基于多基因分析的脊椎动物通用环境DNA metabarcoding检测技术的开发与应用

环境DNA metabarcoding分析是近年来新兴的物种多样性调查技术，具有采样简单、敏感度高、成本低、对调查生物无伤害等特点，在物种分布调查方面表现出了极大的应用潜力。目前环境DNA metabarcoding检测通常只分析1个DNA识别片段，对于调查脊椎动物这样的大型动物类群很难同时兼顾通用性与变异性。针对这一问题，本研究拟为脊椎动物开发4个新的通用DNA识别片段，通过同时分析多个DNA识别片段（复合条形码技术），解决通用性与变异性两难的问题。本项目的目标是开发一种基于多DNA识别片段的脊椎动物通用的环境DNA metabarcoding检测技术，要求实现脊椎动物物种检出率超过95%，使其成为一种简单高效的脊椎动物生物多样性调查手段，并利用该技术检测四川唐家河自然保护区的水样环境DNA，对保护区的脊椎动物多样性进行调查并与历史记录进行比较，对该新技术的可靠性与实用性进行评估。

Environmental DNA (eDNA) metabarcoding analysis of is an emerging technique of species diversity survey in recent years, which has the characteristics of simple sampling, high sensitivity, low cost, and no harm to the investigation organisms, showing great application potential in species survey. Currently, eDNA metabarcoding analysis usually only analyzes one DNA identification fragment, which it is difficult to achieve both universality and variability at the same time when investigating such a large animal groups as vertebrates. To solve this problem, we intend to develop four new universal DNA identification fragments for vertebrates, and solve the dilemma of universality and variability by simultaneously analyzing multiple DNA identification fragments (compound barcode technology).The aim of this study is to develop an highly universal eDNA metabarocding methods for vertebrate survey, requiring detection rate of vertebrate species is over 95%，making it become a kind of simple and efficient means of vertebrate biodiversity survey. We plan to use this new technology to analyze water environmental DNA samples of Sichuan Tangjiahe nature reserve to investigate the vertebrate diversity of reserve. The reliability and applied potential of the new technology will be evaluated by comparing the survey result with historical records of the reserve.