Bertin Sabrina, Bosco Domenico
DIVAPRA-Entomologia e Zoologia applicate all'Ambiente Carlo Vidano, Università degli Studi di Torino, Grugliasco, Italy.
Methods Mol Biol. 2013;938:87-108. doi: 10.1007/978-1-62703-089-2_8.
The correct identification of the insect species involved in phytoplasma transmission is an essential condition for managing phytoplasma diseases and employing control strategies. The taxonomy of leafhoppers, planthoppers, and psyllids traditionally relies on morphological characters. Unfortunately, the identification of the distinctive traits requires skills and experience possessed by only a few specialist entomologists. In this chapter we provide protocols for the molecular identification of phytoplasma-vector species, mainly based on the polymerase chain reaction (PCR) amplification of mitochondrial and ribosomal DNA. Protocols for the application of molecular identification keys to dried specimens stored in insect collections are also provided. The same total DNA preparations can serve as a PCR template for either insect species or phytoplasma identification. The molecular identification methods can be applied not only to males, but also to nymphs and females for which a morphological taxonomic tool is generally unavailable. We suggest that taxonomic databases of planthoppers, leafhoppers, and psyllids should include species-specific DNA sequences as soon as they become available.
正确识别参与植原体传播的昆虫种类是管理植原体病害和采用防治策略的必要条件。叶蝉、飞虱和木虱的分类传统上依赖形态特征。不幸的是,识别这些独特特征需要只有少数专业昆虫学家才具备的技能和经验。在本章中,我们提供了用于植原体传播媒介物种分子鉴定的方案,主要基于线粒体和核糖体DNA的聚合酶链反应(PCR)扩增。还提供了将分子鉴定密钥应用于保存在昆虫标本馆中的干燥标本的方案。相同的总DNA制备物可作为昆虫物种或植原体鉴定的PCR模板。分子鉴定方法不仅可以应用于雄性,也可以应用于若虫和雌性,而对于后者通常没有形态分类工具。我们建议,一旦有可用的飞虱、叶蝉和木虱物种特异性DNA序列,就应将其纳入分类数据库。