生物多样性 ›› 2015, Vol. 23 ›› Issue (3): 306-313.doi: 10.17520/biods.2014185

所属专题: DNA条形码应用进展

• • 上一篇    下一篇

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

王太1, 张艳萍1, *(), 管丽红2, 杜岩岩1, 娄忠玉1, 焦文龙1   

  1. 1 (甘肃省水产研究所, 兰州 730030)
    2 (中国科学院水生生物研究所, 武汉 430072);
  • 收稿日期:2014-09-02 接受日期:2015-04-18 出版日期:2015-06-09
  • 通讯作者: 张艳萍 E-mail:aqhongqi@qq.com
  • 基金项目:
    甘肃省科技创新服务平台建设计划(1309JTPA015)

Current freshwater fish resources and the application of DNA barcoding in species identification in Gansu Province

Tai Wang1, Yanping Zhang1, *, Lihong Guan2, Yanyan Du1, Zhongyu Lou1, Wenlong Jiao1   

  1. 1 Gansu Fishery Research Institute, Lanzhou 730030
    2 Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072
  • Received:2014-09-02 Accepted:2015-04-18 Online:2015-06-09
  • Contact: Zhang Yanping E-mail:aqhongqi@qq.com

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

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

图1

甘肃省鱼类调查采样点分布图"

表1

甘肃省的鱼类多样性"

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

图2

基于COI序列构建的NJ系统发育树"

表2

不同分类阶元的K2P遗传距离值"

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

图3

同属种间与种内遗传距离频率分布图"

附录1

甘肃省鱼类种类组成"

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
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
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
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
Hemibarbus Hemibarbus labeo
Hemibarbus maculatus
Belligobio Belligobio nummifer
马口鱼属 Opsariichthys 马口鱼 Opsariichthys bidens
鱲属 Zacco 宽鳍鱲 Zacco platypus
Phoxinus 拉氏 Phoxinus lagowskii
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
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
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
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
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