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Method for horizontal transfection of exogenesis endosymbiosis bacterium to bemisia tabaci gennadius

A technology of bemisia tabaci and symbiotic bacteria, applied in the field of agricultural biology, can solve the problems of poor fixation and easy death of bemisia tabaci adults, and achieve the effect of simple method, good technical platform and increased infection rate

Inactive Publication Date: 2011-10-19
山东省农业科学院高新技术研究中心
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Problems solved by technology

In addition, when conducting other tests on whitefly at home and abroad, such as the RNAi injection test, paralyzed whitefly adults are used, but the whitefly adults wake up quickly and are not easy to fix, and the fixed whitefly adults are easy to die. It is also a problem that needs to be solved in the process of experimenting with Bemisia tabaci

Method used

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  • Method for horizontal transfection of exogenesis endosymbiosis bacterium to bemisia tabaci gennadius
  • Method for horizontal transfection of exogenesis endosymbiosis bacterium to bemisia tabaci gennadius

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Embodiment 1

[0026] A method for horizontal transfection of exogenous endosymbiotic bacteria to whitefly whitefly, comprising preparation of exogenous endosymbiotic bacteria Wolbachia (Wolbachia), preparation of whitefly pseudopupal receptors, injection, and injection of whitefly whitefly For the cultivation of pseudopupal receptors, the specific steps are as follows:

[0027] With the eggs of the rice moth as the donor of the symbiotic bacteria Wolbachia (Wolbachia), 30 μL of the eggs of the rice moth were placed in a 200 uL sterilized tube, and 100 uL of SPG buffer (218 mM sucrose, 3.8 mM KH 2 PO 4 , 7.2 mM K 2 HPO 4 , 4.9mM L-glutamic acid, pH7.2) rinsed 4 times; In a 0.2mL sterile tube, use 100uL of SPG buffer to homogenize the eggs, centrifuge at 300g for 5min, and transfer the supernatant to a new tube ( Centrifuge at 12000g for 10min in 0.2mL), remove the supernatant after centrifugation, and resuspend the pellet with SPG buffer;

[0028] Centrifuge the suspension at 300g for 5 ...

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Abstract

The invention relates to a method for the horizontal transfection of bemisia tabaci of the exogenous endosymbiotic bacteria, which pertains to the field of agricultural biotechnology. The preparation method includes: the purification of the exogenous endosymbiotic bacteria Wolbachia, the collection and fixation of a bemisia tabaci receptor, the horizontal transfection of the exogenous endosymbiotic bacteria Wolbachia to the bemisia tabaci receptor, the collection and fixation of an injection individual and the detection of the endosymbiotic bacteria. The invention is simple and quick, which can successfully transfect the exogenous endosymbiotic bacteria to a bemisia tabaci individual, and a bemisia tabaci offspring has higher infection rate.

Description

(1) Technical field [0001] The invention relates to a method for horizontal transfection of exogenous endosymbiotic bacteria to whitefly, and belongs to the field of agricultural biotechnology. (2) Background technology [0002] Biological invasion often poses a threat to the economic security, ecological security, and social security of a country or region. The direct economic losses caused by biological invasions in my country amount to hundreds of billions of yuan every year. Among them, 13 alien species such as Eupatorium adenophorum and pine xylophilus cause economic losses of 57.4 billion yuan to agriculture, forestry, animal husbandry and fishery production every year. . At present, Bemisia tabaci (type B) that has invaded our country is breaking out in vast areas. It is the only worldwide invasive insect that has been dubbed a "super pest" in the history of the international scientific and technological circles. It not only directly sucks plant juice, secretes honey ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/89
Inventor 褚栋陶云荔刘国霞方华丛斌
Owner 山东省农业科学院高新技术研究中心
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