Article(id=1241049967259874217, tenantId=1146029695717560320, journalId=1234093305789726721, issueId=1241049962679694215, articleNumber=null, orderNo=null, doi=null, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1718294400000, receivedDateStr=2024-06-14, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773818969788, onlineDateStr=2026-03-18, pubDate=1737302400000, pubDateStr=2025-01-20, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773818969788, onlineIssueDateStr=2026-03-18, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773818969788, creator=13701087609, updateTime=1773818969788, updator=13701087609, issue=Issue{id=1241049962679694215, tenantId=1146029695717560320, journalId=1234093305789726721, year='2025', volume='45', issue='1', pageStart='1', pageEnd='592', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773818968696, creator=13701087609, updateTime=1773819749443, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241053237428671382, tenantId=1146029695717560320, journalId=1234093305789726721, issueId=1241049962679694215, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241053237428671383, tenantId=1146029695717560320, journalId=1234093305789726721, issueId=1241049962679694215, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=477, endPage=486, ext={EN=ArticleExt(id=1241049968744657856, articleId=1241049967259874217, tenantId=1146029695717560320, journalId=1234093305789726721, language=EN, title=High nitrate load in lakeside groundwater around Lake Taihu promotes the enrichment of denitrifying functional bacteria community, columnId=1240689595814302661, journalTitle=China Environmental Science, columnName=Environmental Microbiology, runingTitle=null, highlight=null, articleAbstract=

The denitrifying functional bacteria community plays a crucial role in the degradation of nitrate pollution. However, the understanding of the relationships between its interspecies links and nitrate load is still limited. Groundwater resources in the vicinity of Lake Taihu serve as vital reserves, yet in some areas, nitrate load in groundwater exceeds health thresholds. This paper focused on 14 groundwater samples with varying nitrate concentrations from the Lake Taihu area. With microbial sequencing techniques, the diversity of bacterial communities in groundwater was explored, and the denitrifying functional bacteria community was identified. The response of interspecies links within the denitrifying functional bacteria community to nitrate load was investigated by co-occurrence network analysis. The results indicate: Significant differences were found in bacterial community composition between high and low nitrate samples; The α-diversity and β-diversity were highly influenced by nitrate load; Interspecies links within the denitrifying functional bacteria community in the high nitrate samples were denser than those in the low nitrate samples. This study demonstrates that nitrate load in groundwater significantly influences the interspecies links within the denitrifying functional bacteria community, providing new insights into the interplay between groundwater denitrifying functional bacteria community and nitrate load in the Lake Taihu area.

, correspAuthors=Kou-ping CHEN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Zhuo-qun WANG, Kou-ping CHEN, Ji-chun WU, Bao-zhan WANG, Xu LIU), CN=ArticleExt(id=1241049974927061162, articleId=1241049967259874217, tenantId=1146029695717560320, journalId=1234093305789726721, language=CN, title=太湖周边地下水中高硝酸盐负荷促进反硝化功能菌群富集, columnId=1240689596959346705, journalTitle=中国环境科学, columnName=环境微生物, runingTitle=null, highlight=null, articleAbstract=

反硝化功能菌群在降解硝态氮污染中发挥着重要作用,目前对其内部种间关系与硝酸盐负荷的联系仍了解有限.太湖周边地区地下水资源是重要的储备水源,但部分区域的地下水硝酸盐负荷超出了范围.本文以太湖周边地区14个不同硝酸盐浓度的地下水样品作为研究对象,运用微生物测序技术探究了地下水细菌群落多样性并识别出反硝化功能菌群,通过共现网络分析探究了反硝化功能菌群内部种间关系对硝酸盐负荷的响应.结果表明:高硝态氮组与低硝态氮组间细菌群落组成存在显著差异;细菌群落α多样性、β多样性受到硝酸盐负荷的显著影响;高硝态氮组地下水反硝化功能菌群内部种间关系较低硝态氮组更为密切.本研究表明地下水硝酸盐负荷对反硝化功能菌群内部种间关系具有显著影响,为太湖周边地区地下水反硝化功能菌群与硝态氮的互作机制研究提供了新的见解.

, correspAuthors=陈扣平, authorNote=null, correspAuthorsNote=
*责任作者,副教授,
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王卓群(1999-),女,山东淄博人,硕士研究生,主要从事地下水污染研究..

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王卓群(1999-),女,山东淄博人,硕士研究生,主要从事地下水污染研究..

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王卓群(1999-),女,山东淄博人,硕士研究生,主要从事地下水污染研究..

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***为P<0.001,*为P<0.05

, figureFileSmall=4BiyU/a3C6myNQ7TnRl2Tg==, figureFileBig=YcrEypW120Clu74s2G/kbQ==, tableContent=null), ArticleFig(id=1241049989808452416, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241049967259874217, language=EN, label=Fig.6, caption=The co-occurrence network of groundwater microbial communities at the genus level (a) and the co-occurrence network of denitrifying functional microbial communities at the OTU level (b), figureFileSmall=Da7wXHmCMu6BcEEi5lWkCA==, figureFileBig=fVmmCLRHyzXLCewNthI8Gw==, tableContent=null), ArticleFig(id=1241049990005584720, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241049967259874217, language=CN, label=图6, caption=属水平上的地下水微生物群落共现网络(a)和OTU水平上的反硝化功能菌群共现网络(b), figureFileSmall=Da7wXHmCMu6BcEEi5lWkCA==, figureFileBig=fVmmCLRHyzXLCewNthI8Gw==, tableContent=null), ArticleFig(id=1241049990194328412, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241049967259874217, language=EN, label=Table 1, caption=

Physicochemical properties of groundwater around the Lake Taihu area

, figureFileSmall=null, figureFileBig=null, tableContent=
样品分组T (℃)pH值TDS (mg/L)DO (mg/L)ORP(mV)碱度(mmol/L)
HN(n=7)AVG22.486.90301.712.5660.364.33
MIN19.516.60230.000.2322.601.80
MAX25.707.18407.007.33123.109.30
LN(n=7)AVG25.807.11341.295.5141.645.17
MIN20.486.66151.002.83-5.401.50
MAX29.957.67661.007.9358.809.30
), ArticleFig(id=1241049990471152492, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241049967259874217, language=CN, label=表1, caption=

太湖周边地区地下水理化性质

, figureFileSmall=null, figureFileBig=null, tableContent=
样品分组T (℃)pH值TDS (mg/L)DO (mg/L)ORP(mV)碱度(mmol/L)
HN(n=7)AVG22.486.90301.712.5660.364.33
MIN19.516.60230.000.2322.601.80
MAX25.707.18407.007.33123.109.30
LN(n=7)AVG25.807.11341.295.5141.645.17
MIN20.486.66151.002.83-5.401.50
MAX29.957.67661.007.9358.809.30
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太湖周边地下水中高硝酸盐负荷促进反硝化功能菌群富集
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王卓群 1 , 陈扣平 1, * , 吴吉春 1 , 王保战 2 , 柳旭 2
中国环境科学 | 环境微生物 2025,45(1): 477-486
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中国环境科学 | 环境微生物 2025, 45(1): 477-486
太湖周边地下水中高硝酸盐负荷促进反硝化功能菌群富集
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王卓群1 , 陈扣平1, * , 吴吉春1, 王保战2, 柳旭2
作者信息
  • 1.南京大学地球科学与工程学院,江苏 南京 210023
  • 2.南京农业大学生命科学学院,江苏 南京 210095
  • 王卓群(1999-),女,山东淄博人,硕士研究生,主要从事地下水污染研究..

通讯作者:

*责任作者,副教授,
High nitrate load in lakeside groundwater around Lake Taihu promotes the enrichment of denitrifying functional bacteria community
Zhuo-qun WANG1 , Kou-ping CHEN1, * , Ji-chun WU1, Bao-zhan WANG2, Xu LIU2
Affiliations
  • 1.School of Earth Sciences and Engineering, Nanjing University, Nanjing 210023, China
  • 2.College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China
出版时间: 2025-01-20
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反硝化功能菌群在降解硝态氮污染中发挥着重要作用,目前对其内部种间关系与硝酸盐负荷的联系仍了解有限.太湖周边地区地下水资源是重要的储备水源,但部分区域的地下水硝酸盐负荷超出了范围.本文以太湖周边地区14个不同硝酸盐浓度的地下水样品作为研究对象,运用微生物测序技术探究了地下水细菌群落多样性并识别出反硝化功能菌群,通过共现网络分析探究了反硝化功能菌群内部种间关系对硝酸盐负荷的响应.结果表明:高硝态氮组与低硝态氮组间细菌群落组成存在显著差异;细菌群落α多样性、β多样性受到硝酸盐负荷的显著影响;高硝态氮组地下水反硝化功能菌群内部种间关系较低硝态氮组更为密切.本研究表明地下水硝酸盐负荷对反硝化功能菌群内部种间关系具有显著影响,为太湖周边地区地下水反硝化功能菌群与硝态氮的互作机制研究提供了新的见解.

太湖  /  地下水  /  微生物多样性  /  硝态氮  /  共现网络

The denitrifying functional bacteria community plays a crucial role in the degradation of nitrate pollution. However, the understanding of the relationships between its interspecies links and nitrate load is still limited. Groundwater resources in the vicinity of Lake Taihu serve as vital reserves, yet in some areas, nitrate load in groundwater exceeds health thresholds. This paper focused on 14 groundwater samples with varying nitrate concentrations from the Lake Taihu area. With microbial sequencing techniques, the diversity of bacterial communities in groundwater was explored, and the denitrifying functional bacteria community was identified. The response of interspecies links within the denitrifying functional bacteria community to nitrate load was investigated by co-occurrence network analysis. The results indicate: Significant differences were found in bacterial community composition between high and low nitrate samples; The α-diversity and β-diversity were highly influenced by nitrate load; Interspecies links within the denitrifying functional bacteria community in the high nitrate samples were denser than those in the low nitrate samples. This study demonstrates that nitrate load in groundwater significantly influences the interspecies links within the denitrifying functional bacteria community, providing new insights into the interplay between groundwater denitrifying functional bacteria community and nitrate load in the Lake Taihu area.

Lake Taihu  /  groundwater  /  microbial diversity  /  nitrate  /  co-occurrence network
王卓群, 陈扣平, 吴吉春, 王保战, 柳旭. 太湖周边地下水中高硝酸盐负荷促进反硝化功能菌群富集. 中国环境科学, 2025 , 45 (1) : 477 -486 .
Zhuo-qun WANG, Kou-ping CHEN, Ji-chun WU, Bao-zhan WANG, Xu LIU. High nitrate load in lakeside groundwater around Lake Taihu promotes the enrichment of denitrifying functional bacteria community[J]. China Environmental Science, 2025 , 45 (1) : 477 -486 .
太湖周边地区地下水资源丰富,是当地居民生活用水的重要来源,在资源储备、生态功能维持、水质平衡等方面起到不可忽略的作用[1].然而,受到生活污水、人畜粪便及化肥施用的影响[2],外源硝酸盐通过淋滤、径流等方式入侵地下水[3],导致太湖周边地区地下水硝酸盐负荷在一定程度上超出了健康范围.太湖西苕溪流域部分地区地下水中硝酸盐浓度(以氮记)超过了10mg/L,最大值达到15.31mg/L[4].太湖西南侧长兴县浅层地下水中,硝酸盐浓度超标率为13.95%,浓度最高可达22mg/L,达到了我国《地下水质量标准》GB/T 14848 -2017中所规定的地下水质量Ⅳ类标准[5].地下水中过高的硝酸盐负荷不仅对人体健康存在各类潜在不良影响,而且会通过水力联系引发地表水体的同步恶化[6].
反硝化功能菌群能够将硝酸盐还原,最终以含氮气体的形式排放到大气中.这一过程是地下水中硝酸盐负荷降解的主要途径,因此反硝化功能菌群在降低地下水硝酸盐负荷方面发挥着至关重要的作用[7,8].已有研究表明,硝酸盐作为反硝化功能菌群生命活动的重要氮源,对反硝化功能菌群存在一定程度的影响,如过高的硝酸盐负荷会对脱氮硫杆菌产生抑制作用[911].然而,这些研究往往针对某一特定物种展开,而忽略了反硝化功能菌群作为一个功能整体,其内部种间关系对硝酸盐负荷的响应.
为了对上述问题进行探讨,本文在查明太湖周边地区地下水硝态氮负荷情况的基础上,选取地下水中的反硝化功能菌群为研究对象,探究其群落组成、结构及群落内种间关系对硝酸盐负荷的响应.将目光聚焦于地下水反硝化功能菌群并探究其对硝酸盐负荷的响应能够帮助人们更好地理解地下水微生物与硝态氮负荷之间的关系,进而更好地指导地下水硝态氮污染风险的治理.
选取太湖湖岸周边3km内地区(30°26′~31°56′N,119°53′~120°32′E)为研究区,研究区普遍覆盖着复杂第四系松散沉积物,孔隙水发育丰富,仅西南少部地区出露变质岩、岩浆岩,故导水性较差,常发育有裂隙水.研究区地下水水力坡度极小,且地下径流十分缓慢.属亚热带季风气候区,降水集中在4~10月(丰水期).区域地下水以大气降水入渗补给为主,以侧向补给、地下径流、蒸发等方式排泄.当地稠密的人口、发达的农业,为地下水环境造成了一定的生态压力.
2022年7月沿太湖湖岸周边3km内地区采集14个地下水样品,取样点位置图如图1所示.抽取地下水样品一段时间后,取水样进行后续处理,以确保采集的样品来自含水层而非井中滞留水.经0.45μm孔径滤膜过滤后,收集水样于预冲洗过的聚乙烯瓶中,使用H2SO4ρ=1.84g/mL)酸化至pH<2,封口胶封存好后于4℃保温箱中保存,用于实验室内测定地下水中溶解氮.使用0.22μm孔径滤膜真空抽滤4L左右水样,取下滤膜后立即放入-80℃冰箱保存,用于后续16sRNA分析.
抽取地下水样品一段时间,排空井中滞留水后,使用便携智能多参水质分析仪(HANNA HI9828,意大利)原位测量地下水的水温(WT)、酸碱度(pH值)、总溶解固体(TDS)、氧化还原电位(ORP)、溶解氧(DO)等指标.使用碱度试剂盒(MERCK 1.11109.0001,德国)对地下水中重碳酸根进行原位滴定,滴定精度为0.1mol/L.地下水中溶解氮组分,包括NO3-N、TN-N,由荷兰SKALAR公司生产的连续流动分析仪(SKALAR SAN++,荷兰)测定,检出限分别为0.002mg/L及0.01mg/L.
利用FastDNA SPIN Kit(MP Biomedicals,Santa Ana,CA,USA)从滤膜上提取DNA后,采用NanoDrop 2000测定(Thermo Fisher Scientific,USA)DNA浓度.基于上海美吉生物医药科技有限公司的Illumina MiSeq system(PE300),利用338F(ACTCCTACGG-GAGGCAGCAG)、806R(GGACTACHVGGGTWT-CTAAT)2个引物,对16s rRNA基因的高变V3~V4区进行扩增.使用QIME2对原始数据进行质控、拼接等处理,利用USEARCH去除嵌合体,最终按照相似性97%的有效序列聚类成OTU.比对Silva 16S RNA数据库(v.138),得到每个OTU所对应的物种信息.
使用R语言计算样品的Alpha多样性指数,并基于Bray-Curtis距离进行主坐标分析(PCoA).使用STAMP软件通过Wilcoxon秩和检验分析组间优势物种的丰度差异.基于Spearman相关系数绘制共现网络,选取相关系数大于0.6,且具有统计学意义(P<0.05)的样点,采用R语言进行网络分析,并导入Cytoscape(v.3.10.1)与Gephi(v.0.10.1)进行可视化.
研究区地下水的温度平均为24.14℃,整体呈现中性,所有样品均为淡水.地下水溶解氧DO介于0.23与7.93mg/L之间,氧化还原电位ORP的范围是-5.4~123.1mV.研究区地下水整体呈现氧化性,只有一个样点(TXC-G)ORP为负,呈现还原性.地下水碱度的范围是1.5~9.3mmol/L,平均值为4.75mmol/L.
以NO3-N浓度10mg/L为界限,将地下水样品分为高硝态氮组(HN)和低硝态氮组(LN)两组.由表1可以得知,HN组的ORP整体明显高于LN组,HN组地下水温度、pH值、DO整体低于LN组.高硝酸盐污染往往在氧化环境中出现,常伴有较高的地下水氧化还原电位.此外,硝酸盐倾向于稳定存在于酸性环境中,因此高硝酸盐可能与低pH值同时出现.硝酸盐污染带来的丰富氮源,促进了地下水微生物的生命活动,从而导致了较低的溶解氧.在碱度上,两组之间没有显著差异.
图2(a)展示了研究区地下水硝态氮负荷在HN、LN两组内的分布.总体而言,地下水NO3-N的浓度范围是0.92~25.42mg/L.对比而言,HN组的NO3-N浓度显著高于LN组(p<0.001),浓度范围分别为10.65~25.42mg/L和0.92~9.16mg/L,两组地下水的硝态氮浓度差异显著(p<0.001).图2(b)展示了硝态氮浓度与总氮浓度的关系,可以得知太湖周边地区地下水中硝态氮浓度与总氮浓度之间呈现较好的线性关系(R2=0.9969,p<0.001).意味着硝态氮浓度的变化能够很大程度上解释地下水中总体氮含量的变化.地下水中的溶解氮以各种形式存在,如硝态氮、亚硝态氮、氨氮等[12]. HN组和LN组的硝态氮浓度与总氮浓度比值的平均值分别为92.2%和80.4%,说明研究区地下水中硝态氮在总氮中的占比极高.此外,研究区地下水以微酸性氧化环境为主,为硝酸盐的稳定存在提供了有利条件[13-14].综上所述,研究区地下水中的氮主要以硝态氮的形式存在.我国渭河流域关中地段地下水中,地下水氮污染也同样主要以硝酸盐形式存在[15].
群落Alpha多样性直观体现了微生物群落的丰富度与均匀度,可通过香农多样性指数,辛普森均匀度指数进行衡量.图2(c)、2(d)分别展示了研究区地下水微生物群落的香农多样性指数(Shannon Diversity Index)和辛普森均匀度指数(Simpson Even Index). HN组的香农多样性指数、辛普森均匀度指数显著高于LN组(p<0.05),说明HN组的微生物群落Alpha多样性显著高于LN组,这可能是由HN组较高的硝酸盐负荷所导致.因为较高的硝酸盐负荷为微生物提供额外氮源,使群落内部竞争压力减小,并刺激某些嗜氮菌群的繁殖[16-17],从而导致微生物群落的丰富度和均匀度在一定程度上有所提高.此外,LN组较低的群落Alpha多样性说明一种或几种微生物在群落中具有极大的优势.HN组地下水微生物群落Alpha多样性显著高于LN组,表明太湖周边地区硝酸盐负荷较高的地下水环境中,微生物群落Alpha多样性更高.中国湖北省的一项研究表明,喀斯特洞穴内沉积物中细菌群落Alpha多样性的提升与地下水硝酸盐负荷的升高具有密切联系[18].在澳大利亚污染场地进行的一项研究表明,硝酸盐浓度在0~150mg/L范围内时,地下水微生物群落Alpha多样性随硝酸盐浓度的提高有所上升[19].然而,一项针对华北平原地下水的研究发现,硝酸盐负荷的差异与微生物群落Alpha多样性之间没有显著的相关关系[20].尽管两者都涉及到地下水中硝酸盐浓度微生物群落Alpha多样性的关系,但是却出现了不同的现象.两者的研究区域相差甚远,不同的地理位置、地质条件、营养物质类型等因素都可能导致硝酸盐浓度与Alpha多样性的不同.同时,不同的硝酸盐浓度范围,以及不同的时间跨度、样本数量,也会导致硝酸盐浓度与微生物群落Alpha多样性之间的关系得到不同程度的呈现.此外,不同地区的差异化微生物群落组成对硝酸盐的适应性和抵抗力存在不同,从而导致其对硝酸盐污染的敏感程度及响应程度各不相同.
图3展示了研究区地下水细菌群落PCoA主坐标分析结果,横纵两个坐标轴分别代表着地下水细菌群落的最大变异方向.PCo1轴解释了群落结构变化的31.22%,PCo2轴解释了群落结构变化的15.98%,两轴累积解释了群落结构变化的47.2%.样本在PCo1与PCo2上的投影代表了他们在该方向的主要差异.因此,HN组和LN组样点分别聚成了两个不同的集合,说明HN组与LN组在物种组成上存在显著差异.人为氮源输入能够显著改变微生物群落的Beta多样性[21],地下水理化性质的差异也能使得微生物群落组成发生巨大变化[22]. HN组、LN组之间环境差异主要集中在硝酸盐浓度的高低,因此可以推测PCo1可能代表地下水中硝酸盐的浓度,PCo2可能代表地下水理化性质的线性组合.HN组内样点间距离更大,LN组内间距离更小,说明HN组内各个样品间的群落结构差异更大,LN组内各个样品间的群落结构更为相似.
图4展示了HN组和LN组样品门水平上的微生物群落组成及相对丰度.总体而言,两组内最具优势的门是变形菌门(Proteobacteria),拟杆菌门(Bacteroidota)次之.HN、LN两组对比而言,变形菌门(Proteobacteria)在HN组内的优势显著低于LN组,两组内的平均相对丰度分别为67.13%和82.62%.拟杆菌门(Bacteroidota)在HN组内的优势大于LN组(p<0.05),两组内的平均相对丰度分别为12.57%和2.28%.变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)是地下水体中十分常见的门,在我国莱州湾南部、哈尔滨河岸、华北平原沿海地区地下水微生物群落中均有大量的分布[20].变形菌门(Proteobacteria)在地下水氮循环中起着重要作用,其门内的多种属参与并控制着氨氧化过程、硝化和反硝化过程.变形菌门(Proteobacteria)内含多种氨氧化细菌,参与并控制水体的氨氧化过程,能够将氨转化为亚硝酸盐和硝酸盐[23-24].在这一过程中,变形菌门(Proteobacteria)内的细菌往往对硝酸盐负荷的亲和力较低,因此在高硝酸盐负荷的环境中很难占据优势[25],而在硝酸盐负荷较低的环境中相对丰度较高,即其相对丰度在LN组显著更高.并没有研究明确指出拟杆菌门(Bacteroidota)是否具有氮循环功能,但一项在俄罗斯东乌拉尔地区进行的调查表明,其对硝酸盐污染具有较高的耐受能力,并能够在受硝酸盐污染的地下水中发挥其生物修复作用[26].这表明拟杆菌门(Bacteroidota)对硝酸盐负荷具有一定程度的耐受性,能够适应高硝酸盐负荷的地下水环境,因此在HN组内的更具优势.
图5展示了研究区地下水微生物群落在属水平上的Wilcoxon秩和检验结果,查明了HN、LN两组间属水平上的显著差异物种.不动杆菌属(Acinetobacter)、短波单胞菌属(Brevundimonas)、变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)、沉积物杆状菌属(Sediminibacterium)(P<0.001)以及弯曲杆菌属(Curvibacter)(P<0.05)在HN、LN两组间具有显著差异.前2者在LN组具有显著优势,后3者在HN组具有显著优势.不动杆菌属(Acinetobacter)在LN组具有极大优势,LN组内平均相对丰度占比高达54.2%,在两组间平均相对丰度差值高达近50%,是LN组的标志性物种.变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)在两组间平均相对丰度差值约为5%,是HN组的标志性物种.变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)能够通过反硝化作用和氨化作用代谢环境中的硝酸盐负荷,并具有一定的生物环境修复功能[27].因此,HN组更高的硝酸盐负荷可能为微生物提供了更为丰富的氮源,进而导致变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)在HN组地下水样品中的富集.不动杆菌属(Acinetobacter)内的多种菌株能够代谢水体中的硝酸盐负荷,进行异养硝化-好氧反硝化过程,即在有氧条件下同时进行硝化和反硝化作用[28-29].在这一过程过程中,硝酸盐作为中间产物,累积量很少[30],因此不动杆菌属(Acinetobacter)对硝酸盐的耐受性可能较差,从而更适宜在低硝酸盐环境中生存.此外,LN组地下水中溶解氧整体高于HN组,能够为异养硝化-好氧反硝化过程提供适宜的条件,因此不动杆菌属(Acinetobacter)在LN组内的相对丰度显著更高.
图6(a)展示了研究区属水平上微生物共现网络分析结果,并揭示出地下水微生物群落中可能存在的功能菌群.图6(a)中每一个节点代表一个属,涉及相对丰度前15的优势属.共生网络中66.67%的节点属于变形菌门(Proteobacteria),20%的节点属于拟杆菌门(Bacteroidota),6.67%的节点属于弯曲杆菌门(Campilobacterota),6.67%的节点属于异常球菌门(Deinococcota).节点之间的连线代表相关关系,共有11个正相关关系和8个负相关关系.连线粗细表示相关关系的强弱.根据15个优势属之间的相互关系划分出3个模块,每个模块代表一个可能存在的地下水功能菌群.模块1内含7个属,模块2和模块3各含4个属.模块1、模块3内部节点之间均呈正相关关系,且与模块2之间呈现负相关关系.
尽管模块1中的成员分别属于不同门,但它们均具有参与反硝化过程的潜力.变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)内有众多反硝化细菌成员,例如Comamonas denitrificans sp.等[33-34].黄杆菌属(Flavobacterium)具有多种反硝化基因,包括硝酸还原酶、亚硝酸还原酶、一氧化氮还原酶和一氧化二氮还原酶等,能够利用硝酸盐作为电子受体厌氧生长[33-34].未分类红环菌科(unclassified_f__Rhodocyclaceae)、硫单胞菌属(sulfurimonas)在各类污水处理厂水体中十分常见,他们的反硝化基因表达量较高,往往通过反硝化作用降解水体中的过量的硝酸盐[35-37].沉积物杆状菌属(Sediminibacterium)能够间接参与反硝化过程,在反硝化生物过滤器中起到重要作用[38-39].水杆菌属(Aquabacterium)、弯曲杆菌属(Curvibacter)虽也不能直接参与反硝化过程,但它们参与的铁氧化过程能够间接地降解硝酸盐,减少水体硝酸盐负荷[28,40-41].综上所述,模块1中的所有属都能够直接或间接地参与反硝化过程,并降低水体中的硝酸盐负荷.因此,模块1可能代表着研究区地下水微生物群落的反硝化功能菌群.
此外,结合图5的Wilconxon秩和检验结果可知,模块1中除水杆菌属(Aquabacterium)外,沉积物杆状菌属(Sediminibacterium)、弯曲杆菌属(Curvibacter)、变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)均是HN组的优势属,在HN组的相对丰度高于LN组.因此,模块1在一定程度上能够反映HN组优势微生物的属性.该现象在一定程度上说明,高硝态氮负荷可能会导致地下水反硝化功能菌群的富集.LN组的优势微生物不动杆菌属(Acinetobacter)、短波单胞菌属(Brevundimonas)均位于模块2中,且与模块1之间存在负相关关系.
为进一步探究高硝态氮负荷导致反硝化功能菌群富集的机制,在OTU水平上进行细菌群落共现网络分析,并计算了网络拓扑特征.图6(b)展示了图6(a)中模块1,即地下水反硝化功能菌群,在HN组和LN组的共现网络差异,每个节点代表一个OTU,节点间的连线(边)代表OTU之间的相关关系.在HN、LN两组的反硝化功能菌群共现网络中,网络节点数均为78个,HN组内节点间的边数明显高于LN组,分别为400和240.HN组内的节点加权平均度显著高于LN组,分别为8.68和5.33.HN组和LN组相同数目的节点数说明硝酸盐负荷的提升并没有筛选掉反硝化功能菌群内的物种.与LN组相比,HN组内更多的边数、更高的加权平均度说明,较高的硝酸盐负荷会促进反硝化功能菌群内各物种间的联系.此外,网络分析中,正相关的连接往往反映物种之间的合作关系,负相关关系反映物种间的竞争关系.对比两组可以得知,HN组正相关边的比例略高于LN组,分别为98.0%和93.3%,说明硝酸盐负荷的提高一定程度上提升了反硝化功能菌群物种间的协作关系.
3.1.1 太湖周边地区部分地下水硝酸盐污染达到了我国《地下水质量标准》GB/T 14848 -2017中的Ⅲ类水标准,极少部分达到了Ⅳ类水标准.研究区地下水中的氮主要以硝酸盐形式存在.此外,高硝酸浓度地下水具有氧化还原电位较高,地下水温度、pH值及溶解氧较低的特点.
3.1.2 高浓度硝酸盐地下水中的群落Alpha多样性显著高于低浓度硝酸盐地下水,且两者在群落组成上具有显著差异.变形菌门下未分类的丛毛单胞菌科(unclassified_f__Comamonadaceae)、沉积物杆状菌属(Sediminibacterium)、弯曲杆菌属(Curvibacter)在高硝酸盐浓度地下水中占有优势,而不动杆菌属(Acinetobacter)、短波单胞菌属(Brevundimonas)在低硝酸盐浓度地下水中具有优势.
3.1.3 地下水反硝化功能菌群倾向于在高硝态氮负荷的水体环境中富集.较高的硝酸盐负荷没有筛选掉反硝化功能菌群内的物种,而是使得反硝化功能菌群内各物种间的联系更为紧密,一定程度上提升了反硝化功能菌群物种间的协作关系.硝酸盐负荷是塑造地下水反硝化功能菌群的主要因素.
高硝酸盐浓度与其他水质、微生物指标之间的伴生关系,能够在地下水管理中起到指导作用.一方面,将理化指标与微生物群落多样性指标相结合,建立联合水质指标,可以评估地下水的污染程度以及潜在的健康风险,进一步为饮用水源地保护和优化管理提供依据.同时,这一指标也可以为地下水的长期监测和管理提供依据,从而确保地下水资源的水质稳定性.另一方面,将水质数据与微生物数据共同结合以深度学习方法,有望在污染源识别、预测领域提供新的见解.此外,反硝化功能菌群在地下水硝态氮降解上起着至关重要的作用,在地下水硝酸盐污染治理方面具有广泛的应用前景.探明野外环境中地下水反硝化功能菌群对硝酸盐负荷的响应,能够指导人们利用生物修复技术以解决自然环境中各类氮污染问题.
  • 国家自然科学基金资助项目(42177043)
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  • 接收时间:2024-06-14
  • 首发时间:2026-03-18
  • 出版时间:2025-01-20
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  • 收稿日期:2024-06-14
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国家自然科学基金资助项目(42177043)
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    1.南京大学地球科学与工程学院,江苏 南京 210023
    2.南京农业大学生命科学学院,江苏 南京 210095

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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