Geisenheim Research Center, Department of Phytomedicine, Von-Lade-Str. 1, D-65366 Geisenheim, Germany.
J Econ Entomol. 2010 Jun;103(3):898-907. doi: 10.1603/ec09381.
A couple of different members of the coleopteran genus Otiorhynchus (Coleoptera: Curculionidae) are becoming increasingly important as pests of nursery and ornamental plants in global horticulture. Although adult weevils are morphologically distinguishable by skilled personnel, high potential for misidentification is given for cryptic larval stages. For developing and applying efficient pest management strategies the determination of the respective species is however a prerequisite, because each species may have a different phenology or a varying susceptibility to pesticides. Here, we report on the development of a diagnostic polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method for differentiation among 16 Otiorhynchus and seven other weevil species independent of their developmental stage. An approximately 780-bp fragment of the mitochondrial cytochrome oxidase subunit II was amplified and subsequently digested with at most four restriction enzymes generating species-specific fragment patterns. The assay was validated on a total of 127 individuals and the obtained fragment patterns correctly identified 23 different weevil species. The PCR-RFLP method reported here is cost-effective, robust, and fast and could be used in the future by plant protection services for diagnostic purposes.
几种不同的鞘翅目象甲属（Coleoptera：Curculionidae）成员越来越成为全球园艺中苗圃和观赏植物的重要害虫。虽然成年象甲在形态上可以由熟练的人员区分，但由于隐蔽的幼虫阶段，存在很高的错误鉴定可能性。然而，为了制定和应用有效的害虫管理策略，确定各自的物种是前提条件，因为每个物种可能具有不同的物候或对农药的不同敏感性。在这里，我们报告了一种诊断聚合酶链反应（PCR）-限制性片段长度多态性（RFLP）方法的开发，该方法可在不考虑其发育阶段的情况下，区分 16 种象甲和其他 7 种象甲。扩增了线粒体细胞色素氧化酶亚基 II 的大约 780-bp 片段，然后用最多四种限制酶进行消化，产生种特异性片段模式。该测定在总共 127 个人上进行了验证，并且获得的片段模式正确识别了 23 种不同的象甲。本文报道的 PCR-RFLP 方法具有成本效益、稳健和快速的特点，将来可以由植物保护部门用于诊断目的。
