The European (Melolontha melolontha L.) and Forest (M. hippocastani F.) cockchafer are widespread pests throughout Central Europe. Both species exhibit a 3–5-year life cycle and occur in temporally shifted populations, which have been monitored and documented for more than 100 years. Visual identification of adults and larvae belonging to these morphologically similar species requires expertise and, particularly in the case of larvae, is challenging and equivocal. The goal of the study was the development of an efficient and fast molecular genetic tool for the identification and discrimination of M. melolontha and M. hippocastani. We established a collection of both species from Switzerland, Austria and Northern Italy in 2016, 2017 and 2018. An approximately 1550 bp long fragment of the cytochrome c oxidase subunit 1 (CO1) mitochondrial gene was amplified and sequenced in 13 M. melolontha and 13 M. hippocastani beetles. Alignment of the new sequences with reference sequences (NCBI GenBank and BOLDSYSTEMS databases) and subsequent phylogenetic analysis revealed consistent clustering of the two species. After the identification of M. melolontha and M. hippocastani species-specific single nucleotide polymorphisms (SNPs) in the CO1 alignment, we developed an effective SNP tool based on the ABI PRISM® SNaPshot™ Multiplex Kit for the rapid and accurate species discrimination of adults and larvae.

中文翻译：

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开发基于 SNP 的工具，用于识别和区分 Melolontha melolontha 和 Melolontha hippocastani

欧洲（甜瓜L.) 和森林 (M. hippocastaniF.) 金龟子是整个中欧地区普遍存在的害虫。这两个物种都表现出 3-5 年的生命周期，并出现在时间变化的种群中，这些种群已被监测和记录了 100 多年。视觉识别属于这些形态相似物种的成虫和幼虫需要专业知识，特别是在幼虫的情况下，具有挑战性和模棱两可。该研究的目标是开发一种高效、快速的分子遗传工具，用于识别和区分甜瓜和M. hippocastani. 我们在 2016 年、2017 年和 2018 年建立了来自瑞士、奥地利和意大利北部的这两个物种的集合。细胞色素 c 氧化酶亚基 1 (CO1) 线粒体基因的一个大约 1550 bp 长的片段在 13 中被扩增和测序甜瓜和 13M. hippocastani甲虫。新序列与参考序列（NCBI GenBank 和 BOLDSYSTEMS 数据库）的比对和随后的系统发育分析揭示了这两个物种的一致聚类。鉴定完毕后甜瓜和M. hippocastaniCO1 比对中的物种特异性单核苷酸多态性 (SNP)，我们基于 ABI PRISM® SNaPshot™ Multiplex Kit 开发了一种有效的 SNP 工具，用于快速准确地识别成虫和幼虫的物种。