EAPSI: Innovative Molecular Methods for Low Cost DNA Barcoding of Tropical Arthropods

保护生物多样性对生态系统至关重要？我们的经济能力是抵御气候变化等干扰的能力，是为人类社会提供服务的能力。传统的测量昆虫和其他节肢动物多样性的方法既耗时又昂贵，这阻碍了有效保护政策的制定。与新加坡国立大学的Rudolf Meier博士合作，该项目将改进和分享Meier实验室开发的用于生成昆虫识别DNA条形码的新技术的知识。DNA条形码是一种特定的DNA短序列，可用于识别生物体，就像在杂货店扫描谷物盒背面的条形码一样。与研究人员合作？这项工作将改进生物多样性评估和监测方法，同时揭示节肢动物森林群落中的关系。该项目将使用新的直接PCR方案和下一代测序方法对数千种昆虫标本进行DNA条形码。目前正在利用元条码对肯塔基州（美国）和法属圭亚那的森林进行生物多样性评估和比较。感兴趣的群体，包括寄生虫和毛虫，以及所有节肢动物目的代表将从元条形码样本中删除，用作凭证标本。为了评估元条形码编码过程并发现寄生虫-宿主关系，将使用Rudolf Meier实验室开发的低成本方法生成与这些标本相关的DNA条形码。在元条形码序列数据集中发现的凭证样本序列的比例将允许调整多样性估计值，以考虑测序过程中的偏倚。直接PCR和下一代测序（NGS）方法可以以每个样本不到1美元的价格生产条形码，但需要培训和经验才能成功和一致地执行。梅尔医生的协议是专为使用双翅目，并将优化用于与hyperteran寄生蜂。该项目产生的数据将用于比较热带和温带节肢动物的生物多样性，比较形态学和分子鉴定技术，并发现新的寄生虫-宿主关系。该奖项属于东亚和太平洋夏季研究所计划，支持美国研究生的夏季研究，由NSF和新加坡国家研究基金会共同资助。

Preservation of biodiversity is essential to an ecosystem?s ability to withstand disturbances like climate change and to provide services to human society. Traditional methods of measuring insect and other arthropod diversity are time consuming and expensive which impedes creation of effective conservation policies. In collaboration with Dr. Rudolf Meier at the National University of Singapore, this project will improve and share the knowledge of new techniques developed in the Meier lab for generating DNA barcodes for insect identification. DNA barcodes are a specific short sequence of DNA which can be used to identify an organism, much like how the barcode on the back of a cereal box is scanned at a grocery store. Paired with the researcher?s larger thesis project, this work will improve biodiversity evaluation and monitoring methods while uncovering relationships in arthropod forest communities. This project will use new direct PCR protocols and next-generation sequencing methods to DNA barcode thousands of insect specimens. A biodiversity assessment and comparison between the forests of Kentucky (United States) and French Guiana using metabarcoding is underway. Groups of interest, including parasitoids and caterpillars, as well as representatives of all arthropod orders present will be removed from the metabarcoded samples for use as voucher specimens. To evaluate the metabarcoding process and discover parasitoid-host relationships, DNA barcodes linked to these specimens will be generated using a low cost method developed by Rudolf Meier's lab. The proportion of voucher specimen sequences found within the metabarcoding sequence datasets will allow for the adjustment of diversity estimates to account for biases in the sequencing process. Direct PCR and next-generation sequencing (NGS) methods can produce barcodes at less than $1 per specimen but require training and experience to perform successfully and consistently. Dr. Meier?s protocols were designed for use with Diptera and will be optimized for use with hymenopteran parasitoids. The data generated during this project will be used to compare tropical and temperate arthropod biodiversity, to compare morphological and molecular identification techniques, and discover new parasitoid-host relationships.This award under the East Asia and Pacific Summer Institutes program supports summer research by a U.S. graduate student and is jointly funded by NSF and the National Research Foundation of Singapore.