CN114437943A - High-efficiency pathogenicity biocontrol bacterium fijimycin strain and application thereof - Google Patents

Abstract

The invention discloses a high-efficiency pathogenicity biocontrol bacterium fijimycin strain and application thereof, belonging to the technical field of biological control. The invention relates to a Feijingqu mould (Aspergillus fijiensis) GDIZM-1 which is preserved in Guangdong province microorganism strain preservation center 12-17 th 2021 with the preservation number: GDMCC No: 62135. the results of long-term infection biology and indoor bioassay show that the strain has strong insecticidal effect on the diaphorina citri; and the strain has the characteristics of high growth rate and large spore yield. The fijix aspergillus is a biocontrol fungus, is used as a living biological insecticide, has a brand new action mechanism different from the existing chemical insecticide, has no pollution and residue to the environment, can solve the problems of drug resistance and pesticide residue in the control process of the diaphorina citri, is suitable for the requirements of organic food production, and has very good application potential in the biological control of citrus pests.

Description

High-efficiency pathogenicity biocontrol bacterium fijimycin strain and application thereof

Technical Field

The invention relates to the technical field of microbial control, in particular to a high-efficiency pathogenicity biocontrol strain fijiri aspergillus strain and application thereof.

Background

The Diaphorina citri Kuwayama is an important pest in the young shoot stage of a rutaceae plant, adult nymphs gather on young shoots, young buds and young leaves, the phloem juice of a host plant is sucked by a piercing-sucking mouthpart, the damaged leaves are yellowed, Kawasaki, withered and even dead, and a large amount of white honey dew is discharged after the citrus psylla is eaten, so that the sooty smoke is caused, the photosynthesis of the plant is influenced, and the growth and development of the plant are seriously influenced. And is also an important transmission vector of citrus greening disease. The diaphorina citri is fed and spawned on the citrus greening disease plants for propagation, so that a large amount of bacteria-carrying imagoes can be generated, and the imagoes can be transferred into new harmful plants to spread the greening disease. At present, the main method for preventing and controlling the diaphorina citri still depends on chemical agents, but a series of negative problems of mass death of insects which are natural enemies of citrus orchards, loss of biological diversity, generation of drug resistance in different degrees, pollution to the ecological environment of the orchards and the like are caused by long-term and large-scale use of chemical pesticides, and the problem of overproof pesticide residues of fruits sometimes occurs. In the face of such passive situations, research and application of non-chemical means to control diaphorina citri have been an urgent problem facing the citrus industry. Therefore, under the big background that the country advocates the reduction of both pesticides and fertilizers, the search of an efficient and environment-friendly control means is very important.

Biological control is a control measure in which one type of organism inhibits another type of organism by utilizing the interdependence or restriction relationship between species, and the dynamic equilibrium relationship existing between organisms is inclined toward the favorable direction of agricultural production. Compared with other control methods, the biological control method is safe to human and livestock and ecological environment, has high efficiency and durability, and is the most promising control means in comprehensive control measures of pests. Therefore, the search for effective biological control products and application techniques is one of the main research directions for sustainable pest control in the future. The research and development of the microbial pesticide for safely, efficiently and sustainably controlling the diaphorina citri to replace the chemical pesticide has important social and economic benefits and ecological benefits.

Entomopathogenic fungi are excellent environment-friendly substitutes for chemical pesticides, and are widely used for biological control of agricultural pests. The entomogenous fungi has the same control effect on diaphorina citri with drug resistance and pesticide sensitivity, and the prevention and control prospect is widely seen. Research shows that the pathogenic mechanism of entomopathogenic fungi to pests is complex and different from the insecticidal mechanism of the existing chemical pesticide. No pest with drug resistance to entomopathogenic fungi is discovered so far, so that the application and development potential of the entomopathogenic fungi is huge in the aspect of controlling the drug resistance of the pest.

Disclosure of Invention

The invention aims to overcome the defects of the existing diaphorina citri control technology and provides a biocontrol bacterium with high pathogenicity on diaphorina citri and a control method thereof. Entomogenous fungi have diversity in genetics, ecology, biology and the like. The screening and obtaining of high-yield and high-quality strains are the first premise for obtaining better control effect. The strain screening mainly considers 3 indexes of sporulation quantity, colony growth rate and pathogenicity respectively. The invention selects excellent strains based on the indexes.

The first purpose of the invention is to provide a strain of aspergillus fijiensis (aspergillus fijiensis) GDIZM-1, which has the collection number of: GDMCC No: 62135.

the strain GDIZM-1 of the invention has compact texture on a PDA culture medium, flat bacterial colony and radial furrows on the back. The early stage of the front of the colony is white hypha, and the later stage is yellow brown and is powdery; the color of the back surface is gradually deepened and is earthy yellow. Conidiophores are different in length and smooth in wall, have podocytes and are 1100 Mu & ltx & gt 8 & lt-16 & gt in size, the tops of the conidiophores expand to form top sacs which are approximately spherical and 20-45 Mu & ltx & gt 10 & lt-15 & gt in size, a layer of small peduncles are covered on the top sacs and are arranged in a radial mode and 3-6 Mu & ltx & gt 2 & lt-3 & gt in size, and the surface 3/4 can be cultivated. The peduncle entomogenous spore is nearly spherical, has rough wall, is full of thorns, is arranged in a chain shape, and has the diameter of about 3-5 mu m.

The invention relates to the non-existence of the aspergillus fijiensis GDIZM-1Conidial suspensions (1X 10) at the same concentration⁴、1×10⁵、1×10⁶、1×10⁷、1×10⁸spores/mL) had a mean difference between the 6-year corrected mortality rates for diaphorina citri (1-5 instar nymphs and adults) compared to the control, indicating that aspergillus fijiensis has high pathogenicity for diaphorina citri and adults. Meanwhile, the GDIZM-1 can be used for treating 1-2-year (low age), 3-4-year (high age), 5-year (end age) nymphs and adults of diaphorina citri₅₀And LT₅₀Respectively, are 0.43X 10⁴、1.41×10⁴、2.41×10⁴、3.79×10⁴spores/mL and 2.30, 2.86, 3.43, 4.25 days. The fijix aspergillus is shown to have important application potential in the aspect of biological control of diaphorina citri.

The second object of the present invention is to provide a microbial preparation comprising the culture of GDIZM-1 or the fermentation broth thereof as an active ingredient.

Preferably, GDIZM-1 is attached to an SDAY plate at a temperature of 24-26 ℃ under 14L light: 10D, thereby obtaining a culture of GDIZM-1.

A third object of the present invention is to provide a use of aspergillus fijiri (aspergillus fisjiensis) in any one of the following:

(1) controlling diaphorina citri or diseases caused by the diaphorina citri;

(2) preparing the biological prevention and treatment medicine.

Preferably, the Aspergillus fijiensis is Aspergillus fijiensis GDIZM-1 with the deposit number: GDMCC No: 62135.

preferably, the disease caused by the diaphorina citri is a disease caused by the diaphorina citri as a transmission vector.

Preferably, the diseases caused by the diaphorina citri include, but are not limited to, citrus yellow dragon disease.

Preferably, the fijix aspergillus is used for controlling diaphorina citri nymphs and adults.

Preferably, the biocontrol agent comprises a conidium of aspergillus fijiri or a suspension thereof.

Further, the preparation of the conidia of aspergillus fijiensis comprises the following steps: taking out and activating a strain on a Fiji aspergillus slant, inoculating the strain on an SDAY plate, culturing for 10 days in a constant-temperature incubator, scraping conidia on the surface of a culture medium, and filtering and collecting the conidia by using sterile water containing 0.1% Tween 80; the suspension is prepared by collecting conidia to obtain conidium suspension.

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

(1) the Aspergillus fijiensis GDIZM-1 with high pathogenicity to diaphorina citri is obtained by screening, and LC (inductance-capacitance) of the Aspergillus fijiensis GDIZM-1 to 1-2 years old (low age), 3-4 years old (high age), 5 years old (late age) nymphs and adults of the diaphorina citri is determined through long-term infection biology and indoor bioassay₅₀And LT₅₀Respectively, are 0.43X 10⁴、1.41×10⁴、2.41×10⁴、3.79×10⁴Spores per mL and 2.30, 2.86, 3.43 and 4.25 days show that the strain has strong infection insecticidal effect on the diaphorina citri and has very strong application potential in biological control of the diaphorina citri;

(2) the Aspergillus fijii GDIZM-1 has high spore yield, and the spore yield can reach 3.86 multiplied by 10 days after being cultured on SDAY culture medium⁸spore/mL;

(3) the growth rate of the colonies of the aspergillus fijiensis GDIZM-1 is high, and the diameter of the colonies can reach 83mm after the colonies are cultured on an SDAY culture medium for 5 days;

(4) the fijiaspergillus can be used as a living biological insecticide, has no pollution and residue to the environment, can solve the problems of drug resistance and pesticide residue in the control process of the diaphorina citri, and is suitable for the requirements of organic food production.

The Aspergillus fijiensis GDIZM-1 is preserved in the microbial culture collection center (GDMCC) of Guangdong province in 12 th and 17 th of 2021, is addressed to No. 59 floor of Dazhou No. 100 Yue Jie of the overseas district of Guangzhou city of Guangdong province, and is coded by a postal code: 510070, accession number: GDMCC No: 62135.

drawings

FIG. 1 shows a psyllid nymph of Citrus and an adult infected Aspergillus fijiensis (1X 10)⁷spores/mL), graphic representation A, B, C, D, E, F for 1, 2, 3, 4, 5-year-old nymphs and adults, respectively;the scale values of the graph A, B, C, D, E, F are 100, 200, 500 μm, respectively.

FIG. 2 is a colony morphology of an Aspergillus fijii isolate; FIG. A, B shows day 3 colonies of Aspergillus fijii on the front and back sides; FIG. C, D shows the front and back of an Aspergillus fijii colony on day 10; FIG. E, F, G, H shows a FIJIASCUS colony gradient; the scale values of the graph A, B, C, D are all 10mm, and the scale values of the graph E, F, G, H are 1mm, 500 μm, 200 μm, and 500 μm, respectively.

FIG. 3 is a morphogram of the conidiophore and conidiophore of the strain GDIZM-1 of Aspergillus fijiensis, in which A is the apical vesicle and conidiophore of GDIZM-1 of Aspergillus fijiensis; panel B shows Aspergillus fijii GDIZM-1 podocytes; diagram C is Aspergillus fijii GDIZM-1 conidia; the scale values of the graph A, B, C are all 10 μm.

FIG. 4 shows a molecular characterization phylogenetic tree of the strain GDIZM-1 of Aspergillus fijiri.

FIG. 5 shows different concentrations of Aspergillus fijiri spore suspensions (1X 10)⁴，1×10⁵，1×10⁶，1×10⁷，1×10⁸Individual conidia/ml) mortality of diaphorina citri at different developmental stages.

Detailed Description

The present invention is further illustrated by the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Example 1 isolation, purification and characterization of pathogenic strains of diaphorina citri

1.1 sources of Material

(1) Sample preparation: collecting diaphorina citri cadavers infected by entomogenous fungi in a lemon garden.

(2) Test insects: the diaphorina citri is bred and bred for multiple generations in a net room of an environmental insect research center of the animal research institute of the academy of sciences of Guangdong province.

(3) PDA culture medium: 6g/L of potato extract powder, 20g/L of glucose and 20g/L of agar; SDAY medium: 10g/L yeast extract powder, 40g/L glucose, 10g/L peptone and 20g/L agar; the solvent of the culture medium is water, and the preparation method comprises mixing the above components, and sterilizing.

(4) Sterile operating conditions: all the utensils and appliances need to be sterilized in autoclave (121 ℃, 30min), and the operations of inoculation and the like are all carried out in a clean bench.

1.2 isolation and purification of the Strain

Isolating the target bacteria from diaphorina citri cadaver samples infected with entomogenous fungi. The method comprises the following steps: sterilizing the surface of a sample by using a 5% sodium hypochlorite solution, washing the sterilized sample in sterilized water for 3 times, putting the washed sample into a PDA flat plate, inverting the PDA flat plate into a thermostat at 25 +/-1 ℃ and illuminating (14L:10D) for culturing, transferring the sample into a PDA inclined plane after bacterial colonies are formed, and then transferring the sample into a refrigerator at 4 ℃ for storage. Respectively culturing the strains on PDA culture medium for 10 days, after forming spores, selecting conidia to prepare 1 × 10³Conidium suspension of single spore/mL, dropping the suspension on a glass slide with a cover glass, observing under a biological microscope, inserting a glass slide with only one conidium in one drop into a culture medium, and culturing in an incubator to obtain an isolate.

Under natural conditions, typical coreopsis is grow on diaphorina citri parasites parasitized by fungi. After separation and purification, a fungus strain is obtained and is numbered as GDIZM-1. Through the back grafting verification, the strain can successfully infect the diaphorina citri at all ages, white mycelium can be observed to grow on the body surface after inoculation for 2d, the symptoms such as reaction retardation, slow action, body color change and the like are shown, the typical sporophyte bundles grow out of the insect body after inoculation for 10d, and the infection form is shown in figure 1. The separation result shows that the culture characters and morphological characteristics of thalli grown on the inoculated worm body are the same as those of the original inoculated strain, and the strain is the entomogenous fungus infecting the diaphorina citri.

1.3 identification of the strains

(1) Morphological identification:

the strain is spotted on a PDA flat plate and cultured in a constant temperature incubator at the temperature of 25 +/-1 ℃, and the color, texture, size, height, surface ornamentation, edge, exudate and the like of the front surface and the back surface of the colony are observed every day to take a picture. Selecting a small amount of spores with an inoculating needle, planting the spores on a PDA plate, preparing a glass slide when the spores are mature, selecting a transparent adhesive tape with fine texture, clamping the small adhesive tape cut into a proper size with a pair of tweezers, slightly and slowly adhering from the edge of a bacterial colony to the center, fixing with 95% alcohol, immersing into cotton blue for dyeing, putting the transparent adhesive tape with the spore-forming structures upwards, slightly putting down the cover glass after 2-3min, and carefully avoiding generating bubbles. Observing under a low power lens to find a visual field, then dripping cedar oil or paraffin oil, and observing structural characteristics of conidiophores, spore-forming cells and spores under the oil lens. The photographs were taken at 1000 x with an IMAGING MP 5.0.0-RTV-CLR-10-A camera, and then a finished picture was synthesized.

The strain is compact on a PDA culture medium, the colony is flat, and the back surface of the strain is provided with radial furrows. The colony morphology of the strain on the PDA culture medium is shown in FIG. 2, the early stage of the front of the colony is white hypha, and the later stage is yellow brown and powdery; the color of the back surface is gradually deepened and is earthy yellow. As shown in FIG. 3, conidiophores are different in length, smooth in wall, sufficient in cells and 200-1100 μm × 8-16 μm in size, the top of the conidiophores are expanded to form a top sac which is approximately spherical and 20-45 μm × 10-15 μm in size, a layer of peduncles is covered on the top sac, the peduncles are arranged in a radial shape and 3-6 μm × 2-3 μm in size, and the surface 3/4 is fertile. The peduncle entomogenous spore is nearly spherical, has rough wall, is full of thorns, is arranged in a chain shape, and has the diameter of about 3-5 mu m.

(2) And (3) molecular identification:

the specific DNA sequence of this study consists of three genes, including the transcribed spacer, the elongation factor and the RNA polymerase II large subunit. Taking 100mg of cultured mature strain, extracting the genome DNA of hyphae by adopting a fungus genome DNA extraction kit, using genomic DNA of the strain as a template, ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3'), EF-1983F (5 '-GC (Y) CC (Y) GG (H) CA (Y)) G GTGA (Y) TT (Y) AT-3') and EF-12218R (5 '-ATAC (R) TG (R) GC (R) AC (R) GT (Y) TG-3'), fRPB2-5F (5 '-GA (Y)) GA (Y) GA (M) G (W) GAW (Y) TT TCA (Y) GG-3') and fRPB2-7cR (5 '-CCCAT (R) GCTTG (Y) TT (R) CCCAT-3') were amplified as primers, followed by BLAST alignment analysis in GENBANK. The above symbols (Y), (H), (R), (W) represent degenerate bases, i.e., one symbol in place of some two or more bases, wherein (Y) represents C/T; (H) represents A/T/C; (R) represents A/G; (W) represents A/T.

PCR 50. mu.L reaction: the upstream and downstream primers were 1. mu. L, ddH2O 22. mu. L, DNA template, 1. mu.L, 2 × Master Mix, 25. mu.L, respectively. PCR amplification procedure for amplification of rDNA-ITS sequences: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 30s, and extension at 72 ℃ for 45s for 35 cycles; finally, extension is carried out for 10min at 72 ℃. PCR amplification procedure for amplification of EF-1. alpha. Gene sequence: pre-denaturation at 95 ℃ for 10 min; denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 30s, and extension at 72 ℃ for 1min for 40 cycles; finally, extension is carried out for 10min at 72 ℃. PCR amplification procedure for amplification of fRPB2 gene sequence: pre-denaturation at 95 ℃ for 10 min; denaturation at 94 ℃ for 30s, annealing at 50 ℃ for 30s, and extension at 72 ℃ for 1min for 40 cycles; finally, extension is carried out for 10min at 72 ℃. And (3) carrying out electrophoresis detection on the amplification products in 1.5% agarose gel, observing whether a target band exists or not, and sequencing the amplification products with correct band sizes in Shenzhen Hua Dagen Limited. And splicing the sequencing result by using the MeGA7.0 software to remove part of sequences with uneven head and tail, carrying out gene homology similarity comparison by using an online program BLAST of an NCBI website, and downloading corresponding sequences of related published model strains. Based on ITS, EF-1 alpha and fRPB2 marker genes, the phylogenetic tree of the fungus to be identified is constructed by adopting the evolution analysis software MEGA 7.0. An adjacency method (Neighbor Joining) is selected for building the tree, the step length test (Bootstrapmeter) selection parameter is 1000, and a genetic distance calculation model (Substitutionmodel) selects p-distance.

The PCR products of the GDIZM-1 strain are purified, recovered and sequenced to obtain ITS (SEQ ID NO.1), EF-1 alpha (SEQ ID NO.2) and fRPB2(SEQ ID NO.3) gene fragments which are spliced and connected in series, and then a Neighbor Joining Tree is constructed by an adjacent method (figure 4). The resulting DNA sequences were Blast aligned at NCBI. After comparison, the similarity of the strain GDIZM-1 and the Aspergillus fijii is 99-100%. Meanwhile, the GDIZM-1 strain and the related strains of the aspergillus fijiensis are gathered into one and have a longer genetic distance with the outlier. The strain GDIZM-1 can be determined to be Aspergillus fijiensis (Aspergillus fijiensis) by combining with strain morphological identification of GDIZM-1.

Therefore, in combination with morphology and molecular biology, the strain GDIZM-1 is named as Aspergillus fijii (Aspergillus fijiensis) GDIZM-1, is preserved in the culture Collection of microorganisms of Guangdong province (GDMCC) in 12 months and 17 days in 2021, is located at Lou 5 th of Hao 59, Yao 100, Rexiu Miao, Zuizon, Guangdong province, and is coded by: 510070, accession number: GDMCC No: 62135.

example 2 study of hyphal growth and spore production characteristics of Aspergillus fijii GDIZM-1 Strain

Determination of colony growth rate and spore yield: slant strain of Aspergillus fijii GDIZM-1 is taken out and activated, inoculated on an SDAY plate, cultured for 10 days in a constant temperature incubator, then conidia on the surface of the culture medium are scraped, and the conidia are collected by filtering sterile water containing 0.1% Tween 80 to prepare 1 × 10⁷spores/mL of spore suspension. And (3) sucking 1mL of spore suspension liquid, dripping the spore suspension liquid into the center of the SDAY flat plate, sealing the SDAY flat plate after uniform smearing, culturing in a constant-temperature incubator, observing for 10d, recording the diameter of each bacterial colony every day, and repeating for 5 times. After 10d, conidia were filtered with sterile water containing 0.1% tween 80 and sporulation was determined using a hemocytometer.

The colony growth rate and spore yield of the A.fijiensis isolate are shown in Table 1, and the data are mean. + -. standard error. The study found that after 5 days of incubation of a. fijiensis colony diameter reached 83.7 ± 0.47mm (mean ± sem); after 10 days of cultivation, the spore yield in SDAY medium is 3.86X 10⁸spores/mL.

TABLE 1 FIJIASCUS SEPARATING STRAIN GDIZM-1 colonial growth Rate and spore yield

Example 3 pathogenicity assay of Aspergillus fijiensis GDIZM-1 Strain on Trionychus citri

(1) Preparation of conidium suspension of the strain GDIZM-1 of Aspergillus fijii was activated by taking out the slant strain, inoculated on an SDAY plate, cultured for 10d in a constant temperature incubator, then the conidia on the surface of the medium was scraped off, and the conidia were collected by sterile filtration using sterile water containing 0.1% Tween 80 to prepare 1X 10⁴，1×10⁵，1×10⁶，1×10⁷，1×10⁸Spore suspension/mL spore。

(2) Selecting healthy lemon plants, cutting old leaves, cleaning leaves with clear water, completely soaking 1-5-year-old diaphorina citri nymphs and adults in 5 spore suspensions with different concentrations for 2 minutes, drying by using filter paper, immediately transferring to young leaves of lemon seedlings and bagging, feeding in a climatic chamber with the conditions of 25 +/-2 ℃, 75 +/-5% RH humidity and 14:10 photoperiod (L: D), and repeating for 3 times for 20 times, wherein the humidity is 75 +/-5% RH and the photoperiod is 14:10(L: D). Continuously observing and recording 8d mortality, and evaluating LC of young nymphs of diaphorina citri (1-2 years old), old nymphs (3-4 years old), terminal nymphs (5 years old) and adults (hatching for 3 days)₅₀Value sum LT₅₀The value is obtained. The negative control was sterile water. All the test data are processed on a data processing software SPSS system.

The results of the pathogenicity study are shown in FIG. 5, and 5 conidiospore concentrations of the Aspergillus fijii GDIZM-1 spore suspension (1X 10)⁴、1×10⁵、1×10⁶、1×10⁷、1×10⁸spores/mL) corrected mortality for diaphorina citri (1-5 instar nymphs and adults) at 6 instars differed significantly compared to controls. The concentration is 1X 10⁸Single spore/mL aspergillus fijiensis GDIZM-1 was highly pathogenic to diaphorina citri nymphs (88.67% -98.14%) and adults (87.32%). Meanwhile, the compound preparation can be used for treating 1-2-year (low age), 3-4-year (high age), 5-year (late age) nymphs and adults of diaphorina citri (L) and LC (L-LC)₅₀And LT₅₀Respectively, are 0.43X 10⁴、1.41×10⁴、2.41×10⁴、3.79×10⁴spores/mL (table 2) and 2.30, 2.86, 3.43, 4.25 days (table 3). Wherein X in tables 2 and 3 represents a logarithmic value of the concentration of the spore suspension, and Y represents a value corresponding to the logarithmic value of the concentration of the spore suspension.

TABLE 2 virulence of Aspergillus fijiensis at different developmental stages of diaphorina citri after 8d infection

TABLE 3 infection with Aspergillus fijiensis (1X 10)⁸Individual spores/mL) on Citrus psyllid at different developmental stages

Sequence listing

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aaaactgcgg ctctctggtg ctcaacaaag aacacatccg caaactagaa caagataaag 780

acctgcctcc agacttggat ccagaagacc gccgcgatcg ttactttggt tgggatggcc 840

tggtgaagtc tggtgtagtg gagtacgtcg atgctgaaga agaagagacc atcatgattt 900

ccatgactcc ggaagatctc gaaatttcca agcaactcca ggctggttat gctcttccgg 960

atgaggacta cacgatccga acaagcgtgt acgctccatt ctgagtcaaa aggcgca 1017

Claims (8)

1. A strain of Aspergillus fijiensis (Aspergillus) GDIZM-1 with a deposit number of: GDMCC No: 62135.

2. a microbial preparation comprising the culture product of GDIZM-1 or a fermentation broth thereof as set forth in claim 1 as an active ingredient.

3. The microbial preparation of claim 2, wherein GDIZM-1 is attached to a SDAY plate and irradiated at a temperature of 24-26 ℃ in 14L:10D, thereby obtaining a culture of GDIZM-1.

4. Use of aspergillus fijiri (aspergillus fisriensis) in any one of the following:

(1) controlling diaphorina citri or diseases caused by the diaphorina citri;

(2) preparing the biological prevention and treatment medicine.

5. The use according to claim 4, wherein the Aspergillus fijiensis strain is Aspergillus gdIZM-1 with the deposit number: GDMCC No: 62135.

6. use according to claim 4 or 5, wherein the disease caused by diaphorina citri is citrus yellow dragon.

7. The use according to claim 4 or 5, wherein the biocontrol agent comprises a conidium of Aspergillus fijiri or a suspension thereof.

8. Use according to claim 7, wherein the preparation of conidia of Aspergillus fijiri comprises the steps of: taking out and activating a strain on a Fiji aspergillus slant, inoculating the strain on an SDAY plate, culturing for 10 days in a constant-temperature incubator, scraping conidia on the surface of a culture medium, and filtering and collecting the conidia by using sterile water containing 0.1% Tween 80;

the suspension is prepared by collecting conidia to obtain conidium suspension.

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