There is an increasing demand for salmonid authentication due to the globalization of the salmonid trade.DNA barcoding and mini-DNA barcoding are widely used for identifying fish species based on a fragment of the mit...There is an increasing demand for salmonid authentication due to the globalization of the salmonid trade.DNA barcoding and mini-DNA barcoding are widely used for identifying fish species based on a fragment of the mitochondrial cytochrome c oxidase subunit I(COI)sequence.In this study,rainbow trout(Oncorhynchus mykiss),steelhead trout(O.mykiss),and Atlantic salmon(Salmo salar)collected from two salmonid aquaculture bases in China were authenticated by DNA barcoding(about 650 bp)and mini-DNA barcoding(127 bp)to evaluate the accuracy of the two methods in the identification of different salmonid species.The results revealed that both methods could effectively distinguish O.mykiss and S.salar with 100%accuracy.However,the two methods failed to separate rainbow trout(O.mykiss)and steelhead trout(O.mykiss),which are the same species but cultured in different water environments.Moreover,salmonid samples from three main distribution channels in the Qingdao area(traditional supermarkets,online supermarkets,and sushi bars)were identified by the two methods.Substitution of S.salar with O.mykiss was discovered,and the 27.78%overall substitution rate of salmonids in the Qingdao area was higher than those in other regions reported in previous studies.In addition,the mislabeling rates of salmonids from traditional supermarkets,online supermarkets,and sushi bars were compared in this study.The mislabeling rate was significantly greater in sushi bars(50%)than in the other two channels(16.67%),suggesting that stronger monitoring and enforcement measures are necessary for the aquatic food catering industry.展开更多
目的建立一种高效、精准的DNA条形码技术准确快速鉴定鱼胶物种。方法以细胞色素C氧化酶I(cytochrome C oxidase subunit I,COI)基因为靶标,筛选并优化DNA条形码引物,通过聚合酶链式反应(polymerase chain reaction,PCR)扩增和Sanger测...目的建立一种高效、精准的DNA条形码技术准确快速鉴定鱼胶物种。方法以细胞色素C氧化酶I(cytochrome C oxidase subunit I,COI)基因为靶标,筛选并优化DNA条形码引物,通过聚合酶链式反应(polymerase chain reaction,PCR)扩增和Sanger测序评估其鉴定效果。对市售鱼胶样品进行特异性扩增,结合生命条形码数据系统(barcode of life data systems,BOLD)和美国国家生物技术信息中心(national center for biotechnology information,NCBI)数据库进行物种比对,并采用邻接法(neighbor-joining,NJ)和系统发育树分析验证鉴定结果。结果通过筛选8对COI基因微型DNA条形码引物,最终确定了一组扩增目标片段约190 bp的引物组合,其PCR测序成功率达100%,物种鉴定准确率达98.15%。基于该引物对16份鱼胶样品的鉴定结果显示,仅56.25%的样本与标注物种一致,其中不符样品主要是鳘鱼胶和白花胶,鉴定阈值设为95%。进一步通过NJ分析和系统发育树构建,证实鱼胶样品的系统发育关系与BOLD/NCBI数据库比对结果高度一致,验证了鉴定结果的可靠性。结论本研究成功筛选出适用于鱼胶微型DNA条形码鉴定的引物,为鱼胶产品的真伪鉴别和质量控制提供了高效、可靠的技术手段。展开更多
基金the National Key Research and Development Program of China(No.2019YFD0901000)the Natural Science Foundation of Shandong Pro-vince,China(No.ZR2020MC194).
文摘There is an increasing demand for salmonid authentication due to the globalization of the salmonid trade.DNA barcoding and mini-DNA barcoding are widely used for identifying fish species based on a fragment of the mitochondrial cytochrome c oxidase subunit I(COI)sequence.In this study,rainbow trout(Oncorhynchus mykiss),steelhead trout(O.mykiss),and Atlantic salmon(Salmo salar)collected from two salmonid aquaculture bases in China were authenticated by DNA barcoding(about 650 bp)and mini-DNA barcoding(127 bp)to evaluate the accuracy of the two methods in the identification of different salmonid species.The results revealed that both methods could effectively distinguish O.mykiss and S.salar with 100%accuracy.However,the two methods failed to separate rainbow trout(O.mykiss)and steelhead trout(O.mykiss),which are the same species but cultured in different water environments.Moreover,salmonid samples from three main distribution channels in the Qingdao area(traditional supermarkets,online supermarkets,and sushi bars)were identified by the two methods.Substitution of S.salar with O.mykiss was discovered,and the 27.78%overall substitution rate of salmonids in the Qingdao area was higher than those in other regions reported in previous studies.In addition,the mislabeling rates of salmonids from traditional supermarkets,online supermarkets,and sushi bars were compared in this study.The mislabeling rate was significantly greater in sushi bars(50%)than in the other two channels(16.67%),suggesting that stronger monitoring and enforcement measures are necessary for the aquatic food catering industry.
文摘目的建立一种高效、精准的DNA条形码技术准确快速鉴定鱼胶物种。方法以细胞色素C氧化酶I(cytochrome C oxidase subunit I,COI)基因为靶标,筛选并优化DNA条形码引物,通过聚合酶链式反应(polymerase chain reaction,PCR)扩增和Sanger测序评估其鉴定效果。对市售鱼胶样品进行特异性扩增,结合生命条形码数据系统(barcode of life data systems,BOLD)和美国国家生物技术信息中心(national center for biotechnology information,NCBI)数据库进行物种比对,并采用邻接法(neighbor-joining,NJ)和系统发育树分析验证鉴定结果。结果通过筛选8对COI基因微型DNA条形码引物,最终确定了一组扩增目标片段约190 bp的引物组合,其PCR测序成功率达100%,物种鉴定准确率达98.15%。基于该引物对16份鱼胶样品的鉴定结果显示,仅56.25%的样本与标注物种一致,其中不符样品主要是鳘鱼胶和白花胶,鉴定阈值设为95%。进一步通过NJ分析和系统发育树构建,证实鱼胶样品的系统发育关系与BOLD/NCBI数据库比对结果高度一致,验证了鉴定结果的可靠性。结论本研究成功筛选出适用于鱼胶微型DNA条形码鉴定的引物,为鱼胶产品的真伪鉴别和质量控制提供了高效、可靠的技术手段。
