A shared Fusarium strain and its application

Abstract

The present invention discloses a shared Fusarium and its application, and belongs to the technical field of plant endophytic fungi. The shared Fusarium isolated by the present invention has a deposit number of CGMCC No.41090. Experiments have found that strain G3-29 has excellent inhibitory effects on multiple pathogens of Fusarium graminearum, pseudo-Fusarium graminearum, anthracnose bacteria, sclerotinia, wheat root rot flat navel helminth spores and corn flat navel helminth spores, and has good application prospects. Strain G3-29 has a significant promoting effect on wheat growth, and has a significant biological control effect on wheat scab. In addition, strain G3-29 processes lycopersicon and corn, and can also significantly reduce the occurrence of lycopersicon leaf spot disease and corn leaf spot disease. The present invention provides new strain resources for the prevention and control of gramineous plant diseases, and provides a theoretical basis for the effective prevention and control of multiple gramineous plant diseases.

Description

Technical Field

[0001] The invention relates to the technical field of plant endophytic fungi, in particular to a shared Fusarium and application thereof. Background Art

[0002] Fusarium graminearum complex ( Fusarium graminearum complex ) is a worldwide wheat disease. In recent years, it has become one of the most important diseases threatening my country's safe wheat production. The pandemic of this disease not only caused a large-scale reduction in wheat production, but the greater threat is that the grains infected with Fusarium head blight will accumulate fungal toxins, including zearalenone (ZEN), nivalenol (NIV) and deoxynivalenol (DON), as well as acetylated derivatives 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON), which seriously threaten food safety and human health. Chemical control is still an important measure for the prevention and control of Fusarium head blight, but over-reliance on chemical control leads to resistance of the pathogen to fungicides. (Bipolaris sorokiniana) It is one of the most common and serious pathogens that cause plant fungal diseases. This pathogen can cause serious diseases in many grasses and plants such as barley, wheat, and rice, leading to leaf spots, root rot, mildew, etc. The disease is widely distributed and can cause huge economic losses. (Bipolaris maydis It can cause corn leaf diseases worldwide, often leading to poor growth and affecting yield. In years with severe disease, corn is infected on a large scale, affecting plant growth and reducing yield, affecting high and stable yields of corn and causing serious economic losses.

[0003] Endophytic fungi are a potential biocontrol resource that can control various pathogens and promote plant growth and health. These fungi are interdependent with their hosts without causing any obvious disease symptoms. Endophytic fungi are able to adapt to life inside plants and produce antagonistic secondary metabolites that are essential for growth and disease resistance of host plants. Among endophytic fungi, Penicillium ( Penicillium ), Epicococcus ( Epicoccum )、Trichoderma( Trichoderma ), Chaetomium ( Chaetomium ), Piriformis ( Piriformospora ) and Fusarium ( Fusarium ) can enhance the host's tolerance to biotic and abiotic stresses. Studies have shown that Fusarium spp. Fusarium ) can show biological control activity against a variety of plant diseases. However, the shared Fusarium commune) There are few reports on its application in preventing and controlling fungal diseases and promoting plant growth. Summary of the invention

[0004] The purpose of the present invention is to provide a shared Fusarium and its application to solve the problems existing in the above-mentioned prior art. The shared Fusarium has excellent inhibitory effect on Fusarium graminearum, Pseudocerium graminearum, anthracnose and Sclerotinia sclerotiorum, has a broad-spectrum antibacterial activity, and also has a significant promoting effect on wheat growth, as well as significant biological control effects on wheat ergot, corn leaf spot and Elymus salsa leaf spot.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a shared Fusarium commune G3-29, with a deposit number of CGMCC No.41090 and a deposit date of January 29, 2024. The deposit unit is the General Microbiology Center of China Culture Collection Administration, and the deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0007] The present invention also provides a biocontrol agent, comprising the shared Fusarium.

[0008] The present invention also provides the use of the shared Fusarium in any of the following:

[0009] (1) Application in the prevention and control of grass plant diseases;

[0010] (2) Application in the preparation of microbial agents for preventing and controlling plant diseases;

[0011] Among them, the gramineous plant diseases include wheat fusarium head blight, Elymus salsa leaf spot and corn leaf spot.

[0012] The present invention also provides the use of the shared Fusarium in any of the following:

[0013] (1) Application in inhibiting pathogens of grass plant diseases;

[0014] (2) Application in the preparation of microbial agents for inhibiting pathogenic bacteria of grass plant diseases;

[0015] The pathogens of grass plant diseases include Fusarium graminearum, Fusarium pseudograminearum, Colletotrichum gloeosporioides, Sclerotinia sclerotiorum, Bipolaris sorokiniana and Bipolaris maydis.

[0016] The present invention also provides the use of the bacterial agent in preventing and controlling diseases of gramineous plants, wherein the diseases of gramineous plants include wheat fusarium head blight, lycopersicon esculentum leaf spot and corn leaf spot.

[0017] The present invention also provides the use of the bacterial agent in inhibiting pathogenic bacteria of gramineous plant diseases, wherein the pathogenic bacteria of gramineous plant diseases include Fusarium graminearum, Pseudo-Fusarium graminearum, Anthracnose, Sclerotinia sclerotiorum, Helminthosporium tataricum and Helminthosporium maydis.

[0018] The present invention also provides the use of the shared Fusarium in any of the following:

[0019] (1) Application in promoting wheat growth;

[0020] (2) Application in the preparation of bacterial agents for promoting wheat growth;

[0021] The wheat growth promotion includes promoting the increase of root fresh weight, root length, above-ground fresh weight and plant tillering number of wheat.

[0022] The invention also provides a wheat growth promoter, comprising the shared Fusarium.

[0023] The present invention also provides a method for preventing and controlling diseases of gramineous plants, comprising applying the shared Fusarium or the biocontrol agent to gramineous plants; the gramineous plant diseases include wheat fusarium rust, Elymus salsa leaf spot and corn leaf spot.

[0024] The present invention also provides a method for promoting wheat growth, comprising applying the shared Fusarium or the wheat growth promoter to Gramineae plants; the wheat growth promotion comprises promoting the increase of root fresh weight, root length, above-ground fresh weight and plant tillering number of wheat.

[0025] The present invention discloses the following technical effects:

[0026] The present invention screened and isolated a new plant endophytic fungus: shared Fusarium G3-29. Experiments have found that strain G3-29 has excellent inhibitory effects on multiple pathogens such as Fusarium graminearum, Pseudocerium graminearum, Colletotrichum, Sclerotinia, Helminthosporium oleraceum and Helminthosporium maydis, and has good application prospects. Under greenhouse and field conditions, strain G3-29 has a significant promoting effect on wheat growth; wheat is treated with spore liquid of strain G3-29, and it is found that it has a significant biological control effect on wheat fusarium scab; strain G3-29 is used to treat lycopersicon esculentum and corn, and the occurrence of lycopersicon esculentum leaf spot and corn leaf spot can also be significantly reduced. The present invention provides a new strain resource for the prevention and control of diseases of Gramineae plants, and provides a theoretical basis for the effective prevention and control of various diseases of Gramineae plants. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Figure 1 The colony and conidia morphology of strain G3-29, and the growth conditions under different pH and different culture media, among which A is the top view colony morphology of G3-29 after five days of culture in PDA culture medium, B is the bottom view colony morphology of G3-29 after five days of culture in PDA culture medium, C is the conidia morphology of G3-29, the scale is 20μm, D is the effect of pH value on the growth of strain G3-29 5 days after inoculation, and E is the growth statistics of G3-29 under five different culture media;

[0029] Figure 2 This is the electrophoresis diagram of the PCR amplification of the translation elongation factor-1 α (EF-1α) fragment of the ribosomal DNA (rDNA) of strain G3-29; DNA Marker is the standard DNA molecule, and G3-29 is the amplified product of the ribosomal DNA of strain G3-29;

[0030] Figure 3 is the phylogenetic tree of strain G3-29;

[0031] Figure 4is the broad-spectrum antifungal activity of strain G3-29 against pathogenic fungi, where A is the antifungal activity of strain G3-29 against plant pathogenic fungi on PDA medium, B, C, D, and E are the inhibition rate statistics of G3-29 on Fg, Fp, Cg, and Ss on PDA plates at 3d, 5d, and 7d, respectively; F is the inhibition rate statistics of G3-29 on Bs and Bm on PDA plates; where Fg is Fusarium graminearum, Fp is Pseudomonas graminearum, Cg is Colletotrichum, Ss is Sclerotinia sclerotiorum, Bs is Helminthosporium spp., and Bm is Helminthosporium maydis; error bars represent standard errors of the mean; significant differences with p≤0.05 are indicated by * according to student t test, or significant differences are indicated by different lowercase letters according to one-way ANOVA and Tukey post hoc test analysis;

[0032] Figure 5 The effect of strain G3-29 on wheat growth under greenhouse and field conditions; A is a picture of wheat plants treated with spores of strain G3-29 after 45 days of growth in the greenhouse, B is the fresh weight analysis of wheat roots, C is the fresh weight analysis of wheat aerial parts, D is the analysis of wheat tiller number, and E is the analysis of wheat root length; F is the height of wheat plant; t-test was used, n=20, the displayed values ​​represent the mean ± standard error, the error bars represent the standard error, * indicates significant difference, and ns indicates insignificant difference;

[0033] Figure 6 The resistance of strain G3-29 to wheat fusarium ergot is determined. The five wheat ears in the upper row correspond to A: G3-29, B: G3-29 (2d) + PH-1, C: ddH2O (2d) + PH-1, D: G3-29 (4d) + PH-1, and E: ddH2O (4d) + PH-1, respectively. A: G3-29 represents wheat ears inoculated with G3-29 conidia suspension alone. B: G3-29 (2d) + PH-1 represents wheat ears inoculated with conidia suspension of Fusarium graminearum PH-1 2 days after spraying G3-29 spore solution. C: ddH2O (2d) + PH-1 represents wheat ears sprayed with ddH2O Wheat ears inoculated with conidia suspension of Fusarium graminearum PH-1 2 days later; D: G3-29 (4d) + PH-1, which means wheat ears sprayed with G3-29 spore solution and inoculated with conidia suspension of Fusarium graminearum PH-1 4 days later; E: ddH2O (4d) + PH-1, which means wheat ears sprayed with ddH2O and inoculated with conidia suspension of Fusarium graminearum PH-1 4 days later (* indicates significant difference at p ≤ 0.05 level, ns indicates insignificant difference);

[0034] Figure 7The control effect of strain G3-29 on leaf spot of lycopene grass under greenhouse conditions, wherein A is the control effect of strain G3-29 spore solution on leaf spot pathogen of lycopene grass - Helminthosporium spp. (Bs) after treating lycopene grass plants, and B is the control effect of strain G3-29 spore solution on leaf spot pathogen of lycopene grass - Helminthosporium spp. (Bm) after treating lycopene grass plants;

[0035] Figure 8 The control effect of corn leaf spot disease on corn plants treated with spore liquid of strain G3-29 under greenhouse conditions; A is a phenotypic diagram, and B is a statistical diagram. DETAILED DESCRIPTION

[0036] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0037] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0038] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0039] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0040] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0041] In the present invention, unless otherwise specified, the raw materials and equipment used can be purchased from the market or are commonly used in the art. The methods in the following examples, unless otherwise specified, are all conventional methods in the art.

[0042] (1) Potato dextrose agar medium (PDA medium): Cut 200g of potatoes into small pieces and put them into a pot. Add 1000mL of water and heat to boiling on a heater for 30 minutes. Filter through a measuring cup with two layers of gauze while hot and discard the residue. Add 20g of glucose and 20g of agar to the filtrate and add water to 1000mL. Then put it into a high-temperature and high-pressure sterilizer for sterilization at 121℃ for 15 minutes.

[0043] (2) PDB culture medium: Cut 200g of potatoes into small pieces and put them into a pot. Add 1000mL of water and heat to boiling on a heater for 30 minutes. Filter through a measuring cup with two layers of gauze while hot and discard the residue. Add 20g of glucose to the filtrate and add water to 1000mL. Then put it into a high-temperature and high-pressure sterilizer for sterilization at 121℃ for 15 minutes.

[0044] (3) CMC medium: 15 g sodium carboxymethyl cellulose, 2 g sodium nitrate, 1 g potassium dihydrogen phosphate, 0.5 g magnesium sulfate heptahydrate and 1 g yeast extract, add water to 1000 mL. Then sterilize in a high-temperature and high-pressure autoclave at 121°C for 15 min.

[0045] Example 1 Isolation and identification of strains

[0046] 1. Isolation of strains

[0047] Healthy and symptom-free wheat was collected in Heilongjiang Province, China as wheat samples. The wheat samples were immersed in a 70% ethanol aqueous solution for 1 min for surface disinfection, then transferred to a 2% sodium hypochlorite solution for 2 min, and finally immersed in a 70% ethanol aqueous solution for 30 s. After this treatment, they were washed three times with sterile distilled water and dried on sterile filter paper. The wheat samples were cut into 5 mm small segments and placed on the surface of PDA culture medium. 50 μg / mL ampicillin was added to the PDA culture medium to inhibit the growth of bacteria in the PDA culture medium. 100 μL of sterile distilled water used for washing in the last step of surface disinfection was also added to the surface of the PDA culture medium to evaluate the efficiency of surface sterilization. All PDA culture media were cultured in a dark incubator at 25 °C. After the mycelium grew out, the mycelium at the tip of the colony was taken and transferred to a new PDA plate without ampicillin antibiotics for preliminary purification culture. The colony was subcultured and purified three times, and the obtained strain was named G3-29. The strain was stored on a PDA slant at 4°C and simultaneously placed in 25% glycerol at -80°C for further use.

[0048] 2. Morphological and molecular biological identification of strain G3-29

[0049] 2.1 Morphological identification of G3-29

[0050] The preserved G3-29 strain was activated and cultured in PDA medium at 25°C in the dark for 5 days. Then, a 5 mm diameter bacterial cake was taken from the edge of the activated culture colony with a hole puncher and placed in a freshly prepared PDA medium and cultured for 5 days under the same culture conditions. The front and back of the colony were observed and photographed. Figure 1 A and B show the front (Top view) and back (Bottom view) colony morphology of G3-29 after five days of culture in PDA medium.

[0051] 2.2 Growth of G3-29 on culture media with different pH values

[0052] The strain G3-29 was cultured on PDA medium with pH values ​​of 5, 6, 7, 8, 9, and 10, respectively. The growth rate of the strain was counted after 5 days of culture, and it was found that pH value did not affect the growth of the strain G3-29 (e.g. Figure 1 (shown in D).

[0053] 2.3 DNA extraction and PCR amplification

[0054] After strain G3-29 was cultured in PDA medium for 7 days, genomic DNA was extracted using DNeasy Plant Mini kit (purchased from Qingdao Shengke Biotechnology Co., Ltd.). The concentration of the extracted DNA was determined using a NanoDrop spectrometer. PCR specifically amplified the EF-1α fragment. The total volume of the specific PCR reaction system was 25 μL, which contained 1 μL DNA template, 1 μL of each primer, 12.5 μL of 2×Taq Mix (Quanshijin, Beijing, China) and 9.5 μL ddH2O. The specific PCR primers, reaction system and reaction procedure are shown in Tables 1-3. The PCR products were separated and purified using 1% agarose gel (w / v) containing 0.01% (v / v) GoldView nucleic acid stain, and electrophoresed at 120V, 400A for 30min. The amplified bands were observed under UV light using a gel imaging system (BioRad, Chemi Doc, MP), and the electrophoresis results are shown in Tables 1-3. Figure 2 The single band was sent to Beijing Qingdao Biotechnology Co., Ltd. for purification and sequencing.

[0055] Table 1 PCR primers

[0056]

[0057] Table 2 PCR reaction system

[0058]

[0059] Table 3 PCR reaction procedure

[0060]

[0061] 2.4 Phylogenetic evaluation of endophytic fungus strain G3-29

[0062] According to the sequencing results and NCBI alignment results, strains related to Fusarium were selected (as shown in Tables 4-5) to construct phylogenetic trees. The sequences of EF-1α were aligned using the maximum likelihood method based on the Tamura 3-parameter model for multi-locus linkage, and 1000 repeated bootstrap analyses were performed to estimate evolutionary distances. The phylogenetic relationships showed that, Figure 3 Shown is the phylogenetic tree of strain G3-29 and its closely related species calculated by the maximum likelihood method based on EF-1α sequence. Strain G3-29 and several strains of shared Fusarium commune are classified into one branch, indicating that strain G3-29 belongs to shared Fusarium commune.

[0063] The shared Fusarium commune G3-29 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on January 29, 2024. The deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Building 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No.41090.

[0064] Table 4 GenBank accession numbers of Fusarium used for phylogenetic tree analysis

[0065]

[0066] Note: The bold font represents the strain G3-29 isolated from wheat, and the nodes give a maximum likelihood value greater than 60% (1000 replicates).

[0067] Table 5 GenBank accession numbers of Fusarium used for phylogenetic tree analysis

[0068]

[0069] Note: The nodes give the maximum likelihood value greater than 60% (1000 replicates).

[0070] In addition, the strain G3-29 isolated and identified above was cultured on 5 different solid media. Figure 1As shown in E, the results showed that the growth effect was best on PDA medium, so PDA was used for culture in subsequent experiments.

[0071] The above five solid culture media are prepared by adding different components to PDA culture medium, specifically: sodium dodecyl sulfate (SDS, 0.01%), Congo red (congo red, 0.3 g / L), and NaCl (0.9 mmol / L) are added to PDA culture medium respectively, and then sterilized with high-temperature steam and poured into plates for use; and hydrogen peroxide (H2O2, 0.069%) is added after the temperature of PDA culture medium is lower than that of hot hands, mixed well, poured into plates for use, and then added to PDA culture medium to prepare.

[0072] Example 2 Determination of the antibacterial effect of strain G3-29

[0073] The inhibitory effect of strain G3-29 on Fusarium graminearum, Fusarium graminearum, Colletotrichum gloeosporioides, Sclerotinia sclerotiorum, Bipolaris sorokiniana and Bipolaris maydis was determined by plate confrontation, as follows:

[0074] The radial growth of colonies, inhibition zone and inhibition rate of fungal colony growth were determined by the confrontation culture method. A 5 mm diameter cake was taken from the edge of the pathogenic fungal colony after 5 days of activation culture and inoculated in the center of the PDA plate. A 5 mm diameter cake was taken from the edge of the G3-29 colony that was also activated and inoculated on both sides close to the edge of the plate. The inoculated G3-29 cake was in a straight line with the plant pathogenic fungal cake. All culture dishes were placed in a 25 ° C incubator for 7 days in the dark. The experiment was carried out 3 times, and each treatment was repeated 3 times. The inhibitory effect of endophytes on pathogenic fungi was determined by observing and counting the growth of pathogenic fungi. Error bars represent the standard error of the mean. According to the student t test, significant differences with p ≤ 0.05 are indicated by *, or according to the one-way analysis of variance and Tukey post hoc test, significant differences are indicated by different lowercase letters.

[0075] like Figure 4 As shown in Figure 2, on PDA medium, the endophytic fungus strain G3-29 had a strong inhibitory effect on six pathogenic fungi including Fusarium graminearum ( Figure 4 A), radial growth of colonies and inhibition zones were recorded in Figure 4The inhibition rate of endophytic fungus G3-29 against Fusarium graminearum was 76%, against Fusarium pseudograminearum was 72%, against Colletotrichum gloeosporioides was 90%, against Sclerotinia sclerotiorum was 68%, against Bipolarissorokiniana was 45.79%, and against Bipolaris maydis was 61.09%. In summary, the inhibition rate of G3-29 against pathogenic fungi can reach up to 90%.

[0076] Wherein, Fusarium graminearum is Fusarium graminearum PH_1: This strain has been disclosed in the literature (Wang S, Kondo H, Liu L, Guo L, Qiu D. A novel virus in the family Hypoviridae from the plantpathogenic fungus Fusarium graminearum. Virus Res. 2013 Jun; 174 (1_2): 69_77. doi: 10. 1016 / j. virusres. 2013. 03. 002. Epub 2013 Mar 14. PMID: 23499998.), and the public can obtain the above biological materials from the applicant. The obtained above biological materials are only used for repeating the experiment of the present invention and cannot be used for other purposes. Other pathogens are all common pathogens and can be obtained through commercial channels or other regular channels. The other pathogens used in the present invention are all stored in the Institute of Plant Protection, Chinese Academy of Agricultural Sciences.

[0077] Example 3 Preparation of conidia solution of strain G3-29

[0078] Take 2cm of the activated strain G3-29 2 The bacterial block was cut into small pieces with a sterile scalpel and placed in 50 mL CMC liquid medium, cultured in the dark at 25°C and 180 rpm for 3-5 days, filtered with 3 layers of sterile filter cloth, and the spore suspension was centrifuged at 8000 rpm for 5 minutes to collect the spores. The spore precipitate was resuspended with sterile water to obtain a concentrated spore solution. The spore concentration was then measured with a hemocytometer and the spore concentration was adjusted to a working concentration (10 6 Spores / mL) to obtain a spore suspension, which was stored. The spore morphology was observed as follows: Figure 1 As shown in C.

[0079] Example 4 Effect of strain G3-29 on promoting wheat growth

[0080] Wheat variety Jimai 22 seeds were rinsed with tap water, surface disinfected with 5% NaClO solution for 5 min, and then rinsed with sterile distilled water for 3 times. The surface-sterilized seeds were cultured in a petri dish containing sterile filter paper soaked in ddH2O and cultured at 25°C for 3 days until germination. At the same time, the bacterial cake of strain G3-29 that had been cultured in PDA medium for 5 days was placed in a triangular flask containing 100 mL of CMC culture medium, and cultured at 175 rpm in a shaker at 25°C for 5-7 days. The spores were filtered with 3 layers of sterile filter cloth, and the spore suspension was centrifuged at 8000 rpm for 5 min. The spores were collected and resuspended with sterile water to the use concentration (10 6 spores / mL) to obtain a conidia suspension of the endophytic fungus G3-29.

[0081] Then, the newly germinated seedlings of Jimai 22 were treated with the conidia suspension of strain G3-29, and the root injury method was as follows:

[0082] After 3 days of culture, the germinated seedlings of Jimai 22 were taken out from the culture dish containing filter paper, and the root tips were cut off by 2 mm. The roots of the germinated seedlings of Jimai 22 were immersed in the conidia suspension of strain G3-29 for 6 hours (seedlings immersed in PDB culture medium were used as control). The inoculated seedlings were transplanted into sterilized soil and cultured in a greenhouse with a photoperiod of 16h / 8h at a relative humidity of 65%. After 45 days of growth, the wheat growth parameters were measured and statistical analysis was performed.

[0083] The results showed that wheat plants treated with G3-29 grew more vigorously than those in the control plants (e.g. Figure 5 The average fresh root weight of wheat roots treated with G3-29 strain was 2.65±0.79 g, which was significantly greater than that of untreated control plants (1.53±0.49 g, P≤0.05). Figure 5 As shown in Figure B, the average fresh weight of the aboveground part of wheat treated with strain G3-29 was 4.51±0.87 g, which was significantly increased (p≤0.05) compared with 3.19±0.86 g of the untreated control plants (as shown in Figure 2B). Figure 5 The tiller number of wheat plants treated with G3-29 was 4.06±1.03, which was significantly greater than that of the untreated control plants (2.8±0.86 (p≤0.05)). Figure 5 The average root length of the plants treated with strain G3-29 was 25.01±3.29 cm, which was significantly increased (p≤0.05) compared with 21.18±3.34 cm of the untreated control plants (as shown in D). Figure 5The average plant height of strain G3-29 treated plants was 38.56±2.95 cm, while that of untreated plants was 36.91±2.99 cm, with no significant difference (as shown in Figure E). Figure 5 The experiment was repeated three times, and the results were consistent, with significant differences between the treatment group and the control group. t test, n=20.

[0084] Example 5 Functional determination of strain G3-29 in controlling wheat scab

[0085] The spore liquid prepared in Example 3 was used for treatment, specifically as follows:

[0086] Inoculation: G3-29 spore suspension (1×10 6 Spore ml -1 ) was inoculated on individual wheat spikelets, and the wheat ears were covered with plastic bags for the first two days to maintain humidity. Fifteen days after inoculation, the number of diseased spikelets on each inoculated wheat ear was counted, recorded, and photographed.

[0087] Spray inoculation: Use a handheld atomizer to spray approximately 5 mL of G3-29 spore suspension (1×10 6 Spore ml -1 ) was sprayed on different wheat ears until a large number of droplets were deposited on the wheat ears. After 2 or 4 days of application, the treated wheat ears were inoculated with a conidia suspension of Fusarium graminearum PH-1 (1×10 6 Spore ml -1 )10μL. Sterile distilled water was used instead of G3-29 spore suspension or culture filtrate as control. Wheat ears were covered with plastic bags for the first two days to maintain humidity. 15 days after inoculation, the number of diseased spikelets on each inoculated wheat ear was counted, recorded and photographed.

[0088] like Figure 6 As shown in Table 6, the control effect of inoculating the spore suspension of strain G3-29 2 days in advance on wheat fusarium head blight was significant, at 84.6% (p ≤ 0.05). In addition, the control effect of spraying the spore suspension of strain G3-29 4 days in advance was 77.2% (p ≤ 0.05), see Table 6.

[0089] Table 6 Statistics on the effect of spraying strain G3-29 on the number of diseased wheat spikelets

[0090]

[0091] Note: Different letters indicate significant differences at the p≤0.05 level, and the same letters indicate insignificant differences; “-” indicates no effect.

[0092] Example 6 Functional determination of strain G3-29 in controlling leaf spot of Elymus salsa

[0093] The spore solution prepared in Example 3 was used to treat Elymus dahliae, specifically as follows:

[0094] Spray inoculation: Use a handheld atomizer to spray approximately 5 mL of G3-29 spore suspension (1×10 7 Spore ml -1 ) was sprayed on the leaves of different Elymus dahliae until a large number of droplets were deposited on the leaves of Elymus dahliae. Two days after application, the treated leaves of Elymus dahliae were sprayed with a suspension of conidia of Helicoverpa spp. (1×10 4 Spore ml -1 ). Sterile distilled water was used instead of G3-29 spore suspension as control. Black plastic bags were used to cover the leaves of Elymus dahliae for the first two days to maintain humidity and dark conditions. 15 days after inoculation, the number and area of ​​diseased leaf spots on each inoculated Elymus dahliae leaf were counted, recorded and photographed.

[0095] The results are as follows Figure 7 As shown in A, spraying the spore suspension of strain G3-29 2 days in advance can significantly reduce the number of diseased leaves and leaf spot area of ​​Elymus dahliae (p≤0.05), with a prevention efficiency of 65.65%.

[0096] Example 7 Functional determination of strain G3-29 in controlling Elymus leaf spot disease

[0097] The spore solution prepared in Example 3 was used to treat Elymus dahliae, specifically as follows:

[0098] Spray inoculation: Use a handheld atomizer to spray approximately 5 mL of G3-29 spore suspension (1×10 7 Spore ml -1 ) was sprayed on different leaves of Elymus dahliae until a large number of droplets were deposited on the leaves of Elymus dahliae. Two days after application, the treated leaves of Elymus dahliae were sprayed with a suspension of conidia of Helicoverpa maydis (1×10 4 Spore ml -1 ). Sterile distilled water was used instead of G3-29 spore suspension as control. Black plastic bags were used to cover the leaves of Elymus dahliae for the first two days to maintain humidity and dark conditions. 15 days after inoculation, the number and area of ​​diseased leaf spots on each inoculated Elymus dahliae leaf were counted, recorded and photographed.

[0099] The results are as follows Figure 7 As shown in B, spraying the spore suspension of strain G3-29 2 days in advance can significantly reduce the number of diseased leaves and leaf spot area of ​​Elymus dahliae (p≤0.05), with a prevention efficiency of 64.61%.

[0100] Example 8 Functional determination of strain G3-29 in controlling corn leaf spot

[0101] The spore liquid prepared in Example 3 was used to treat corn, specifically as follows:

[0102] Spray inoculation: Use a handheld atomizer to spray approximately 5 mL of G3-29 spore suspension (1×10 7 Spore ml -1 ) was sprayed on different corn leaves until a large number of droplets were deposited on the corn leaves. Two days after application, the treated corn leaves were sprayed with a suspension of conidia of Helicoverpa maydis (1×10 4 Spore ml -1 ). Sterile distilled water was used instead of G3-29 spore suspension as a control. The corn leaves were covered with black plastic bags for the first two days to maintain humidity and dark conditions. 15 days after inoculation, the number and area of ​​leaf spots on each inoculated corn leaf were counted, recorded and photographed.

[0103] The results are as follows Figure 8 As shown, spraying the spore suspension of strain G3-29 2 days in advance can significantly reduce the number of diseased leaves and leaf spot area of ​​corn (p≤0.05), with a prevention efficiency of 67.80%.

[0104] The embodiments described are only descriptions of the preferred modes of the present invention and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A shared strain of Fusarium ( Fusarium commune ), characterized in that, The deposit number of the shared Fusarium is CGMCC No.41090.

2. A biocontrol agent, characterized in that: Comprising the shared Fusarium spores as described in claim 1.

3. Use of the shared Fusarium according to claim 1 in any of the following: (1) Application in the prevention and control of grass plant diseases; (2) Application in the preparation of microbial agents for preventing and controlling diseases of gramineous plants; in, The gramineous plant diseases include wheat fusarium head blight, lycopersicon esculentum leaf spot and corn leaf spot.

4. Use of the shared Fusarium according to claim 1 in any of the following: (1) Application in inhibiting pathogens of grass plant diseases; (2) Application in the preparation of microbial agents for inhibiting pathogenic bacteria of grass plant diseases; in, The grass plant disease pathogens include Fusarium graminearum ( Fusarium graminearum ), Pseudomonas graminearum ( Fusarium pseudograminearum )、Anthrax ( Colletotrichum gloeosporioides ), Sclerotinia sclerotiorum ( Sclerotinia sclerotiorum )、Helicoverpa spp. Bipolaris sorokiniana ) and Helminthosporium maydis ( Bipolaris maydis ).

5. The use of the microbial agent as claimed in claim 2 in preventing and controlling diseases of Gramineae plants, characterized in that: The gramineous plant diseases include wheat fusarium head blight, lycopersicon esculentum leaf spot and corn leaf spot.

6. Use of the microbial agent according to claim 2 in inhibiting pathogenic bacteria of gramineous plant diseases, characterized in that: The pathogenic bacteria of grass plant diseases include Fusarium graminearum, Pseudo-Fusarium graminearum, Anthracnose, Sclerotinia sclerotiorum, Helminthosporium tataricum and Helminthosporium maydis.

7. Use of the shared Fusarium according to claim 1 in any of the following: (1) Application in promoting wheat growth; (2) Application in the preparation of bacterial agents for promoting wheat growth; The wheat growth promotion includes promoting the increase of root fresh weight, root length, above-ground fresh weight and plant tillering number of wheat.

8. A wheat growth promoter, characterized in that Including the shared Fusarium as described in claim 1.

9. A method for preventing and controlling diseases of grass plants, characterized in that: The method comprises applying the shared Fusarium of claim 1 or the biocontrol agent of claim 2 to gramineous plants; the diseases of the gramineous plants include wheat fusarium rust, lycopersicon erythrorhizium leaf spot and corn leaf spot.

10. A method for promoting wheat growth, characterized in that: The method comprises applying the shared Fusarium described in claim 1 or the wheat growth promoter described in claim 8 to Gramineae plants; the wheat growth promotion comprises promoting the increase of the root fresh weight, root length, above-ground fresh weight and plant tiller number of wheat.

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