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Strain with strong pathogenicity on euphorbia juglandis and application of strain

A technology of pathogenicity and strains, applied in the direction of application, fungi, plant growth regulators, etc., can solve the problems of food safety, heavy environmental pollution, time-consuming and labor-intensive, etc., and achieve the effects of drug resistance, rapid growth, and environmental friendliness

Pending Publication Date: 2022-05-13
贵州省核桃研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above methods are time-consuming, laborious, produce food safety problems, and cause serious environmental pollution. Therefore, it is urgent to explore an efficient, stable, and pollution-free prevention and control method.

Method used

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  • Strain with strong pathogenicity on euphorbia juglandis and application of strain
  • Strain with strong pathogenicity on euphorbia juglandis and application of strain
  • Strain with strong pathogenicity on euphorbia juglandis and application of strain

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Experimental program
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Effect test

Embodiment 1

[0018] Embodiment 1: Isolation and identification of Beauveria bassiana

[0019] (1) Separation, purification and storage

[0020] Dip the walnut pedigree in 70% ethanol for 30 seconds, then sterilize it with 0.1% mercuric chloride for 1.5 minutes, rinse it with sterilized water for 3 times, use sterilized filter paper to dry the water on the adult walnut pedactid, and then place it in in the PDA tablet. Cultivate in a constant temperature incubator at 27°C for 2-3 days. After mycelium grows around the insect body, pick the mycelium and transfer it to a new PDA plate. Repeat this for 3 times, and then use the dilution purification method to isolate the single colony of the pathogen. , to obtain rejuvenated, highly pathogenic strains of Beauveria bassiana. The obtained highly virulent strains were transferred to fresh PDA slant medium, and then stored at 4°C. On September 23, 2021, it was deposited in the General Microbiology Center of China Microbiological Culture Collectio...

Embodiment 2

[0023] The molecular identification of embodiment 2 bacterial strains

[0024] (1) Extraction of fungal genomic DNA

[0025] Use TSINGKE Plant DNA Extraction Kit (General Type) to extract the genomic DNA of Beauveria bassiana strains, the specific steps are:

[0026] S1: Put SpinCoulumn in CollectionTube, add 250μL BufferBL, centrifuge at 12000rpm / min for 1min to activate the silica gel membrane;

[0027] S2: Grind the dried tissue (≤20mg) with liquid nitrogen thoroughly, put it into a 1.5ml centrifuge tube after grinding, add 400μL BuffergP1, vortex for 1min, and put it in a water bath at 65℃ for 10-30min. ;

[0028] S3: Add 150 μL BuffergP2, vortex for 1 min, and bathe in ice water for 5 min;

[0029] S4: centrifuge at 12000rpm / min for 5min, transfer the supernatant to a new centrifuge tube;

[0030] S5: Add an equal volume of absolute ethanol to the supernatant, shake and mix immediately, transfer all the liquid into the SpinCoulumn, centrifuge at 12000rpm / min for 30s, ...

Embodiment 3

[0047] The virulence test of embodiment 3 bacterial strains

[0048] (1) Preparation of fungal spore suspension

[0049] Inoculate the preserved Beauveria bassiana strain (CGMCCNO: 23261) into PDA medium, culture at a constant temperature of 28°C for 10 days, scrape the conidia, and dilute them with sterile water to 1×10 8 Spores / mL, 1×10 7 Spores / mL, 1×10 6 Spores / mL, 1×10 5 Spores / mL, 1×10 4 Spores / mL of spore suspension, set aside.

[0050] (2) Indoor toxicity test

[0051] Toxicity test was carried out by immersion method. Each treatment concentration picks about 30 strong adults of the walnut pedigree respectively (about 10 for each repetition, a total of 3 repetitions), and respectively adopts the spore suspension of the corresponding concentration to soak the strong walnut pedactid in the suspension for 8s, , put fresh walnut branches, leaves and walnut fruit that were clipped to cultivate. The branches and walnut fruit are placed in a glass bottle and the top su...

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Abstract

The invention relates to the technical field of biological prevention and control, and discloses a strain with strong pathogenicity to euphorbia juglandis and application of the strain, the strain is beauveria bassiana and is preserved in China General Microbiological Culture Collection Center on September 23, 2021, and the strain preservation number is CGMCC NO: 23261. The beauveria bassiana strain is separated from the bombyx batryticatus collected from the forest for the first time, the strain is easy to culture, rapid in growth, large in sporulation quantity and high in spore germination rate, and experiments show that the strain has relatively strong pathogenicity to the adult euphorbia juglandis, is environment-friendly, is not easy to generate drug resistance, and has a good application prospect. The method can be widely applied to biological prevention and control of the euphorbia juglandis.

Description

technical field [0001] The invention relates to the technical field of biological control, in particular to a bacterial strain with strong pathogenicity to walnut mammoth and application thereof. Background technique [0002] Walnut is one of the important economic tree species in our country. Its wood, fruit, branches and leaves, and fruit shell all have high utilization and development value. The walnut industry has been established as one of the leading industries in the development of regional economy and new rural economy in many areas. The walnut industry can increase farmers' income, stimulate regional economic development, and improve the ecological environment at the same time. It has been promoted in many provinces of our country. However, due to the blind introduction of new varieties, good and bad varieties are mixed, coupled with the continuous expansion of cultivation area, fruit farmers relax management, and walnut pests and diseases occur seriously. Among the...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/02A01N63/30A01P7/04C12R1/645
CPCC12N1/02A01N63/30Y02A50/30
Inventor 赵玉雪朱佳敏杨霞吴柳燕苏州
Owner 贵州省核桃研究所
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