西藏东北部热泉水中硫氧化菌的多样性及分布特征

doi:10.16258/j.cnki.1674-5906.2023.05.016

生态环境学报 ›› 2023, Vol. 32 ›› Issue (5): 989-1000.DOI: 10.16258/j.cnki.1674-5906.2023.05.016

西藏东北部热泉水中硫氧化菌的多样性及分布特征

寇祝¹^,²(), 卿纯¹^,², 袁昌果¹^,², 李平¹^,²^,^*()

1.中国地质大学（武汉）/生物地质与环境地质国家重点实验室，湖北武汉 430074
2.长江流域环境水科学湖北省重点实验室，湖北武汉 430074

收稿日期:2023-02-27 出版日期:2023-05-18 发布日期:2023-08-09
通讯作者: *李平（1975年生），女，研究员，主要从事与环境地质成因的砷、硫、铁循环密切相关的微生物群落结构和功能及其生物地球化学方面的研究。E-mail: pli@cug.edu.cn
作者简介:寇祝（1997年生），女，硕士研究生，主要从事环境微生物方面的研究。E-mail: kouzhu_kz@163.com
基金资助:
国家自然科学基金项目(41772260)

Diversity and Distribution of Sulfur Oxidizing Bacteria in Hot Springs of Northeast Tibet, China

KOU Zhu¹^,²(), QING Chun¹^,², YUAN Changguo¹^,², LI Ping¹^,²^,^*()

1. State Key Laboratory of Biogeology and Environmental Geology/China University of Geosciences, Wuhan 430074, P. R. China
2. Hubei Key Laboratory of Yangtze Catchment Environmental Aquatic Science/China University of Geosciences, Wuhan, 430074, P. R. China

Received:2023-02-27 Online:2023-05-18 Published:2023-08-09

摘要/Abstract

摘要：

热泉中存在大量的硫氧化菌，而西藏东北部丰富的热泉资源中硫氧化菌的相关研究较少。为探究西藏东北部热泉水中不同类型硫氧化菌（Sulfur-Oxidizing Bacteria，SOB）的多样性、群落结构及其分布特征，采集西藏东北部5个地热区总共13个热泉的地球化学及微生物样品，构建硫氧化基因dsrA和soxB的功能基因克隆文库、qPCR定量分析dsrA和soxB基因丰度，结合水化参数对比分析热泉水中dsrA型SOB和soxB型SOB的群落结构及其分布特征的差异。结果表明，西藏东北部热泉水中的硫氧化菌主要为β变形菌纲（Betaproteobacteria）、α变形菌纲（Alphaproteobacteria）、γ变形菌纲（Gammaproteobacteria）和嗜氢菌纲（Hydrogenophilalia），其中dsrA型SOB和soxB型SOB的优势类群在纲水平上均为Betaproteobacteria，但目水平上则存在差异，即soxB型SOB优势菌目为亚硝化单胞菌目（Nitrosomonadales）（30.0%-91.7%）；dsrA型SOB优势菌目为红环菌目（Rhodocyclales）（33.3%-96.0%）。冗余分析（redundancy analysis，RDA）表明，SO₄^2-、S^2-、HCO₃^-、pH、温度等是影响硫氧化菌群落分布的关键环境因子。soxB基因相对丰度主要与SO₄^2-浓度、氧化还原电位（Oxidation-Reduction Potential，ORP）以及S^2-浓度相关，dsrA基因相对丰度受总砷（As^T）浓度和HCO₃^-浓度影响较大。这些结果表明，soxB基因型SOB主要分布于偏氧化环境，而dsrA基因型SOB主要分布于砷浓度、碱度相对偏高的热泉。该研究进一步完善了关于西藏热泉水环境中SOB多样性和分布规律的认识，促进了对热泉硫的生物地球化学循环的理解。

关键词: 西藏, 热泉, 硫氧化菌, soxB基因, dsrA基因, 群落结构, 环境因子

Abstract:

Sulfur (S) -oxidizing bacteria (SOB) widely inhabit hot springs, but the diversity and distribution of SOB in hot springs of northeast Tibet are not well known. In this study, the diversity and distribution of SOB in hot springs of northeast Tibet were explored with gene clone libraries and qPCR of different types of SOB based on sulfur-oxidizing genes dsrA and soxB. Samples from 13 hot springs in 5 geothermal areas in northeast Tibet were collected. The results showed that the SOB in the hot springs mainly belonged to Betaproteobacteria, Alphaproteobacteria, Gammaproteobacteria and Hydrogenophilalia, and the dominant groups of dsrA-type SOB and soxB-type SOB were Betaproteobacteria at the class level. At the order level, soxB-type SOB were mainly composed of Nitrosomonadales (30.0%-91.7%), while dsrA-type SOB were predominant by Rhodocyclales (33.3%-96.0%). Redundancy analysis (RDA) showed that SO₄^2-, S^2-, HCO₃^-, pH, and temperature were the key environmental factors shaping the distribution of SOB communities. The relative abundance of soxB genes was mainly related to SO₄^2-, S^2-, and oxidation-reduction potential, while the relative abundance of dsrA genes was greatly affected by the concentrations of total arsenic and HCO₃^-, indicating that soxB-type SOB was mainly distributed in the oxidizing environment, while dsrA-type SOB mainly existed in hot springs with relatively high arsenic concentration and alkalinity. This study improved the understanding of the distribution and diversity of SOB and biogeochemistry cycle of sulfur in hot springs.

Key words: Tibet, hot spring, sulfur-oxidizing bacteria, soxB gene, dsrA gene, community structure, environmental factors

中图分类号:

X172

寇祝, 卿纯, 袁昌果, 李平. 西藏东北部热泉水中硫氧化菌的多样性及分布特征[J]. 生态环境学报, 2023, 32(5): 989-1000.

KOU Zhu, QING Chun, YUAN Changguo, LI Ping. Diversity and Distribution of Sulfur Oxidizing Bacteria in Hot Springs of Northeast Tibet, China[J]. Ecology and Environment, 2023, 32(5): 989-1000.

图/表 12

参考文献 46

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采样点	温度 t/℃	pH	氧化还原电位 ORP/mV	ρ_{(五价砷As5+)}/ (µg·L^-1)	ρ_(总砷AsT)/ (µg·L^-1)	ρ_{(硫酸根SO42-)}/ (mg·L^-1)	ρ_{(硫离子S2-)}/ (mg·L^-1)	ρ_(总铁FeT)/ (mg·L^-1)	ρ_{(碳酸氢根HCO3-)}/ (mg·L^-1)
DB1	56.0±0.1e¹⁾	7.19±0.10g	-253.4±0.3g	8331.27±2.02b	8331.27±2.02b	615.1±1.5c	1.92±0.03e	0.010±0.006h	2221.08±1.35a
DB41	69.8±0.2a	7.27±0.15e	-301.0±0.4j	9298.58±1.08a	9298.58±1.08a	643.3±0.9a	1.55±0.04j	0.057±0.025g	ND
DB45	37.7±0.1i	8.37±0.01a	-105.8±0.5b	2891.80±1.15f	2891.80±1.15g	455.6±2.1h	1.52±0.01k	ND²⁾	1763.48±1.95c
DB5	51.4±0.1f	7.21±0.15fg	-157.5±0.8e	1279.50±1.37h	6053.57±1.10d	557.3±1.4f	2.00±0.03d	0.186±0.002e	1434.05±1.11d
DB6	50.2±0.1g	7.01±0.01j	-307.3±0.5k	7415.97±1.54c	7415.97±1.54c	604.2±0.9d	1.76±0.02g	0.144±0.002f	1385.16±1.83e
DB7	36.7±0.2j	7.35±0.02d	-126.6±0.3d	2331.15±1.06g	2331.15±1.06h	536.1±1.8g	1.62±0.02i	0.495±0.003c	2105.17±1.33b
DB8	28.5±0.2l	8.01±0.01b	-109.9±0.3c	4211.16±2.22d	4211.16±2.22e	578.1±1.4e	1.67±0.03h	0.200±0.002d	1113.62±1.99h
DB10	35.0±0.1k	7.05±0.04i	-229.5±0.4f	3445.68±0.97e	3445.68±0.97f	47.7±1.0l	2.08±0.02c	0.144±0.003f	1354.56±1.93f
ZM1	69.2±0.1b	7.11±0.03h	-301.3±0.4j	24.57±2.32k	29.27±2.33l	54.4±1.1k	2.56±0.02a	0.495±0.003c	1113.60±1.71h
ZM2	61.0±0.0d	7.66±0.03c	-277.3±0.8h	ND	ND	31.7±1.2m	2.16±0.03b	0.200±0.001d	988.50±2.28j
BDW	38.2±0.1h	6.59±0.01k	-45.5±0.3a	0.00±0.00l	43.17±2.08k	620.7±1.3b	0.80±0.04l	3.315±0.003b	1025.11±1.55i
QS	66.0±0.0c	7.23±0.01f	-282.5±0.4i	235.89±2.00j	235.89±2.00j	125.4±1.5i	1.82±0.02f	0.183±0.002e	1189.93±0.87g
GN	25.0±0.1m	7.07±0.01i	ND	346.66±1.51i	346.66±1.51i	109.6±1.8j	ND	4.294±0.005a	ND

采样点	温度 t/℃	pH	氧化还原电位 ORP/mV	ρ_{(五价砷As5+)}/ (µg·L^-1)	ρ_(总砷AsT)/ (µg·L^-1)	ρ_{(硫酸根SO42-)}/ (mg·L^-1)	ρ_{(硫离子S2-)}/ (mg·L^-1)	ρ_(总铁FeT)/ (mg·L^-1)	ρ_{(碳酸氢根HCO3-)}/ (mg·L^-1)
DB1	56.0±0.1e¹⁾	7.19±0.10g	-253.4±0.3g	8331.27±2.02b	8331.27±2.02b	615.1±1.5c	1.92±0.03e	0.010±0.006h	2221.08±1.35a
DB41	69.8±0.2a	7.27±0.15e	-301.0±0.4j	9298.58±1.08a	9298.58±1.08a	643.3±0.9a	1.55±0.04j	0.057±0.025g	ND
DB45	37.7±0.1i	8.37±0.01a	-105.8±0.5b	2891.80±1.15f	2891.80±1.15g	455.6±2.1h	1.52±0.01k	ND²⁾	1763.48±1.95c
DB5	51.4±0.1f	7.21±0.15fg	-157.5±0.8e	1279.50±1.37h	6053.57±1.10d	557.3±1.4f	2.00±0.03d	0.186±0.002e	1434.05±1.11d
DB6	50.2±0.1g	7.01±0.01j	-307.3±0.5k	7415.97±1.54c	7415.97±1.54c	604.2±0.9d	1.76±0.02g	0.144±0.002f	1385.16±1.83e
DB7	36.7±0.2j	7.35±0.02d	-126.6±0.3d	2331.15±1.06g	2331.15±1.06h	536.1±1.8g	1.62±0.02i	0.495±0.003c	2105.17±1.33b
DB8	28.5±0.2l	8.01±0.01b	-109.9±0.3c	4211.16±2.22d	4211.16±2.22e	578.1±1.4e	1.67±0.03h	0.200±0.002d	1113.62±1.99h
DB10	35.0±0.1k	7.05±0.04i	-229.5±0.4f	3445.68±0.97e	3445.68±0.97f	47.7±1.0l	2.08±0.02c	0.144±0.003f	1354.56±1.93f
ZM1	69.2±0.1b	7.11±0.03h	-301.3±0.4j	24.57±2.32k	29.27±2.33l	54.4±1.1k	2.56±0.02a	0.495±0.003c	1113.60±1.71h
ZM2	61.0±0.0d	7.66±0.03c	-277.3±0.8h	ND	ND	31.7±1.2m	2.16±0.03b	0.200±0.001d	988.50±2.28j
BDW	38.2±0.1h	6.59±0.01k	-45.5±0.3a	0.00±0.00l	43.17±2.08k	620.7±1.3b	0.80±0.04l	3.315±0.003b	1025.11±1.55i
QS	66.0±0.0c	7.23±0.01f	-282.5±0.4i	235.89±2.00j	235.89±2.00j	125.4±1.5i	1.82±0.02f	0.183±0.002e	1189.93±0.87g
GN	25.0±0.1m	7.07±0.01i	ND	346.66±1.51i	346.66±1.51i	109.6±1.8j	ND	4.294±0.005a	ND

采样点	序列数	OTU数	覆盖度/ %	Chao 1 指数	Shannon指数	Simpson_ 1-D指数
DB1	25	4	96	4	1.4	0.5
DB45	16	5	94	5	2.1	0.7
DB5	23	5	96	5	2.0	0.7
DB6	25	3	96	3	1.0	0.4
DB7	22	6	95	6	2.4	0.8
DB8	17	4	100	4	1.9	0.7
DB10	29	4	93	5	1.2	0.5
BDW	23	4	95	4	1.2	0.4
QS	27	5	92	6	1.6	0.6
GN	20	7	95	7	2.6	0.8

采样点	序列数	OTU数	覆盖度/ %	Chao 1 指数	Shannon指数	Simpson_ 1-D指数
DB1	25	4	96	4	1.4	0.5
DB45	16	5	94	5	2.1	0.7
DB5	23	5	96	5	2.0	0.7
DB6	25	3	96	3	1.0	0.4
DB7	22	6	95	6	2.4	0.8
DB8	17	4	100	4	1.9	0.7
DB10	29	4	93	5	1.2	0.5
BDW	23	4	95	4	1.2	0.4
QS	27	5	92	6	1.6	0.6
GN	20	7	95	7	2.6	0.8

采样点	序列数	OTU数	覆盖度/ %	Chao 1指数	Shannon指数	Simpson_ 1-D指数
DB1	20	3	95	3.0	1.1	0.4
DB41	41	8	95	12.0	2.9	0.8
DB5	41	5	97	5.5	1.9	0.7
DB6	40	8	95	9.0	2.3	0.7
DB7	37	7	94	8.0	2.2	0.7
DB8	28	9	93	10.0	2.9	0.8
DB10	38	6	95	7.0	1.6	0.5
ZM1	25	7	92	6.5	2.2	0.7
ZM2	24	4	92	5.0	0.9	0.3
BDW	20	4	95	4.5	1.5	0.6

西藏东北部热泉水中硫氧化菌的多样性及分布特征

Diversity and Distribution of Sulfur Oxidizing Bacteria in Hot Springs of Northeast Tibet, China

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环境因子	Chao1指数		Shannon指数		Simpson_1-D指数
环境因子	r	P	r	P	r	P
pH	0.244	0.497	0.591	0.072	0.642	0.045
ORP	0.280	0.432	0.289	0.418	0.327	0.356
As⁵⁺	-0.476	0.165	-0.154	0.671	-0.099	0.786
T	-0.500	0.141	-0.474	0.166	-0.543	0.105
As^T	-0.530	0.115	-0.098	0.787	-0.105	0.773
SO₄^2-	-0.854	0.002	-0.455	0.186	-0.451	0.191
S^2-	-0.177	0.625	-0.302	0.397	-0.358	0.310
Fe^T	0.468	0.173	0.457	0.184	0.433	0.211
HCO₃^-	-0.146	0.687	0.068	0.853	0.012	0.973

样点名称	soxB基因拷贝数/ (copies·mL^-1)		dsrA基因拷贝数/ (copies·mL^-1)		16S rRNA基因拷贝数/ (copies·mL^-1)		soxB基因相对丰度/ %	dsrA基因相对丰度/ %
样点名称	soxB	SD	dsrA	SD	16S²⁾	SD	soxB/16S	dsrA/16S
DB1	4.53E+04	1.16E+04	2.24E+05	2.83E+03	7.41E+05	4.22E+05	6.12	30.25
DB41	3.62E+03	7.85E+02	ND	ND	2.31E+05	4.88E+04	1.57	ND
DB45	ND¹⁾	ND	2.18E+02	1.48E+02	9.69E+02	6.36E+00	ND	22.50
DB5	2.14E+05	3.61E+03	8.42E+04	3.99E+03	4.66E+06	2.51E+05	4.59	1.80
DB6	3.72E+03	1.12E+03	2.07E+04	1.59E+03	2.50E+05	7.35E+04	1.49	8.28
DB7	1.37E+05	3.17E+04	6.55E+04	7.06E+03	3.43E+06	4.24E+04	4.00	1.91
DB8	4.89E+04	1.07E+04	1.36E+04	1.46E+03	2.23E+06	1.06E+05	2.19	0.61
DB10	3.31E+04	2.25E+03	1.69E+04	2.19E+03	1.84E+06	3.25E+05	1.80	0.92
ZM1	1.77E+03	5.02E+02	ND	ND	5.33E+05	3.61E+04	0.33	ND
ZM2	2.00E+03	2.28E+02	ND	ND	5.17E+05	6.79E+04	0.39	ND
BDW	7.88E+05	2.05E+04	9.76E+04	0.00E+00	6.68E+06	2.83E+04	11.80	1.46
QS	ND	ND	2.00E+04	5.40E+03	2.35E+06	3.82E+05	ND	0.85
GN	ND	ND	4.38E+03	1.63E+02	1.95E+05	1.02E+05	ND	2.24

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