植食性昆虫食物网鉴别中DNA宏条形码的高通量测序研究

生态中的食物网络决定了生态系统的功能和稳定性。目前DNA条形码技术在食物网络的研究中已有较广泛的应用，但在食物网分子鉴定中的关键问题之一-DNA宏条形码的选择及高通量测序建库仍了解甚少。本项目以昆虫纲第二大目植食性的鳞翅目幼虫作为研究材料，尝试采用多重PCR对DNA宏条形码进行一次性扩增，并结合DNA宏条形码的选择、引物设计、DNA文库构建及后续生物信息分析，以已知食性的鳞翅目幼虫作为参考验证方法的便捷性和准确性。阐明高通量测序中的多重PCR，DNA宏条形码长度（>600bp或<500bp）及后续生物信息分析对单食和多食的植食性昆虫食物网鉴定准确性的影响。研究结果可简化食物网的分子鉴定过程，提高DNA宏条形码技术构建食物网的鉴定效率。为了解昆虫食物网络结构及构建食物网数据库奠定基础。同时，本研究基于新方法拟分析不同食性鳞翅目幼虫分布特点和植物多样性的关系。

Ecological food network determines the functioning and stability of the ecosystem. At present, DNA barcoding technology has been widely used in the study of food networks. However, some problems still exist in the molecular identification by High troughput sequencing of food network, further study are required. In this project, we attempt to use multiplex PCR for one-time amplification, and combined with the selection of DNA metabarcodes, primer design, library construction and bioinformation analysis, the known feeding larva of Lepidopteran will be used to verify the convenience and accuracy of the method. We will further investigate the effects of multiple PCR, universal full length or <500bp DNA metabarcodes and subsequent bioinformatics analysis on the identification of monophagous and polyphagous herbivorous insects. The new method may simplify the molecular identification of food webs and improve the identification efficiency of food network reconstructed by DNA metabarcoding technology.