基于环境DNA的长江中华鲟分布特征探究

周权; 杜浩; 王洁; 邵芸; 闫振广

doi:10.12153/j.issn.1674-991X.20230203

基于环境DNA的长江中华鲟分布特征探究

doi: 10.12153/j.issn.1674-991X.20230203

周权^1,,
杜浩²,
王洁¹,
邵芸¹,
闫振广^1, ,

1.
环境基准与风险评估国家重点实验室，中国环境科学研究院
2.
中国水产科学研究院长江水产研究所

基金项目: 国家重点研发计划项目(2021YFC3201005)；长江生态环境保护修复联合研究（第二期）（2022-LHYJ-02-0101）

详细信息

作者简介:
周权（1992—），男，硕士研究生，主要从事环境DNA研究，1067174951@qq.com

通讯作者:
闫振广（1972—），男，研究员，博士，主要从事环境组学与水质基准研究，zgyan@craes.org.cn

中图分类号: X52；S932.4
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- 文章访问数: 131
- HTML全文浏览量: 49
- PDF下载量: 32
- 被引次数: 0
出版历程
- 收稿日期: 2023-03-15

Distribution characteristics of Chinese sturgeon in the Yangtze River based on environmental DNA

ZHOU Quan^1
,,
DU Hao²,
WANG Jie¹,
SHAO Yun¹,
YAN Zhenguang^{1
, ,}

1.
State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy of Environmental Sciences
2.
Yangtze River Fisheries Research Institute of Chinese Academy of Fishery Sciences

摘要

摘要:
中华鲟（Acipenser sinensis）是我国长江流域的旗舰物种，在长江十年禁渔的大背景下，研究中华鲟无损化检测技术具有重要意义。环境DNA（eDNA）技术是一种环境友好型生物监测技术，可以在不直接观察或捕获生物体的情况下对物种进行检测。从文献中筛选出可以用于检测中华鲟eDNA的特异性引物，于2020年9月在长江中下游选取4个中华鲟常出现的区域，进行各断面立体式采样；提取16个点位的eDNA，使用筛选得到的引物对中华鲟进行eDNA的检测，以探究中华鲟的分布特征。结果显示，成功筛选到1组可以检测中华鲟eDNA的引物，使用该引物成功检测到包括中华鲟在内的长江4种鲟类的eDNA，共计测得约300万条鲟类序列。依据测序结果分析不同断面检测到的中华鲟eDNA的差异，发现宜昌江段断面的中华鲟eDNA最多，洞庭湖口断面最少，且表层和底层水体的中华鲟eDNA检出也有显著差异。筛选得到的引物可以用于中华鲟eDNA的检测，中华鲟eDNA的检测结果与中华鲟的历史调查和洄游特征较为吻合。不同水深条件中华鲟eDNA的检出量有显著差异，表明在今后的调查中采用混合或者立体采样可以更加全面地进行中华鲟eDNA的检测。
- 中华鲟 /
- 特异性引物 /
- 环境DNA /
- 操作分类单元 /
- 长江
Abstract:
Chinese sturgeon (Acipenser sinensis) is a flagship species in the Yangtze River basin. Under the background of the 10-year fishing ban in the Yangtze River, it is of great significance to study the nondestructive testing technology of Chinese sturgeon. Environmental DNA (eDNA) technology is an environmentally friendly biological monitoring technology, which can detect species without direct observation or capture of the organism. It was tried to screen out the specific primers that could be used to detect the eDNA of Chinese sturgeon from the literature. In September 2020, four areas where Chinese sturgeon often appeared were selected in the middle and lower reaches of the Yangtze River, and three-dimensional sampling was performed on each section. eDNA of 16 sites was extracted, and the screened primers were used to detect the eDNA of Chinese sturgeon, in order to try to explore the distribution characteristics of Chinese sturgeon. A group of primers that could detect the eDNA of Chinese sturgeon was successfully screened. By using this primer, the eDNA of 4 species of sturgeon in the Yangtze River including the Chinese sturgeon was successfully detected, and about 3 million sequences of sturgeon were measured. Based on the sequencing results, the differences in the eDNA of Chinese sturgeon detected in different sections were analyzed. It was found that the eDNA in Yichang River section was the most, and the eDNA in Dongting Lake mouth section was the least, and there were significant differences in the eDNA detection between the surface and bottom waters. The study found that the screened primers could be used for the detection of eDNA of Chinese sturgeon, and the detection results of eDNA were consistent with the historical investigation and migration characteristics of Chinese sturgeon. There were significant differences in the amount of Chinese sturgeon eDNA detected under different water depths, which indicated that mixed or three-dimensional sampling may be more comprehensive for the detection of Chinese sturgeon eDNA in the future investigation.
- Chinese sturgeon /
- specific primers /
- environmental DNA(eDNA) /
- operational taxonomic units(OTUs) /
- the Yangtze River

HTML全文

图 1 4个采样区域特征和采样点位分布

Figure 1. Characteristics of four sampling areas and distribution of sampling points

下载: 全尺寸图片幻灯片

图 2 长江中华鲟引物特异性评估2号引物凝胶电泳图谱

1—中华鲟肌肉；2—中华鲟肝脏；3—达氏鲟肌肉；4—达氏鲟肝脏；5—江豚；6—青鱼；7—草鱼；8—鲢； 9—鳙。

Figure 2. Primer specificity evaluation gel electrophoresis pattern of primer No. 2 of Chinese Sturgeon

下载: 全尺寸图片幻灯片

图 3 不同点位eDNA测序样品的Alpha指数稀释性曲线

Figure 3. Alpha exponential dilution curves of sequenced samples at different sampling points

下载: 全尺寸图片幻灯片

图 4 各采样点位检测获得的鲟类OTUs所含reads数

Figure 4. Number of reads contained in sturgeon OTUs detected at each site

下载: 全尺寸图片幻灯片

图 5 不同点位表层和底层水样检出中华鲟eDNA情况

Figure 5. eDNA of Chinese Sturgeon detected in the surface and bottom water samples at different sampling points

下载: 全尺寸图片幻灯片

表 1 文献发表的中华鲟基因扩增引物

Table 1. Literature published on Acipenser sinensis gene amplification primers

引物编号	序列	扩增方式	产物大小/bp	数据来源
1	DL： CAAGAACACAAGATTAATGAG	PCR	462	文献[25]
1	H740： GATCAAGGTATGTCGATGACA	PCR	462	文献[25]
2	GAS4-F： CTACTAAAACTTGGCGGATACG	巢氏PCR	第1轮313	文献[22]
	GAS4-R：TGTGGAGGCGTTCATAGTTAGGA		第1轮313
	GAS4N-F： GACCGGGTCAATTTGTCTACG		第2轮113
	GAS4N-R： AGCCTCATGGGGTTTGGATG		第2轮113
3	Forward： GGCAATTTTAATCTGGGT TTCCA	ddPCR	132	文献[24]
3	Reverse： TGGATGTTAGATATATGT CCTTG	ddPCR	132	文献[24]
4	L15925： TCAAAGCTTACACCAGTCTTGTAAACC	PCR	747	文献[26]
4	H740： GATCAAGGTATGTCGATGACA	PCR	747	文献[26]

下载: 导出CSV

表 2 各点位检测到的鲟类OTUs所含reads数

Table 2. Number of reads contained in sturgeon OTUs detected at each site 个　

断面	点位编号	中华鲟	杂交鲟	达氏鲟	史氏鲟
宜昌江段	Y1	52 726	141 238	0	201
	Y2	48 546	138 097	1	126
	Y3	42 129	139 960	1	142
	Y4	43 667	121 332	1	139
洞庭湖湖口	D1	42 637	134 637	0	434
	D2	3 435	193 462	1	53
	D3	4 319	194 798	0	47
	D4	3 609	189 642	0	37
鄱阳湖湖口	P1	33 934	118 113	0	73
	P2	42 939	132 712	1	396
	P3	28 481	120 312	2	54
	P4	37 073	124 948	1	133
长江入海口	S1	35 083	109 090	0	118
	S2	10 924	186 367	0	97
	S3	31 145	120 048	0	117
	S4	36 825	145 978	2	183

下载: 导出CSV

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