甘肃省鱼类资源现状及DNA条形码在鱼类物种鉴定中的应用

doi:10.17520/biods.2014185

摘要/Abstract

摘要：

为了摸清甘肃省土著鱼类资源与分布现状, 探索DNA条形码在鱼类辅助物种鉴定中的适用性, 2012年6-9月对甘肃境内黄河水系、嘉陵江水系和河西内陆河水系进行了较全面的鱼类调查。共采集鱼类标本3,087尾, 隶属于5目10科38属64种, 以鲤科种类最多, 为30种, 占总种数的46.88%。物种多样性分析表明, 在黄河水系的夏河和庄浪河多样性指数是所有调查点中最低的, 分别为1.38和1.09。嘉陵江水系各河段的多样性指数较高(H = 2.15-3.27), 其次为河西内陆河水系(H = 2.01-2.83)。在河西内陆河水系中, 疏勒河的均匀度指数最高, 为1.10, 黑河最低(0.68)。庄浪河的优势度指数最高, 为0.34, 而嘉陵江干流两当段的优势度指数在所有调查点中最低, 为0.04。利用DNA条形码分析了49种662尾标本的COI基因部分序列, 大部分种类在neighbor-joining系统树中形成各自的单系, 种内平均遗传距离0.88%, 种间平均遗传距离为9.99%, 在种内和种间COI序列遗传距离之间形成明显的条形码间隙, 斯氏高原鳅(Triplophysa stoliczkae)与达里湖高原鳅(T. dalaica), 甘肃高原鳅(T. robusta)与似鲇高原鳅(T. siluroides), 嘉陵裸裂尻鱼(Schizopygopsis kialingensis)与黄河裸裂尻鱼(S. pylzovi)之间的遗传距离低于2%, 甘肃高原鳅与似鲇高原鳅不能通过COI基因片段区分开, 其他两对物种可以采用核苷酸诊断法来进一步区分。斯氏高原鳅和拉氏鱼岁(Phoxinus lagowskii)种内遗传分歧较大, 揭示种内可能存在隐存种。结果表明, 对某些近缘种和不同地理种群差异较大的物种, 要将分子、形态和地理分布特点结合起来才能准确鉴定。

关键词: 淡水鱼类, 物种多样性, DNA条形码, COI基因, 物种鉴定, 隐存种

Abstract:

Between June and September in 2012, we investigated fish resources from the Yellow River, Jialing rivers and Hexi inland rivers in Gansu Province, China. Three thousand specimens were collected, belonging to 64 species, 38 genera, 10 families and 5 orders. Cyprinidae was the predominant species, accounting for 45% of the total species. Diversity analysis showed that fish diversity in the Jialing River basin was the highest (H = 2.15-3.27), followed by the inland rivers (H = 2.01-2.83). Two tributaries of the Yellow River, the Xiahe River and the Zhuanglang River had relatively lower diversity levels with the H values of 1.38 and 1.09, respectively. The Pielou index of Shule River was the highest with a value of 1.10, while that of the Heihe River was the lowest with a value of 0.68. The Simpson index of the Zhuanglang River was 0.34, the highest of the investigated rivers, while the trunk streams of Jialing River was the lowest. A total of 662 specimens of 49 species were randomly selected for DNA barcoding analysis using a partial sequence of the COI gene. The results indicated that most species formed monophyletic groups in the neighbor-joining tree. The average K2P genetic divergence was 0.88% within species and 9.99% among species. A significant barcode gap was found between intraspecific genetic distance and interspecific genetic distance. A nucleotide diagnostic method was needed for further identification because the K2P genetic distance of three species pairs (between Triplophysa stoliczkae and T. dalaica, T. robusta and T. siluroides, Schizopygopsis kialingensis and S. pylzovi) was less than 2%. Cryptic species may exist within Triplophysa stoliczkae and Phoxinus lagowskii. Triplophysa robusta and T. siluroides cannot be distinguished through COI gene sequences. The results showed that molecular methods, morphological characteristic and geographical distribution of species should be combined in order to comprehensively and accurately identify closely related species and geographic populations.

Key words: freshwater fish, species diversity, DNA barcoding, COI, species identification, cryptic species

王太, 张艳萍, 管丽红, 杜岩岩, 娄忠玉, 焦文龙. 甘肃省鱼类资源现状及DNA条形码在鱼类物种鉴定中的应用[J]. 生物多样性, 2015, 23(3): 306-313.

Tai Wang,Yanping Zhang,Lihong Guan,Yanyan Du,Zhongyu Lou,Wenlong Jiao. Current freshwater fish resources and the application of DNA barcoding in species identification in Gansu Province[J]. Biodiv Sci, 2015, 23(3): 306-313.

图/表 6

参考文献 11

12	Tang WJ (唐文家), Chen YF (陈毅峰), Ding CZ (丁城志) (2013) The current situation and protection of fish resources in Huangshui River in Qinghai Province.Journal of Dalian Ocean University(大连海洋大学学报), 28, 307-313. (in Chinese with English abstract)
13	Tang WJ (唐文家), Cui YX (崔玉香), Zhao X (赵霞) (2012) Hydrobiontes resource survey of Lancang River in Qinghai Province.Journal of Hydroecology(水生态学杂志), 33, 20-28. (in Chinese with English abstract)
14	Tang WJ (唐文家), Zhao X (赵霞), Zhang MT (张妹婷) (2012) Investigation on aquatic organisms in the upper stream of Heihe River in Qinghai Province, China. Journal of Dalian Ocean University(大连海洋大学学报), 27, 477-482. (in Chinese with English abstract)
15	Wang XT (王香亭) (1991) Gansu Vertebrate Fauna (甘肃省脊椎动物志). Gansu Science and Technology Press, Lanzhou. (in Chinese)
16	Wang ZD, Guo YS, Liu XM, Fan UB, Liu CW (2012) DNA barcoding South China Sea fishes.Mitochondrial DNA, 23, 405-410.
17	Ward RD, Zemlak TS, Innes BH, Last PR, Hebert PDN (2005) DNA barcoding of Australia’s fish species.Philosophical Transactions of the Royal Society of London Series B-Biological Sciences, 360, 1847-1857.
18	Yue PQ (乐佩琦) (2000) Fauna Sinica, Osteichthyes, Cypriniformes Ⅲ (中国动物志·硬骨鱼纲·鲤形目(下卷)). Science Press, Beijing. (in Chinese)
19	Zhao K (赵凯), Duan ZY (段子渊), Yang GS (杨公社), Peng ZG (彭作刚), He SP (何舜平), Chen YY (陈宜瑜) (2007) The origin of Gymnocypris przewalskii and Gymnocypris eckloni population evolution.Progress in Natural Science(自然科学进展), 17, 320-328. (in Chinese with English abstract)
20	Zhu SQ (朱松泉) (1989) The Loaches of the Subfamily Nemacheilinae in China (Cypriniformes: Cobitidae) (中国条鳅志). Jiangsu Science and Technology Press, Nanjing. (in Chinese)
1	April J, Mayden RL, Hanner R, Bernatchez L (2011) Genetic calibration of species diversity among North America’s freshwater fishes. Proceedings of the National Academy of Sciences, USA, 26, 10602-10607.
2	Chen YY (陈宜瑜) (1998) Fauna Sinica, Osteichthyes, Cypriniformes Ⅱ (中国动物志·硬骨鱼纲·鲤形目(中卷)). Science Press, Beijing. (in Chinese)
3	Chu XL (褚新洛), Zheng BS (郑葆珊), Dai DY (戴定远) (1999) Fauna Sinica, Osteichthyes, Siluriformes (中国动物志·硬骨鱼纲·鲇形目). Science Press. Beijing. (in Chinese)
4	He DK (何德奎), Chen YF (陈毅峰) (2007) Molecular phylogenetic and biogeography of the highly specialized grade Schizothoracinae fishes.Chinese Science Bulletin(科学通报), 52, 303-312. (in Chinese)
5	Hebert N, Cywinska A, Ball SL, Ward JR (2003) Biological identifications through DNA barcodes.Proceedings of the Royal Society of London B: Biological Sciences, 270, 313-321.
6	Hubert N, Hanner R, Holm E, Mandrak NE, Taylor E, Burridge M, Watkinson D, Dumont P, Curry A, Bentzen P, Zhang J, April J, Bernatchez L (2008) Identifying Canadian freshwater fishes through DNA barcodes. PLoS ONE, 3, e2490.
7	Hubert N, Meyer CP, Bruggemann HJ, Guérin F, Komeno RJL, Espiau B, Causse R, Williams JT, Planes S (2012) Cryptic diversity in Indo-Pacific coral-reef fishes revealed by DNA-barcoding provides new support to the centre-of- overlap hypothesis. PLoS ONE, 7, e28987.
8	Li XJ (李学军), Li SF (李思发), Yang HQ (杨和荃) (1996) A comparative study on the diversity of aquatic organisms in Tiane Zhou Oxbow and Laohe Oxbow of Yangtze River.Chinese Biodiversity(生物多样性), 4, 211-216. (in Chinese with English abstract)
9	Pereira LHG, Hanner R, Foresti F, Oliveira C (2013) Can DNA barcoding accurately discriminate megadiverse Neotropical freshwater fish fauna?BMC Genetics, 14, 20.
10	Rach J, DeSalle R, Sarkar IN, Schierwater B, Hadrys H (2008) Character-based DNA barcoding allows discrimination of genera, species and populations in Odonata.Proceedings of the Royal Society B: Biological Sciences, 275, 237-247.
11	Shen YB (沈玉帮), Zhang JB (张俊彬), Feng BB (冯冰冰), Li JL (李家乐) (2011) Genetic diversity of blue mussel Mytilus galloprovincialis in China based on sequence analyses of mitochondrial COI gene.Marine Science Bulletin(海洋通报), 30, 435-440. (in Chinese with English abstract)

采样点 Sites		物种数 No. of species	Shannon-Wiener指数 Shannon-Wiener index (H)	均匀度指数 Pielou index (J′)	优势度指数 Simpson index (λ)
黄河 Yellow River	玛曲段 Maqu section	11	2.32	0.97	0.10
	兰州段 Lanzhou section	14	2.37	0.90	0.11
	夏河 Xiahe River	5	1.38	0.99	0.25
	庄浪河 Zhuanglang River	4	1.09	0.99	0.34
	洮河上游 Upper reaches of Taohe River	7	1.78	0.92	0.20
	洮河中游 Middle reaches of Taohe River	7	1.77	0.91	0.20
	洮河下游 Lower reaches of Taohe River	9	2.08	0.94	0.14
	渭河上游 Upper reaches of Weihe River	8	2.03	0.98	0.14
	籍河 Xihe River	9	1.98	0.90	0.16
	葫芦河 Hulu River	8	1.91	0.92	0.17
	泾河平凉段 Pingliang section of Jinghe River	14	2.47	0.93	0.09
	蒲河茹河 Pu River and Ru River	8	1.93	0.93	0.15
	马莲河 Malian River	14	2.40	0.91	0.11
嘉陵江 Jialing River	白龙江 Bailong River	12	2.17	0.87	0.14
	西汉水 Xihan River	12	2.30	0.92	0.12
	永宁河 Yongning River	15	2.53	0.93	0.09
	燕子河 Yanzi River	27	3.14	0.95	0.05
	杜家河 Dujia River	9	2.15	0.98	0.12
	干流两当段 Liangdang section	32	3.27	0.94	0.04
河西内陆河 Hexi inland rivers	石羊河 Shiyang River	12	2.19	0.88	0.14
	黑河 Heihe River	19	2.01	0.68	0.05
	疏勒河 Shule River	13	2.83	1.10	0.19

采样点 Sites		物种数 No. of species	Shannon-Wiener指数 Shannon-Wiener index (H)	均匀度指数 Pielou index (J′)	优势度指数 Simpson index (λ)
黄河 Yellow River	玛曲段 Maqu section	11	2.32	0.97	0.10
	兰州段 Lanzhou section	14	2.37	0.90	0.11
	夏河 Xiahe River	5	1.38	0.99	0.25
	庄浪河 Zhuanglang River	4	1.09	0.99	0.34
	洮河上游 Upper reaches of Taohe River	7	1.78	0.92	0.20
	洮河中游 Middle reaches of Taohe River	7	1.77	0.91	0.20
	洮河下游 Lower reaches of Taohe River	9	2.08	0.94	0.14
	渭河上游 Upper reaches of Weihe River	8	2.03	0.98	0.14
	籍河 Xihe River	9	1.98	0.90	0.16
	葫芦河 Hulu River	8	1.91	0.92	0.17
	泾河平凉段 Pingliang section of Jinghe River	14	2.47	0.93	0.09
	蒲河茹河 Pu River and Ru River	8	1.93	0.93	0.15
	马莲河 Malian River	14	2.40	0.91	0.11
嘉陵江 Jialing River	白龙江 Bailong River	12	2.17	0.87	0.14
	西汉水 Xihan River	12	2.30	0.92	0.12
	永宁河 Yongning River	15	2.53	0.93	0.09
	燕子河 Yanzi River	27	3.14	0.95	0.05
	杜家河 Dujia River	9	2.15	0.98	0.12
	干流两当段 Liangdang section	32	3.27	0.94	0.04
河西内陆河 Hexi inland rivers	石羊河 Shiyang River	12	2.19	0.88	0.14
	黑河 Heihe River	19	2.01	0.68	0.05
	疏勒河 Shule River	13	2.83	1.10	0.19

	平均值 Average (%)	最小值 Minimum (%)	最大值 Maximum (%)	标准误 SE
种内 Species	0.88	0.00	11.46	0.07
同属种间 Among species within genus	9.99	0.00	15.15	0.29
同科属间 Among genera within family	17.95	2.81	24.95	0.29
同目科间 Among families within order	24.13	20.33	27.52	0.53

	平均值 Average (%)	最小值 Minimum (%)	最大值 Maximum (%)	标准误 SE
种内 Species	0.88	0.00	11.46	0.07
同属种间 Among species within genus	9.99	0.00	15.15	0.29
同科属间 Among genera within family	17.95	2.81	24.95	0.29
同目科间 Among families within order	24.13	20.33	27.52	0.53

目 Order	科 Family	属 Genus	种 Species
鲑形目 Salmoniformes	鲑科 Salmonidae	细鳞鲑属 Brachymystax	秦岭细鳞鲑 Brachymystax lenok tsinlingensis
		公鱼属 Hypomesus	池沼公鱼 Hypomesus olidus
鲇形目 Siluriformes	鲇科 Siluridae	鲇属 Silurus	鲇鱼 Silurus asotus
鲇形目 Siluriformes	鲇科 Siluridae		怀头鲶 Silurus soldatovi
	鮡科 Sisoridae	鮡属 Pareuchiloglanis	前臀鮡 Pareuchiloglanis anteanalis
		纹胸鮡属 Glyptothorax	中华纹胸鮡 Glyptothorax sinense
	鲿科 Bagridae	拟鲿属 Pseudobagrus	切尾拟鲿 Pseudobagrus truncatus
		鱯属 Mystus	大鳍鱯 Mystus macropterus
鲈形目 Perciformes	鰕虎鱼科 Gobiidae	吻鰕虎鱼属 Rhinogobius	褐吻鰕虎鱼 Rhinogobius brunneus
	塘鳢科 Hypseleotris	黄黝鱼属 Hypseleotris	黄黝鱼 Hypseleotris swinhonis
合鳃鱼目 Symbranchiformes	合鳃鱼科 Synbranchidae	黄鳝属 Monopterus	黄鳝 Monopterus albus
鲤形目 Cypriniformes	鲤科 Cyprinidae	鰟鮍属 Rhodeus	高体鰟鮍 Rhodeus ocellatus
		棒花鱼属 Abbottina	棒花鱼 Abbottina rivularis
		颌须鮈属 Gnathopogon	嘉陵颌须鮈 Gnathopogon herzensteini
		颌须鮈属 Gnathopogon	短须颌须鮈 Gnathopogon imberbis
		胡鮈属 Huigobio	清徐胡鮈 Huigobio chinssuensis
		鮈属 Gobio	黄河鮈 Gobio huanghensis
		鮈属 Gobio	似铜鮈 Gobio coriparoides
		蛇鮈属 Saurogobio	蛇鮈 Saurogobio dabryi
		似鮈属 Pseudogobio	似鮈 Pseudogobio vaillanti
		片唇鮈属 Platysmacheilus	裸腹片唇鮈 Platysmacheilus nudiventris
		麦穗鱼属 Pseudorasbora	麦穗鱼 Pseudorasbora parva
		属 Hemibarbus	唇 Hemibarbus labeo
		属 Hemibarbus	花 Hemibarbus maculatus
		似属 Belligobio	似 Belligobio nummifer
		马口鱼属 Opsariichthys	马口鱼 Opsariichthys bidens
		鱲属 Zacco	宽鳍鱲 Zacco platypus
		属 Phoxinus	拉氏 Phoxinus lagowskii
		属 Phoxinus	尖头 Phoxinus oxycephalus
		白甲鱼属 Onychostoma	多鳞白甲鱼 Onychostoma macrolepis
		鲫属 Carassius	鲫鱼 Carassius auratus
		重唇鱼属 Gymnodiptychus	渭河裸重唇鱼 Gymnodiptychus pachycheilus weiheensis
			厚唇裸重唇鱼 Gymnodiptychus pachycheilus
		裸鲤属 Gymnocypris	花斑裸鲤 Gymnocypris eckloni
		裸鲤属 Gymnocypris	青海湖裸鲤 Gymnocypris przewalskii
			祁连裸鲤 Gymnocypris chilianensis
		裸裂尻鱼属 Schizopygopsis	嘉陵裸裂尻鱼 Schizopygopsis kialingensis
			黄河裸裂尻鱼 Schizopygopsis pylzovi
		裂腹鱼属 Schizothorax	重口裂腹鱼 Schizothorax (Racoma) davidi
			齐口裂腹鱼 Schizothorax (schizothorax.) prenanti
		扁咽齿鱼属 Platypharodon	极边扁咽齿鱼 Platypharodon extremus
	平鳍鳅科 Balitoridae	后平鳅属 Metahomaloptera	峨眉后平鳅 Metahomaloptera omeiensis
	鳅科 Cobitidae	泥鳅属 Misgurnus	泥鳅 Misgurnus anguillicaudatus
		副泥鳅属 Paramisgurnus	大鳞副泥鳅 Paramisgurnus dabryanus
		副鳅属 Paracobitis	红尾副鳅 Paracobitis variegatus
		花鳅属 Cobitis	北方花鳅 Cobitis granoci
		花鳅属 Cobitis	中华花鳅 Cobitis sinensis
		鼓鳔鳅属 Hedinichthys	大鳍鼓鳔鳅 Hedinichthys yarkandensis macroptera
		高原鳅属 Triplophysa	短尾高原鳅 Triplophysa brevviuda
		高原鳅属 Triplophysa	酒泉高原鳅 Triplophysa hsutschouensi
			岷县高原鳅 Triplophysa minxianensis
			赛丽高原鳅 Triplophysa sellaefer
			甘肃高原鳅 Triplophysa robusta
			似鲇高原鳅 Triplophysa siluroides
			安氏高原鳅 Triplophysa angeli
			武威高原鳅 Triplophysa wuweiensis
			小眼高原鳅 Triplophysa microps
			黑体高原鳅 Triplophysa obscura
			细尾高原鳅 Triplophysa stenura
			长身高原鳅 Triplophysa tenuis
			硬刺高原鳅 Triplophysa scleroptera
			东方高原鳅 Triplophysa orientalis
			斯氏高原鳅 Triplophysa stoliczkae
			达里湖高原鳅 Triplophysa dalaica