鲟鱼分子鉴定方法的整合应用

doi:10.17520/biods.2022034

生物多样性 ›› 2022, Vol. 30 ›› Issue (6): 22034. DOI: 10.17520/biods.2022034

鲟鱼分子鉴定方法的整合应用

牛晓锋¹^,², 王晓梅²^,³, 张研², 赵志鹏²^,^*(), 樊恩源²^,^*()

1.上海海洋大学水产与生命学院, 上海 201306
2.中国水产科学研究院, 北京 100141
3.中国水产科学研究院长岛增殖实验站, 山东烟台 265800

收稿日期:2022-01-15 接受日期:2022-04-19 出版日期:2022-06-20 发布日期:2022-06-20
通讯作者: 赵志鹏,樊恩源
作者简介:zhaozhipeng@cafs.ac.cn
* E-mail: ecofan@foxmail.com;
基金资助:
中国水产科学研究院本级基本科研业务费(2020B001);中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金(2020TD11);国家自然科学基金(32101239);农业农村部物种品种资源保护费-水生野生动物保护专项研究(21301350001007)

Integration and application of sturgeon identification methods

Xiaofeng Niu¹^,², Xiaomei Wang²^,³, Yan Zhang², Zhipeng Zhao²^,^*(), Enyuan Fan²^,^*()

1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306
2. Chinese Academy of Fishery Sciences, Beijing 100141
3. Changdao Enhancement and Experiment Station, Chinese Academy of Fishery Sciences, Yantai, Shandong 265800

Received:2022-01-15 Accepted:2022-04-19 Online:2022-06-20 Published:2022-06-20
Contact: Zhipeng Zhao,Enyuan Fan

1. 附录1 本文涉及的SNP引物和微卫星引物.pdf(173KB)

摘要/Abstract

摘要：

鲟形目物种是国家重点保护水生野生动物和CITES附录物种。其人工养殖种群数量众多, 种类丰富, 产品贸易量大, 但种类鉴定困难。本文在厘清当前鲟鱼商业类群的基础上, 通过分析现有种类鉴定方法, 整合了线粒体DNA遗传分析、SNP分析和微卫星DNA分析的鉴定方法, 探讨其鉴定国际贸易所涉鲟鱼的可行性。结果表明: 上述3种方法整合应用可在11种纯种鲟鱼及其正反杂交产生的杂交鲟范围内进行盲检。当前共有贸易鲟鱼36种, 其中杂交鲟14种, 杂交鲟的亲本共涉及9种鲟鱼。整合方法可准确鉴定小体鲟(Acipenser ruthenus)、达氏鳇(Huso dauricus)、施氏鲟(Acipenser schrenckii)、欧洲鳇(Huso huso)、闪光鲟(Acipenser stellatus)、高首鲟(A. transmontanus)两两杂交所产生的杂交鲟, 小体鲟为母本与纳氏鲟(Acipenser naccarii)或富氏鲟(A. fulvescens)或中华鲟(A. sinensis)产生的杂交鲟, 纯种的达氏鳇、高首鲟、富氏鲟和中华鲟, 但无法准确鉴定纯种的西伯利亚鲟(Acipenser baerii)和俄罗斯鲟(A. gueldenstaedti)以及父母本涉及此两种鲟鱼的杂交鲟。由于已开发的分子标记仍有限, 上述结果是对当前CITES贸易所涉鲟鱼鉴定的最大范围, 可以满足一些鲟鱼野生种群保护、贸易产品检测、种质资源管理等情景下的鉴定需求。

关键词: 鲟鱼, 种类鉴定, SNP, mtDNA, 微卫星DNA, 物种保护, 种质资源管理

Abstract

Aims: Sturgeons are important economic fishes with high market value. However, with the overfishing and the habitat destruction, the natural population of sturgeon have been declined worldwide. To address the challenges, all sturgeon species has been included in the appendices of CITES in 1997, besides, all species in natural waters of China are also included in the China’s protection catalogue under the Wildlife Protection Law. The identification of sturgeons still faces some difficulties. There are many identification methods for sturgeons, but most of them are applicable conditionally within range of species, respectively. This study aims to integrate available methods to meet more identification requirement in judicial authentication, wildlife supervision and germplasm resource management.
Methods: This study analyzed the identification methods and search the trade situation of sturgeons from the CITES database. Based on this, an integrated identification method which include mtDNA genetic analysis, Single Nucleotide Polymorphism (SNP) analysis and microsatellite DNA was established and analyzed to evaluate the effectiveness in identifying the 11 kinds of commercial sturgeon species and their hybrids.
Results: A total of 36 kinds of aquaculture sturgeons can be found from the CITES sturgeon trade database, including 14 hybrids and 22 purebred species. Although there are many methods to identify sturgeon species, molecular biological methods are proved to be effective identification methods. By integrating three identification methods above, blind detection is applicable to identify 11 purebred sturgeons and their hybrid sturgeons referring to the current international trade. Among them, two kinds of hybrid sturgeons can be accurately identified: (1) The hybrid sturgeons with parents from Acipenser ruthenus, Huso dauricus, A. schrenckii, Huso huso, A. stellatus or A. transmontanus. (2) The hybrid sturgeons whose female parent is A. ruthenus and male parent is A. naccarii or A. fulvescens or A. sinensis. Besides, the purebred H. dauricus, A. transmontanus, A. fulvescens and A. sinensis can be identified only under the current trade background. However, the hybrid sturgeons with parents from A. baerii or A. gueldenstaedti still cannot be accurately identified. Meanwhile, it is difficult to distinguish some purebred sturgeons among several of their hybrid sturgeons (A. baerii or A. gueldenstaedti as male parent).
Conclusion & suggestion: The identifiable sturgeon species are still limited based on the literature, the integration of the method above could satisfy the needs of sturgeon identification in some scenarios and is important for sturgeon conservation, trade product detection and germplasm resources management. Since many species still lack available markers for identification, developing new specific markers and further verifying the scope of existing specific markers should be encouraged with the international cooperation for the diverse sturgeon materials, especially for A. baerii and A. gueldenstaedti as parents of many hybrid sturgeons.

Key words: sturgeon, species identification, SNP, mtDNA, microsatellite DNA, sturgeon conservation, germplasm resources management

牛晓锋, 王晓梅, 张研, 赵志鹏, 樊恩源 (2022) 鲟鱼分子鉴定方法的整合应用. 生物多样性, 30, 22034. DOI: 10.17520/biods.2022034.

Xiaofeng Niu, Xiaomei Wang, Yan Zhang, Zhipeng Zhao, Enyuan Fan (2022) Integration and application of sturgeon identification methods. Biodiversity Science, 30, 22034. DOI: 10.17520/biods.2022034.

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链接本文: https://www.biodiversity-science.net/CN/10.17520/biods.2022034

https://www.biodiversity-science.net/CN/Y2022/V30/I6/22034

图/表 4

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种类^aSpecies	拉丁名 Latin name			鉴定可行性^bIdentification feasibility
西伯利亚鲟(BAE)		Acipenser baerii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
西伯利亚鲟 × 俄罗斯鲟 (BAE × GUE)		Acipenser baerii × Acipenser gueldenstaedtii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
西伯利亚鲟 × 纳氏鲟 (BAE × NAC)		Acipenser baerii × Acipenser naccarii	N		正反杂交种均不可鉴定, 但贸易中实际为NAC♀ × BAE♂ Reciprocal hybrids cannot be identified, but in trade it is NAC♀ × BAE♂
西伯利亚鲟 × 施氏鲟 (BAE × SCH)		Acipenser baerii × Acipenser schrenckii	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
西伯利亚鲟 × 小体鲟 (BAE × RUT)		Acipenser baerii × Acipenser ruthenus	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
俄罗斯鲟(GUE)		Acipenser gueldenstaedtii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
俄罗斯鲟 × 西伯利亚鲟 (GUE × BAE)		Acipenser gueldenstaedtii × Acipenser baerii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
俄罗斯鲟 × 闪光鲟 (GUE × STE)		Acipenser gueldenstaedtii × Acipenser stellatus	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
纳氏鲟(NAC)		Acipenser naccarii	N		无法区分NAC和NAC × BAE、NAC × GUE NAC, NAC × BAE and NAC × GUE could not be distinguished
纳氏鲟 × 俄罗斯鲟 (NAC × GUE)		Acipenser naccarii × Acipenser gueldenstaedtii	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
小体鲟(RUT)		Acipenser ruthenus	N		无法区分RUT和RUT × BAE RUT and RUT × BAE could not be distinguished
小体鲟 × 欧洲鳇 (RUT × HUS)		Acipenser ruthenus × Huso huso	Y		正反杂交均可鉴定, 但贸易中实际为HUS♀ × RUT♂ Both positive and negative hybrids could be identified, but in trade it is HUS♀ × RUT♂
施氏鲟(SCH)		Acipenser schrenckii	N		无法区分SCH和SCH × BAE SCH and SCH × BAE could not be distinguished
闪光鲟(STE)		Acipenser stellatus	N		无法区分STE和STE × GUE STE and STE × GUE could not be distinguished
高首鲟(TRA)		Acipenser transmontanus	Y		尽管贸易中没有TRA × BAE、TRA × GUE, 但无法区分TRA与两种杂交鲟, 仅可在现有贸易情况下判定其为纯种TRA TRA, TRA × BAE and TRA × GUE could not be distinguished although the two hybrids did not exist in trade, so TRA only be considered pure under the current trade conditions
高首鲟 × 欧洲鳇 (TRA × HUS)		Acipenser transmontanus × Huso huso	Y		正反杂交均可鉴定 Reciprocal hybrids could be identified
达氏鳇(DAU)		Huso dauricus	Y		尽管贸易中没有DAU × BAE、DAU × GUE, 但无法区分DAU与两种杂交鲟, 仅可在现有贸易情况下判定其为纯种DAU DAU, DAU × BAE and DAU × GUE could not be distinguished although the two hybrids did not exist in trade, so DAU only be considered pure under the current trade conditions
达氏鳇 × 施氏鲟 (DAU × SCH)		Huso dauricus × Acipenser schrenckii	Y		正反杂交均可鉴定, 但贸易中实际为DAU♀ × SCH♂ Reciprocal hybrids could be identified, but in trade it is DAU♀ × SCH♂
欧洲鳇(HUS)		Huso huso	N		无法区分HUS和HUS × BAE HUS and HUS × BAE could not be distinguished
欧洲鳇 × 纳氏鲟 (HUS × NAC)		Huso huso × Acipenser naccarii	Y		正反杂交种均可鉴定 Reciprocal hybrids could be identified
欧洲鳇 × 西伯利亚鲟 (HUS × BAE)		Huso huso × Acipenser baerii	N		正反杂交种均不可鉴定 Reciprocal hybrids can not be identified
欧洲鳇 × 施氏鲟 (HUS × SCH)		Huso huso × Acioenser schrenckii	Y		正反杂交种均可鉴定 Reciprocal hybrids could be identified
欧洲鳇 × 闪光鲟 (HUS × STE)		Huso huso × Acipenser stellatus	Y		正反杂交种均可鉴定 Reciprocal hybrids could be identified
富氏鲟(FUL)		Acipenser fulvescens	Y		尽管贸易中没有FUL × BAE、FUL × GUE、FUL × RUT, 但无法区分FUL与三种杂交鲟, 仅可在现有贸易情况下判定其为纯种FUL FUL, FUL × BAE, FUL × GUE and FUL × RUT could not be distinguished although the three hybrids did not exist in trade, so FUL only be considered pure under the current trade conditions
中华鲟(SIN)		Acipenser sinensis	Y		尽管贸易中没有SIN × BAE、SIN × GUE、SIN × RUT, 但无法区分SIN与三种杂交鲟, 仅可在现有贸易情况下判定其为纯种SIN SIN, SIN × BAE, SIN × GUE and SIN × RUT could not be distinguished although the three hybrids did not exist in trade, so SIN only be considered pure under the current trade conditions
尖吻鲟(OXY)		Acipenser oxyrhynchus	Y		贸易中暂无其杂交鲟, 仅可通过现有引物扩增判断其是否与11种鲟鱼中除BAE和GUE之外的种类杂交产生杂交鲟; 仅可在现有贸易情况下判定其为纯种 These species have not produced hybrid sturgeon at present, and the existing primers can only judge whether these species produce hybrid sturgeon with other species except BAE and GUE among the 11 sturgeon species. So these species can only be considered pure under current trade conditions
铲鲟(PLA)		Scaphirhynchus platorynchus				Y
匙吻鲟(SPA)	Polyodon spathula			Y
短吻鲟(BRE)	Acipenser brevirostrum			Y
波斯鲟(PER)	Acipenser persicus			Y
中吻鲟(MED)	Acipenser medirostris			Y
大西洋鲟(STU)	Acipenser sturio			Y
裸腹鲟(NUD)	Acipenser nudiventris			Y
白铲鲟(ALB)	Scaphirhynchus albus			Y
太平洋鲟(MIK)	Acipenser mikadoi			Y
阿姆河大拟铲鲟(KAU)	Pseudoscaphirhynchus kaufmanni			Y

种类^aSpecies	拉丁名 Latin name			鉴定可行性^bIdentification feasibility
西伯利亚鲟(BAE)		Acipenser baerii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
西伯利亚鲟 × 俄罗斯鲟 (BAE × GUE)		Acipenser baerii × Acipenser gueldenstaedtii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
西伯利亚鲟 × 纳氏鲟 (BAE × NAC)		Acipenser baerii × Acipenser naccarii	N		正反杂交种均不可鉴定, 但贸易中实际为NAC♀ × BAE♂ Reciprocal hybrids cannot be identified, but in trade it is NAC♀ × BAE♂
西伯利亚鲟 × 施氏鲟 (BAE × SCH)		Acipenser baerii × Acipenser schrenckii	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
西伯利亚鲟 × 小体鲟 (BAE × RUT)		Acipenser baerii × Acipenser ruthenus	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
俄罗斯鲟(GUE)		Acipenser gueldenstaedtii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
俄罗斯鲟 × 西伯利亚鲟 (GUE × BAE)		Acipenser gueldenstaedtii × Acipenser baerii	N		缺乏可用于种类鉴定的基因标记 Lack of markers for species identification
俄罗斯鲟 × 闪光鲟 (GUE × STE)		Acipenser gueldenstaedtii × Acipenser stellatus	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
纳氏鲟(NAC)		Acipenser naccarii	N		无法区分NAC和NAC × BAE、NAC × GUE NAC, NAC × BAE and NAC × GUE could not be distinguished
纳氏鲟 × 俄罗斯鲟 (NAC × GUE)		Acipenser naccarii × Acipenser gueldenstaedtii	N		正反杂交种均不可鉴定 Reciprocal hybrids cannot be identified
小体鲟(RUT)		Acipenser ruthenus	N		无法区分RUT和RUT × BAE RUT and RUT × BAE could not be distinguished
小体鲟 × 欧洲鳇 (RUT × HUS)		Acipenser ruthenus × Huso huso	Y		正反杂交均可鉴定, 但贸易中实际为HUS♀ × RUT♂ Both positive and negative hybrids could be identified, but in trade it is HUS♀ × RUT♂
施氏鲟(SCH)		Acipenser schrenckii	N		无法区分SCH和SCH × BAE SCH and SCH × BAE could not be distinguished
闪光鲟(STE)		Acipenser stellatus	N		无法区分STE和STE × GUE STE and STE × GUE could not be distinguished
高首鲟(TRA)		Acipenser transmontanus	Y		尽管贸易中没有TRA × BAE、TRA × GUE, 但无法区分TRA与两种杂交鲟, 仅可在现有贸易情况下判定其为纯种TRA TRA, TRA × BAE and TRA × GUE could not be distinguished although the two hybrids did not exist in trade, so TRA only be considered pure under the current trade conditions
高首鲟 × 欧洲鳇 (TRA × HUS)		Acipenser transmontanus × Huso huso	Y		正反杂交均可鉴定 Reciprocal hybrids could be identified
达氏鳇(DAU)		Huso dauricus	Y		尽管贸易中没有DAU × BAE、DAU × GUE, 但无法区分DAU与两种杂交鲟, 仅可在现有贸易情况下判定其为纯种DAU DAU, DAU × BAE and DAU × GUE could not be distinguished although the two hybrids did not exist in trade, so DAU only be considered pure under the current trade conditions
达氏鳇 × 施氏鲟 (DAU × SCH)		Huso dauricus × Acipenser schrenckii	Y		正反杂交均可鉴定, 但贸易中实际为DAU♀ × SCH♂ Reciprocal hybrids could be identified, but in trade it is DAU♀ × SCH♂
欧洲鳇(HUS)		Huso huso	N		无法区分HUS和HUS × BAE HUS and HUS × BAE could not be distinguished
欧洲鳇 × 纳氏鲟 (HUS × NAC)		Huso huso × Acipenser naccarii	Y		正反杂交种均可鉴定 Reciprocal hybrids could be identified
欧洲鳇 × 西伯利亚鲟 (HUS × BAE)		Huso huso × Acipenser baerii	N		正反杂交种均不可鉴定 Reciprocal hybrids can not be identified
欧洲鳇 × 施氏鲟 (HUS × SCH)		Huso huso × Acioenser schrenckii	Y		正反杂交种均可鉴定 Reciprocal hybrids could be identified
欧洲鳇 × 闪光鲟 (HUS × STE)		Huso huso × Acipenser stellatus	Y		正反杂交种均可鉴定 Reciprocal hybrids could be identified
富氏鲟(FUL)		Acipenser fulvescens	Y		尽管贸易中没有FUL × BAE、FUL × GUE、FUL × RUT, 但无法区分FUL与三种杂交鲟, 仅可在现有贸易情况下判定其为纯种FUL FUL, FUL × BAE, FUL × GUE and FUL × RUT could not be distinguished although the three hybrids did not exist in trade, so FUL only be considered pure under the current trade conditions
中华鲟(SIN)		Acipenser sinensis	Y		尽管贸易中没有SIN × BAE、SIN × GUE、SIN × RUT, 但无法区分SIN与三种杂交鲟, 仅可在现有贸易情况下判定其为纯种SIN SIN, SIN × BAE, SIN × GUE and SIN × RUT could not be distinguished although the three hybrids did not exist in trade, so SIN only be considered pure under the current trade conditions
尖吻鲟(OXY)		Acipenser oxyrhynchus	Y		贸易中暂无其杂交鲟, 仅可通过现有引物扩增判断其是否与11种鲟鱼中除BAE和GUE之外的种类杂交产生杂交鲟; 仅可在现有贸易情况下判定其为纯种 These species have not produced hybrid sturgeon at present, and the existing primers can only judge whether these species produce hybrid sturgeon with other species except BAE and GUE among the 11 sturgeon species. So these species can only be considered pure under current trade conditions
铲鲟(PLA)		Scaphirhynchus platorynchus				Y
匙吻鲟(SPA)	Polyodon spathula			Y
短吻鲟(BRE)	Acipenser brevirostrum			Y
波斯鲟(PER)	Acipenser persicus			Y
中吻鲟(MED)	Acipenser medirostris			Y
大西洋鲟(STU)	Acipenser sturio			Y
裸腹鲟(NUD)	Acipenser nudiventris			Y
白铲鲟(ALB)	Scaphirhynchus albus			Y
太平洋鲟(MIK)	Acipenser mikadoi			Y
阿姆河大拟铲鲟(KAU)	Pseudoscaphirhynchus kaufmanni			Y

鲟鱼分子鉴定方法的整合应用

Integration and application of sturgeon identification methods

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