was then diluted to 10 6 CFU / mL, reserve for future use.

[0082] 2) Preparation of biocontrol bacteria Monascus suspension

[0083] The colonies grown on the PDA plate were inoculated into 200 mL of CYA medium and cultured in a shaker at 28°C for 3 days. The number of spores in the suspension was counted using a hemocytometer to obtain the concentration of the spore suspension, which was then diluted to 10 9 , 10 8 , 10 6 , 10 4 CFU / mL, reserve for future use.

[0084] 3) Potted plant experiment

[0085] Select plump and similar-sized cowpea seeds and sow them in order in natural soil that has been sterilized twice at 121°C for 30 minutes and then naturally cooled. There are 15 seeds in each pot, for a total of 6 pots. Water them and place them in the sun to wait for germination. Each pot is treated with the following: (1) pathogen treatment + biocontrol bacteria 10 4 CFU / mL(F+10 4 S), (2) pathogen treatment + biocontrol bacteria 10 6 CFU / mL(F+10 6 S), (3) pathogen treatment + biocontrol bacteria 10 8 CFU / mL(F+10 8 S), (4) pathogen treatment + biocontrol bacteria 10 9 CFU / mL(F+10 9S), (5) water control (CK), (6) pathogen control (F). Plant height, root length, and chlorophyll content of different treatments were measured, the disease incidence was observed and recorded, and the incidence rate, disease index, and control effect were calculated. Disease index reference (POTTORFF M, WANAMAKER S, MA YQ, et al. Genetic and physical mapping of candidate genes for resistance to Fusarium oxysporum f.sp.tracheiphilum race 3 in cowpea [Vigna unguiculata (L.) Walp] [J]. PLoS One, 2012, 7 (7): e41600). Incidence rate = number of diseased plants / total number of plants × 100%; disease index = ∑(disease grade of each plant) / (total number of plants × highest disease grade) × 100; control effect = (CK disease index - treatment group disease index) / CK disease index × 100% (Xie Haipeng, Lin Yingtao, Wu Xiaoyan, et al. Screening and identification of biocontrol bacteria for cowpea wilt and preliminary study on disease resistance mechanism [J]. Journal of Tropical Crops, 2023, 44(06): 1224-36).

[0086] The experimental results are shown in Table 3, Table 4 and Figure 9 The results showed that the application of different concentrations of biocontrol bacteria can effectively control the invasion of cowpea wilt disease. 4 The leaves of the diseased plants treated with S turned yellow obviously, and the control effect was poor. 6 The disease of the diseased plants treated with S was significantly improved, the incidence rate and disease index were significantly reduced, and the control effect reached 42.35%, which was relatively high.

[0087] Comprehensively considering the incidence rate, disease index and control effect, it is believed that the optimal application concentration of the biocontrol fungus Monascus is 10 6 CFU / mL, which has the best prevention and control effect and does not cause harm to the growth of plants.

[0088] Table 3. Effects of different concentrations of the biocontrol agent Monascus 6-6 on the growth promotion of cowpea in potted plants

[0089]

[0090] Note: Different lowercase letters in the same column indicate significant differences (P<0.05).

[0091] Table 4 Disease index and control effect of different biocontrol bacteria concentrations

[0092]

[0093]

[0094] 4. Further analysis of the antibacterial activity of Monascus strains

[0095] The above experimental operations were adopted to analyze the antibacterial activity of Monascus strains, and strains 6-4 and 6-6 with better antagonistic effects were selected from the five target strains for further antibacterial activity experiments to analyze the antagonistic effects of the above two Monascus strains on cowpea Corynespora, wheat fusarium, rice sheath blight, asparagus phomopsis, dahliae verticillium, and rice blast pathogens.

[0096] Among the six newly acquired plant pathogens, Monascus had little to no antibacterial effect against Gibberella fusca and Rhizoctonia solani. Gibberella fusca grows very quickly, so by the third day, its growth had already covered the entire culture medium, showing no antibacterial results at all. Monascus strains showed some inhibition against Coryneformis spp., Verticillium dahliae, and Rice blast fungi. Strain 6-6 had a more pronounced inhibitory effect against Verticillium dahliae and Rice blast fungi. The inhibition rate against Coryneformis spp. was higher, but the range of inhibition was limited, so the effect was not obvious from the images. The most pronounced antibacterial effect was against Phomopsis officinale, with a wide range of inhibition and an inhibition rate of 20%.

[0097] Table 5. Further antibacterial experiments of strains 6-4 and 6-6

[0098]

[0099] 3. Conclusion

[0100] Based on the results of plate standoff experiments, Monascus extracted from the rainforest exhibits a moderate inhibitory effect against common plant pathogens, indirectly demonstrating its potential for plant biocontrol applications. Monascus was fermented using liquid media (MEA, CYA, and PDB), as well as solid media (rice and industrial media), and its fermentation products were extracted. Crude extracts of Monascus secondary metabolites were obtained, and the antibacterial properties, activity, and abundance of these products were investigated under different fermentation media conditions. The results demonstrated that Monascus secondary metabolites exhibited significant antibacterial activity against common plant pathogens and were abundant in solid media.

[0101] The pot experiment showed that the target Monascus had a certain preventive effect on cowpea wilt according to the chlorophyll content of leaves, plant height, root length and disease situation. Although there was no very obvious regular growth-promoting effect, it would not cause harm to the growth of the plant. The best control effect was achieved at a concentration of 10 6 CFU / mL.

[0102] The results of this experiment show that the secondary metabolites of Monascus have a good antibacterial effect on common plant pathogens, and can produce abundant secondary metabolites in solid culture medium. It can be seen that Monascus isolated from tropical rainforest can achieve an inhibitory effect on some plant pathogens represented by cowpea wilt virus at an appropriate concentration, and has the potential and ability to be used in plant biocontrol.

Claims

1. A Monascus strain, characterized by: The name of the Monascus sanguineus is Monascus sanguineus BNMZJ6-6, the classification name is Monascus sanguineus, and the preservation number in Guangdong Provincial Microbiological Culture Collection Center (GDMCC) is GDMCC No. 65369.

2. A biological preparation containing the Monascus strain according to claim 1, wherein the biological preparation is used to control plant pathogens, and the plant pathogens are one or more of Corynespora vulgaris, Gibberella zeae, Sheath blight of rice, Phomopsis officinalis, Verticillium dahliae and Rice blast pathogen.

3. Use of the Monascus strain according to claim 1 in preventing and controlling plant pathogens.

4. The use according to claim 3, characterized in that The plant pathogenic bacteria are one or more of Corynespora vulgaris, Gibberella fusca, Sheath blight of rice, Phomopsis officinalis, Verticillium dahliae and Rice blast pathogens.

5. The method for culturing the Monascus strain according to claim 1, wherein The Monascus strain is cultured using a rice culture medium or an industrial fermentation culture medium; the rice culture medium is a culture medium prepared by adding 50 mL of water to every 50 g of rice; the industrial culture medium is a culture medium prepared by mixing 25 g of corn flour with 25 g of wheat bran and 50 mL of water.

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