基于DNA条形码技术的虹鳟分子鉴定

doi:10.14012/j.cnki.fjsc.2023.04.002

摘要/Abstract

摘要：

为保护消费者权益,强化市场监管,确保冷水鱼养殖产业健康可持续发展,找到鲑鳟鱼类种属鉴定的有效方法十分必要。本试验利用细胞色素氧化酶Ⅰ(COⅠ)基因开发一种简便、经济、高效的分子鉴定方法,旨在实现对虹鳟(Oncorhynchus mykiss)的鉴定。结果表明,利用COⅠ基因能够很好地区分二倍体虹鳟、大西洋鲑(Salmo salar)、大麻哈鱼(Oncorhynchus keta)和高白鲑(Coregonus peled)四个物种,但无法对变种金鳟和三倍体虹鳟进行区分,后续研究中将尝试其他方法进行近缘物种的鉴定。本研究表明生产实践中可采用DNA条形码技术提供一种基因标签对虹鳟进行鉴别,为其物种鉴定及产业的健康发展提供技术支撑。

关键词: 虹鳟, DNA条形码, COⅠ, 分子鉴定

Abstract:

In order to ensure the healthy and sustainable development of cold-water fish culture industry, and to ensure consumers’ clear consumption and strengthen the market supervision, it is necessary to find an effective method for species identification of salmon and trout. In this study, cytochrome oxidase I gene (COⅠ) was used to develop a simple, economical and efficient molecular identification method, aiming at identifying O.mykiss. The results showed that COⅠ gene could distinguish diploid rainbow trout, Salmo salar, Oncorhynchus keta and Coregonus peled, but could not distinguish golden trout and triploid rainbow trout. In the follow-up study, we will try to identify related species according to other methods. In production practice, this method could provide a gene tag to identify O.mykiss, and provide technical support for its species identification and healthy development of industry.

Key words: Oncorhynchus mykiss, DNA barcode, COⅠ, molecular identification

中图分类号:

李昭楠, 李长忠, 保长虹, 贺彩霞, 金文杰, 陈艳霞. 基于DNA条形码技术的虹鳟分子鉴定[J]. 渔业研究, 2023, 45(4): 331-340.

LI Zhaonan, LI Changzhong, BAO Changhong, HE Caixia, JIN Wenjie, CHEN Yanxia. Molecular identification of Oncorhynchus mykiss based on DNA barcode technology[J]. Journal of Fisheries Research, 2023, 45(4): 331-340.

图/表 9

参考文献 42

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物种 Species	登录号 Accession number	长度/bp Length
帝王鲑 Oncorhynchus tshawytscha	KX958414.1	1 204
金腹大麻哈鱼 Oncorhynchus chrysogaster	JX960908.1	1 247
美洲红点鲑 Salvelinus fontinalis	HQ167687.1	1 551
钝吻鲑 Salmo obtusirostris	JX960948.1	1 262
多瑙哲罗鱼 Hucho hucho	JX960904.1	1 247
细鳞鲑 Brachymystax lenok	JX227988.1	1 581
突唇白鲑 Coregonus lavaretus	JX960890.1	1 246
北鲑 Stenodus leucichthys	KT630716.1	1 201
普伦白鲑 Coregonus pollan	JX960898.1	1 262
茴鱼 Thymallus thymallus	JX960975.1	1 262

物种 Species	登录号 Accession number	长度/bp Length
帝王鲑 Oncorhynchus tshawytscha	KX958414.1	1 204
金腹大麻哈鱼 Oncorhynchus chrysogaster	JX960908.1	1 247
美洲红点鲑 Salvelinus fontinalis	HQ167687.1	1 551
钝吻鲑 Salmo obtusirostris	JX960948.1	1 262
多瑙哲罗鱼 Hucho hucho	JX960904.1	1 247
细鳞鲑 Brachymystax lenok	JX227988.1	1 581
突唇白鲑 Coregonus lavaretus	JX960890.1	1 246
北鲑 Stenodus leucichthys	KT630716.1	1 201
普伦白鲑 Coregonus pollan	JX960898.1	1 262
茴鱼 Thymallus thymallus	JX960975.1	1 262

引物名称 Name of primers		引物序列 Sequence of primers	产物长度/bp Length of product
COⅠ-1(DNA barcode)	COⅠ-F24	5’-GGGATGACCAAATCTATAACGTGA-3’	681
	COⅠ-R24	5’-CTCAGACCATTCCTATATACCCGAAG-3’	681
	COⅠ-F25	5’-GATTAATTCCCCTAATAATCGGAGC-3’	678
	COⅠ-R25	5’-ATGTAAAGTAAGCACGAGTGTCCA-3’	678
COⅠ-2(Mini barcode)	TEYI-0F	5’-AACCTCCAGCCATCTCTCAG-3’	153
	TEYI-0R	5’-CCGGGTCAAAGAAAGTGGTG-3’	153
	TEYI-1F	5’-CCGCCCTGAGTCTACTGATT-3’	230
	TEYI-1R	5’-TGGAGGAAGGAGTCAGAAGC-3’	230

引物名称 Name of primers		引物序列 Sequence of primers	产物长度/bp Length of product
COⅠ-1(DNA barcode)	COⅠ-F24	5’-GGGATGACCAAATCTATAACGTGA-3’	681
	COⅠ-R24	5’-CTCAGACCATTCCTATATACCCGAAG-3’	681
	COⅠ-F25	5’-GATTAATTCCCCTAATAATCGGAGC-3’	678
	COⅠ-R25	5’-ATGTAAAGTAAGCACGAGTGTCCA-3’	678
COⅠ-2(Mini barcode)	TEYI-0F	5’-AACCTCCAGCCATCTCTCAG-3’	153
	TEYI-0R	5’-CCGGGTCAAAGAAAGTGGTG-3’	153
	TEYI-1F	5’-CCGCCCTGAGTCTACTGATT-3’	230
	TEYI-1R	5’-TGGAGGAAGGAGTCAGAAGC-3’	230

种名 Species		登录号 Accession number	长度/bp Length
帝王鲑 Oncorhynchus tshawytscha		FJ999371.1	652
银鲑 Oncorhynchus kisutch		MG951604.1	652
虹鳟 Oncorhynchus mykiss		MN850431.1	653
大麻哈鱼 Oncorhynchus keta		MN850432.1	653
大西洋鲑 Salmo salar		MN850430.1	651
花羔红点鲑 Salvelinus malma		EU522414.1	652
北极红点鲑 Salvelinus alpinus		KJ128606.1	648
远东红点鲑 Salvelinus leucomaenis		MF503661.1	697
多瑙哲罗鱼 Hucho hucho		KJ553640.1	652
哲罗鱼 Hucho taimen		MG951558.1	652
细鳞鲑 Brachymystax lenok		JX261990.1	633
远东哲罗鱼 Parahucho perryi		HQ693237.1	655
秋白鲑 Coregonus autumnalis	EU202649.1		650
高白鲑 Coregonus peled	MF632325.1		637
欧白鲑 Coregonus albula	KX457959.1		704
茴鱼 Thymallus thymallus	HQ961016.1		652
黑龙江茴鱼 Thymallus grubii	MG951577.1		652