基于环境DNA宏条形码测序的季节性热带珊瑚礁区鱼类多样性及其相对丰度变动研究

The coral reef fish diversity and their relative abundance are important health indicators of coral reef ecosystem, but it is hard to find a non-invasive or environmental friendly, fast and accuracy research method to acquire these data. In the past five years, an eDNA-based meta-barcoding technology provides us a potential tool to monitor the marine fish diversity and their relative abundance, although it still have some technical problems need to be sorted out in practical applications. This study will use several species-specific primer pairs and absolute quantitative PCR technology to determine a suitable high quality eDNA capture method for coral reef ecosystem. After several rounds of eDNA-based high throughput sequencing and bioinformatics analysis, a set of meta-barcoding primer pairs with the best library construction condition will be developed. Ultimately, eDNA-based meta-barcoding as a coral reef fish diversity monitoring tool developed by our study will be implemented into Sanya National Coral Reef Conservation Zone for evaluation seasonal variation of the coral reef fish diversity and their relative abundance. We aim to establish a reliable eDNA-based meta-barcoding workflow for evaluation of the coral reef fish diversity, which can improve the understanding of eDNA spatial and temporal distribution pattern and provide fundamental data for constructing an eDNA-based coral reef fish biomass model in the future. Therefore, this study has important theoretical significance and application prospects.

珊瑚礁鱼类多样性及其相对丰度是评价珊瑚礁生态系统健康度的重要指标之一，但长期以来缺乏一种环境友好且快速准确的研究方法对其进行准确评估。近五年来，基于环境DNA的宏条形码技术为我们提供了一个潜在的海水鱼类多样性及生物量评价的工具，尽管其在实际应用时仍存在众多亟待解决的技术问题。本研究拟利用种间特异性引物进行绝对定量PCR的方法来确定针对珊瑚礁生态系统的高质量环境DNA的提取方案，并通过多轮环境样本的高通量测序与生物信息学分析等方法开发出一套具有最适文库构建条件的宏条形码引物簇，最终将其应用于三亚珊瑚礁保护区季节性鱼类多样性及其相对丰度检测。本研究成果将建立一套便捷可靠的珊瑚礁水域鱼类多样性评估技术方案，有望增进对珊瑚礁鱼类环境DNA时空分布规律的认识并为未来利用该技术构建基于环境DNA拷贝数的珊瑚礁鱼类生物量评估模型提供高质量的基础数据，具有重要的理论意义和应用前景。