雅鲁藏布江中游拉萨裸裂尻鱼的环境DNA检测及生物量评估
Detection and Biomass Assessment of Schizopygopsis younghusbandi younghusbandi Based on Environmental DNA in the Middle Reach of Yarlung Zangbo River
投稿时间:2022-01-12  修订日期:2022-02-05
DOI:10.15928/j.1674-3075.202201120016
中文关键词:环境DNA  微滴式数字PCR  拉萨裸裂尻鱼  物种检测  雅鲁藏布江中游
英文关键词:environmental DNA  droplet digital PCR  Schizopygopsis younghusbandi  species detection  the middle reach of Yarlung Zangbo River
基金项目:第二次青藏高原综合科学考察研究项目(2019QZKK0304、2019QZKK0501);中国科学院战略性先导科技专项(XDA2005020403)
作者单位
李 冰 中国科学院水生生物研究所湖北 武汉 430072中国科学院大学北京 100049 
冯 秀 中国科学院水生生物研究所湖北 武汉 430072 
朱 仁 中国科学院水生生物研究所湖北 武汉 430072 
隋晓云 中国科学院水生生物研究所湖北 武汉 430072 
贾银涛 中国科学院水生生物研究所湖北 武汉 430072 
陈毅峰 中国科学院水生生物研究所湖北 武汉 430072 
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中文摘要:
      建立快速和准确的环境DNA(eDNA)检测方法,可为鱼类的长期监测提供便利,为鱼类资源保护和管理提供技术支撑。2019年在雅鲁藏布江中游干流及拉鲁湿地和茶巴朗湿地采集了20种鱼类和水样,针对拉萨裸裂尻鱼(Schizopygopsis younghusbandi younghusbandi)设计Cytb基因的特异性引物和探针,利用微滴式数字PCR(ddPCR)检测水样中eDNA拷贝数,并分析其与生物量和丰度之间的相关性。结果显示,正反向引物及探针序列与其他19种鱼类之间的平均错配碱基数分别为6.8、6.9和3.6,且对其基因组DNA进行ddPCR扩增没有荧光信号。通过对比ddPCR和网捕2种方法,70%样点中检测结果相同,剩余30%样点都是网捕未捕获但ddPCR检出的情况。在拉鲁湿地和雅鲁藏布江干流的样点中,水样eDNA拷贝数与拉萨裸裂尻鱼生物量和丰度之间均具有显著的相关性,前者的相关系数分别为0.987和0.647,后者的相关系数分别为0.786和0.756,表明eDNA技术是鱼类分布和生物量监测的有效方法。但是,eDNA检测易受到近缘物种[如高原裸鲤(Gymnocypris waddelli)等]和水体理化性质的影响。
英文摘要:
      Establishing an effective and accurate environmental DNA (eDNA) method can facilitate long-term fish monitoring and provide technical support for the conservation and management of fish resources. Schizopygopsis younghusbandi younghusbandi is the primary economic fish species of the Xizang Autonomous Region, naturally distributed in the stem and tributaries of the upper and middle Yarlung Zangbo River. In 2019, 20 fish species and water samples were collected from 23 sites in the middle reach of Yarlung Zangbo River, and the Lalu and Chabalang wetlands. Species-specific primers and a probe of the Cytb gene for S. younghusbandi younghusbandi were then designed to estimate eDNA concentrations in water samples using droplet digital PCR (ddPCR) and the relationship of eDNA concentration with species biomass and abundance was analyzed. Results show that the average number of base pair mismatches for Cytb gene probe sequences, and the forward and reverse primers between S. younghusbandi younghusbandi and the 19 other fish species were, respectively, 6.8, 6.9 and 3.6. No fluorescence signal was observed in the ddPCRs with the genomic DNA of the other 19 fish species. Species detection by ddPCR showed the same results in 70% of the sampling sites compared with the netting survey. In the remaining 30% of the sampling sites, S. younghusbandi younghusbandi were detected by ddPCR, but not captured in the netting survey, demonstrating that the eDNA method is more effective in detecting fish species than capture by netting. For sampling sites from the Lalu wetland and the mainstem of Yarlung Zangbo River, significant correlations were observed between the eDNA concentration and species biomass and abundance, with respective correlation coefficients of 0.987 and 0.647 for the Lalu wetland sites, and 0.786 and 0.756 for the Yarlung Zangbo River sites. The linear equations between eDNA concentrations and species biomass and abundance were, respectively, y1= 2.0408x + 1.4452 and y2 = 0.0279x + 1.8000 for the Lalu wetland sites, and y1 = 878.9849x - 659.9753 and y2 = 5.314 9x - 2.8281 for the Yarlung Zangbo River sites. These results indicate that the eDNA method is an effective and useful approach for monitoring fish occurrence and biomass. However, the results of eDNA species detection can be influenced by closely related species and the physical and chemical properties of water.
李 冰,冯 秀,朱 仁,隋晓云,贾银涛,陈毅峰.2024.雅鲁藏布江中游拉萨裸裂尻鱼的环境DNA检测及生物量评估[J].水生态学杂志,45(2):84-91.
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