Research on fish diversity in Xijiang Rare Fish Provincial Nature Reserve based on environmental DNA technology
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摘要:
为更好地保护西江珍稀鱼类省级自然保护区的鱼类资源，利用基于COI和12S rRNA 2个基因标记的环境DNA技术 (Environmental DNA, eDNA)，对保护区江段的左岸、中间和右岸共计18个采样站位开展了鱼类种类组成及河段不同横向断面的鱼类多样性分析。结果发现，COI和12S rRNA 2个基因共检测出53种鱼类，隶属于4目12科48属；12S rRNA基因检测出47种鱼类，COI基因检测出19种鱼类，2个基因同时检测出的鱼类有13种。基于序列丰度的Alpha多样性分析结果显示，2个基因的结果一致支持保护区江段左岸的鱼类物种数和Chao1丰富度最高，说明左岸具有较高的鱼类多样性。基于12S rRNA基因的Beta多样性结果发现，保护区的左岸、中间和右岸的鱼类组成存在显著性差异。研究结果可为保护区鱼类生物多样性的保护和动态监测提供基础资料与技术支持。
Abstract:To better protect the fish resources in the Xijiang Rare Fish Provincial Nature Reserve, we used the environmental DNA (eDNA) technology to analyze the fish species composition and fish diversity at different cross sections of 18 sampling stations on the left, middle and right banks of the river section. Two molecular markers, i.e. COI and 12S rRNA, were used in this study. The results show that the two genes of 12S rRNA and COI were detected in 53 fish species, belonging to 48 genera, 12 families and 4 orders. Forty-seven and nineteen fish species were examined based on 12S rRNA and COI, respectively. In addition, 13 shared fish species were discovered by using the two gene markers. Alpha diversity analysis based on sequence abundance shows that the two markers unanimously supported the highest number of fish species and the highest richness of Chao 1 index on the left bank of the river section, indicating that the left bank appears to have higher fish diversity than the middle and right banks. Beta diversity analysis based on 12S rRNA shows obvious differences in the fish composition on the left, middle and right banks. The research provides basic data and technical support for the protection and dynamic monitoring of fish biodiversity in this nature reserve.
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Keywords:
- Environmental DNA /
- Fish diversity /
- Nature reserve /
- Xijiang rare fish
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图 1 采样站位示意图
注：L1—L6 为左岸区域；M1—M6 为中间区域；R1—R6 为右岸区域。
Figure 1. Information of sampling stations
Note: L1–L6 are the left bank area; M1–M6 are the middle area; R1–R6 are the right bank area.
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图 3 不同横向采样断面平均物种数与Alpha多样性指数
注：a和b表示12S rRNA的鉴定结果；c和d表示COI的鉴定结果。
Figure 3. Average species number and Alpha diversity indexes of different transverse sections
Note: a and b represent identification results based on 12S rRNA; c and d represent identification results based on COI.
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图 4 基于12S rRNA (左) 和COI基因 (右) 的各区域eDNA检测鱼类组成相似性
Figure 4. Composition similarity of fish detected in each area based on 12S rRNA (Left) and COI gene (Right) of eDNA
表 1 基于12S rRNA和COI基因检测的物种名录及序列数
Table 1 List of fish species and sequence number of detected based on 12S rRNA and COI genes
种类
Species12S rRNA COI 左岸
Left bank中间
Middle bank右岸
Right bank左岸
Left bank中间
Middle bank右岸
Right bank鲤形目 Cypriniformes 鲤科 Cyprinidae 宽鳍鱲 Zacco platypus 54 93 8 171 0 0 0 马口鱼 Opsariichthys bidens● 0 0 0 1 308 1 403 1 677 南方波鱼 Rasbora steineri 0 0 0 709 1 155 693 青鱼 Mylopharyngodon piceus 14 043 436 1 699 0 0 0 草鱼 Ctenopharyngodon idella 4 639 22 577 6 209 594 270 442 鳤 Ochetobius elongatus 6 521 78 12 0 0 0 鳡 Elopichthys bambusa 35 532 12 799 10128 1 699 1 437 498 赤眼鳟 Squaliobarbus curriculus 90 047 28 123 23 347 197 1 0 银飘鱼 Pseudolaubuca sinensis 2 781 20 893 275 0 0 0 鳊 Parabramis pekinensis 4 774 487 494 0 0 0 䱗 Hemiculter leucisculus 853 176 224 336 29 961 0 0 0 细鳊 Metzia formosae● 3 0 0 0 0 0 南方拟䱗 Pseudohemiculter dispar● 0 0 0 543 6 0 海南似鱎 Toxabramis houdemeri 4 0 0 0 0 0 广东鲂 Megalobrama terminalis 3 268 50 479 9 171 12 54 1 海南鲌 Chanodichthys erythropterus 0 0 1 0 0 0 黄尾鲴 Xenocypris davidi 953 229 383 550 20 524 4 070 1 059 95 圆吻鲴 Distoechodon tumirostris● 5 2 2 0 0 0 鲢 Hypophthalmichthys molitrix 36 452 379 14 407 74 406 46 鳙 Aristichthys nobilis 29 337 16 072 22 872 0 0 0 银鮈 Squalidus argentatus 1 6 433 4 0 100 0 福建小鳔鮈 Microphysogobio fukiensis● 1 1 866 0 0 0 0 兴凯鱊 Acanthorhodeus chankaensis● 3 260 0 0 0 0 0 越南鱊 Acheilognathus tonkinensis● 0 0 482 6 52 0 高体鳑鲏 Rhodeus ocellatus 5 521 78 071 1 760 0 25 0 倒刺鲃 Spinibarbus denticulatus 37 1 3 098 0 0 0 虹彩光唇鱼 Acrossocheilus iridescens● 0 23 6 0 0 0 鲮 Cirrhinus molitorella 60 257 55 395 26 860 0 0 0 纹唇鱼 Osteochilus salsburyi 0 18 581 0 0 0 0 鲤 Cyprinus carpio 62 807 63 992 53 396 145 13 158 鲫 Carassius cuvieri 12 1 0 0 0 0 平鳍鳅科 Homalopteridae 平舟原缨口鳅 Vanmanenia pingchowensis● 0 0 9 0 0 0 鲇形目 Siluriformes 鲇科 Siluridae 鲇 Silurus asotus 831 0 0 0 0 0 大口鲇 Silurus meridionalis● 565 0 0 0 0 0 胡子鲇科 Clariidae 革胡子鲇 Clarias gariepinus▲● 1 0 0 0 0 0 甲鲶科 Loricariidae 豹纹翼甲鲇 Pterygoplichthys pardalis▲● 4 853 8 315 0 0 0 长臀鮈科 Cranoglanididae 长臀鮈 Cranoglanis bouderius 0 0 191 0 0 0 鲿科 Bagridae 黄颡鱼 Pelteobagrus fulvidraco 0 0 905 0 0 0 瓦氏黄颡鱼 Pseudobagrus vachelli 0 0 0 1 000 388 376 粗唇鮈 Pseudobagrus crassilabris 947 0 1 303 25 0 0 条纹鮈 Tachysurus virgatus 863 0 0 0 0 0 斑鳠 Hemibagrus guttatus★ 178 4 233 49 0 0 0 鲈形目 Perciformes 鮨鲈科 Percichthyidae 斑鳜 Siniperca scherzeri 1 106 0 0 0 0 0 波纹鳜 Siniperca undulata● 626 2 0 0 0 0 大眼鳜 Siniperca knerii 0 0 0 2 0 0 花鲈 Lateolabrax japonicus 0 0 0 27 133 0 刺臀鱼科 Centrarchidae 大口黑鲈 Micropterus salmoides▲● 1 075 0 0 0 0 0 丽鱼科 Cichlidae 齐氏罗非鱼 Coptodon zillii▲● 8 762 6 784 5 0 530 0 尼罗罗非鱼 Oreochromis niloticus▲ 3 927 3 5 660 641 1 192 296 鰕虎鱼科 Gobiidae 金黄舌鰕虎 Glossogobius aureus● 356 2 291 4 0 0 0 粘皮鲻鰕虎 Mugilogobius myxodermus● 1 10 3 0 0 0 波氏吻鰕虎 Rhinogobius cliffordpopei● 6 11 266 82 0 0 0 鲀形目 Tetraodontiformes 鲀科 Tetraodontidae 弓斑东方鲀Takifugu ocellatus 2 013 27 400 10 454 0 0 0 注：▲. 外来物种；★. 国家重点保护水生野生动物；●. 与李捷等[21]2006—2008年保护区调查相比的新增种类。 Note: ▲. Alien species; ★. National key protected aquatic wildlife; ●. New species compared with the conservation area survey conducted by Li et al[21] from 2006 to 2008. 
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表 2 各区域Alpha多样性指数
Table 2 Alpha diversity index of each area
区域
Area物种数
Species indexChao1指数
Chao1 index香农指数
Shannon index辛普森指数
Gini-Simpson index覆盖度
Coverage12S rRNA COI 12S rRNA COI 12S rRNA COI 12S rRNA COI 12S rRNA COI L1 26 10 1.94 1.20 1.60 1.60 0.70 0.69 1.00 1.00 L2 28 8 2.11 0.98 1.67 0.34 0.72 0.13 1.00 1.00 L3 31 9 2.32 1.08 1.20 0.51 0.60 0.20 1.00 1.00 L4 27 3 2.02 0.46 1.66 0.41 0.71 0.21 1.00 1.00 L5 24 3 1.79 0.41 1.36 0.70 0.67 0.48 1.00 0.99 L6 32 8 2.41 1.01 1.65 1.17 0.72 0.64 1.00 0.99 M1 28 4 2.22 0.63 2.18 0.89 0.85 0.55 1.00 0.99 M2 27 5 2.16 0.70 2.17 1.15 0.86 0.61 1.00 1.00 M3 21 4 1.63 0.45 1.61 0.79 0.77 0.49 1.00 1.00 M4 24 3 1.86 0.28 1.53 0.55 0.66 0.34 1.00 1.00 M5 29 3 2.27 0.45 1.78 0.71 0.75 0.48 1.00 0.99 M6 23 3 1.81 0.37 1.83 0.71 0.75 0.50 1.00 0.99 R1 25 4 2.35 0.52 1.40 0.89 0.60 0.55 0.99 0.99 R2 29 5 2.54 0.92 1.83 0.88 0.76 0.45 1.00 0.98 R3 24 6 2.12 0.87 1.97 0.54 0.80 0.25 0.99 0.99 R4 24 5 2.11 0.91 1.66 1.11 0.67 0.60 0.99 0.99 R5 21 1 1.72 0 2.24 0 0.85 0 1.00 0.97 R6 17 2 1.39 0.91 1.55 0.64 0.73 0.67 1.00 0.98 注：L1—L6为左岸区域；M1—M6为中间区域；R1—R6为右岸区域。 Note: L1–L6 are the left bank area; M1–M6 are the middle area; R1–R6 are the right bank area.
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