CN111363685B - Fusarium verticillatum and application thereof - Google Patents
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Abstract
The invention discloses fusarium verticillium and application thereof. The Fusarium verticillium FCPC-L01 strain (Fusarium concentricum FCPC-L01) has the preservation number of: GDMCC No: 60971, respectively; the optimal liquid fermentation conditions are as follows: the temperature of the fermentation culture of the sucrose potato culture solution PS is 25-28 ℃, the initial pH is 4-6, and the rotating speed of a shaking table is 180-210 rpm; and transferring the fermentation liquor to rice for solid fermentation, wherein the produced conidia has stronger toxicity to the larvae of the cabbage moth. Compared with the chemical prevention method aiming at the tip roller moths at the present stage, the biological prevention strain and the fermentation method for preventing and treating the major pest of the fir wood, namely the tip roller moths, are more environment-friendly, nontoxic and safe, and have important market application and popularization values.
Description
Technical Field
The invention belongs to the technical field of prevention and treatment of forestry and agricultural pests by utilizing entomogenous fungi, and particularly relates to fusarium verticillium and application thereof.
Background
China fir is a special species in China, has the advantages of fast growth, easy survival and good material property, and is a main fast-growing wood species in southern provinces. The China fir seed garden is a base for providing excellent seed sources. According to statistics, the area of the improved variety base of the Chinese fir in 1987 exceeds 4000 mu, and the method plays a positive promoting role in improved variety, fast growth and high yield of the Chinese fir. The outbreak and damage of fir diseases and pests are one of important factors influencing the seed yield and the seeding quality, so the pest control is one of important contents of the management and the management of the fir seed orchard. The Dendrolimus cunninghamiacola is an insect belonging to the Lepidoptera Torlidae, and recently has developed a disaster in North Guangdong. The insect is distributed in the main fir producing areas of Guangdong, Guangxi, Fujian, Jiangxi, Zhejiang, Guizhou, Sichuan and the like, and young insects are eaten into the top buds of the tender tips of the fir, so that the phenomena of multiple heads, no heads or partial crowns and the like are caused, and the dry shape can be distorted, thereby seriously influencing the growth and the material of the trees. At present, the chemical pesticide is mainly used for preventing and controlling the cabbage moth. However, because the planting area of the fir is large, a large amount of high-toxicity chemical pesticide is used, so that the ecological environment is polluted and destroyed, the safety of forest products is influenced, natural enemies are killed, and pests are easy to generate drug resistance. Along with the increasing social demand of pollution-free agricultural and forestry products and toward beautiful ecological environment, the nation increasingly emphasizes the concept of green, coordinated and sustainable economic development, more attaches importance to the reduction of the use of chemical pesticides and vigorously advocates the development of biological pesticides.
Entomogenous fungi are fungi which can parasitize and kill insects, and play an important role in the aspects of comprehensive pest control, natural enemy protection, ecological balance and the like. Entomogenous fungi invade the body wall of the insect through spores, and then a large amount of hyphae grow to absorb host nutrition and destroy host tissues so as to kill the insect.
Therefore, there is a need to develop biocontrol bacteria and methods for controlling cabbage caterpillar.
Disclosure of Invention
The invention aims to provide a strain of fusarium verticillata which can sustainably and naturally control pests and an application thereof, aiming at the problems that the existing fir wood planting area is enlarged, the harm of the hemsleya fusca is increasingly rampant, the prevention and control of the hemsleya fusca in the prior art are difficult, the manual pesticide application in the chemical pesticide prevention and control process is difficult, the spraying cost of an airplane is high, and the pollution damages the ecological environment.
The invention aims to provide a strain of fusarium verticillium with the preservation number as follows: GDMCC No: 60971.
the excellent strain is separated from a field Chinese fir tip diamond back moth larva body, is determined to be Fusarium verticillarum through indoor separation and morphological and molecular identification, and is named as Fusarium verticillarum FCPC-L01(Fusarium concentricum FCPC-L01) strain.
The mycological characteristics of the Fusarium verticillarum FCPC-L01(Fusarium concentricum FCPC-L01) are as follows:
morphological characteristics: conidia of the strain have two forms of megaspore and microspore, and the microspore has a large quantity, is not separated and is oval. The large conidia are small in number, crescent in shape, slightly bent, and 1-5 partitions, most of which are 3 partitions and about 2 × 17um in size. According to the morphological observation results, the strain is preliminarily judged to belong to fusarium.
Molecular biological characteristics: and (3) identifying by using a TEF-1 alpha gene sequence alignment method. Using primer pair F: 5'-ATGGGTAAGGAGGACAAGAC-3' and R: 5'-GGAAGTACCAGTGATCATGTT-3' A partial sequence fragment of the TEF-1 alpha gene was amplified by PCR from the genomic DNA of the strain. The amplified fragment is sequenced, and the sequencing result shows that the sequence of the fragment is 678bp, and the sequence is shown as SEQ ID NO. 1.
The phylogenetic tree is constructed by three different methods of Mega5, the strain always gathers into one branch with the Fusarium verticillii and has high score, and the strain is determined to be Fusarium verticillium concentricum by combining the morphological observation result and is numbered as FCPC-L01 strain.
The invention also provides a microbial inoculum which takes the fusarium verticillium as an active ingredient.
The invention also provides a fermentation method of the fusarium verticillium, which comprises the following steps:
a. inoculating the fusarium verticillium into a liquid culture medium for fermentation culture to obtain a fermentation liquid;
b. and pouring the fermentation liquor into sterile rice, uniformly stirring, and culturing to produce spores.
Preferably, the liquid culture medium is sucrose potato culture solution PS, and the preparation method comprises the following steps: according to the weight portion, 200 portions of potatoes are taken, washed, peeled and cut up, water is added for boiling for 30min, gauze is used for filtering, 40 portions of cane sugar are taken out of filtrate, and water is used for supplementing to 1000 portions; then sterilized at 121 ℃ for 20 min.
Preferably, the liquid culture medium is culture fluid DY, and the preparation method comprises the following steps: taking 40 parts of glucose and 20 parts of yeast extract by weight, and supplementing water to 1000 parts; then sterilized at 121 ℃ for 20 min.
Preferably, the temperature of the fermentation culture in the step a is 25-28 ℃, the initial pH is 4-6, the rotation speed of a shaking table is 180-210rpm, and the fermentation culture is performed for 3 days to obtain the fermentation broth.
Preferably, the culture sporulation conditions of the step b are 25 +/-1 ℃ and 75% of relative humidity.
The invention also provides application of the fusarium verticillatum in prevention and treatment of forestry pests.
Preferably, the forestry pests are hemsleya amabilis.
The fusarium of the invention belongs to fusarium verticillium, and the liquid-solid fermentation condition of the fusarium is groped through a single-factor experiment. Conidia generated by the strain of the invention has stronger insecticidal toxicity to the tip roller moths. Therefore, compared with the existing chemical prevention and control method for the tip roller moths, the biological prevention and control strain and the fermentation method for the tip roller moths are more environment-friendly, nontoxic and safe, so that the biological prevention and control method has important market application and popularization values.
Preservation description:
fusarium verticillium FCPC-L01(Fusarium concentricum FCPC-L01) of the present invention was deposited in Guangdong province culture Collection of microorganisms (GDMCC) at 09/03/2020, addresses: the preservation number of the Guangzhou city, Jielizhou 100 college No. 59 building 5, Guangdong province microbiological research institute: GDMCC No: 60971.
drawings
FIG. 1 shows larvae of Sequoia xylostella lethal by Fusarium infestation; a: a whole figure; b: a partial enlarged view.
FIG. 2 is a diagram showing the shapes of fusarium hyphae, spore-forming structures and conidia separated from the larva of a muscardine insect; a: big and small spores; b: hyphae; c and d: and (4) spore-forming structures.
FIG. 3 is a molecular phylogenetic tree of strains constructed from the TEF-1. alpha. gene sequence.
FIG. 4 is a comparison of spore yields under different broth fermentation conditions.
FIG. 5 is a comparison of the difference in spore yields under different temperature culture conditions.
FIG. 6 is a graph showing the comparison of the difference in spore yields under different conditions of the shaking table rotation speed.
FIG. 7 is a comparison of the difference in spore yields under different pH culture conditions.
FIG. 8 shows the results of the toxicity of Fusarium verticillatum to the larvae of Trichoplusia sequoyii.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
The Fusarium of the embodiment of the invention is Fusarium verticillium FCPC-L01 strain (Fusarium concentricum FCPC-L01).
Example 1: separation and identification of fusarium of parasitoid spruce budworm moth
The detailed methods for obtaining and identifying fusarium strains are as follows:
s1, obtaining fusarium strains: the dead larva of the tip of the fir which is infected and killed by fusarium is brought back to a laboratory under the natural condition in a small pit forest field fir seed garden in the Yangjiang region of Shaoguan city (figure 1), and the fungus on the body surface of the dead body is separated and cultured in time.
S2, strain laboratory culture and morphological identification: preparing a PDA culture medium: namely 200g of potatoes, are cleaned, peeled, cut up, added with water, boiled for 30min, filtered by gauze, added with 40g of glucose and 18g of agar, fixed to 1L by deionized water, split into 250mL triangular bottles when the materials are hot after being fully dissolved, about 100mL of each bottle, added with a silica gel plug, sealed, sterilized at 121 ℃ for 20min, taken out, cooled, inoculated with the preserved strains, and cultured on a PDA culture medium for 7 days. Transferring part of hypha and spore into 2mL centrifuge tube under aseptic condition, adding 20% -50% glycerol, and sealing. Storing in an ultra-low temperature refrigerator at-80 deg.C, taking out strain before test, activating, inoculating and culturing on slant, and storing in a refrigerator at 4 deg.C for use. And a small amount of hyphae and spores were picked with an inoculating needle and placed on a glass slide, and the morphology of the strain was observed under a microscope by lightly pressing and covering the glass slide (FIG. 2). The result shows that the conidium of the strain has two forms of megaspore and microspore. The number of the small conidia is large, no separation exists, and the small conidia are oval. The large conidia are less in number, crescent in shape, slightly bent, and contain 1-5 partitions, most of which are 3 partitions, and the size of the conidia is about 2 multiplied by 17 um. According to the morphological observation results, the strain is preliminarily judged to belong to fusarium.
S3, strain DNA extraction and molecular identification: the fusarium genome DNA is extracted by adopting a polymeric magnesium M5 Fungal Genomic DNA Kit, and the specific operation steps are as follows:
(1) culturing the inoculated PDA plate in a constant-temperature illumination (14L/10D) incubator at 25 ℃ for 14D, after the hyphae grow over the culture dish, gently scraping the hyphae on the culture dish by using a sterilized scalpel in a super-clean workbench, and placing the culture dish in double-layer filter paper.
(2) Adding liquid nitrogen into fresh hypha tissue of fungus, and grinding.
(3) The ground powder was collected in a centrifuge tube, and 400. mu.l Buffer LP1 and 6. mu.l RNase were added thereto, vortexed for 1 minute, and left at room temperature for 10 minutes to lyse.
(4) Add 130. mu.l Buffer LP2, mix well and vortex for 1 min.
(5) Centrifuge at 12000rpm for 5 minutes and transfer the supernatant to a new centrifuge tube.
(6) 1.5 volumes of Buffer LP3 were added and mixed well.
(7) Adding all the solution and precipitate into adsorption column filled with collecting tube, and transferring for several times if the solution can not be added at one time. Centrifuge at 12000rpm for 1 min, decant supernatant from the collection tube, and replace the adsorption column back into the collection tube.
(8) Mu.l Buffer GW2 was added to the adsorption column, centrifuged at 12000rpm for 1 min, the supernatant in the collection tube was decanted, and the adsorption column was replaced in the collection tube.
(9) And repeating the step 8.
(10) Centrifuge at 12000rpm for 2 minutes and pour off the supernatant from the collection tube. The column was left at room temperature for several minutes to dry thoroughly.
(11) The adsorption column is placed into a new centrifuge tube, 50-100 mul of Buffer GE or sterile water is suspended and dripped into the middle part of the adsorption membrane, the adsorption membrane is placed at room temperature for 2-5 minutes and centrifuged at 12000rpm for 1 minute, and DNA solution is collected. Storage of DNA at-20 ℃
(12) Carrying out PCR amplification on the TEF-1 alpha gene sequence of the strain, wherein the primer pair is F: 5'-ATGGGTAAGGAGGACAAGAC-3' and R: 5'-GGAAGTACCAGTGATCATGTT-3' are provided. The amplified fragment is sequenced, and the sequencing result shows that the sequence of the fragment is 678bp, and the sequence is shown as SEQ ID NO. 1. The sequence was phylogenetically constructed using MEGA5, and the results are shown in FIG. 3, which indicates that the strain belongs to Fusarium verticillium concentricum, and thus the strain was named as Fusarium verticillium FCPC-L01(Fusarium concentricum FCPC-L01) strain.
Example 2: preparation of spore suspension of Fusarium verticillium FCPC-L01 strain
The preparation method of the spore suspension of the fusarium verticillium FCPC-L01 strain comprises the following steps:
taking fusarium verticillium FCPC-L01 strain, inoculating the strain on a PDA plate by using an inoculating loop under the aseptic condition for activation, and irradiating at the temperature of 25 ℃: cultured in an incubator under dark (12L: 12D) conditions for 7D. After hyphae overgrow the whole culture dish, 20mL of sterile water containing Tween-80 with the mass fraction of 0.03% is added, hyphae and spores are scraped lightly by using an inoculating needle, the bacterial liquid is poured into a beaker, the bacterial liquid is filtered by using double-layer medical gauze, and the mixture is stirred for 10min by using a magnetic stirrer, so that conidium suspension is obtained. The spore concentration of the spore suspension was calculated using a hemocytometer and diluted to 1X 10 with sterile water having a mass fraction of 0.05% Tween-806spores/mL, as used in the examples below.
Example 3: screening of optimal formula of liquid culture medium
The fusarium verticillium FCPC-L01 strain prepared in example 2 was fermented in the following five media using the spore suspension as seed liquid, and the spore production capacities of the strain in the different media were compared:
culture solution No.1 (glucose potato culture solution PD): taking 200g of potatoes, cleaning, peeling, chopping, adding water, boiling for 30min, filtering with gauze, taking filtrate, adding 40g of glucose, fixing the volume to 1L with deionized water, subpackaging in 250mL triangular bottles when the filtrate is hot after being fully dissolved, adding 100mL of each bottle, adding a silica gel plug, sealing, naturally adjusting the pH value, sterilizing at 121 ℃ for 20min, taking out, cooling and storing for later use.
Culture solution No. 2 (sucrose potato culture solution PS): taking 200g of potatoes, cleaning, peeling, chopping, adding water, boiling for 30min, filtering by using gauze, taking filtrate, adding 40g of cane sugar, and fixing the volume to 1L by using deionized water; and after the materials are fully dissolved, the materials are split into 250mL triangular bottles while the materials are hot, each bottle is 100mL, a silica gel plug is added for sealing, the pH value is natural, sterilization is carried out at 121 ℃ for 20min, and then the materials are taken out, cooled and stored for later use.
Culture solution No. 3 (saxas culture solution SDY): taking 40g of glucose, 10g of peptone and 20g of yeast extract, and using deionized water to fix the volume to 1L; and after the materials are fully dissolved, the materials are split into 250mL triangular bottles while the materials are hot, each bottle is 100mL, a silica gel plug is added for sealing, the pH value is natural, sterilization is carried out at 121 ℃ for 20min, and then the materials are taken out, cooled and stored for later use.
Culture solution No. 4 (DY): taking 40g of glucose and 20g of yeast extract, and fixing the volume to 1L by using deionized water; and after the materials are fully dissolved, the materials are split into 250mL triangular bottles while the materials are hot, each bottle is 100mL, a silica gel plug is added for sealing, the pH value is natural, sterilization is carried out at 121 ℃ for 20min, and then the materials are taken out, cooled and stored for later use.
Culture solution No. 5 (DP): taking 40g of glucose and 10g of peptone, and fixing the volume to 1L by using deionized water; and after the materials are fully dissolved, the materials are split into 250mL triangular bottles while the materials are hot, each bottle is 100mL, a silica gel plug is added for sealing, the pH value is natural, sterilization is carried out at 121 ℃ for 20min, and then the materials are taken out, cooled and stored for later use.
The sample loading amount in the fermentation process is 100mL/250mL, the inoculation amount is 5mL, the fermentation temperature is set to be 25 ℃, the rotating speed of a shaking table is 150rpm, the initial pH value is the natural pH value of the culture solution, and the fermentation culture is carried out for 3 days. Then 5mL of fermentation broth was withdrawn and diluted 10-fold for spore number determination, each treatment was repeated 4 times. Selecting 5 middle squares by using a blood counting plate counting method, wherein 16 cells are arranged in each middle square, namely the number N of spores in 80 cells is a biological repetition, and the corresponding calculation formula of the spore concentration is as follows: spore concentration N/80 × 400 × 10000 × dilution factor.
The results show that different culture medium formulas have significant influence on the sporulation quantity of the strain F4,15＝184.48,p<0.001 (FIG. 4), wherein the number of spores in culture solution No. 2 (sucrose potato culture solution PS) was the highest, the number of spores reached 221.75. + -. 16.15 (mean. + -. standard deviation), and the corresponding spore concentration was (1.11. + -. 0.08). times.108spore/mL, and number 4 spore yield in turn>Culture solution No.1 PD>No. 3 culture solution SDY>Culture solution No. 5 DP.
Example 4: influence of fermentation culture temperature on spore yield of Fusarium moniliforme strain FCPC-L01
The seed solution was fermented with the spore suspension of the strain Fusarium verticillarum FCPC-L01 prepared in example 2. The fermentation culture solution contains sucrose potato culture solution PS. The sample loading amount is 100mL/250mL, and the inoculation amount is 5 mL. During the shake culture process, the principle of single factor variable is followed. The initial pH was the natural pH of the broth and the shaker speed was 150 rpm. The temperature conditions are set to be 22 ℃, 25 ℃, 28 ℃ and 31 ℃, the spore production amount of the strain under different culture temperatures is measured, the strain is cultured for 3d in a shaking table, each temperature treatment is repeated for 4 times, and the optimal fermentation culture temperature of the fusarium verticillium FCPC-L01 is obtained according to the spore production amount.
The results show that the spore yield difference of the strains is obvious under the fermentation conditions of different temperatures (F)3,153.47, p 0.051, fig. 5). The spore yield is highest at 25 deg.C and 28 deg.C, the spore number is 101.5 + -25.83 and 100.25 + -15.99, and the corresponding spore concentration is (5.08 + -1.29) × 107spore/mL and (5.01. + -. 0.79). times.107spores/mL. The sporulation yield is lower under the conditions of 22 ℃ and 31 ℃, and is respectively (3.47 +/-0.77) multiplied by 107spore/mL and (3.52. + -. 0.87). times.107spores/mL. It can be seen that the optimum temperature for sporulation is in the range of 25 ℃ to 28 ℃.
Example 5: influence of shaking table rotating speed on spore yield of fusarium verticillium FCPC-L01 strain
The seed solution was fermented with the spore suspension of the strain Fusarium verticillarum FCPC-L01 prepared in example 2. The fermentation culture solution contains sucrose potato culture solution PS. The sample loading amount is 100mL/250mL, and the inoculation amount is 5 mL. Setting different rotating speed gradients of a shaking table on the basis that the initial pH value is the natural pH value of the culture solution and the optimal fermentation temperature of the screened spore yield is 28 ℃: 90. 120, 150, 180 and 210rpm, the effect of different shaker speeds on the sporulation of the strain was determined, and the other conditions and the determination methods were as described above, in order to obtain optimum sporulation of the shaker speeds.
The results show that the spore production of the strains has significant difference under different shaker rotation speed conditions (F)4,197.75, p 0.001, fig. 6). With the increase of the rotating speed of the shaking table, the spore yield is gradually increasedAnd reaches the highest value under the condition of 180rpm, the number of spores reaches 90 +/-25.03, and the corresponding spore concentration is (4.50 +/-1.25) multiplied by 107spores/mL.
Example 6: influence of initial pH value of fermentation liquor on spore yield of Fusarium verticillarum FCPC-L01 strain
The fermentation was carried out using the spore suspension of the F.verticillata strain FCPC-L01 prepared in example 2 as seed liquid. The fermentation culture solution contains sucrose potato culture solution PS. The sample loading volume is 100mL/250mL, and the inoculation volume is 5 mL. Setting 5 different initial pH values of fermentation liquor on the basis of the optimal fermentation temperature of 28 ℃ and the shaking table rotation speed of 180rpm for screening the sporulation quantity, wherein the initial pH values are respectively 4, 5, 6, 7 and 8, measuring the influence of the different initial pH values on the sporulation quantity of the strain, and obtaining the optimal initial pH value of the sporulation by other conditions and measuring methods as described above.
The results show that there is a significant difference between the sporulation of the strains at different initial pH values (F)4,195.32, p 0.007, fig. 7). At 4 and 6, the spore yield is high, the spore number reaches 109.75 + -19.14 and 90.75 + -12.84 respectively, and the corresponding spore concentration is (5.49 + -0.96) × 107spore/mL and (4.54. + -. 0.64). times.107spores/mL.
Example 7: solid fermentation of fusarium liquid fermentation liquid on rice and conidium insecticidal activity determination
The fermentation was carried out using the spore suspension of the F.verticillata strain FCPC-L01 prepared in example 2 as seed liquid. The fermentation culture solution contains sucrose potato culture solution PS. The sample loading amount is 100mL/250mL, and the inoculation amount is 5 mL. The fermentation temperature was 28 ℃, the rotational speed of the shaker was 180rpm, the initial pH was 4, and the fermentation was carried out for 3 days. Stirring the fermentation broth obtained by fermentation and sterile rice, placing on sterilized stainless steel tray for solid fermentation, and culturing in incubator at constant temperature and humidity of 25 + -1 deg.C and 75% RH for 7 days. After sporulation, fusarium conidia prepared by solid fermentation are prepared into 5 multiplied by 107And (4) measuring the toxicity of the spore/mL bacterial solution to the tip roller moth. The specific method comprises carefully picking 30 heads of 3-4 instar fir tip diamond back moth larvae with soft forceps,at 5X 107The spores/mL of the culture solution were immersed for 5 seconds, and 30 of the spores/mL of the culture solution were immersed for 5 seconds in a control solution containing 0.05% by mass of Tween-80. Placing fresh fir young shoots in a test box, wrapping the bottoms of the young shoots tightly with absorbent cotton dipped in water, then placing larvae on the young shoots for free feeding, placing the test box in an incubator with constant temperature and humidity at 25 +/-1 ℃ and 85% RH, and testing the death rate of the larvae of the fir young cabbage moth at different times. The results are shown in fig. 8, the death rate of the larvae of the spruce budworm is gradually increased with the time being prolonged, the death rate is up to 76.8% on the 8 th day and is obviously higher than that of the control, and the conidia of the FCPC-L01 strain has stronger infection capacity on the larvae of the spruce budworm.
Sequence listing
<110> Guangdong province forestry scientific research institute
<120> fusarium verticillium and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 678
<212> DNA
<213> Fusarium verticillatum FCPC-L01(Fusarium concentricum FCPC-L01)
<400> 1
ccttataccg ttcgtcatcg ggcacgtcga ctctggcaag tcgaccactg tgagtactac 60
ccttgacgat gagcttatct gccatcgtaa tcctgaccaa gatctggcgg ggtgtatctc 120
aaaagacaac atgctgacat agcttcacag accggtcact tgatctacca gtgcggtggt 180
atcgacaagc gaaccatcga gaagttcgag aaggttagtc tcccttcgag cgcgcgtcct 240
ttgcccatcg attttcccct acgactcgaa acgtgcccgc taccccgctc gagaccaaaa 300
attttgcgat atgaccgtaa ttttttttgg tggggcattt accccgccac tcgagcgatg 360
ggcgcgtttg ccctctccac aatctcaatg agcgcatcgt cacgtgccaa gcagtcacta 420
accatccgac aataggaagc cgctgagctc ggtaagggtt ccttcaagta cgcctgggtt 480
cttgacaagc tcaaggccga gcgtgagcgt ggtatcacca tcgatatcgc tctctggaag 540
ttcgagactc ctcgctacta tgtcaccgtc attggtatgt tgtcgcctat gcttcattct 600
tcttcctcgt actaacatat cactcagacg ctcccggtca ccgtgatttc atcaagaaca 660
tgatcactgg tacttcca 678
Claims (8)
1. Fusarium rhizomorph (F. rhizomorpha)Fusarium concentricum) It is characterized in that the preservation number is as follows: GDMCC No: 60971.
2. a bacterial preparation comprising the Fusarium verticillium of claim 1 as an active ingredient.
3. A fermentation process of Fusarium moniliforme of claim 1 comprising the steps of:
a. inoculating the fusarium verticillium into a liquid culture medium for fermentation culture to obtain a fermentation liquid;
b. and pouring the fermentation liquor into sterile rice, uniformly stirring, and culturing to produce spores.
4. The fermentation method according to claim 3, wherein the liquid culture medium is sucrose potato broth PS, and the preparation method comprises: according to the weight portion, 200 portions of potatoes are taken, washed, peeled and cut up, water is added for boiling for 30min, gauze is used for filtering, 40 portions of cane sugar are taken out of filtrate, and water is used for supplementing to 1000 portions; then sterilized at 121 ℃ for 20 min.
5. The fermentation process according to claim 3, wherein the liquid medium is DY, and the production process comprises: taking 40 parts of glucose and 20 parts of yeast extract by weight, and supplementing water to 1000 parts; then sterilized at 121 ℃ for 20 min.
6. The fermentation method according to claim 3, wherein the temperature of the fermentation culture in step a is 25-28 ℃, the initial pH is 4-6, the rotation speed of the shaking table is 180-.
7. The fermentation process of claim 3, wherein the spore-forming conditions of step b are 25 ± 1 ℃ and 75% relative humidity.
8. The use of fusarium verticillatum of claim 1 for the control of cabbage caterpillar.
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