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Isolated culture method for nilaparvata lugens in-vivo yeast-like symbiotic bacteria and special culture medium

An in vitro culture, brown planthopper technology, applied in the field of microbial culture, can solve the problems of easy to cause pollution, difficult to remove pollution, no mention of medium formula, etc., and achieve the effect of reasonable compatibility and reasonable design.

Inactive Publication Date: 2011-04-13
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the fusion of yeast-like symbionts and host N. lugens, it is difficult to exclude the contamination from the environment of the insects by isolating the fat body cells of the host. The in vitro culture of yeast-like symbionts in insects was carried out in N. lugens (Socho et al., 1981) and N. lugens. Although there are reports on the tobacco beetle (Noda et al., 1996), both of them adopt the method of cultivating worms, which is easy to cause pollution, and the specific medium formula is not mentioned, and the in vitro subculture of symbiotic bacteria cannot be obtained. Purpose

Method used

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  • Isolated culture method for nilaparvata lugens in-vivo yeast-like symbiotic bacteria and special culture medium
  • Isolated culture method for nilaparvata lugens in-vivo yeast-like symbiotic bacteria and special culture medium
  • Isolated culture method for nilaparvata lugens in-vivo yeast-like symbiotic bacteria and special culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Composition and ratio of a special medium for cultivating yeast-like symbionts in brown planthopper

[0030] L-aspartic acid 10mg / L, L-arginine-hydroxide 50mg / L, L-asparagine 50 mg / L, L-alanine 30 mg / L, B-alanine 10mg / L, L-cystine-hydroxide 5 mg / L, L-glutamic acid 10mg / L, L-glutamine 100 mg / L, L-aminoacetic acid 10 mg / L, L-group Amino acid 300 mg / L, L-isoleucine 1mg / L, L-leucine 5 mg / L, L-lysine-hydroxide 100 mg / L, L-methionine 5 mg / L L, L-proline 50 mg / L, L-phenylalanine 10 mg / L, DL-serine 100 mg / L, L-tyrosine 10mg / L, L-tryptophan 5mg / L, L -threonine 50 mg / L, L-valine 50mg / L, glucose 10mg / L, streptomycin sulfate 17.5 mg / L, malic acid 50 mg / L, α-ketoglutaric acid 50mg / L, Succinic acid 1mg / L, fumaric acid 10 mg / L, riboflavin 0.5mg / L, calcium pantothenate 0.001mg / L, folic acid 1mg / L and niacin 0.25mg / L.

[0031]

Embodiment 2

[0032] Example 2: Components and proportions of a special medium for cultivating yeast-like symbionts in brown planthopper

[0033] L-aspartic acid 15mg / L, L-arginine-hydroxide 10mg / L, L-asparagine 18mg / L, L-alanine 10 mg / L, B-alanine 30mg / L, L-cystine-hydroxide 1 mg / L, L-glutamic acid 50 mg / L, L-glutamic acid 40 mg / L, L-aminoacetic acid 75 mg / L, L-histamine Acid 100 mg / L, L-isoleucine 5mg / L, L-leucine 10 mg / L, L-lysine-hydroxide 50 mg / L, L-methionine 1mg / L, L-proline 30 mg / L, L-phenylalanine 25 mg / L, DL-serine 50 mg / L, L-tyrosine 5mg / L, L-tryptophan 25.5mg / L, L- Threonine 30 mg / L, L-valine 25 mg / L, glucose 10 mg / L, streptomycin sulfate 25 mg / L, malic acid 75 mg / L, α-ketoglutarate 40 mg / L, succinic acid 3.5mg / L, fumaric acid 5 mg / L, riboflavin 0.001mg / L, calcium pantothenate 0.5mg / L, folic acid 0.01mg / L and niacin 0.1mg / L.

[0034]

Embodiment 3

[0035] Example 3: Composition and ratio of a special medium for cultivating yeast-like symbiotic bacteria in brown planthopper

[0036] L-aspartic acid 50mg / L, L-arginine-hydroxide 100mg / L, L-asparagine 10mg / L, L-alanine 50 mg / L, B-alanine 50mg / L, L-cystine-hydroxide 3 mg / L, L-glutamic acid 100 mg / L, L-glutamic acid 25 mg / L, L-aminoacetic acid 100 mg / L, L-histamine Acid 500 mg / L, L-isoleucine 10mg / L, L-leucine 1 mg / L, L-lysine-hydroxide 60 mg / L, L-methionine 10mg / L, L-proline 10 mg / L, L-phenylalanine 50 mg / L, DL-serine 150 mg / L, L-tyrosine 1mg / L, L-tryptophan 50mg / L, L-threo Amino acid 10 mg / L, L-valine 5 mg / L, glucose 100 mg / L, streptomycin sulfate 50 mg / L, malic acid 100 mg / L, α-ketoglutaric acid 25 mg / L, succinic acid 10 mg / L, fumaric acid 1 mg / L, riboflavin 1 mg / L, calcium pantothenate 0.1 mg / L, folic acid 0.25 mg / L and niacin 0.01 mg / L.

[0037]

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Abstract

The invention discloses an isolated culture method for nilaparvata lugens in-vivo yeast-like symbiotic bacteria and a special culture medium, and belongs to the technical field of microculture. The special culture medium comprises 31 nutrient substances needed by nilaparvata lugens in-vivo yeast-like symbiotic bacteria, and the culture method comprises the following steps of: feeding adult nilaparvata lugen; treating ovaries of nilaparvata lugens; preparing culture solution, performing isolated culture and the like. By employing the culture method, two yeast-like strains are isolated from the nilaparvata lugens in vivo so as to provide basis for further researching genetic background of the yeast-like symbiotic bacteria and the interavailable nutrient relation between the yeast-like symbiotic bacteria and host nilaparvata lugens, and modifying the symbiotic bacteria to control the harm of the nilaparvata lugens. Meanwhile, the special culture medium is reasonable in components and provides necessary nutrition basis for isolated subculture of the nilaparvata lugens in-vivo yeast-like symbiotic bacteria. The method also provides technical support for isolated culture of other homoptera insect in-vivo yeast-like symbiotic bacteria.

Description

technical field [0001] The invention belongs to the technical field of microorganism culture, and in particular relates to an in vitro culture method and a special medium for yeast-like symbiotic bacteria in brown planthoppers. Background technique [0002] brown planthopper Nilaparvata lugens St?l is an important long-distance migratory pest in rice production in my country and Southeast Asian countries, which seriously threatens the safe production of rice in my country. The annual occurrence area in my country is about 13-20 million hm 2 , accounting for about 50% of the rice planting area, with an average annual loss of 1.0-1.5 billion kg of rice. Planting insect-resistant varieties and using chemical insecticides are the most economical and effective control methods to control the damage of BPH. medicinal production. Studies have shown that there are common symbiotic bacteria in homopteran insects, such as planthoppers, aphids, leafhoppers, etc. These symbiotic bact...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12R1/72
Inventor 张珏锋陈建明陈列忠何月平俞晓平
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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