CN108624514A - One plant of m5-2 Pei Lijin Bipolaris Bipolaris peregianensis and its application - Google Patents

Abstract

The invention relates to an m5-2 strain of Bipolaris peregianensis and its application. The strain was deposited in the Guangdong Microbial Culture Collection Center on July 11, 2016, with a preservation number of GDMCC.NO:60057. The m5‑2 Bipolaris peregianensis obtained from bermudagrass disease strains provided by the present invention can effectively control grass weeds, and the herbicide made from it is environmentally friendly and can inhibit the drug resistance and resistance of weeds. The development of medicinal properties is conducive to the promotion of green food and organic agriculture, and it has low cost, no pollution, and is safe for crops.

Description

A strain of m5-2 Bipolaris peregianensis and its application

technical field

The invention belongs to the field of biological control, and in particular relates to an m5-2 strain of Bipolaris peregianensis and its application.

Background technique

Chemical herbicides have developed rapidly in my country because of their rapid action and ease of use. However, with the widespread use of chemical agents, the huge environmental pollution and pesticide residue problems have seriously affected the sustainable development of agricultural ecology. Therefore, the development and utilization of biopesticides play an important role in environmental protection agriculture and integrated weed management (IPM). Microbial herbicides are a class of herbicides developed from living microorganisms and their metabolites. Because of their low environmental hazards, they have become an important research direction in the field of herbicides. Recent studies have found that living phytopathogenic fungi and their secondary metabolites have herbicidal activity. The development of microbial herbicides using weed pathogenic fungi and their secondary metabolites generally has the characteristics of diverse action sites and modes of action. Weeds are not easy to develop resistance, low development costs, and harmless to the environment, showing good development. Application prospects.

Bermudagrass ( Cynodon dactylon (Linn.) Pers.), also known as Bermuda grass, is widely distributed in various parts of the world. It is a perennial herb and is a good embankment and soil protection plant. It is often used to pave lawns or courts; but when it grows in orchards or farmland, it is a harmful weed that is difficult to get rid of. Bermudagrass has strong reproductive ability, fast growth speed, long growth period and strong adaptability, and can grow in various soil environments. Bermudagrass in farmland often competes with crops for water and nutrients, which seriously affects crop growth, quality and yield. In severe cases, it may even form a dominant population and densely cover the field, resulting in severe crop yield reduction.

At present, the methods of controlling bermudagrass weeds mainly rely on artificial or chemical methods. Manual weeding is labor-intensive and time-consuming, while chemical weeding causes environmental pollution and leads to the generation of weed population resistance. Therefore, we are looking for methods other than the above two methods. The Bermudagras approach has become increasingly urgent. At present, the research and development of biological herbicides has been paid more and more attention, and the use of microorganisms to control weed damage is becoming an effective new way and has a huge market prospect.

Contents of the invention

The object of the present invention is to overcome the defects of the prior art that the prevention and control of bermudagrass weeds is labor-intensive and time-consuming, and it is easy to cause environmental pollution and lead to the generation of weed population resistance, and to provide a m5-2 Bipolaris peregianensis . The m5-2 Bipolaris peregianensis provided by the invention can effectively control grass weeds.

Another object of the present invention is to provide the application of the m5-2 Bipolaris peregianensis in weed control.

Another object of the present invention is to provide a herbicide.

In order to realize the above-mentioned purpose of the invention, the present invention adopts following technical scheme:

A m5-2 strain of Bipolaris peregianensis , which was deposited in the Guangdong Provincial Microbial Culture Collection Center on July 11, 2016, with the preservation number: GDMCC.NO:60057.

The new strain m5-2 ( Bipolaris peregianensis ) of the present invention has been deposited in the Guangdong Microbial Culture Collection Center on July 11, 2016, with the preservation number: GDMCC.NO: 60057. The strain was identified by morphology and molecular biology as Umbilocorium pelikini , belonging to the genus Cochliobolus Drechsler [asexual stage: Bipolaris Shoemarker], belonging to the family Cochliobolus Drechsler ( Pleosporaceae Nitschke ), Pleosporales LuttrellBarr , Pleosporomycetidae , Dothideoetes , Ascomycota , Fungi .

The inventor obtained and identified the strain in the following ways, investigated grass diseases in many areas of my country, found bermudagrass disease, recorded the disease symptoms in detail and photographed the field damage symptoms, and collected the diseased weed leaves and stems , taken back to the laboratory for microscopic examination.

The isolation and purification process of pathogenic bacteria is as follows:

Brown to dark purple oval and fusiform spots appear on the infected leaves of bermudagrass. As the spots expand, the leaf sheaths, stems, and roots are also infected. Under humid conditions, black moldy matter is formed on the surface of the leaves, which are conidiophores. When the disease is severe, most plants wither and die, and dark brown withered grass spots are formed. After washing the leaves of typical diseased weeds in lesion spots, a small piece of tissue of 3 mm × 3 mm was cut from the disease-healthy junction of the diseased site. After the sample was cut, it was placed in 2.5% sodium hypochlorite solution for disinfection for 1 min. Then wash with sterile water for 3 times, transfer to potato dextrose agar [PDA, add gentamicin (1mL/L) to inhibit bacterial growth] medium, and incubate at 25°C. Pathogens were purified and stored on PDA slant medium, and stored in a 4°C refrigerator for later use.

Preferably, the rDNA-ITS sequence of the strain is shown in SEQ ID NO:1.

Preferably, the bacterial strain is 70 mm in diameter after being cultivated on the PDA medium for 5 days, black, felt-like, and the back of the bacterial colony is taupe; Spore peduncles are solitary, a few clustered, cylindrical or knee-shaped, brown, 94.5-147 μm long, 4-9 μm wide; conidia are curved, spindle-shaped, broadly elliptic, dark brown, with 6 ~10 pseudosepta, size 58.5~84.5×13.5~18.5 μm.

The morphological identification of the pathogen was as follows:

The isolated fungi were purified and cultured for single spores, and the strains were inoculated on PDA medium, and after 5 days of constant temperature cultivation at 25 °C, the morphology of each colony was observed and recorded. Continue to observe the production of conidia of pathogenic bacteria colonies, observe their morphology and sporulation structure characteristics under an optical microscope, measure the size of 50 pathogenic bacteria spores and sporophymes, and refer to relevant monographs for identification. After growing on PDA medium at 25°C for 5 days, the cultured colony reached 70 mm in diameter, black, felt-like, and grayish brown on the back of the colony. The colony is not easy to sporulate on PDA medium, mycelium is yellowish brown, branched, with many septa, conidiophores are solitary, a few clusters, cylindrical or knee-shaped, brown, up to 94.5-147 μm, wide 4~9 μm. Conidia are curved, spindle-shaped, broadly elliptic, dark brown, with 6-10 pseudosepta, most of which are 9, and the size is 58.5-84.5×13.5-18.5 μm. Observation of morphological and cultural traits, and literature review [Manamgoda, DS, Rossman, AY, Castlebury, LA, Crous, PW, Madrid, H., Chukeatirote, E., & Hyde, KD (2014). The genus bipolaris . Studies in mycology, 79 , 221-288.]. The pathogen was preliminarily identified as Bipolaris peregianensis .

The molecular bioassay of the pathogen was as follows:

The tested strains were transferred to fresh PDA medium and cultured for 10 days, the hyphae were scraped off with a sterilized inoculation needle, and the genome of the pathogenic bacteria was extracted according to the instructions of the DNA secure new plant genome DNA extraction kit (Takara, Dalian). Universal primers ITS-1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS-4 (5'-TCCTCCGCTTATTGATATGC-3') (synthesized by Shanghai Sangon Biological Co., Ltd.) were designed by White et al. The pathogenic bacteria genome was used as a template for PCR amplification, and the expected product was about 580 bp. PCR amplification system: 2×Taq PCR MasterMix 25 μL (Takara, Dalian), 20 μmol/L primers 1 μL each, template DNA 1 μL, add sterilized double distilled water to 50 μL. PCR reaction conditions: 94°C preheating for 4 minutes; 94°C for 40 s, 62°C for 45 s, 72°C for 1 min, a total of 32 cycles; 72°C extension for 10 min. All PCR products were added to 0.1% agarose gel for electrophoresis, stained with ethidium bromide, and detected on a gel imager, and the PCR products were sequenced by Shanghai Sangon Co., Ltd. (Sangon, Shanghai). The ITS sequence of the isolated pathogenic bacteria was compared with the Gen-Bank nucleic acid database, and the pathogenic bacteria were identified according to the morphological characteristics, cultural characteristics and pathogenicity of the pathogen, combined with sequence analysis. And MEGA5.0 was used for phylogenetic analysis and phylogenetic tree construction, and the Neighbor-joining method was used to construct the phylogenetic tree, and the number of self-extension was 1000 times. After amplifying the rDNA-ITS sequence of the pathogenic bacterial strain m5-2, the PCR products were purified and sequenced, and the rDNA-ITS sequence fragments of about 580 bp were obtained by sequencing, which were consistent with the expected fragment size. Using the BLAST software in the NCBI website, the homology comparison of the sequences of the tested strains was carried out: the homology of the pathogenic bacteria of the strains and Helminthes umbilicali reached 99%. The result of sequence alignment was used to assist in proving that the bermudagrass pathogen was B. peregianensis . From the constructed phylogenetic tree, it can be seen that Pelikinpinus umbilicalis isolated from Bermudagrass and the existing B. peregianensis in the NCBI database are clustered in the same taxon, while Bipolaris cannot be clustered in one branch, thus proving that The strain of the present invention is Pelikinping umbilicalsporium.

The present invention also claims the application of the above-mentioned m5-2 Bipolaris peregianensis in weed control.

Preferably, the weeds are gramineous weeds.

More preferably, the grass weed is bermudagrass, goosegrass, crabgrass or bony grass.

The m5-2 Bipolaris peregianensis can be applied to farmland and gardens in non-grass planting areas, and is preferably used for preventing and controlling gramineous weeds in dicotyledons, orchards and garden environments.

A kind of herbicide, said herbicide comprises the mixture of crude toxin, living cell hyphae and conidia of the above bacterial strain m5-2 Bipolaris peregianensis .

The crude toxin referred to in the present invention is the secondary metabolite of the strain.

The herbicide provided by the invention is friendly to the environment, can inhibit the drug resistance and drug resistance development of weeds, is beneficial to the popularization of green food and organic agriculture, and is low in cost, pollution-free and safe to crops.

The mixture of crude toxin, living cell hyphae and conidia in the herbicide can be prepared by the following method: take the edge of the pathogenic bacteria colony of the bacterial strain m5-2 Bipolaris peregianensis after culturing for 5 days Five pieces of bacteria cake were inoculated in potato flour agar liquid medium, cultured in a shaker at 25 °C for 10 days, and then filtered to obtain the crude toxin extract of the strain and the mixture of live cell hyphae and conidia.

The herbicide can be processed into various dosage forms, such as liquid, emulsion, suspension, powder, granule, wettable powder, water dispersible granule, etc., preferably processed into a dosage form suitable for spraying application. The carrier used in the above dosage form is a pesticide acceptable carrier, preferably a carrier commonly used in the field of pesticide and biologically inert, and may be in solid or liquid form. Examples of solid carriers include minerals such as clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica and diatomaceous earth; plant materials such as corn flour, soybean flour or starch; high molecular compounds such as polyvinyl alcohol, polyglycol. Examples of liquid carriers include various organic solvents such as decane and dodecane, vegetable oil, mineral oil and water.

According to needs, the herbicide of the present invention can use surfactants, adhesives, stabilizers and other co-surfactants, such as Tween 20, Tween 80, etc., and stabilizers can use antioxidants or pH regulators. The number of conidia in the herbicides provided by the present invention varies with the form of the formulation and the method of application.

Preferably, when the herbicide is powder, granule, wettable powder or water-dispersible granule (i.e. solid preparation), the number of conidia in the herbicide is 1×10 ⁵ to 1×10 ⁸ Spores/g.

More preferably, the number of conidia in the herbicide is 1×10 ⁶ -1×10 ⁷ spores/g.

Preferably, when the herbicide is liquid, emulsion or suspension, the content of conidia in the spray liquid of the herbicide is 1×10 ⁴ -1×10 ⁶ spores/mL.

More preferably, the content of conidia in the spray liquid of the herbicide is 1×10 ⁵ -1×10 ⁶ spores/mL.

The herbicide of the present invention can be used in conjunction with other suitable chemical herbicides, thereby reducing the consumption of chemical herbicides and reducing environmental pollution.

Compared with the prior art, the present invention has the following beneficial effects:

The m5-2 Bipolaris peregianensis obtained from the bermudagrass disease strain provided by the present invention can effectively prevent and control grass weeds, and the herbicide made from it is environmentally friendly and can inhibit the drug resistance and resistance of weeds. The development of medicinal properties is conducive to the promotion of green food and organic agriculture, and it has low cost, no pollution, and is safe for crops.

Description of drawings

Figure 1 is a picture of conidia of m5-2 Bipolaris peregianensis .

Detailed ways

The present invention is further set forth below in conjunction with embodiment. These examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following example embodiment, usually according to the conventional conditions in this field or according to the conditions suggested by the manufacturer; used raw materials, reagents, etc., if no special instructions, are available from commercial channels such as conventional markets Raw materials and reagents obtained. Any insubstantial changes and substitutions made by those skilled in the art on the basis of the present invention fall within the scope of the present invention.

Example 1 Isolation, purification and identification of pathogenic bacteria

(1) Isolation and purification of pathogenic bacteria

Brown to dark purple oval and fusiform spots appear on the infected leaves of bermudagrass. As the spots expand, the leaf sheaths, stems, and roots are also infected. Under humid conditions, black moldy matter is formed on the surface of the leaves, which are conidiophores. When the disease is severe, most plants wither and die, and dark brown withered grass spots are formed. After washing the leaves of typical diseased weeds in lesion spots, a small piece of tissue of 3 mm × 3 mm was cut from the disease-healthy junction of the diseased site. After the sample was cut, it was placed in 2.5% sodium hypochlorite solution for disinfection for 1 min. Then wash with sterile water for 3 times, transfer to potato dextrose agar [PDA, add gentamicin (1ml/L) to inhibit bacterial growth] medium, and culture at 25°C. Pathogens were purified and stored on PDA slant medium, and stored in a 4°C refrigerator for later use.

(2) Morphological identification of pathogenic bacteria

The isolated fungi were purified and cultured for single spores, and the strains were inoculated on PDA medium, and after 5 days of constant temperature cultivation at 25 °C, the morphology of each colony was observed and recorded. Continue to observe the production of conidia of pathogenic bacteria colonies, observe their morphology and sporulation structure characteristics under an optical microscope, measure the size of 50 pathogenic bacteria spores and sporophymes, and refer to relevant monographs for identification. After growing on PDA medium at 25°C for 5 days, the cultured colony reached 70 mm in diameter, black, felt-like, and grayish brown on the back of the colony. The colony is not easy to sporulate on PDA medium, mycelium is yellowish brown, branched, with many septa, conidiophores are solitary, a few clusters, cylindrical or knee-shaped, brown, up to 94.5-147 μm, wide 4~9 μm. Conidia (as shown in Figure 1) are curved, spindle-shaped, broadly elliptical, dark brown, with 6-10 pseudosepta, most of which are 9, and the size is 58.5-84.5×13.5-18.5 μm. Through observation of morphological and cultural traits, and literature review [Manamgoda, DS, Rossman, AY, Castlebury, LA, Crous, PW, Madrid, H., Chukeatirote, E., & Hyde, KD (2014). Thegenus bipolaris . Studies in mycology, 79 , 221-288.]. The pathogen was preliminarily identified as Bipolaris peregianensis .

(3) Molecular biological identification of pathogenic bacteria

The tested strains were transferred to fresh PDA medium and cultured for 10 days, the hyphae were scraped off with a sterilized inoculation needle, and the genome of the pathogenic bacteria was extracted according to the instructions of the DNA secure new plant genome DNA extraction kit (Takara, Dalian). Universal primers ITS-1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS-4 (5'-TCCTCCGCTTATTGATATGC-3') (synthesized by Shanghai Sangon Biological Co., Ltd.) The root pathogenic bacteria genome was used as a template for PCR amplification, and the expected product was about 580 bp. PCR amplification system: 2×Taq PCR MasterMix 25 μL (Takara, Dalian), 20 μmol/L primers 1 μL each, template DNA 1 μL, add sterilized double distilled water to 50 μL. PCR reaction conditions: 94°C preheating for 4 minutes; 94°C for 40 seconds, 62°C for 45 seconds, 72°C for 1 minute, a total of 32 cycles; 72°C extension for 10 minutes. All the PCR products were added to 0.1% agarose gel for electrophoresis, stained with ethidium bromide, and detected by a gel imager, and the PCR products were sequenced by Shanghai Sangon Co., Ltd. (Sangon, Shanghai). The ITS sequence of the isolated pathogenic bacteria was compared with the Gen-Bank nucleic acid database, and the pathogenic bacteria were identified according to the morphological characteristics, cultural characteristics and pathogenicity of the pathogen, combined with sequence analysis. And MEGA5.0 was used for phylogenetic analysis and phylogenetic tree construction, and the Neighbor-joining method was used to construct the phylogenetic tree, and the number of self-extension was 1000 times. After the rDNA-ITS sequence of the pathogenic strain M5-2 was amplified, the PCR product was purified and then sequenced. The rDNA-ITS sequence fragments of about 580 bp were obtained by sequencing, which were consistent with the expected fragment size. Using the BLAST software in the NCBI website, the homology comparison of the sequences of the tested strains was carried out: the homology of the pathogenic bacteria of the strains and Helminthes umbilicali reached 99%. The result of sequence alignment was used to assist in proving that the bermudagrass pathogen was B. peregianensis . From the constructed phylogenetic tree, it can be seen that Pelikinpinus umbilicalis isolated from Bermudagrass and the existing B. peregianensis in the NCBI database are clustered in the same taxon, while Bipolaris cannot be clustered in one branch, thus proving that The strain of the present invention is Pelikinping umbilicalsporium. .

Specifically, the rDNA-ITS sequence is as follows:

TCCGTAGGTGAACCTGCGGAGGGATCATTACACAACAAAATATGAAGGCCTGGCTTCGCGGCCGGCTGAAATATTTTTTTTCACCCGTGTCTTTTGCGCACTTGTTGTTTCCTGGGCGGGCTCGCCCGCCACCAGGACCAAACCATAAACCTTTTTCTTTTGCAGTTTTCATCAGCGTCAGTAAAAAACAATGTAATTATTACAACTTTCAACAACGGATCTCTTGGTTCTGGCATCGATGAAGAACGCAGCGAAATGCGATACGTAGTGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTTTGGTATTCCAAAGGGCATGCCTGTTCGAGCGTCATTTGTACCTTCAAGCTTTGCTTGGTGTTGGGCGTTTTTTGTCTCTCCTTGCGGGAGACTCGCCTTAAAACGATTGGCAGCCGGCCTACTGGTTTCGGAGCGCAGCACATTTTTTGCGCTTTGTATCAGGAGAAAAGGACGGTACTCCATCAAGACGTTTACATTTTTAACTTTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCAATAAGCGGAGGA。

Example 2 Bipolaris peregianensis Bipolaris peregianensis Bipolaris peregianensis m5-2

The pathogenicity of the m5-2 strain to common weeds and crop seedlings was determined by artificial inoculation to understand its host range.

The seeds of the tested plants (see Table 1 for the list) were sown in plastic flower pots with a diameter of 10 cm. Nine pots were sown for each plant, and they were cultivated at 25°C with a photoperiod of 12 h. Inoculate when the test plants reach 2-4 true leaves, and keep 5-10 plants per pot according to the plant size of the test plants.

Two inoculation methods were adopted: (1) Inoculation by spore suspension spraying method: after the bacterial strain m5-2 was cultured in PDA medium for 10 days, the spores were scraped with a sterile cover glass and prepared into ¹ ×106 spores/ml of aqueous solution. The mixed spore suspension was evenly sprayed on the test plants with a micro-sprayer, and each plant was inoculated in 3 pots. (2) Spray inoculation of secondary metabolites: the bacterial strain m5-2 was inoculated at the edge of the pathogenic bacteria colony after 5 days of cultivation, and 5 pieces of bacterial cakes were inoculated in the potato flour agar liquid medium, and cultured in a shaker at 25 °C After 10 days, it was filtered to obtain the crude toxin of the strain, and the mixed spore suspension was evenly sprayed on the test plants with a micro-sprayer, and each plant was inoculated in 3 pots. The other 3 pots were sprayed with sterile water as a control. The inoculated plants were placed in an airtight container at room temperature for 24 hours, and then moved to an artificial climate box for cultivation. The light/dark alternated for 12 hours every day, the temperature was 28°C/25°C (light/dark), and the relative humidity was 90%. One week after the inoculation, the incidence was recorded according to the following criteria. Disease grading standard: Grade 0 means no plant disease; Grade 1: 1%~10% of diseased leaves; Grade 2: 10%~30% of diseased leaves; Grade 3: 30%~50% of diseased leaves; Grade 4: Diseased leaves are 50%~70%; Grade 5: diseased leaves are 70%~100%.

Pathogenicity assay results

The herbicidal activity of 9 kinds of weeds and 5 kinds of crops was tested with the spores and crude toxin of the strain, and it was found that the conidia and crude toxin of the strain had certain lethal activity on the tested weeds. Among them, the lethality rate of mushroom cake to grass weeds such as bony grass, crabgrass, bermudagrass, and goosegrass reached level 4 or above, and the lethality rate of crude toxin to several kinds of weeds reached level 5. The pathogenicity to centella asiatica, wood sorrel, and hollow lotus is strong, reaching grade 2-3, but the lethality to a little red is weak, and the pathogenicity of fungus cake and crude toxin are both grade 1. It has weak pathogenicity to rice and corn, and its curative intensity is 1-2, but it has no pathogenicity to soybean, watermelon and radish. The control leaves had no disease symptoms.

Table 1 Pathogenicity of conidia and toxins of strain m5-2 to different plants

Test plants conidia crude toxin control Bone Grass 4 5 0 Centella 3 2 0 Sorrel 3 3 0 crabgrass 4 5 0 a little red 1 1 0 Bermudagrass 5 5 0 Hollow lotus 2 3 0 Ghost needles 2 1 0 goosegrass 5 5 0 rice 2 1 0 corn 1 2 0 soybean 0 0 0 watermelon 0 0 0 radish 0 0 00

Note: Disease grading standard: Disease grading standard: Grade 0 means no plant disease; Grade 1: 1%~10% of diseased leaves; Grade 2: 10%~30% of diseased leaves; Grade 3: 30%~ 50%; Grade 4: 50%~70% of diseased leaves; Grade 5: 70%~100% of diseased leaves.

sequence listing

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<120> A strain of m5-2 Bipolaris peregianensis and its application

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Claims (10)

1. one plant of m5-2 Pei Lijin BipolarisBipolaris peregianensis, which is characterized in that the bacterial strain in On July 11st, 2016, it is preserved in Guangdong Province's Culture Collection, preserving number：GDMCC.NO:60057.

2. m5-2 Pei Lijin Bipolaris according to claim 1Bipolaris peregianensis, which is characterized in that The bacterial strain rDNA-ITS sequences such as SEQ ID NO：Shown in 1.

3. m5-2 Pei Lijin Bipolaris according to claim 1Bipolaris peregianensis, which is characterized in that The bacterial strain is that the colony diameter that is formed is 70 mm after cultivating 5d in PDA culture medium, black, felted, and bacterium colony back side ash is brown Color；The mycelia yellowish-brown of the bacterial strain has branch, more diaphragms, conidiophore Dan Sheng, minority collection life, cylindrical shape or shape of going down on one's knees, Brown, it is 4 ~ 9 μm wide up to 94.5 ~ 147 μm；Conidium is bent, spindle, wide ellipse, crineous, has 6 ~ 10 vacations Diaphragm, 58.5 ~ 84.5 × 13.5 ~ 18.5 μm of size.

4. Pei Lijin Bipolaris m5-2 Pei Lijin Bipolaris described in claim 1Bipolaris peregianensis Application in cutting weeds.

5. applying according to claim 4, which is characterized in that the weeds are grassy weed.

6. applying according to claim 5, which is characterized in that the grassy weed is Bermuda grass, eleusine indica, lady's-grass or hard Bone grass.

7. a kind of herbicide, which is characterized in that the herbicide includes any m5-2 Pei Lijinping navels of claim 1 ~ 3 Compacted sporeBipolaris peregianensisRaw toxin, active somatic cell mycelia and conidial mixture.

8. herbicide according to claim 7, which is characterized in that the dosage form of the weeding is liquor, emulsion, suspension Agent, pulvis, granule, wettable powder or water dispersible granules.

9. herbicide according to claim 8, which is characterized in that when the herbicide is pulvis, granule, wettable powder Or when water dispersible granules, conidial quantity is 1 × 10 in the herbicide formulations⁵~1×10⁸A spore/gram.

10. herbicide according to claim 8, which is characterized in that when the herbicide is liquor, emulsion or suspending agent, Conidial content is 1 × 10 in the flushing liquor of the herbicidal formulations⁴~1×10⁶ A spore/mL.