Germain Jean-François, Chatot Catherine, Meusnier Isabelle, Artige Emmanuelle, Rasplus Jean-Yves, Cruaud Astrid
ANSES-Laboratoire de la Santé des Végétaux, Unité Entomologie et Plantes Invasives, CBGP, Campus international de Baillarguet, CS 30016, 34988 Montferrier-sur Lez, France.
Bull Entomol Res. 2013 Jun;103(3):354-62. doi: 10.1017/S000748531200079X. Epub 2013 Mar 1.
Epitrix species (Coleoptera: Chrysomelidae) feed mostly on plants from the family Solanaceae and some of them are major pests of potato crops. All Epitrix species are morphologically highly similar, which makes them difficult to identify and limits their study and management. Identification of species is mostly based on the observation of the genitalia and requires a high level of expertise. Here, we propose a tool to reliably identify all developmental stages of the most economically important Epitrix species feeding on potato in Europe and North America (Epitrix cucumeris, Epitrix similaris, Epitrix tuberis, Epitrix subcrinita and Epitrix hirtipennis). We first sequenced two DNA markers (mitochondrial cytochrome c oxidase I (COI) and nuclear internal transcribed spacer 2 (ITS2)) to test their effectiveness in differentiating among six Epitrix species (126 specimens). Morphospecies of Epitrix were well-differentiated by both DNA barcodes and no mitochondrial introgression was detected. Then, we developed an RFLP-based diagnostic method and showed that unambiguous species discrimination can be achieved by using the sole restriction enzyme TaqI on COI polymerase chain reaction products. The tool proposed here should improve our knowledge about Epitrix species biology, distribution and host range, three capacities that are particularly important in the detection and management of these pest species. Specifically, this tool should help prevent the introduction of E. tuberis and E. subcrinita in Europe and limit the spread of the recently introduced E. cucumeris and E. similaris, with minimal disruption to Solanaceae trade.
马铃薯叶甲属昆虫（鞘翅目：叶甲科）主要以茄科植物为食，其中一些是马铃薯作物的主要害虫。所有马铃薯叶甲属昆虫在形态上高度相似，这使得它们难以鉴定，并限制了对它们的研究和管理。物种鉴定主要基于生殖器的观察，需要高水平的专业知识。在此，我们提出一种工具，用于可靠地鉴定在欧洲和北美以马铃薯为食的最具经济重要性的马铃薯叶甲属物种的所有发育阶段（黄瓜马铃薯叶甲、相似马铃薯叶甲、块茎马铃薯叶甲、亚细马铃薯叶甲和多毛马铃薯叶甲）。我们首先对两个DNA标记（线粒体细胞色素c氧化酶I（COI）和核内转录间隔区2（ITS2））进行测序，以测试它们在区分六种马铃薯叶甲属物种（126个标本）方面的有效性。通过这两种DNA条形码都能很好地区分马铃薯叶甲属的形态物种，并且未检测到线粒体基因渗入。然后，我们开发了一种基于限制性片段长度多态性的诊断方法，并表明通过对COI聚合酶链反应产物仅使用限制性内切酶TaqI就可以实现明确的物种区分。这里提出的工具应该能增进我们对马铃薯叶甲属物种生物学、分布和寄主范围的了解，这三个方面在检测和管理这些害虫物种时尤为重要。具体而言，该工具应有助于防止块茎马铃薯叶甲和亚细马铃薯叶甲传入欧洲，并限制最近传入的黄瓜马铃薯叶甲和相似马铃薯叶甲的传播，同时对茄科贸易的干扰最小。
