A multiple PCR‐LDR assay for identification of two invasive cryptic species of whiteflies Bemisia tabaci
A multiple PCR‐LDR assay for identification of two invasive cryptic species of whiteflies Bemisia tabaci
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DOI：
10.1111/jen.12226
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发表时间：
2015-04
影响因子：
1.9
通讯作者：
Hongzhi Wang;J. Guo;L. Qi;Kai Li
中科院分区：
文献类型：
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作者：
Hongzhi Wang;J. Guo;L. Qi;Kai Li
烟粉虱的MEAM 1和MED种是重要的入侵害虫，在世界范围内造成巨大的作物损失。一个快速和高度可靠的分子技术是必要的，以确定这些物种，因为它们是形态上难以区分。因此，基于B的线粒体细胞色素氧化酶I（mtCOI）基因多态性的多重聚合酶链反应结合连接酶检测反应（PCR-LDR）。烟粉虱是用来区分这两个神秘物种的。对该方法的评估表明，PCR-LDR在区分MEAM 1（SHB）和MED（SHQ）粉虱方面具有较高的特异性和灵敏度。在田间试验中，在一个96孔板中进行PCR-LDR基因分型，以鉴定来自上海郊区8个区的93个个体。在PCR-LDR和测序方法之间观察到完全一致性。用该方法对两个地区进行了MEAM 1和MED的检测，而在其他6个地区只检测到MED。PCR-LDR是一种可移植的平台，为B的种属鉴定提供了一种替代方法。烟粉虱成本低。
The MEAM1 and MED species of the cryptic species complex Bemisia tabaci are important invasive pests that cause tremendous crop losses worldwide. A rapid and highly reliable molecular technique is necessary to identify these species because they are morphologically indistinguishable. Therefore, a multiple polymerase chain reaction coupled with a ligase detection reaction (PCR‐LDR) that was based on polymorphisms in the mitochondrial cytochrome oxidase I (mtCOI) gene of B. tabaci was developed to distinguish the two cryptic species. An assessment of the method indicated that PCR‐LDR provided high specificity and sensitivity in discriminating MEAM1 (SHB) and MED (SHQ) whiteflies. In field tests, PCR‐LDR genotyping was performed in one 96‐well plate to identify 93 individuals collected from 8 districts in the suburbs of Shanghai. Complete concordance was observed between PCR‐LDR and sequencing methods. The method was used to confirm that MEAM1 and MED were found in two districts, but only the MED was found in the other six districts. PCR‐LDR, which is a transplantable platform, provides an alternative method for species identification of B. tabaci at low cost.