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| 基于沉积物eDNA评估三疣梭子蟹资源量的方法初探 |
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张凡瑶1, 张再美1, 陈萌琦1, 韦东升1, 徐严欢1, 姚冰1, 孙振宁2, 李慧1, 杨艳艳2, 周兴3, 李凡2, 陈翠霞1
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1.中国石油大学(华东)化学化工学院, 山东 青岛 266580;2.山东省海洋资源与环境研究院, 山东省海洋生态修复重点实验室, 山东 烟台 264006;3.山东省青岛市黄岛区海洋发展局, 山东 青岛 266499
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| 摘要: |
| 本文旨在通过沉积物eDNA (environmental DNA)的分析,建立一种快速评估三疣梭子蟹资源量的方法。采用柱状采泥器对莱州湾同一地点(37°30″E、119°20″N)不同深度(0~24 cm,7个深度)进行样品采集,提取不同深度地层沉积物中的eDNA,并以此为模板,运用三疣梭子蟹COI基因通用型引物和特异性引物分别进行普通PCR和实时荧光定量PCR (quantitative PCR,qPCR)扩增,评估莱州湾近年来三疣梭子蟹的资源量变化。结果显示:7个样品的eDNA中均能扩增出700 bp长度的DNA产物,表明在不同深度沉积物所代表的年份均有三疣梭子蟹的存在;通过Clustal X与MEGA 7.0软件比对各序列碱基之间的差异性,发现不同柱深沉积物中的COI基因共出现11个变化位点,包括8个转换位点、2个颠换位点和1个插入位点;qPCR结果显示,8~9 cm处的三疣梭子蟹COI基因拷贝数要大于其他深度地层沉积物,说明此柱深代表的年份中三疣梭子蟹的资源量高于其他年份。利用沉积物eDNA评估莱州湾海域内的三疣梭子蟹资源量的方法,可以打破时间和空间的限制,检测不同年份三疣梭子蟹的资源量的变化。 |
| 关键词: eDNA 三疣梭子蟹 COI基因 qPCR 资源量 |
| DOI：10.11759/hykx20220520001 |
| 分类号:S932.5＋2 |
| 基金项目:烟台市科技创新发展计划(2020MSGY056);山东省自然科学基金重点项目(ZR2020KE050);烟台市科技创新发展计划(2021XDHZ053);山东省教学改革重点项目(Z2022114) |
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| A preliminary method to assess the Portunus trituberculatus resource based on sedimentary eDNA |
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ZHANG Fan-yao1, ZHANG Zai-mei1, CHEN Meng-qi1, WEI Dong-sheng1, XU Yan-huan1, YAO Bing1, SUN Zhen-ning2, LI Hui1, YANG Yan-yan2, ZHOU Xing3, LI Fan2, CHEN Cui-xia1
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1.College of Chemistry and Chemical Engineering, China University of Petroleum, Qingdao 266580, China;2.The Key Laboratory of Restoration for Marine Ecology of Shandong Province, Shandong Marine Resources and Environment Research Institute, Yantai 264006, China;3.Qingdao Municipal Marine Development Bureau, Qingdao 266499, China
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| Abstract: |
| This study aimed to establish a rapid method to assess the Portunus trituberculatus resource by analyzing sedimentary environmental DNA (eDNA). Samples were collected at different sediment depths (0~24 cm, 7 depths) from the same location (37°30″ E, 119°20″ N) in Laizhou Bay using a column mud collector. The eDNA samples extracted from the ocean sediments served as the template to amplify the cytochrome c oxidase subunit 1 (COI) gene by polymerase chain reaction (PCR) and quantitative PCR (qPCR) using universal and specific primers for the P. trituberculatus COI gene. The amplified products were used to assess the P. trituberculatus resource in Laizhou Bay. The results showed that the 700 bp COI gene fragments were amplified from all extracted eDNA, indicating the presence of P. trituberculatus in the years represented by the sediments at different depths. Genetic consistency was analyzed by Clustal X and MEGA 7.0 software, and 11 variable sites, including 8 conversion sites, 2 inversion sites, and 1 insertion site, were found for the COI gene at the different depths, suggesting high homology of P. trituberculatus in the years represented by the 7 sediments. The qPCR results showed that the P. trituberculatus COI gene copy number at 8–9 cm of soil depth was higher than that at the other depths, suggesting that the P. trituberculatus resource was richer in the years represented by the 8~9 cm depth than the other years. Using sedimentary eDNA saved time and space when assessing the P. trituberculatus resource in Laizhou Bay. |
| Key words: eDNA Portunus trituberculatus COI gene qPCR resource |