Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan c...Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.展开更多
目的运用DNA条形码技术对常见石首鱼科鱼胶进行物种鉴定。方法通过对26份鱼胶样品基因组DNA提取,聚合酶链式反应(polymerase chain reaction,PCR)扩增细胞色素C氧化酶Ⅰ(cytochrome c oxidase,COI)基因、测序,用生命条形码数据(barcode ...目的运用DNA条形码技术对常见石首鱼科鱼胶进行物种鉴定。方法通过对26份鱼胶样品基因组DNA提取,聚合酶链式反应(polymerase chain reaction,PCR)扩增细胞色素C氧化酶Ⅰ(cytochrome c oxidase,COI)基因、测序,用生命条形码数据(barcode of life data,BOLD)物种鉴定系统,与数据库中已有鱼类序列进行比对分析,鉴定出各鱼胶的物种;根据Kimura双参数模型计算样品序列遗传距离,并将所得序列使用邻接法(neighbor-joining,NJ)和最大简约法(maximum parsimony,MP)构建系统发育树,进行聚类分析。结果26份鱼胶样品通过鉴定引物“Fish-F”“Fish-R”均可实现扩增,条带清晰,扩增和测序成功率均为100%;BOLD鉴定结果显示,26份鱼胶样品中23份能够确定物种来源(相似性达98%以上),包括石首鱼科12属15种鱼类,且多数为外来物种,另外3份鱼胶可推测其近缘物种。此外,系统发育树聚类分析结果与物种鉴定结果一致。结论目前石首鱼类鱼胶来源物种较多,且多为外来基原鱼种。DNA条形码技术与BOLD鉴定系统相结合,可对大部分鱼胶进行准确的物种鉴定。展开更多
基金Supported by the Zhejiang Provincial Key Research and Development Program (No.2021C02047)。
文摘Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.
文摘目的运用DNA条形码技术对常见石首鱼科鱼胶进行物种鉴定。方法通过对26份鱼胶样品基因组DNA提取,聚合酶链式反应(polymerase chain reaction,PCR)扩增细胞色素C氧化酶Ⅰ(cytochrome c oxidase,COI)基因、测序,用生命条形码数据(barcode of life data,BOLD)物种鉴定系统,与数据库中已有鱼类序列进行比对分析,鉴定出各鱼胶的物种;根据Kimura双参数模型计算样品序列遗传距离,并将所得序列使用邻接法(neighbor-joining,NJ)和最大简约法(maximum parsimony,MP)构建系统发育树,进行聚类分析。结果26份鱼胶样品通过鉴定引物“Fish-F”“Fish-R”均可实现扩增,条带清晰,扩增和测序成功率均为100%;BOLD鉴定结果显示,26份鱼胶样品中23份能够确定物种来源(相似性达98%以上),包括石首鱼科12属15种鱼类,且多数为外来物种,另外3份鱼胶可推测其近缘物种。此外,系统发育树聚类分析结果与物种鉴定结果一致。结论目前石首鱼类鱼胶来源物种较多,且多为外来基原鱼种。DNA条形码技术与BOLD鉴定系统相结合,可对大部分鱼胶进行准确的物种鉴定。
