Separation and purification method of beauveria bassiana

A technology for separation and purification of Beauveria bassiana, applied in the field of application, can solve the problems of limited practical application, high application cost, carcinogenicity of cycloheximide, etc., and achieve good practical application prospects, simple method, and cost reduction effect

Active Publication Date: 2014-02-12
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, although a lot of research has been done on selective media in the prior art, only the expensive dodine or cycloheximide can be used as a bacteriostatic agent to separate better, but the actual application cost is too high ; and cycloheximide is carcinogenic and affects human health
The deficiencies in the existing technology limit the practical application of the above technology, and cannot solve the current general demand for the separation and purification of Beauveria bassiana
[0006] At present, there is a lack of a simple, safe, efficient, and low-cost method for isolating and purifying Beauveria bassiana to meet the needs of practical applications

Method used

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  • Separation and purification method of beauveria bassiana
  • Separation and purification method of beauveria bassiana
  • Separation and purification method of beauveria bassiana

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Treat the collected pathogens: use 75% (v / v) ethanol cotton balls to disinfect the surface of the silkworm body of the naturally infected Ambrysalis larvae (which can also be artificially infected by conventional means), and then put it into the sterile Put a group of cotton balls soaked with sterile water in the dish, and culture it at 26°C for 3-5 days (d). After the silkworm body surface grows white fluffy hyphae and conidia For strain isolation;

[0044] S1. Preparation of spore liquid: use a sterile inoculation loop to pick 2 to 3 rings of conidia powder from the surface of the silkworm body, put them in a triangular flask filled with 50 mL of sterile water (containing sterile glass beads), and vortex for 5 min, to fully disperse the spores;

[0045] S2. Prepare the plate:

[0046] S21. Prepare the following reagents: the concentration of streptomycin sulfate is 1000U / mL (prepared with sterile water, without autoclaving, and added before use); the mass percentage...

Embodiment 2

[0053] Treat the collected pathogens: sterilize the surface of silkworm chrysalis infected with natural infection or artificial infection of Beauveria bassiana with 75% (v / v) ethanol cotton balls, then put it into a sterile petri dish, and put it in the dish for a period of time. Group cotton balls soaked in sterile water, place them at 26°C and incubate them for 3-5 days (d), and isolate the strains after the white fluff mycelia and conidia grow on the surface of the silkworm body;

[0054] S1. Preparation of spore liquid: use a sterile inoculation loop to pick 2 to 3 rings of conidia powder from the surface of the silkworm body, put them in a triangular flask filled with 50 mL of sterile water (containing sterile glass beads), and vortex for 5 min, to fully disperse the spores;

[0055] S2. Prepare the plate:

[0056] S21. Prepare the following reagents: the concentration of streptomycin sulfate is 10000U / mL (prepared with sterile water, without autoclaving, and added befor...

Embodiment 3

[0063] S1. Dilute the biopesticide containing Beauveria bassiana spores for biological control (randomly selected by Guangzhou Biological Control Station, those skilled in the art can also use similar biopesticides containing Beauveria bassiana spores for experiments) to a mass Dilutions with a percentage concentration of 1 or 10%.

[0064] S2. preparing a plate;

[0065]S21. Prepare the following reagents: the concentration of streptomycin sulfate is 100000U / mL (prepared with sterile water, without autoclaving, and added before use); the mass percentage concentration of sodium deoxycholate is 2.2%, and the mass percentage of Bengal red Concentration is 0.25%, autoclave for standby;

[0066] S22. Peel 200g of potatoes, cut them into small pieces, add 1000mL of water to boil until rotten (boil for 20-30min, it can be pierced with a glass rod), filter with 4 layers of gauze, add 20g of agar and 20g of glucose Afterwards, add water to 1000 mL, adjust the pH of the medium to 6.0...

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Abstract

The invention relates to the technical field of identification, separation and purification application of beauveria bassiana strains, and discloses a separation and purification method of beauveria bassiana. By adopting the separation and purification method, a culture medium is scientifically optimized and improved; simple, quick and effective separation and purification of the beauveria bassiana are achieved by combining improvement of cultivation device coverings on the basis of the culture medium with a scientific formula. The method disclosed by the invention is simple and quick to operate, wide in samples applicable to separation and purification of the beauveria bassiana, low in operation cost and free of pollution on environment; the targets of separation and purification can be well achieved.

Description

technical field [0001] The invention relates to the application technical fields of identification of Beauveria bassiana species, biological control, prevention and control of Beauveria bassiana, etc., and more specifically relates to a separation and purification method of Beauveria bassiana. Background technique [0002] Beauveria bassiana ( Beauveria bassiana ) are now classified as Cordyceps ( Cordycipitaceae ), Cordyceps ( Cordyceps ) important fungus, as early as 1816, the Italian scientist Bassi (1835) isolated it from the silkworm Bombyx mori. [0003] Beauveria bassiana can invade more than 700 species of insects and mites belonging to 15 orders, 149 families and 521 genera (Li Zengzhi. Research and Application of Entomogenic Fungi in China (Vol. 1). Beijing: Academic Journal Press, 1988). Because Beauveria bassiana has the advantages of meeting some basic requirements of biological agents, that is, harmless to warm-blooded animals and plants, easy to cultivate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12R1/645
Inventor 刘吉平李香霖
Owner SOUTH CHINA AGRI UNIV
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