Laboratory of Applied Entomology, Graduate School of Horticulture, Chiba University 648 Matsudo, 271-8510 Chiba, Japan.
J Econ Entomol. 2011 Aug;104(4):1280-5. doi: 10.1603/ec10442.
Three plusiine species, Autographa nigrisigna, Macdunnoughia confusa, and Thysanoplusia intermixta (Lepidoptera: Noctuidae), are commonly found together in lettuce, Lactuca sativa L., fields in Japan. Given the marked morphological similarities between these species and the difficulty associated with discriminating between them using only visual cues, we used multiplex polymerase chain reaction (PCR) assay to distinguish between the three target species. Multiplex PCR uses four primers to simultaneously amplify a specific region of the mitochondrial DNA and produce species-specific banding patterns. The stringency of the method was tested using specimens of different sex, location, and developmental stage, and consistent results were obtained for all samples. Indeed, our method has the potential to clarify the species structure of plusiine species in lettuce fields.
三种夜蛾科（Lepidoptera: Noctuidae）的 plusiine 物种，Autographa nigrisigna、Macdunnoughia confusa 和 Thysanoplusia intermixta，在日本的生菜（Lactuca sativa L.）田中常同时出现。鉴于这些物种之间存在明显的形态相似性，并且仅使用视觉线索很难将它们区分开来，我们使用多重聚合酶链反应（PCR）检测来区分这三个目标物种。多重 PCR 使用四个引物同时扩增线粒体 DNA 的特定区域，并产生物种特异性的带型。该方法的严格性通过不同性别、地点和发育阶段的标本进行测试，所有样本均获得一致的结果。事实上，我们的方法有可能阐明生菜田中 plusiine 物种的物种结构。
