| 王金娜,王相晶,张友军,王少丽,2012,温室白粉虱SCAR分子鉴定技术的建立及应用[J].环境昆虫学报,34(3):295-301 |
| 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 |
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| 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 |
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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.vaporariorumand 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.vaporariorumpopulations 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.vaporariorumadult was diluted to 103fold, showing very high susceptibility.This specific SCAR primers can only amplify the genomic DNA of T.vaporariorumwith 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.vaporariorumin the field. |
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