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Metarhizium anisopliae strain with strong pathogenicity to curculio nucum and applications of metarhizium anisopliae strain

A technology of Metarhizium anisopliae and pathogenicity, which is applied in the field of biological control, can solve the problem of lack of strong pathogenic strains of Hazel weevil, and achieve the effects of not easy drug resistance, simple cultivation and large spore production.

Inactive Publication Date: 2017-11-21
JILIN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

At present, there is a lack of strong pathogenic strains of weevil weevil at home and abroad

Method used

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  • Metarhizium anisopliae strain with strong pathogenicity to curculio nucum and applications of metarhizium anisopliae strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Embodiment 1: Isolation and identification of pathogenic bacteria

[0012] 1.1 Materials

[0013] In Siping City, Jilin Province, in early June, the carcasses of dead weevil adults infected by entomogenic fungi were collected for the initial isolation and purification of the pathogenic bacteria.

[0014] 1.2 Isolation and purification of pathogenic microorganisms of the hazel weevil

[0015] Select the hazel beetle larvae covered with white mycelium layer and green powdery spores on the surface of the insect body, wash it twice with sterile water on the ultra-clean workbench, and then dry the surface water with sterile absorbent paper. Soak the carcass of the weevil weevil in 75% ethanol for 30 seconds, rinse it with sterile water three times, cut out the tissue block from the middle part of the abdomen of the insect body and inoculate it in PDA medium, the diameter of the tissue block is about 2 mm. The formula of the PDA medium is as follows: 200 g of peeled potatoe...

Embodiment 2

[0021] Example 2: Molecular identification of bacterial strains

[0022] 2.1 Extraction of fungal genomic DNA

[0023] Genomic DNA of Metarhizium anisopliae CoCn01 was extracted with the Genomic DNA Quick Extraction Kit produced by Beijing Aidelai Biotechnology Co., Ltd., and the concentration, purity and integrity of DNA were detected by SimpliNano micro-spectrophotometer.

[0024] 2.2 PCR reaction system

[0025] The nucleotide universal primers for the ITS region of ribosomal DNA were synthesized by Dalian Bao Biotechnology Co., Ltd. The primer series are: ITS1 5′-TCCGTAGGTGAACCTGCGG-3′, ITS4 5′-TCCTCCGCTTATTGATATGC-3′. 20 μL PCR reaction system: 1.0 μL each of 10 μM primers ITS1 and ITS4, 2.0 μL 10×PCR Buffer, Mg 2+ (2.5mmol / L) 1.0μL, dNTP (10mmol / L) 1.0μL, 5UμL -1 0.25 μl of Taq DNA polymerase, ddH 2 O make up to 20 μL. PCR amplification conditions were: 94°C pre-denaturation for 4 min, 1 cycle; 94°C denaturation for 1 min, 56°C annealing for 1 min, 72°C extension fo...

Embodiment 3

[0030] Embodiment 3: the virulence test of bacterial strain

[0031] 3.1 Preparation of fungal spore suspension

[0032] There are three kinds of strains for the test: the above-mentioned Metarhizium anisopliae strain CoCn01, the Metarhizium anisopliae strain 3.7986 preserved by the China Microorganism Culture Conservation Center, and the Metarhizium anisopliae strain 3.4607 preserved by the China Microbiology Culture Conservation Center. Inoculate them in PDA medium respectively, culture them at 28°C for 10 days, scrape their conidia, and dilute them with sterile water to 1.0×10 8 Spores / mL of spore suspension.

[0033] 3.2 Indoor toxicity test

[0034] Toxicity test was carried out by immersion method. On August 20, 2014, about 1,200 larvae of the weevil larvae were collected before the larvae entered the soil. Each strain was treated as 1 treatment, and distilled water was used as the control. There were 4 treatments in total; each treatment was about 100 Head, repeat 3...

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Abstract

The invention discloses a metarhizium anisopliae strain with strong pathogenicity to curculio nucum and applications of the metarhizium anisopliae strain. The metarhizium anisopliae strain provided by the invention is Metarhizium anisopliae CoCn01, is preserved in the China General Microbiological Culture Collection Center (CGMCC) on April 12, 2016 through a patent preservation method, and is assigned with the accession number of CGMCC No.12167. The strain is high in growth speed and strong in spore production capacity, and is separated from the adult corpse of curculio nucum suffering from natural death for the first time, a culture method is simple, the propagation speed is high, the metarhizium anisopliae strain has relatively strong pathogenicity to both the adult and larva curculio nucum, is harmless to human beings and the livestock, and is free of pollution to the environment, the pests have difficulty in generating drug resistance, and thus the metarhizium anisopliae strain can be widely used for preventing and treating curculio nucum, and belongs to a biological control strain which has an ultrahigh application value in the prevention and treatment for curculio nucum.

Description

technical field [0001] The invention belongs to the field of biological control, and relates to a strain of Metarhizium anisopliae and its application, in particular to a strain of Metarhizium anisopliae with strong pathogenicity to the hazel weevil and its application. Background technique [0002] Hazel belongs to the genus Corylus (Corylus L.) of the family Corylacea. It is one of the four major dry fruits (almonds, walnuts, hazelnuts, and cashews) in the world. It is of great significance to improve the mountain economy and increase farmers' income. The area of ​​my country's hazel forest is about 1 million hectares, ranking first in the world, but there is a huge gap between the output level and foreign countries. The average yield of Italian hazelnuts is 1550kg / hm 2 , about 2.5 to 9.0 times the average yield per unit area in my country. The Northeast is the traditional main producing area of ​​hazelnuts. The problem of low income due to low yields is quite common an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/04A01P7/04C12R1/645
CPCA01N63/30C12N1/145C12R2001/645
Inventor 刘剑锋程云清李华郭海勇栾剑刘挺赵一欣
Owner JILIN NORMAL UNIV
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