A metarhizium anisopliae strain with strong pathogenicity to the hazel weevil and its application

A technology of Metarhizium anisopliae and pathogenicity, which is applied in the field of biological control, can solve the problems of lack of strong pathogenic strains of hazel weevil, differences in pathogenicity of pests, etc., and achieves difficult drug resistance, high spore germination rate, specificity strong effect

Inactive Publication Date: 2021-02-26
JILIN NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, although Metarhizium anisopliae has a wide range of hosts, its targeting is strong, and its pathogenicity to different pests is very different.
At present, there is a lack of strong pathogenic strains of weevil weevil at home and abroad

Method used

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  • A metarhizium anisopliae strain with strong pathogenicity to the hazel weevil and its application

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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 Rapid 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 to 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...

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, more than 1,200 larvae of the 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 ti...

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Abstract

The invention discloses a strain of Metarhizium anisopliae with strong pathogenicity to the weevil hazel and its application. The Metarhizium anisopliae strain provided by the present invention is Metarhizium anisopliae CoCn01, which has been preserved in the General Microorganism Center (CGMCC) of the China Committee for Microbiological Culture Collection on April 12, 2016 through patent preservation. The number is CGMCC No.12167. The bacterial strain has fast growth rate and strong spore-producing ability. The present invention is isolated and obtained for the first time from the adult corpse of Curculio nucum (Curculio nucum), and its culture method is relatively simple, and the reproduction speed is fast. The larvae have strong pathogenicity, are harmless to humans and animals, have no pollution to the environment, and pests are not easy to develop drug resistance. They can be widely used in the control of Hazel Weevil. It is a plant with super high application in the control of Hazel Weevil. valuable biocontrol strains.

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 Patents(China)
IPC IPC(8): C12N1/14A01N63/30A01P7/04C12R1/645
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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