白洋淀抗重金属细菌多样性及其与生态因子相关性

doi:10.3969/j.issn.1000-1565.2018.06.016

摘要/Abstract

摘要： 为了给白洋淀重金属污染治理提供理论支持和菌种资源, 采用分离培养的方法调查了白洋淀抗重金属细菌的系统发育多样性,并分析了抗重金属细菌分布与生态因子之间的关系. 从各采样点的样品中分离筛选得到抗80 mg/L Pb²⁺细菌54株、抗80 mg/L Cu²⁺细菌46株, 抗100 mg/L Cr(Ⅵ)细菌39株. 综合考虑各菌株的抗性及16S rRNA基因序列差异,将139株抗重金属菌去除重复后合并为69株, 分属于肠杆菌属(Enterobacter)、丛毛单胞菌属(Comamonas)、戴尔福特菌属(Delftia)、短波单胞菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)、假单胞菌属(Pseudomonas)和赖氨酸芽孢杆菌属(Lysinibacillus)共7个属13个种. 进一步采用PCA分析的方法对抗重金属细菌与生态因子相关性进行研究, 发现在菌群组成和分布范围方面抗Pb²⁺和Cu²⁺菌大体相似, 且假单胞菌属和寡养单胞菌属2个属容限最高, 表明菌株对于重金属具有交叉抗性;抗100 mg/L Cr(Ⅵ)细菌中赖氨酸杆菌属为特有属, 且分布广泛;白洋淀水域唯一能够同抗Pb²⁺、Cu²⁺和Cr(Ⅵ)3种重金属的细菌是寡养单胞菌属.

关键词: 白洋淀, 抗重金属细菌, 系统发育多样性, 生态因子

Abstract: In order to provide theoretical support and bacterial resources for the treatment of heavy metal pollution in Baiyangdian Lake, the phylogenetic diversity of heavy metal-resistant bacteria in Baiyangdian was studied using isolation and culture method, and the relationship between the distribution of heavy metal-resistant bacteria and ecological factors was analyzed. 54 strains of anti-80 mg/L Pb²⁺ bacteria, 46 strains of anti-80 mg/L Cu²⁺ bacteria, and 39 strains of anti-100 mg/L Cr(Ⅵ)bacteria were isolated- DOI:10.3969/j.issn.1000-1565.2018.06.016白洋淀抗重金属细菌多样性及其与生态因子相关性陈思雨¹, 王丽娜¹, 杨惠琪¹, 陈玉¹, 张瑾¹, 吕志堂^1,2(1.河北大学生命科学学院河北省微生物多样性研究与应用实验室, 河北保定 071002;2.药物化学与分子诊断教育部重点实验室, 河北保定 071002)摘要:为了给白洋淀重金属污染治理提供理论支持和菌种资源, 采用分离培养的方法调查了白洋淀抗重金属细菌的系统发育多样性,并分析了抗重金属细菌分布与生态因子之间的关系. 从各采样点的样品中分离筛选得到抗80 mg/L Pb²⁺细菌54株、抗80 mg/L Cu²⁺细菌46株, 抗100 mg/L Cr(Ⅵ)细菌39株. 综合考虑各菌株的抗性及16S rRNA基因序列差异,将139株抗重金属菌去除重复后合并为69株, 分属于肠杆菌属(Enterobacter)、丛毛单胞菌属(Comamonas)、戴尔福特菌属(Delftia)、短波单胞菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)、假单胞菌属(Pseudomonas)和赖氨酸芽孢杆菌属(Lysinibacillus)共7个属13个种. 进一步采用PCA分析的方法对抗重金属细菌与生态因子相关性进行研究, 发现在菌群组成和分布范围方面抗Pb²⁺和Cu²⁺菌大体相似, 且假单胞菌属和寡养单胞菌属2个属容限最高, 表明菌株对于重金属具有交叉抗性;抗100 mg/L Cr(Ⅵ)细菌中赖氨酸杆菌属为特有属, 且分布广泛;白洋淀水域唯一能够同抗Pb²⁺、Cu²⁺和Cr(Ⅵ)3种重金属的细菌是寡养单胞菌属.关键词:白洋淀;抗重金属细菌;系统发育多样性;生态因子中图分类号:Q938.1+1 文献标志码:A 文章编号:1000-1565(2018)06-0664-09Phylogenetic Diversity of heavy metal-resistant bacteria in Baiyangdian Lake and its relationship with ecological factors CHEN Siyu¹, WANG Lina¹, YANG Huiqi¹, CHEN Yu¹, ZHANG Jin¹, LÜ Zhitang^1,2(1. Key Laboratory of Microbial Diversity Research and Application of Hebei Province,College of Life Sciences, Hebei University, Baoding 071002, China;2. Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, Hebei University, Baoding 071002, China)Abstract: In order to provide theoretical support and bacterial resources for the treatment of heavy metal pollution in Baiyangdian Lake, the phylogenetic diversity of heavy metal-resistant bacteria in Baiyangdian was studied using isolation and culture method, and the relationship between the distribution of heavy metal-resistant bacteria and ecological factors was analyzed. 54 strains of anti-80 mg/L Pb²⁺ bacteria, 46 strains of anti-80 mg/L Cu²⁺ bacteria, and 39 strains of anti-100 mg/L Cr(Ⅵ)bacteria were isolated- 收稿日期:2018-05-20 基金项目:河北省生物学强势特色学科项目(1050-5030004);河北省生物工程重点学科项目(1050-5030023) 第一作者:陈思雨(1994—),女,河北安新人,河北大学在读硕士研究生. E-mail:331255943@qq.com 通信作者:吕志堂(1973—),男,河北曲周人,河北大学教授,博士,主要从事微生物资源与系统学研究. E-mail:lzt325@126.com第6期陈思雨等:白洋淀抗重金属细菌多样性及其与生态因子相关性and screened from the sampling sites. A total of 69 strains were combined after repeated strains were removed based on heavy metal-resistant characteristics sequence difference, which belonged to 13 species distributed in 7 genera Enterobacter, Comamonas, Delftia, Brevundimonas, Stenotrophomonas, Pseudomonas, and Lysinibacillus. Further PCA analysis was used to study the relationship between heavy metal resistant bacteria and ecological factors. In terms of group composition and distribution, the anti-Pb²⁺ and Cu²⁺ bacteria were generally similar, and Pseudomonas and Stenotrophomonas had the highest abundance, indicating that the strain had cross-resistance to heavy metals. Lysinibacillus is the unique 100 mg/L Cr(Ⅵ)resistant genus, and it is widely distributed. Stenotrophomonas is the only genus which can survive in three kinds of heavy metals Pb²⁺, Cu²⁺ and Cr(Ⅵ)within Baiyangdian Lake.

Key words: Baiyangdian Lake, heavy metal-resistant bacteria, phylogenetic diversity, ecological factors

中图分类号:

Q938.1+1

陈思雨, 王丽娜, 杨惠琪, 陈玉, 张瑾, 吕志堂. 白洋淀抗重金属细菌多样性及其与生态因子相关性[J]. 河北大学学报(自然科学版), 2018, 38(6): 664-672.

CHEN Siyu, WANG Lina, YANG Huiqi, CHEN Yu, ZHANG Jin, LÜ Zhitang. Phylogenetic Diversity of heavy metal-resistant bacteria in Baiyangdian Lake and its relationship with ecological factors[J]. Journal of Hebei University (Natural Science Edition), 2018, 38(6): 664-672.

参考文献

[1] 毛美洲, 刘子慧, 董惠茹.府河-白洋淀水及沉积物的污染研究[J].环境科学, 1995, 16(增刊):1-6. DOI: 10.13227/j.hjkx. 1995. s1.001. MAO M Z, LIU Z H, DONG H R. Study on the pollution of Fuhe-Baiyang Lake water and sediments[J]. Environmental Science, 1995, 16(增刊): 1-6.DOI:10.13227/j.hjkx.1995.s1.001
[2] 环境保护部. 2016年中国环境状况公报 [R]. 2017.06.05. MINISTRY OF ENVIRONMENTAL PROTECTION. 2016 China environmental status bulletin[R]. 2017.06.05.
[3] 杨卓, 王殿武, 李贵宝, 等. 白洋淀底泥重金属污染现状调查及评价研究[J]. 河北农业大学学报, 2006, 28(5): 20-26. YANG Z, WANG D W, LI G B, et al. Investigation and assessment on pollution status of heavy metal in sediment of Baiyangdian Lake [J]. Journal of Hebei Agricultural University, 2006, 28(5): 20-26.
[4] 李玉, 俞志明, 宋秀贤.运用主成分分析( PCA)评价海洋沉积物中重金属污染来源[J].环境科学, 2006, 27(1): 137-141. DOI:10.13227/j.hjkx.2006.01.026 LI Y, YU Z M, SONG X X. Application of principal component analysis(PCA)for the estimation of source of heavy metal contamination in marine sediments [J]. Environmental Science, 2006, 27(1): 137-141. DOI:10.13227/j.hjkx.2006.01.026
[5] BIBI M H, AHMED F, ISHIGA H. Assessment of metal concentrations in lake sediments of southwest Japan base on sediment quality guidelines[J].Environmental Geology, 2007, 52(4): 625-639.DOI:10.1007/s00254-006-0492-x.
[6] DEXTER K S, WARD N I. Mobility of heavy metals within freshwater sediments affected by motorway stormwater[J]. Science of The Total Environment, 2004, 334 /335: 271-277.
[7] 李必才,何连生,杨敏, 等.白洋淀底泥重金属形态及竖向分布[J]. 环境科学, 2012, 33(7):2376-2383. LI B C, HE L S, YANG M, et al. Speciation and vertical distribution of heavy metals in sediments of Baiyangdian Lake[J]. Environmental Science, 2012,33(7):2376-2383.
[8] LI X D, WAI O W H, LI Y S, et al. Heavy metal distribution in sediment profiles of the Pearl River estuary, South China[J]. Applied Geochemistry, 2000, 15( 5): 567-581.
[9] 李鸣, 刘琪璟.鄱阳湖水体和底泥重金属污染特征与评价[J]. 南昌大学学报(理科版), 2010, 34(5): 486-494.DOI:10.3969/j.issn.1006-0464.2010.05.018. LI M, LIU Q J. Characteristics and assessment of heavy metals pollution in Poyang Lake[J]. Journal of Nanchang University(Natural Science), 2010, 34(5):486-494.DOI:10.3969/j.issn.1006-0464.2010.05.018.
[10] 范文宏,张博,陈静生, 等.锦州湾沉积物中重金属污染的潜在生物毒性风险评价[J].环境科学学报,2006,26(6):1000-1005. DOI:10.3321/j.issn:0253-2468.2006.06.021. FAN W H, ZHANG B, CHEN J S, et al. Pollution and potential biological toxicity assessment using heavy metals from surface sediments of Jinzhou Bay[J]. Act a Scientiae Circumstantiae, 2006, 26(6):1000 -1005 DOI:10.3321/j.issn:0253-2468. 2006.06.021.
[11] 宁建凤, 邹献中, 杨少海, 等.广东大中型水库底泥重金属含量特征及潜在生态风险评价[J].生态学报, 2009, 29( 11): 6059-6067.DOI:10.3321/j.issn:1000-0933.2009.11.038
[12] NING J F, ZOU X Z, YANG S H, et al. Heavy metal contents analysis and potential ecological risk appraisal to sediments of large and medium-scaled reservoirs in Guangdong Province[J]. Acta Ecologica Sinica, 2009, 29(11): 6059-6067.DOI:10.3321/j.issn:1000-0933.2009.11.038.
[13] CODD R, DILLON C T, LEVINA T, et al. Studies on the genotoxicity of chromium: from the test tube to the cell[J]. Coord Chem Rev, 2001,(216-217):537-582.
[14] CERVANTES C, CAMPOS-GRACIA J, DEVARS S, et al. Interactions of chromium with microorganisms and plants[J]. FEMS Microbiol Rev, 2001, 25:335-347. https://doi.org/10.1111/j.1574-6976.2001.tb00581.x
[15] 陈程, 陈明. 环境重金属污染的危害与修复[J]. 环境保护, 2010,3(3): 55-57. DOI:10.3969/j.issn.0253-9705.2010.03.017. CHEN C, CHEN M. The harm and repair of environmental heavy metal pollution[J]. Environmental Protection, 2010, 3(3): 55-57. DOI:10.3969/j.issn.0253-9705.2010.03.017.
[16] 水质总磷的测定流动注射-钼酸铵分光光度法: HJ 671-2013[S]. 北京:中国环境科学出版社, 2013. Water quality-determination of total phosphorus-flow injection Analysis(FIA)and Ammonium molybdate spectrophotometry: HJ 671-2013[S]. Beijing:China Environmental Science Press, 2013.
[17] 水质总氮的测定碱性过硫酸钾消解紫外分光光度法: HJ 636-2012[S]. 北京:中国环境科学出版社, 2012. Water quality-determination of total nitrogen-alkaline potassium persulfate digestion UV spectrophotometric method: HJ 636-2012[S]. Beijing:China Environmental Science Press, 2012.
[18] 李长华, 李镇. 苯基萤光酮-溴化十六烷基三甲铵分光光度法测定微量的铅[J]. 冶金分析与测试(冶金分析分册), 1992, 3(2): 16-17. DOI:10.13228/j.issn.1000-7571.1985.04.011 LI C H, LI Z. Spectrophotometric determination of trace lead with phenyl fluorone-hexadecyltrimethylammonium bro-mide[J]. Metallurgical Analysis and Testing(Metallurgical Analysis Volume), 1992, 3(2):16-17. DOI:10.13228/j.issn.1000-7571.1985.04.011
[19] 吴德武, 钟春龙, 吴粦华. 双环己酮草酰二腙分光光度法测定水样中铜(Ⅱ)[J]. 理化检验: 化学分册, 2009, 45(8): 1003-1004 WU D W, ZHONG C L, WU L H. Spectrophotometric determination of copper(II)in water samples with dicyclohexanoneoxaly dihydrazone [J].PTCA(PART B:CHEM.ANAL.)2009, 45(8): 1003-1004
[20] 张世芝, 杨晓琴, 张明锦, 等. 双波长等吸收点法同时测定水样中Cr(III)和Cr(VI)的含量[J]. 重庆理工大学学报(自然科学), 2012, 26(8): 21-25. DOI:10.3969/j.issn.1674-8425-B.2012.08.005. ZHANG S Z, YANG X Q, ZHANG M J, et al. Simultaneous determination of Cr(Ⅲ)and Cr(Ⅵ)by using dual-wavelength isobestic point spectrophotometry[J]. Journal of Chongqing University of Technology( Natural Science). 2012, 26(8): 21-25. DOI:10.3969/j.issn.1674-8425-B.2012.08.005.
[21] MARMUR J. A procedure for the isolation of deoxyribonucleic acid from microorganisms[J]. Journal of molecular biology, 1961, 3(2): 208-218. DOI: 10.1016/S0022-2836(61)80047-8
[22] GAO B, GUPTA R S. Conserved indels in protein sequences that are characteristic of the phylum Actinobacteria[J]. Int J Syst Evol Microbiol, 2005, 55(5):2401-2412.
[23] SAITOU N, NEI M. The neighbor-joining method: a new method for reconstructing phylogenetic trees [J]. Molecular Biology and Evolution, 1987, 4(4): 406-425. DOI: 10.1093/oxfordjournals.molbev.a040454
[24] 地表水环境质量标准: GB3838-2002[S]. 北京:中国环境科学出版社, 2002. Surface water environment quality standard: GB3838-2002[S]. Beijing: China Environmental Science Press, 2002.