鱼类环境DNA metabarcoding片段的近缘物种识别差异

陈治; 马春来; 叶乐; 杨超杰; 王海山

doi:10.12284/hyxb2022150

鱼类环境DNA metabarcoding片段的近缘物种识别差异

doi: 10.12284/hyxb2022150

陈治^{1, 2,},
马春来^{1, 2},
叶乐^{1, 2},
杨超杰^{1, 2},
王海山^{1, 2, ,}

1.
海南热带海洋学院热带海洋生物资源利用与保护教育部重点实验室，海南三亚 572022
2.
海南热带海洋学院海南省热带海洋渔业资源保护与利用重点实验室，海南三亚 572022

基金项目: 国家自然科学基金（32002389）；海南省自然科学基金(422RC717)；海南热带海洋学院引进人才科研启动资助项目（RHDRC201907）。

详细信息

作者简介:
陈治（1990-），男，山东省泰安市人，讲师，博士，主要从事海洋鱼类分类学及其资源保护研究。E-mail：change@139.com

通讯作者:
王海山（1983-），男，讲师，博士，主要从事海洋生物多样性及其系统地理学研究。E-mail：jazz-123@163.com

中图分类号: S931；Q523
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出版历程
- 收稿日期: 2022-02-17
- 修回日期: 2022-03-25
- 网络出版日期: 2022-04-24
- 刊出日期: 2022-08-15

Differences of eDNA metabarcoding fragments in relative fish species resolution

Chen Zhi^{1, 2
,},
Ma Chunlai^{1, 2},
Ye Le^{1, 2},
Yang Chaojie^{1, 2},
Wang Haishan^{1, 2
, ,}

1.
Key Laboratory of Utilization and Conservation for Tropical Marine Bioresources, Ministry of Education, Hainan Tropical Ocean University, Sanya 572022, China
2.
Hainan Key Laboratory for Conservation and Utilization of Tropical Marine Fishery Resources, Hainan Tropical Ocean University, Sanya 572022, China

摘要

摘要: 已知的鱼类环境DNA（eDNA）metabarcoding片段均未被针对性考察其对近缘物种的适用性，实际使用过程中存在“物种丢失”风险。为筛选出物种识别率最高的片段，本研究比较了15个主流片段对106属（共935种）鱼类的识别差异。研究结果如下：（1）蛋白质编码基因（COI，片段15）的物种识别率最高，但其引物通用性最差；片段09、片段11、片段07、片段03、片段12的引物序列总平均遗传距离也较大，均存在eDNA低效扩增的风险；（2）片段长度影响物种识别率，核糖体基因中片段05、片段06、片段01、片段02及片段13的物种识别率较高；（3）非度量多维尺度分析（NMDS）显示，不同基因、同一基因不同片段的识别结果存在较大差异，应考虑多片段、多基因组合应用；片段01与片段02、片段05与片段06等在NMDS图上距离较近，存在相互替代性；（4）物种类群影响识别结果，eDNA研究仍需要进一步开发高识别率片段。综合物种识别率、引物通用性、NMDS分析等多方面因素，本研究推荐2×150 bp测序平台使用片段01（MiFish-U）、2×250 bp测序平台使用片段05（Ac12S），辅以片段13（Vert-16S-eDNA）等进行近缘鱼类多样性调查。本研究旨在为提高鱼类eDNA调查结果准确性提供一定技术支撑。
- 环境DNA metabarcoding /
- 近缘鱼类识别 /
- 12S /
- 扩增长度 /
- 多片段
Abstract: The applicability of environmental DNA (eDNA) metabarcoding fragments to relative fish species had not been compared. There was a risk of “species loss” in diversity surveys. In order to screen out the best fragments, we compared the resolution rate differences of 15 eDNA metabarcoding fragments in 106 genera (a total of 935 species). The results were as follows: (1) the protein-coding gene (COI, fragment 15) had the highest resolution rate, but the universality of its corresponding primer pairs was the worst; the overall mean distance based on primer pair sequence of fragment 09, fragment 11, fragment 07, fragment 03 and fragment 12 were obviously large, suggesting their eDNA amplification efficiency were possibly low. (2) The resolution rates were significantly affected by the length of fragments, and the fragment 05, fragment 06, fragment 01, fragment 02 and fragment 13 of ribosomal genes had a higher resolution rate except fragment 15. (3) Non-metric multidimensional scaling analysis (NMDS) showed that there were great differences among different genes and different fragments belonging to the same gene. Therefore, the combination application of multi-fragment and multi-gene should be considered; besides, fragment 01 and fragment 02, and fragment 05 and fragment 06 were close to each other in the NMDS plot. They function of fish resolution were overlapped. (4) Species groups affected the resolution results, and eDNA studies stilled need to develop fragments with higher resolution rates. Based on the resolution rate of relative species, the universality of primer pairs and NMDS analysis, this study recommended fragment 01 (Mifish-U) for 2×150 bp sequencing platform and fragment 05 (Ac12S) for 2×250 bp sequencing platform, supplemented by fragment 13 (Vert-16S-eDNA) to investigate the diversity of relative fish. This study provided some support for improving the accuracy of fish eDNA survey results.
- eDNA metabarcoding /
- relative fish species resolution /
- 12S /
- amplicon length /
- multi-fragments

HTML全文

图 1 本研究中15个鱼类eDNA metabarcoding片段和对应引物在目标基因上的位置

以花鳗鲡（Anguilla marmorata）线粒体基因序列（GenBank登录号：NC006540）为标注模板

Fig. 1 Locations of the 15 fish eDNA metabarcoding fragments and primer pairs on the target mitochondrial genes

Sequence of the marbled eel (Anguilla marmorata) (GenBank accession number: NC006540) was used as template

下载: 全尺寸图片幻灯片

图 2 本研究中15个鱼类eDNA metabarcoding片段的物种识别率及对应引物序列总平均遗传距离

Fig. 2 Fish species resolution rates of 15 eDNA metabarcoding fragments and overall mean distances of primer pairs in this study

下载: 全尺寸图片幻灯片

图 3 本研究中106属鱼类的物种识别率

Fig. 3 Fish species resolution rates of 106 genera in this study

下载: 全尺寸图片幻灯片

图 4 不同metabarcoding片段的非度量多维尺度（NMDS）分析

F01表示片段01，其余依次类推

Fig. 4 Analysis of non-metric multidimensional scaling (NMDS) for different metabarcoding fragments

F01 represents fragment 01, and the rest are followed by analogy

下载: 全尺寸图片幻灯片

表 1 本研究中15个鱼类eDNA metabarcoding片段简介

Tab. 1 Summary of 15 fish eDNA metabarcoding fragments analyzed in this study

metabarcoding片段	对应引物名称	目标基因	长度/bp	引物序列（5'-3'）	设计者
片段01	MiFish-U	12S	169	For-GTCGGTAAAACTCGTGCCAGC	Miya等^[4]
片段01	MiFish-U	12S	169	Rev-CATAGTGGGGTATCTAATCCCAGTTTG	Miya等^[4]
片段02	Tele02	12S	165	For-AAACTCGTGCCAGCCACC	Taberlet等^[16]
片段02	Tele02	12S	165	Rev-GGGTATCTAATCCCAGTTTG	Taberlet等^[16]
片段03	Am12S	12S	242	For-AGCCACCGCGGTTATACG	Evans等^[17]
片段03	Am12S	12S	242	Rev-CAAGTCCTTTGGGTTTTAAGC	Evans等^[17]
片段04	12S-V5	12S	107	For-ACTGGGATTAGATACCCC	Riaz等^[18]
片段04	12S-V5	12S	107	Rev-TAGAACAGGCTCCTCTAG	Riaz等^[18]
片段05	Ac12S	12S	392	For-ACTGGGATTAGATACCCCACTATG	Evans等^[17]
片段05	Ac12S	12S	392	Rev-GAGAGTGACGGGCGGTGT	Evans等^[17]
片段06	AcMDB07	12S	282	For-GCCTATATACCGCCGTCG	Bylemans等^[19]
片段06	AcMDB07	12S	282	Rev-GTACACTTACCATGTTACGACTT	Bylemans等^[19]
片段07	NeoFish_3	12S	196	For-CGCCGTCGCAAGCTTACCCT	Milan等^[20]
片段07	NeoFish_3	12S	196	Rev-AGTGACGGGCGGTGTGTGC	Milan等^[20]
片段08	Tele01	12S	64	For-ACACCGCCCGTCACTCT	Valentini等^[8]
片段08	Tele01	12S	64	Rev-CTTCCGGTACACTTACCATG	Valentini等^[8]
片段09	Ac16S	16S	339	For-CCTTTTGCATCATGATTTAGC	Evans等^[[17]
片段09	Ac16S	16S	339	Rev-CAGGTGGCTGCTTTTAGGC	Evans等^[[17]
片段10	L2513/H2714	16S	206	For-GCCTGTTTACCAAAAACATCAC	Kitano等^[21]
片段10	L2513/H2714	16S	206	Rev-CTCCATAGGGTCTTCTCGTCTT	Kitano等^[21]
片段11	Fish16S	16S	84	For-CGAGAAGACCCTWTGGAGCTTIAG	Shaw等^[22]
片段11	Fish16S	16S	84	Rev-GGTCGCCCCAACCRAAG	Shaw等^[22]
片段12	Ve16S	16S	335	For-CGAGAAGACCCTATGGAGCTTA	Evans等^[17]
片段12	Ve16S	16S	335	Rev-AATCGTTGAACAAACGAACC	Evans等^[17]
片段13	Vert-16S-eDNA	16S	284	For-AGACGAGAAGACCCYDTGGAGCTT	Vences等^[5]
片段13	Vert-16S-eDNA	16S	284	Rev-GATCCAACATCGAGGTCGTAA	Vences等^[5]
片段14	Fish16S F/D-2R	16S	222	For-GACCCTATGGAGCTTTAGAC	DiBattista等^[23]
片段14	Fish16S F/D-2R	16S	222	Rev-CGCTGTTATCCCTADRGTAACT	DiBattista等^[23]
片段15	PS1	COI	199	For-ACCTGCCTGCCGTATTTGGYGCYTGRGCCGGRATAGT	Balasingham等^[6]
片段15	PS1	COI	199	Rev-ACGCCACCGAGCCARAARCTYATRTTRTTYATTCG	Balasingham等^[6]
注：片段长度以花鳗鲡（Anguilla marmorata）（GenBank登录号：NC006540）为参考。

下载: 导出CSV

表 2 metabarcoding片段对各类群的识别率

Tab. 2 The species resolution rates of metabarcoding fragments for different groups

物种类群	物种数	识别率/%
物种类群	物种数	片段 01	片段 02	片段 03	片段 04	片段 05	片段 06	片段 07	片段 08	片段 09	片段 10	片段 11	片段12	片段 13	片段 14	片段15	全部片段
鱊属 Acheilognathus	14	71.43	71.43	85.71	71.43	85.71	85.71	71.43	57.14	71.43	85.71	57.14	85.71	85.71	71.43	100	77.14±11.83
鲟属 Acipenser	15	13.33	20	26.67	13.33	46.67	33.33	40	6.67	33.33	0	33.33	6.67	13.33	6.67	73.33	24.44± 19.46
光唇鱼属 Acrossocheilus	16	56.25	50	62.5	50	87.5	75	62.5	6.25	68.75	43.75	50	62.5	50	43.75	100	57.92± 21.45
黑头鱼属 Alepocephalus	7	57.14	14.29	28.57	0	28.57	28.57	28.57	0	28.57	14.29	14.29	28.57	42.86	28.57	100	29.52± 24.43
西鲱属 Alosa	6	50	0	0	0	33.33	33.33	33.33	0	0	66.67	16.67	16.67	16.67	16.67	100	25.56± 28.78
双锯鱼属 Amphiprion	10	70	50	60	20	60	50	30	10	60	60	30	80	60	50	40	48.67± 19.22
鳗鲡属 Anguilla	17	29.41	35.29	64.71	0	76.47	70.59	76.47	41.18	64.71	47.06	35.29	76.47	64.71	52.94	88.24	54.9± 23.5
细鲫属 Aphyocypris	5	60	60	60	20	60	40	40	0	100	80	60	100	80	100	100	64± 30.43
旗鳉属 Aphyosemion	8	100	100	100	100	100	100	100	12.5	100	100	100	100	100	100	100	94.17± 22.59
深海鳐属 Bathyraja	8	12.5	25	50	12.5	25	12.5	12.5	0	12.5	12.5	0	25	25	37.5	50	20.83±15.43
鲫属 Carassius	5	0	0	0	0	60	20	20	0	60	40	0	40	0	0	60	20±25.07
真鲨属 Carcharhinus	14	64.29	71.43	100	78.57	100	100	100	50	85.71	35.71	14.29	50	64.29	71.43	100	72.38±26.57
刺尻鱼属 Centropyge	16	75	75	87.5	31.25	62.5	56.25	62.5	12.5	87.5	37.5	0	50	50	62.5	100	56.67±27.8
九棘鲈属 Cephalopholis	5	100	100	100	40	100	60	60	0	40	40	60	100	100	100	100	73.33±32.66
鳢属 Channa	8	62.5	62.5	62.5	62.5	62.5	62.5	62.5	25	75	75	75	75	75	75	50	64.17±13.25
红鳍鲌属 Chanodichthys	5	20	20	20	20	60	60	0	0	100	60	0	100	60	60	100	45.33±36.62
燕鳐属 Cheilopogon	8	37.5	37.5	25	0	25	50	50	0	62.5	25	12.5	12.5	25	12.5	75	30±22.06
鲮属 Cirrhinus	5	60	60	60	40	100	100	100	0	60	60	20	60	40	40	60	57.33±28.15
胡子鲇属 Clarias	6	66.67	16.67	33.33	33.33	83.33	33.33	33.33	0	66.67	33.33	0	66.67	66.67	33.33	100	44.44±29.32
花鳅属 Cobitis	11	81.82	81.82	81.82	81.82	72.73	81.82	81.82	9.09	63.64	81.82	63.64	81.82	81.82	81.82	100	75.15±20.17
鲚属 Coilia	6	16.67	50	50	16.67	66.67	100	66.67	0	50	66.67	33.33	33.33	16.67	16.67	100	45.56±30.52
白鲑属 Coregonus	14	7.14	7.14	0	0	21.43	35.71	21.43	7.14	0	0	0	0	0	0	7.14	7.14±10.8
兵鲶属 Corydoras	6	33.33	33.33	33.33	16.67	50	50	66.67	16.67	50	50	16.67	0	0	0	66.67	32.22±23.12
杜父鱼属 Cottus	12	50	75	100	50	83.33	50	41.67	8.33	83.33	83.33	41.67	83.33	66.67	100	100	67.78±26.7
舌鳎属 Cynoglossus	12	100	100	100	83.33	100	100	100	83.33	100	83.33	83.33	83.33	83.33	83.33	100	92.22±8.61
长吻鳐属 Dipturus	8	0	12.5	37.5	12.5	50	37.5	37.5	0	50	12.5	0	25	25	25	62.5	25.83±19.75
盘属 Discogobio	5	40	40	60	40	60	40	60	0	20	40	0	40	60	40	60	40±20
塘鳢属 Eleotris	6	66.67	66.67	100	100	100	100	100	0	100	100	66.67	66.67	66.67	66.67	100	80±27.6
Enteromius属	5	100	100	100	100	100	100	100	40	100	100	100	100	80	100	100	94.67±15.98
石斑鱼属 Epinephelus	22	90.91	90.91	90.91	50	90.91	90.91	90.91	36.36	72.73	90.91	22.73	81.82	68.18	63.64	90.91	74.85±22.52
扁鳉属 Epiplatys	7	71.43	71.43	71.43	71.43	71.43	71.43	71.43	0	71.43	71.43	71.43	71.43	71.43	71.43	71.43	66.67±18.44
镖鲈属 Etheostoma	10	100	100	100	100	100	100	100	40	100	70	90	100	100	100	100	93.33±16.76
底鳄鳉属 Fundulopanchax	5	100	100	100	100	100	100	100	20	100	100	60	100	100	100	100	92±22.42
底鳉属 Fundulus	5	60	60	60	20	100	100	100	0	100	100	100	100	100	100	100	80±32.95
南乳鱼属 Galaxias	5	100	100	100	60	100	100	100	0	100	60	60	100	100	100	100	85.33±28.75
墨头鱼属 Garra	10	100	100	100	50	100	100	100	30	80	100	50	100	100	100	100	87.33±23.74
骨尾鱼属 Gila	5	40	20	60	20	60	40	40	0	60	60	20	60	60	60	100	46.67±24.69
露齿鲨属 Glyphis	8	25	12.5	25	0	37.5	37.5	37.5	0	25	25	25	25	25	37.5	25	24.17±12.01
纹胸属 Glyptothorax	11	18.18	18.18	63.64	18.18	36.36	27.27	36.36	9.09	54.55	9.09	27.27	54.55	54.55	63.64	81.82	38.18±22.59
颌须属 Gnathopogon	6	33.33	66.67	100	100	100	66.67	66.67	0	100	50	16.67	33.33	33.33	33.33	66.67	57.78±32.65
鳅鮀属 Gobiobotia	6	100	50	100	50	100	100	100	0	66.67	66.67	16.67	83.33	83.33	66.67	100	72.22±31.91
裸鲤属 Gymnocypris	8	37.5	37.5	37.5	0	12.5	12.5	0	0	12.5	37.5	12.5	12.5	12.5	0	25	16.67±14.69
海猪鱼属 Halichoeres	5	100	100	100	100	100	100	100	20	100	100	100	100	100	100	100	94.67±20.66
鳠属 Hemibagrus	5	60	60	60	80	60	40	60	0	60	60	20	60	60	60	60	53.33±19.52
䱻属 Hemibarbus	7	42.86	42.86	42.86	28.57	42.86	28.57	14.29	0	42.86	28.57	14.29	42.86	42.86	42.86	100	37.14±22.13
海马属 Hippocampus	18	88.89	88.89	88.89	55.56	66.67	66.67	61.11	27.78	100	77.78	55.56	100	100	100	100	78.52±22.11
刺蝶鱼属 Holacanthus	5	40	40	60	40	60	60	60	0	40	20	20	40	40	40	60	41.33±17.67
下鱵属 Hyporhamphus	5	100	100	100	100	100	100	100	40	100	100	100	100	100	100	100	96±15.49
野鲮属 Labeo	20	70	70	80	35	90	80	75	10	60	30	25	55	40	50	80	56.67±24.1
兔头鲀属 Lagocephalus	8	100	100	100	75	100	100	100	25	100	75	75	62.5	75	62.5	100	83.33±21.99
太阳鱼属 Lepomis	6	100	100	100	50	100	100	100	0	100	100	66.67	100	100	100	100	87.78±28.5
薄鳅属 Leptobotia	6	50	16.67	66.67	50	66.67	50	33.33	0	33.33	16.67	50	33.33	33.33	33.33	50	38.89±18.55
䱀属 Liobagrus	9	66.67	77.78	77.78	11.11	77.78	55.56	66.67	0	33.33	33.33	33.33	100	100	100	77.78	60.74±31.95
笛鲷属 Lutjanus	13	84.62	84.62	100	100	100	100	84.62	23.08	100	69.23	30.77	100	100	100	100	85.13±25.42
狼绵鳚属 Lycodes	5	60	60	60	40	60	60	60	0	100	40	20	60	60	60	100	56±25.3
虹银汉鱼属 Melanotaenia	6	100	100	50	0	66.67	33.33	33.33	0	66.67	0	50	33.33	100	66.67	100	53.33±36.84
小鳔属 Microphysogobio	14	64.29	64.29	85.71	64.29	71.43	50	35.71	7.14	85.71	35.71	35.71	57.14	71.43	57.14	100	59.05±23.76
泥鳅属 Misgurnus	5	100	100	100	20	60	60	40	0	60	60	0	60	100	60	100	61.33±35.02
假鳃鳉属 Nothobranchius	8	100	100	100	100	100	100	100	12.5	100	100	100	100	100	100	100	94.17±22.59
美洲鱥属 Notropis	10	100	80	100	60	100	100	80	10	100	60	70	100	100	100	100	84±25.58
沙塘鳢属 Odontobutis	6	100	100	100	100	100	100	66.67	33.33	100	100	100	100	100	100	100	93.33±18.69
太平洋鲑属 Oncorhynchus	8	37.5	62.5	100	37.5	100	75	75	0	37.5	0	0	50	50	50	100	51.67±34.68
白甲鱼属 Onychostoma	11	81.82	63.64	45.45	18.18	63.64	63.64	54.55	27.27	54.55	72.73	0	81.82	54.55	45.45	63.64	52.73±22.85
马口鱼属 Opsariichthys	5	100	120	100	40	60	60	20	0	60	60	40	100	100	100	100	70.67±35.35
罗非鱼属 Oreochromis	9	66.67	66.67	66.67	33.33	44.44	11.11	44.44	0	55.56	22.22	22.22	44.44	33.33	22.22	77.78	40.74±22.88
青鳉属 Oryzias	10	80	80	80	80	80	80	80	80	100	100	100	100	100	100	100	89.33±10.33
纹唇鱼属 Osteochilus	5	80	100	100	80	100	100	100	0	100	100	80	100	100	100	100	89.33±26.04
鲳属 Pampus	7	28.57	57.14	28.57	14.29	28.57	28.57	28.57	0	28.57	28.57	14.29	14.29	14.29	14.29	42.86	24.76±13.73
属 Pareuchiloglanis	5	60	60	60	20	60	100	60	0	60	100	60	60	60	60	100	61.33±26.69
鱥属 Phoxinus	7	100	100	100	57.14	100	71.43	71.43	0	71.43	71.43	14.29	100	100	100	100	77.14±32.31
花鳉属 Poecilia	5	20	20	20	20	60	20	40	20	20	20	20	20	20	20	60	26.67±14.47
多鳍鱼属 Polypterus	10	100	100	100	100	100	100	100	40	100	80	60	100	100	100	100	92±18.21
原唇齿脂鲤属 Prochilodus	5	20	20	20	0	20	0	40	0	0	0	0	40	40	40	20	17.33±16.68
拟鲿属 Pseudobagrus	12	41.67	58.33	75	33.33	58.33	58.33	58.33	16.67	66.67	8.33	8.33	33.33	25	25	83.33	43.33±24.03
拟腹吸鳅属 Pseudogastromyzon	9	33.33	55.56	55.56	0	55.56	55.56	55.56	0	77.78	55.56	44.44	100	77.78	55.56	77.78	53.34±26.96
多刺鱼属 Pungitius	7	71.43	100	100	28.57	100	57.14	57.14	0	28.57	28.57	0	71.43	71.43	42.86	100	57.14±34.99
鳐属 Raja	6	16.67	16.67	33.33	16.67	66.67	16.67	16.67	0	50	66.67	16.67	66.67	66.67	66.67	66.67	38.89±25.72
波鱼属 Rasbora	7	100	100	100	71.43	100	71.43	100	14.29	71.43	100	57.14	71.43	71.43	71.43	100	80±24.03
吻鰕虎鱼属 Rhinogobius	5	100	100	100	100	100	100	100	0	100	100	100	100	100	100	100	93.33±25.82
鳑鲏属 Rhodeus	10	80	80	80	30	80	80	60	20	100	80	50	100	100	80	100	74.67±24.75
红点鲑属 Salvelinus	12	8.33	8.33	25	8.33	25	25	25	0	16.67	8.33	0	8.33	0	8.33	58.33	15±15.17
鳈属 Sarcocheilichthys	9	55.56	44.44	55.56	11.11	55.56	55.56	33.33	0	55.56	33.33	0	44.44	33.33	44.44	55.56	38.52±20.08
沙丁鱼属 Sardinella	6	100	100	100	66.67	100	50	66.67	33.33	50	66.67	66.67	100	100	66.67	100	77.78±23.29
蛇属 Saurogobio	7	100	100	100	71.43	100	71.43	71.43	0	71.43	71.43	14.29	100	100	100	100	78.1±31.88
南鳅属 Schistura	11	100	100	100	100	100	100	100	54.55	100	100	63.64	100	100	100	100	94.55±14.5
裸裂尻鱼属 Schizopygopsis	7	14.29	14.29	14.29	0	14.29	28.57	14.29	0	28.57	0	0	42.86	42.86	42.86	71.43	21.91±20.82
裂腹鱼属 Schizothorax	26	11.54	11.54	15.38	7.69	23.08	23.08	11.54	3.85	15.38	7.69	0	11.54	3.85	11.54	30.77	12.56±8.15
马鲛鱼属 Scomberomorus	6	66.67	66.67	66.67	33.33	66.67	66.67	66.67	33.33	100	66.67	100	66.67	66.67	66.67	100	68.89±19.79
平鲉属 Sebastes	19	31.58	31.58	31.58	10.53	36.84	47.37	26.32	0	10.53	15.79	0	26.32	15.79	15.79	68.42	24.56±18.09
瓢鳍虾虎鱼属 Sicyopterus	18	33.33	33.33	44.44	22.22	50	66.67	50	33.33	83.33	33.33	33.33	66.67	55.56	66.67	88.89	50.74±20.13
篮子鱼属 Siganus	6	33.33	33.33	33.33	16.67	100	66.67	66.67	0	66.67	16.67	0	16.67	0	0	33.33	32.22±30.52
鱚属 Sillago	6	100	100	100	100	100	100	100	50	100	100	66.67	100	100	100	100	94.44±15
鲶属 Silurus	6	66.67	66.67	66.67	50	100	100	66.67	16.67	66.67	50	50	66.67	66.67	100	100	68.89±23.46
鳜属 Siniperca	7	57.14	42.86	100	28.57	42.86	28.57	28.57	0	42.86	28.57	14.29	14.29	14.29	14.29	28.57	32.38±23.82
金线鲃属 Sinocyclocheilus	14	85.71	85.71	85.71	35.71	78.57	78.57	64.29	28.57	50	57.14	21.43	64.29	57.14	78.57	85.71	63.81±21.84
银属 Squalidus	7	42.86	28.57	57.14	42.86	100	57.14	57.14	14.29	57.14	28.57	14.29	42.86	28.57	28.57	100	46.67±26.16
扁鲨属 Squatina	5	60	60	60	20	60	60	60	0	20	40	40	60	60	60	100	50.67±23.74
疯鲿属 Tachysurus	8	75	75	75	25	100	100	100	0	75	37.5	0	75	50	50	75	60.83±33.03
东方鲀属 Takifugu	16	0	6.25	6.25	6.25	37.5	25	12.5	12.5	12.5	18.75	6.25	31.25	25	31.25	62.5	19.58±16.34
田中鳑鲏属 Tanakia	6	100	100	100	100	100	100	100	16.67	100	100	33.33	100	100	100	100	90±26.58
鲀属 Tetraodon	5	100	100	100	60	60	60	60	0	100	100	0	100	60	60	100	70.67±34.53
棱鳀属 Thryssa	6	100	100	100	66.67	100	100	100	0	100	100	100	100	100	100	100	91.11±26.63
金枪鱼属 Thunnus	9	0	11.11	11.11	0	0	0	0	0	0	0	0	0	0	0	0	1.48±3.91
茴鱼属 Thymallus	10	20	20	40	0	30	50	20	0	30	20	10	20	30	10	60	24±16.82
结鱼属 Tor	7	71.43	71.43	100	0	71.43	42.86	57.14	0	28.57	57.14	28.57	71.43	57.14	28.57	100	52.38±30.86
高原鳅属 Triplophysa	21	47.62	47.62	80.95	42.86	71.43	71.43	61.9	4.76	66.67	52.38	19.05	47.62	47.62	47.62	80.95	52.7±21.1
全部物种	935	62.3±31.84	61.95±32.54	68.9±30.81	42.92±33.46	71.62±27.3	64.34±29.13	60.89±29.48	11.79±17.79	64.6±30.65	54.58±32.28	36.71±32.27	64.2±31.8	61.6±32.35	59.71±32.78	82.76±24.66

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