A molecular identification key for economically important thrips species (Thysanoptera: Thripidae) using direct sequencing and a PCR-RFLP-based approach
A molecular identification key for economically important thrips species (Thysanoptera: Thripidae) using direct sequencing and a PCR-RFLP-based approach
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DOI：
10.1046/j.1461-9563.2002.00132.x
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发表时间：
2002-05-01
影响因子：
1.6
通讯作者：
Frey, J. E.
中科院分区：
文献类型：
--
作者：
Brunner, P. C.;Fleming, C.;Frey, J. E.
1 针对病原体或害虫的治疗通常非常具体，作为基本的第一步，需要能够正确、明确地识别分类群。我们使用 PCR 扩增技术成功建立了重要经济蓟马物种的分子鉴定关键。2 通过自动直接测序和 RFLP 分析了线粒体 COI 编码基因的 PCR 扩增 433 bp 长片段。对代表 10 个命名物种的 264 个蓟马个体进行测序，检测到 17 个单倍型。物种内变异较低，而物种间变异较高，导致平均序列差异为 18.6%，平均成对物种分化（计算为 FsT 值）为 0.9896.3 两种限制性内切酶（Alul、Sau3A1）产生的模式可以明确识别所有蓟马物种。4 对密钥质量的统计支持通过 (i) 高度显着的排列方法提供，分配个体单倍型正确的物种组和 (ii) 分层 NJ 聚类分析，其中所有同种个体序列在最大 (100%) 引导支持下聚类在一起。5 这项研究表明，遗传标记的使用代表了一种有价值的替代方案，例如流行病学研究，其中使用经典形态学进行正确识别
1 Treatments against pathogens or pests are often very specific and, as a fundamental first step, require the ability to identify taxa correctly and unambiguously. We used PCR amplification techniques to successfully establish a molecular identification key for economically important thrips species.2 A PCR amplified 433 bp long fragment of the mitochondrial COI coding gene was analysed by automated direct sequencing and RFLP. Sequencing of 264 individual thrips representing 10 named species detected 17 haplotypes. Variation within species was low, whereas among species variation was high resulting in an average sequence divergence of 18.6% and an average pairwise species differentiation (calculated as FsT-value) of 0.9896.3 Two restriction enzymes (Alul, Sau3A1) produced patterns that allowed unambiguous identification of all thrips species.4 Statistical support for the quality of the key was given by (i) a highly significant permutation approach, assigning individual haplotypes to the correct species groups and (ii) a hierarchical NJ cluster analysis in which all conspecific individual sequences clustered together with maximal (100%) bootstrap support.5 This study has shown that the use of genetic markers represents a valuable alternative for situations, such as epidemiological research, in which correct identification with classical morphological