| 利用TaqMan等位基因技术鉴定烟粉虱MEAM1和MED隐种 |
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| 引用本文： | 姚晶,郭晓军,王甦,王昱超,罗晨,张帆,李绍勤.利用TaqMan等位基因技术鉴定烟粉虱MEAM1和MED隐种[J].昆虫学报,2013,56(1):98-103. |
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| 作者姓名： | 姚晶 郭晓军 王甦 王昱超 罗晨 张帆 李绍勤 |
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| 作者单位： | （1. 华中农业大学植物科学技术学院, 湖北省昆虫资源利用与害虫可持续治理重点实验室, 武汉 430070; 2. 北京市农林科学院植物保护环境保护研究所, 北京 100097） |
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| 基金项目： | 国家重点基础研究发展规划项目(2009CB119200);“十二五”科技支撑计划课题(2012BAD19B06) |
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| 摘 要： | MEAM1和MED是烟粉虱Bemisia tabaci两种重要的外来入侵隐种, 在我国部分地区常混合发生, 对我国农业生产造成了不同程度的危害和损失。尤其是MED隐种危害寄主范围更广, 对多种杀虫剂具有较高抗性, 防治上更为困难。因此, 如何快速鉴定烟粉虱MEAM1和MED隐种, 对于烟粉虱防治策略的选择具有十分重要的意义。本研究选择线粒体细胞色素氧化酶I(mitochondrial cytochrome oxidase I, mtDNA COI)基因保守区域内的单核苷酸多态性（SNP）为靶标, 应用等位基因聚合酶链式反应技术, 借助TaqMan-MGB荧光染色标记探针, 建立了一种鉴定烟粉虱MEAM1和MED隐种的等位基因选择性PCR方法, 并对北京11个区县的14个烟粉虱种群进行了隐种鉴定。结果表明, 北京地区14个烟粉虱种群样本与已知烟粉虱MED隐种种群在荧光值分布上聚为一簇, 为MED隐种。该鉴定方法具备SNP基因分型的优点, 可快速、 可靠、 高通量地鉴定烟粉虱MEAM1和MED, 为烟粉虱隐种鉴定及遗传分化研究提供了新的可选途径。
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| 关 键 词： | 烟粉虱 隐种 mtDNA COI SNP 等位基因选择性PCR 荧光定量 |
Discrimination of cryptic species MEAM1 and MED of Bemisia tabaci (Hemiptera: Aleyrodidae) by using TaqMan allele-selective PCR |
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| YAO Jing,GUO Xiao-Jun,WANG Su,WANG Yu-Chao,LUO Chen, ZHANG Fan,LI Shao-Qin.Discrimination of cryptic species MEAM1 and MED of Bemisia tabaci (Hemiptera: Aleyrodidae) by using TaqMan allele-selective PCR[J].Acta Entomologica Sinica,2013,56(1):98-103. |
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| Authors: | YAO Jing GUO Xiao-Jun WANG Su WANG Yu-Chao LUO Chen ZHANG Fan LI Shao-Qin |
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| Affiliation: | (1. Hubei Insect Resource Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; 2. Institute of Plant and Environmental Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China) |
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| Abstract: | The whitefly Bemisia tabaci is considered taxonomically as a species complex which contains some destructive pests worldwide. Two of the most prevalent cryptic species are B. tabaci Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED). They are invasive cryptic species and often occur in some areas in China. The MED particularly has strong resistance to some insecticides and is more difficult to be controlled. In this study TaqMan allele-selective PCR was used for high-throughput allelic discrimination of MEAM1 and MED. A single nucleotide polymorphism (SNP) in a conservative region of the mitochondrial cytochrome oxidase I (mtDNA COI) gene was selected as the amplification target. The TaqMan-MGB fluorescent dye-labeled was used in real-time quantitative PCR. Fourteen populations of B. tabaci collected from Beijing were identified with the method. The results showed that all samples of B. tabaci clustered with the known MED species as shown by the higher fluorescence of FAM dye in scatter plot analysis of fluorescence data, suggesting that all Beijing populations examined are MED. The results prove that the method has the advantage of SNP genotyping, with which MEAM1 and MED can be identified rapidly, reliably and high-throughput, and this provides a new alternative way for identification of cryptic species and important information for management strategy selection. |
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| Keywords: | Bemisia tabaci" target="_blank">Bemisia tabaci')" href="#">Bemisia tabaci cryptic species mtDNA COI SNP allele-selective PCR fluorescent quantitation |
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