| 长江中下游干流环境DNA样本鱼类物种检测的初步研究 |
| Preliminary Study on Fish Species Detection in the Middle and Lower Yangtze River Using Environmental DNA |
| 投稿时间：2016-07-08 修订日期：2016-08-05 |
| DOI：10.15928/j.1674-3075.2016.05.007 |
| 中文关键词:环境DNA 物种检测 生物多样性 鱼类 |
| 英文关键词:environmental DNA species detection biodiversity fish species |
| 基金项目:2015年中华鲟当年产卵场调查 |
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| 中文摘要: |
| 为开发适用于长江鱼类的环境DNA检测体系，在长江中下游干流24个采样点同步采集水样，抽滤后提取环境DNA，利用线粒体细胞色素B简并引物进行PCR扩增，扩增产物克隆测序得到419条序列，通过在GenBank核酸序列数据库进行BLAST序列比对确定物种信息，从来源于17个采样点的115条匹配成功的序列中，共检测到10种鱼类序列,代表15种鱼类。 |
| 英文摘要: |
| Environmental DNA analysis is an emerging technology for detecting species based on DNA extracted from environmental samples, and it is a good tool for monitoring fish species composition in freshwater ecosystems. The middle and lower Yangtze River provides important spawning and feeding grounds for many rare migratory and economic fish species, including the endangered Chinese sturgeon and the natural populations of the four domesticated carp species. In this study, fish species were detected in the Yangtze River by environmental DNA using mitochondrial DNA markers. The aim was to explore the feasibility of fish species detection by environmental DNA and, if feasible, to employ the method to gather data on the fish community. A total of 24 sampling sites were selected in the main stem of the Yangtze River, from Yichang to Nanjing, with the interval of 50 km between every two adjacent sampling sites, and all 24 sites from upstream to downstream （Site 1-Site 24） were divided into two groups for synchro-sampling. Water samples were collected synchronously at odd-numbered sampling sites at 10:30 on November 24, 2015 and at even-numbered sampling sitesat 10:30 on November 25, 2015. The samples were vacuum-filtered with polyethersulfone membrane filters (0.45-μm) and DNA was extracted using the PowerWater DNA Isolation kit. The mitochondrial cytochrome b gene was used for amplification and PCR products were sequenced after random cloning. Highly similar DNA sequences were found using BLAST (Basic Local Alignment Search Tool) searches and sequences with no more than a single mismatch in the non-primer region were considered. Of the 419 sequences obtained, 115 significant sequence alignments for 17 sites were found in the Genbank nucleotide database and 15 fish species were detected. The results demonstrate that fish species detection is feasible using environmental DNA from the Yangtze River. This paper lays a foundation for species monitoring and species diversity evaluation using environmental DNA in the Yangtze River. By using high-throughput sequencing of DNA extracted from water samples, more species or entire faunas can be detected. Specific molecular markers can be used to detect target species, both for monitoring rare and endangered species and for early detection of invasive species. |
| 徐念,常剑波.2016.长江中下游干流环境DNA样本鱼类物种检测的初步研究[J].水生态学杂志,37(5):49-55. |
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