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首页> 外文期刊>Journal of Economic Entomology >Improved DNA Barcoding Method for Bemisia tabaci and Related Aleyrodidae: Development of Universal and Bemisia tabaci Biotype-Specific Mitochondrial Cytochrome c Oxidase I Polymerase Chain Reaction Primers
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Improved DNA Barcoding Method for Bemisia tabaci and Related Aleyrodidae: Development of Universal and Bemisia tabaci Biotype-Specific Mitochondrial Cytochrome c Oxidase I Polymerase Chain Reaction Primers

机译：烟粉虱和相关粉虱的改进的DNA条形码编码方法：通用和烟粉虱生物型特异性线粒体细胞色素c氧化酶I聚合酶链反应引物的开发

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摘要

Whiteflies, heteropterans in the family Aleyrodidae, are globally distributed and severe agricultural pests. The mitochondrial cytochrome c oxidase I (mtCOI) sequence has been used extensively in whitefly phylogenetic comparisons and in biotype identification of the agriculturally important Bemisia tabaci (Gennadius) whitefly. Because of the economic importance of several whitefly genera, and the invasive nature of the B and the Q biotypes of Bemisia tabaci, mtCOI sequence data are continually generated from sampled populations worldwide. Routine phylogenetic comparisons and biotype identification is done through amplification and sequencing of an __800-bp mtCOI DNA fragment. Despite its routine use, published primers for amplification of this region are often inefficient for some B. tabaci biotypes and especially across whitefly species. Through new sequence generation and comparison to available whitefly mtCOI sequence data, a set of polymerise chain reaction (PCR) amplification primers (Btab-Uni primers) were identified that are more efficient at amplifying__748 bp of the __800-bp fragment currently used. These universal primers amplify an mtCOI fragment from numerous B. tabaci biotypes and whitefly genera by using a single amplification profile. Furthermore, mtCOI PCR primers specific for the B, Q, and New World biotypes of B. tabaci were designed that allow rapid discrimination among these biotypes. These primers produce a 478-, 405-, and 303-bp mtCOI fragment for the B, New World, and Q biotypes, respectively. By combining these primers and using rapid PCR and electrophoretic techniques, biotype determination can be made within 3 h for up to 96 samples at a time.
机译：粉虱是无翅目科中的异翅类动物，是全球分布和严重的农业害虫。线粒体细胞色素C氧化酶I（mtCOI）序列已广泛用于粉虱的系统发育比较和农业重要烟粉虱（Gennadius）粉虱的生物型鉴定中。由于几个粉虱属的经济重要性，以及烟粉虱的B型和Q型生物型的侵袭性，mtCOI序列数据不断从全球抽样人群中产生。通过对__800 bp mtCOI DNA片段进行扩增和测序，可以进行常规的系统发育比较和生物型鉴定。尽管已常规使用，但已发布的用于扩增该区域的引物对于某些烟粉虱生物型尤其是粉虱种而言通常效率低下。通过新的序列生成并与可用的粉虱mtCOI序列数据进行比较，鉴定出了一组聚合链反应（PCR）扩增引物（Btab-Uni引物），它们在扩增目前使用的__800 bp片段的__748 bp时效率更高。这些通用引物通过使用单个扩增图谱扩增来自许多烟粉虱生物型和粉虱属的mtCOI片段。此外，设计了针对烟粉虱的B，Q和New World生物型特异的mtCOI PCR引物，可以快速区分这些生物型。这些引物分别为B，New World和Q生物型产生478、405和303 bp的mtCOI片段。通过组合这些引物并使用快速PCR和电​​泳技术，可以在3小时内一次确定多达96个样品的生物型。

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