长江上游鱼类环境DNA通用引物的选择与验证

S932.4; 为了选择出适合使用eDNA技术对长江上游鱼类多样性进行研究的通用引物,本研究选择10对扩增序列位于12SrRNA、16SrRNA、Cytb和CO Ⅰ基因片段的常用引物,分别为 Mifish-U、AcMDB07、Teleo、12SPv、Fish16S1、Ve16Sl、PSI、G、VeCB1、FishCB,对长江上游常见的32种鱼类和3种其他水域鱼类肌肉组织提取的DNA扩增.结果显示,有6对引物均能扩增出全部35种鱼类,但引物Mifish-U的扩增效果最好.进一步使用引物Mifish-U对长江上游屏山县、涪陵区和巫山县3个采样点的水样eDNA进行高通量测序,共检测到80种鱼类,包括...

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Published in水产学报 Vol. 48; no. 6; pp. 98 - 110
Main Authors 吕宏森, 王安香, 董智玲, 闫卉果, 龚志韬, 刘嘉豪, 姚维志, 何文平
Format Journal Article
LanguageChinese
Published 西南大学,淡水鱼类资源与生殖发育教育部重点实验室,重庆 400715 2024
西南大学水产学院,重庆 400715%西南大学水产学院,重庆 400715
Subjects
universal primers
environmental DNA
通用引物
fish
Mifish-U
the upper Yangtze River
长江上游
鱼类
eDNA
Online AccessGet full text
ISSN1000-0615
DOI10.11964/jfc.20220813650

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Abstract S932.4; 为了选择出适合使用eDNA技术对长江上游鱼类多样性进行研究的通用引物,本研究选择10对扩增序列位于12SrRNA、16SrRNA、Cytb和CO Ⅰ基因片段的常用引物,分别为 Mifish-U、AcMDB07、Teleo、12SPv、Fish16S1、Ve16Sl、PSI、G、VeCB1、FishCB,对长江上游常见的32种鱼类和3种其他水域鱼类肌肉组织提取的DNA扩增.结果显示,有6对引物均能扩增出全部35种鱼类,但引物Mifish-U的扩增效果最好.进一步使用引物Mifish-U对长江上游屏山县、涪陵区和巫山县3个采样点的水样eDNA进行高通量测序,共检测到80种鱼类,包括本研究使用的32种长江上游鱼类,其辨别度较高.使用引物Mifish-U对室内养殖3种鱼类的水样eDNA进行高通量测序后定性定量分析,结果显示黄颡鱼和鲤的生物量与序列数相关性显著,引物Mifish-U进行eDNA定量分析的潜力较大.研究表明,引物Mifish-U更适合作为eDNA研究长江上游鱼类多样性的通用引物.本研究可为利用eDNA技术监测长江上游鱼类多样性的引物选择提供参考.
AbstractList S932.4; 为了选择出适合使用eDNA技术对长江上游鱼类多样性进行研究的通用引物,本研究选择10对扩增序列位于12SrRNA、16SrRNA、Cytb和CO Ⅰ基因片段的常用引物,分别为 Mifish-U、AcMDB07、Teleo、12SPv、Fish16S1、Ve16Sl、PSI、G、VeCB1、FishCB,对长江上游常见的32种鱼类和3种其他水域鱼类肌肉组织提取的DNA扩增.结果显示,有6对引物均能扩增出全部35种鱼类,但引物Mifish-U的扩增效果最好.进一步使用引物Mifish-U对长江上游屏山县、涪陵区和巫山县3个采样点的水样eDNA进行高通量测序,共检测到80种鱼类,包括本研究使用的32种长江上游鱼类,其辨别度较高.使用引物Mifish-U对室内养殖3种鱼类的水样eDNA进行高通量测序后定性定量分析,结果显示黄颡鱼和鲤的生物量与序列数相关性显著,引物Mifish-U进行eDNA定量分析的潜力较大.研究表明,引物Mifish-U更适合作为eDNA研究长江上游鱼类多样性的通用引物.本研究可为利用eDNA技术监测长江上游鱼类多样性的引物选择提供参考.
Abstract_FL Environmental DNA(eDNA)technology has emerged as a revolutionary tool for biodiversity monitor-ing in aquatic ecosystems,offering a non-invasive,cost-effective,and highly sensitive method for detecting spe-cies presence and abundance,particularly in complex and dynamic environments like the upper Yangtze River.In order to identify universal primers that are suitable for investigating fish diversity in the upper Yangtze River using eDNA technology,this study selected ten frequently used primers that amplify gene fragments from the 12S rRNA,16S rRNA,Cytb,and CO Ⅰ genes.The primers investigated were Mifish-U,AcMDB07,Teleo,12SPv,Fish16Sl,Ve16Sl,PSI,G,VeCBl,and FishCB.DNA samples extracted from muscle tissues of 32 commonly encountered fish species in the upper Yangtze River and three additional aquatic species not from the upper Yangtze River were subjected to amplification using these primers.The results indicated that six primer pairs could amplify all 35 fish species,with Mifish-U showing the highest amplification efficiency.In addition,the eDNA samples from three sampling sites(Pingshan County,Fuling District,and Wushan County)in the upper Yangtze River were further subjected to high-throughput sequencing using the Mifish-U primer.A total of 80 fish species were detected,including the 32 fish species used in this study,with high discriminatory power.Moreover,through high-throughput sequencing,the Mifish-U primer was utilized to qualitatively and quantitatively analyze eDNA samples from indoor farms for three fish species.The results indicated a significant correlation(P＜0.05)between the biomass of Pelteobagrus fulvidraco and Carassius auratus and the number of sequences,highlighting the great potential of the Mifish-U primer for eDNA quantitative analysis.In summary,our findings suggest that Mifish-U is more suitable as a universal primer for investigating fish diversity in the upper Yangtze River using eDNA techno-logy.This study offers important insights into the selection of primers for monitoring fish diversity in the upper Yangtze River using eDNA technology.
Author 吕宏森
闫卉果
王安香
董智玲
刘嘉豪
何文平
龚志韬
姚维志
AuthorAffiliation 西南大学水产学院,重庆 400715%西南大学水产学院,重庆 400715;西南大学,淡水鱼类资源与生殖发育教育部重点实验室,重庆 400715
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Author_FL DONG Zhiling
LIU Jiahao
Lü Hongsen
YAN Huiguo
HE Wenping
WANG Anxiang
YAO Weizhi
GONG Zhitao
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Keywords universal primers
environmental DNA
通用引物
fish
Mifish-U
the upper Yangtze River
长江上游
鱼类
eDNA
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Snippet S932.4; 为了选择出适合使用eDNA技术对长江上游鱼类多样性进行研究的通用引物,本研究选择10对扩增序列位于12SrRNA、16SrRNA、Cytb和CO Ⅰ基因片段的常用引物,分别为...
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Title 长江上游鱼类环境DNA通用引物的选择与验证
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