温室白粉虱SCAR分子鉴定技术的建立及应用
Establishment and application of SCAR marker for rapid identification of Trialeurodes vaporariorum
  
DOI:10.3969/j.issn.1674-0858.2012.03.05
中文关键词:  温室白粉虱  RAPD  SCAR标记  分子鉴定
英文关键词:Trialeurodes vaporariorum  RAPD  SCAR marker  molecular identification
基金项目:国家自然科学基金项目(31171857);现代农业产业技术体系建设专项资金 (CARS-26-10);农业部园艺作物生物学与种质创制重点实验室项目经费
Author NameAffiliation
WANG Jin-Na 1.Department of Life Science, Northeast Agricultural University, Harbin 150030,China2.Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081,China 
WANG Xiang-Jing Department of Life Science, Northeast Agricultural University, Harbin 150030,China 
ZHANG You-Jun Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081,China 
WANG Shao-Li Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081,China 
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中文摘要:
      温室白粉虱Trialeurodes vaporariorum是为害我国蔬菜作物的重大害虫之一。本研究采用随机扩增多态性DNA(Random Amplified Polymorphic DNA, RAPD)技术对温室白粉虱进行研究,筛选出一条620 bp的特异性片段(GenBank登录号为JQ690764),据此片段的测序结果设计一对特异性的序列特异性扩增区(Sequence characterized Amplified Region,SCAR)标记(Tv-F和Tv-R),可成功从室内饲养的和从不同地区田间采集的温室白粉虱的基因组DNA中扩增出一条412 bp的特异性条带,而不能从供试的其他粉虱类害虫中扩增出该相应条带。单头温室白粉虱成虫的基因组DNA稀释1000倍时,该SCAR标记仍可成功扩增出预期条带,显示出极高的灵敏度。该SCAR标记对温室白粉虱的基因组DNA扩增重复性和稳定性好,且操作简便,灵敏度高,可用于样品的大规模检测,为田间温室白粉虱的快速识别鉴定及其有效防治提供了技术基础。
英文摘要:
      Greenhouse whitefly, Trialeurodes vaporariorum, is an important insect pest in vegetable crops in China. In the present study, Random Amplified Polymorphic DNA (RAPD) technique was used to amplify the T.vaporariorumand one specific fragment of 620 bp was screened and sequenced with the accession number JQ690764 in GenBank. One pair of SCAR (Sequence Characterized Amplified Regions) primers (Tv-F/Tv-R) which are specific to T. vaporariorum was designed on the basis of sequencing results of this fragment. The SCAR primers could be used to amplify a 412 bp fragment successfully from T.vaporariorumpopulations in laboratory and those collected from 6 different sites in China, but not from other familiar whitefly species in this study. The SCAR marker could amplify the 412 bp band when the genomic DNA of a single T.vaporariorumadult was diluted to 103fold, showing very high susceptibility.This specific SCAR primers can only amplify the genomic DNA of T.vaporariorumwith high reproducibility and susceptibility, and are easy to operate, which can be applied to detect the large scale specimen and provide a technical basis for rapid identification and effective control for T.vaporariorumin the field.
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