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Abstract：
The fruit fly Drosophila melanogaster is a model organism to study several aspects of metazoan biology. Most of the work has been conducted in adult fruit flies, including laboratory and field-derived specimens, but Drosophila melanogaster larvae recently became a valuable model to better understand animal physiology, development, or host-microbe interactions. While adult flies can be easily assigned to a given Drosophila species based on morphological characteristics, such visual identification is more intricate at the larval stage. This could explain the limited number of studies focusing on larvae, especially field-derived samples. Here, we developed a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay that discriminates D. melanogaster from other ecologically relevant Drosophila species at the larval stage. The method, which targets the cytochrome oxidase I (COI) gene, was validated using laboratory-derived larvae from seven D. melanogaster populations originating from different geographic areas as well as six Drosophila species. We further validated this PCR-RFLP assay in a natural context, by identifying wild larvae collected in two locations in France. Notably, among all PCR-RFLP profiles that matched the D. melanogaster species, 100% were correctly identified, as confirmed by COI sequencing. In summary, our work provides a rapid, simple, and accurate molecular tool to identify D. melanogaster from field-collected larvae.
摘要：
果蝇果蝇是研究后生生物学几个方面的模型生物。大部分工作是在成年果蝇中进行的，包括实验室和野外标本，但是果蝇幼虫最近成为更好地了解动物生理学的有价值的模型，发展,或宿主-微生物相互作用。虽然成年果蝇可以根据形态特征轻松地分配给给定的果蝇物种，这种视觉识别在幼虫阶段更加复杂。这可以解释为什么关注幼虫的研究数量有限，尤其是场衍生样本。这里,我们开发了一种聚合酶链反应-限制性片段长度多态性（PCR-RFLP）测定法，该测定法可在幼虫期将黑腹果蝇与其他与生态相关的果蝇区分开。方法,靶向细胞色素氧化酶I（COI）基因，使用来自不同地理区域的七个D.melanogaster种群以及六个果蝇物种的实验室幼虫进行了验证。我们在自然环境中进一步验证了这种PCR-RFLP测定，通过识别在法国两个地方收集的野生幼虫。值得注意的是,在所有与黑腹D.melanogaster物种匹配的PCR-RFLP图谱中，100%被正确识别，如COI测序所证实。总之,我们的工作提供了一个快速，简单,和精确的分子工具，从野外采集的幼虫中鉴定黑腹。
