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Extraction method of Tobacco whitefly endosymbiont mycetocytes

A technology for Bemisia tabaci and bacterial cells, which is applied in the field of extraction of endosymbiotic bacterial cells of Bemisia tabaci, and can solve problems such as interference and inability to exclude exogenous microorganisms in a host.

Active Publication Date: 2013-11-13
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0008] The invention relates to a method for extracting whitefly endosymbiotic bacterial cells, which aims to solve the problem that in the existing research on insect symbiotic bacteria, the conventional method of extracting symbiotic bacteria cannot eliminate the interference of exogenous microorganisms in the host body, and obtain high-purity, high-concentration symbiotic bacteria. Sufficiently large insect symbiotic bacterial cells

Method used

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  • Extraction method of Tobacco whitefly endosymbiont mycetocytes
  • Extraction method of Tobacco whitefly endosymbiont mycetocytes
  • Extraction method of Tobacco whitefly endosymbiont mycetocytes

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Embodiment 1

[0040] 1) Collect 1000-2000 whitefly adults, soak them in a 5ml sterilized centrifuge tube containing 2ml 70% ethanol for a short time, let it stand for 1 minute, after the insects settle, remove the upper layer of alcohol, and collect the bottom of the centrifuge tube Worm, repeat the above steps 3 times to disinfect the surface of the worm;

[0041] 2) Repeatedly rinse with 2 ml of sterilized double distilled water for at least 3 times, each time for 5-10 minutes;

[0042] 3) Place the sterilized worm under a dissecting microscope, remove the head, thorax, midgut and other tissues of the worm with micro-dissecting forceps, and collect the remaining tissue into a clean 2 ml centrifuge tube;

[0043] 4) Put the collected worms in 0.5 ml of PBS buffer, use a micro electric tissue homogenizer and grind into a coarse sample, wherein, the composition of the PBS buffer is 10mM phosphate-buffered saline, pH 7.4; 137mM NaCl ; 2.7mM KCl; 10mM Na 2 HPO 4 ; 2mM KH 2 PO 4 , when gri...

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Abstract

The invention relates to an extraction method of insect endosymbiont mycetocytes, and concretely relates to an extraction method of Tobacco whitefly endosymbiont mycetocytes. The method mainly comprises the steps of dissecting, filtration and discontinuous density gradient centrifugation, and the interferences of exogenous microbes can be fully removed layer by layer through the steps to obtain pure and adequate mycetocytes, so the influences of hosts and in-vivo exogenous microbes thereof are eliminated in the subsequent researches of the functions of endosymbionts, thereby the Tobacco whitefly endosymbiont mycetocytes can be used in the subsequent tests of the sequencing and active substance separation of the whole genome of the endosymbionts to comprehensively and deeply understand the functions of the Tobacco whitefly endosymbionts.

Description

technical field [0001] The invention relates to a method for extracting endosymbiotic bacterial cells of insects, in particular to a method for extracting endosymbiotic bacterial cells of whitefly whitefly. Background technique [0002] Whitefly (Bemisia tabac Gennadaius) belongs to the subfamily A. Due to its morphological variability, many synonyms have been produced, such as Tobacco whitefly, Cotton whitefly or Sweetpotato whitefly. Bemisia tabaci is a polyphagous pest with a wide range of hosts. There are more than 500 species of host plants in 74 families, such as Fabaceae, Compositae, Malvacaceae, Solanaceae, Euphorbiaceae, Cucurbitaceae, Convolvulaceae and Cruciferae, etc. , mainly harm cotton, vegetables, flowers and other economic crops, including cotton, kenaf, peanut, pigeonpea, chickpea, Shoukai, kidney bean, pea, broad bean, pepper, tomato, nicotine, nightshade, Sweet potato, cantaloupe, zucchini, cassava, rubber, sweet cassava, bitter cassava, cassava, brassi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 郭建洋万方浩赵瑞华龙楚云
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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