Method for preventing and treating bemisa babaci by utilizing RNA (Ribose Nucleic Acid) interference technology

A technology of RNA interference and whitefly, applied in the field of agricultural biology, can solve the problems of death, cannot be directly fed to whitefly, cannot be used in large quantities to control whitefly, and achieves the effect of broad application prospects

Inactive Publication Date: 2011-10-12
山东省农业科学院高新技术研究中心
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Problems solved by technology

[0005] Because Bemisia tabaci is a piercing-sucking insect and sucks plant juice, it cannot be directly fed with dsRNA to Bemisia tabaci; RNA interference can also be performed by injection, but it cannot be widely used in the control of Bemisia tabaci, and because the body of Bemisia tabaci Smaller, the injection easily damages the whitefly, leading to its death

Method used

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  • Method for preventing and treating bemisa babaci by utilizing RNA (Ribose Nucleic Acid) interference technology
  • Method for preventing and treating bemisa babaci by utilizing RNA (Ribose Nucleic Acid) interference technology
  • Method for preventing and treating bemisa babaci by utilizing RNA (Ribose Nucleic Acid) interference technology

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Embodiment 1

[0041] A method utilizing RNA interference technology to prevent and control whitefly, the steps are as follows:

[0042] (1) According to Annex 1 in Leshkowitz et al. (2006) Whitefly (Bemisia tabaci) genome project, select the target sequence IAP gene of RNAi, and design PCR amplification primers for the target sequence, and introduce BglII digestion at the 5' end of the upstream primer Site, the 5' end of the downstream primer introduces the Xho I restriction site.

[0043] The primer sequence is as follows: IAP F: 5'- GAAGATCTTC GGCGATGTTCCTTGTTTAG-3', IAP R: 5- CCGCTCGAGCGG ACTGAGATGATTCCGAGAC-3', BglII and XhoI sites (underlined) were introduced into the 5' ends of the upstream and downstream primers.

[0044] (2) Collect Bemisia tabaci, press TaKaRa RNAiso TM Plus instructions to extract its RNA, with the reverse transcription kit of MBI company (RevertAid TM First Strand cDNA Synthesis Kit, #K1622) was reverse transcribed to synthesize the first strand of cDNA. ...

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Abstract

The invention provides a method for preventing and treating bemisa babaci by utilizing an RNA (Ribose Nucleic Acid) interference (RNAi) technology, belonging to the technical field of agriculture and biology. The method provided by the invention comprises the following steps of: (1) selecting a target sequence IAP (Inhibitor Of Apoptosis Protein) gene of RNAi according to a gene sequence of the bemisa babaci and designing a PCR (Polymerase Chain Reaction) amplification primer of the target sequence; (2) extracting the RNA of the bemisa babaci and carrying out transcription to obtain a cDNA (Complementary Deoxyribonucleic Acid); then carrying out PCR amplification on the cDNA and purifying to prepare a target gene; (3) inserting the target gene into an expression vector to prepare the expression vector containing the target gene; (4) and transforming the expression vector containing the target gene into a crop to obtain a transgenic crop for preventing and treating bemisa babaci; and planting the crop. The method provided by the invention has important guiding value and theoretical significance on researching the gene function of the bemisa babaci by utilizing the RNAi technology, preventing and treating the bemisa babaci by utilizing the RNAi technology, and preventing and treating the bemisa babaci by utilizing the transgenic crop.

Description

technical field [0001] The invention relates to a method for controlling Bemisia tabaci by using RNA interference technology, and belongs to the field of agricultural biotechnology. Background technique [0002] Bemisia tabaci (Gennadius) is widely distributed in all continents except Antarctica, and is one of the main pests of cotton, vegetables, garden flowers and other plants and field crops in tropical, subtropical and adjacent temperate regions (Brown et al., 1995). It not only feeds on the juice of host plants, but also can transmit more than 100 kinds of viruses, and secretes a large amount of honeydew, which induces soot and affects photosynthesis (Liu et al., 2006; Jiu et al., 2007). Bemisia tabaci is also the only insect that has been dubbed a "super bug" (superbug) in the history of science and technology. It has caused serious economic losses to agriculture in many countries in the world in the past 20 years. Bemisia tabaci has broken out and caused serious dam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N15/11A01H5/00
Inventor 郭强褚栋夏晗王兴军毕玉平安立国范仲学刘国霞陶云荔
Owner 山东省农业科学院高新技术研究中心
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