Article(id=1226855189949367181, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226855188863038235, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20240836, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1734796800000, receivedDateStr=2024-12-22, revisedDate=null, revisedDateStr=null, acceptedDate=1740844800000, acceptedDateStr=2025-03-02, onlineDate=1770434671151, onlineDateStr=2026-02-07, pubDate=1748966400000, pubDateStr=2025-06-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770434671151, onlineIssueDateStr=2026-02-07, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770434671151, creator=13701087609, updateTime=1770434671151, updator=13701087609, issue=Issue{id=1226855188863038235, tenantId=1146029695717560320, journalId=1192105938417971205, year='2025', volume='65', issue='6', pageStart='2321', pageEnd='2769', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1770434670891, creator=13701087609, updateTime=1770435273893, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1226857718103851267, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226855188863038235, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1226857718103851268, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226855188863038235, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2499, endPage=2513, ext={EN=ArticleExt(id=1226855191228629903, articleId=1226855189949367181, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Community structure and enzyme activity characteristics of halophilic bacteria in sulfate- and carbonate-type salt lakes in Xinjiang, China, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

[Objective] To study the distribution characteristics and enzyme potential of halophilic bacteria in two distinct types of salt lakes located in Xinjiang, China. [Methods] Soil samples were collected from sulfate-type (Qijiaojing) and carbonate-type (Nanhu Alkaline Lake) salt lakes, and their physicochemical properties were analyzed. The diversity, dominant taxa, and enzyme activities of halophilic bacteria were compared between the two salt lakes by Illumina MiSeq and culture experiments. [Results] The physicochemical properties of soil differed significantly between the two salt lakes, and the soil salinity of Qijiaojing salt lake (227.15 mg/g) was higher than that of Nanhu Alkaline Lake (158.61 mg/g). Significant differences were also observed in pH, HCO3-, Cl-, Mg2+, and K+ content. Spearman correlation analysis revealed positive correlations between Cl- and Mg2+ content and the relative abundance of dominant bacterial genera such as Pontibacter and Bacteroides. Illumina MiSeq results of bacterial 16S rRNA genes indicated that the Simpson and Shannon indexes of Nanhu Alkaline Lake were significantly higher than those of Qijiaojing. Halophilic bacteria belonging to 590 genera of 37 phyla were identified, including Bacteroidota (33.41%), Bacillota (24.71%), Actinomycetota (14.64%), and Pseudomonadota (10.58%). The dominant phylum was Bacteroidota (35.05%) in Nanhu Alkaline Lake, while it was Bacillota (44.66%) in Qijiaojing. The richness of halophilic bacteria in Nanhu Alkaline Lake exceeded that in Qijiaojing, with Pontibacter identified as the dominant genus in both lakes. A total of 1 130 strains were obtained from two salt lakes, belonging to 9 genera, 7 families of 4 phyla, among which Bacillota, Actinomycetota, and Pseudomonadota accounted for 40.53%, 36.81%, and 21.15%, respectively. The results of culture experiments with seven different media indicated that the F6 medium exhibited the highest selectivity towards halophilic microorganisms. Culture experiments demonstrated similar dominant species in both lakes, primarily comprising low-abundant bacteria, such as Nocardiopsis and Bacillus. Enzyme activity screening results revealed that 46.81%, 44.07%, and 20.88% of halophilic bacteria produced esterase, cellulase, and amylase, respectively, with Bacillus exhibiting the highest overall enzyme production capability. [Conclusion] There are significant differences in the halophilic bacterial diversity between sulfate- and carbonate-type salt lakes in Xinjiang. The halophilic bacteria in the carbonate-type Nanhu Alkaline Lake salt lake have higher diversity and exhibit stronger enzyme activities. This investigation contributes valuable insights for the advancement and sustainable utilization of microbial resources and the ecological preservation in salt lakes.

, correspAuthors=Yun WANG, authorNote=null, correspAuthorsNote=
*E-mail:
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These authors contributed equally to this work.

, authorsList=Lin GAO, Zhuobin XIE, Yun WANG, Gangqiang JIANG, Yanyan HAN, Xueying CHEN, Peng SUN), CN=ArticleExt(id=1226855192491115424, articleId=1226855189949367181, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=新疆典型硫酸盐和碳酸盐型盐湖嗜盐细菌群落特征及酶活特性, columnId=1192149544164012138, journalTitle=微生物学报, columnName=研究报告, runingTitle=null, highlight=null, articleAbstract=

【目的】 探究嗜盐细菌在中国新疆2种不同类型盐湖中的分布特征及其酶学特性。【方法】 采集代表性硫酸盐型(七角井)和碳酸盐型(南湖碱湖)盐湖土壤样本,分析其理化性质,并结合培养实验比较2个盐湖的微生物多样性、嗜盐细菌优势类群及其酶活特性。【结果】 2种类型盐湖的理化性质存在显著差异,七角井盐湖盐度高达227.15 g/kg,高于南湖碱湖的158.61 g/kg,其余pH、HCO3-、Cl-、Mg2+和K+含量也均有显著差异。Spearman相关性分析发现,Cl-和Mg2+含量与多数优势菌属丰度呈正相关,如海洋杆菌属(Pontibacter)和拟杆菌属(Bacteroides)。细菌16S rRNA基因Illumina MiSeq测序结果显示,南湖碱湖的Simpson和Shannon指数均显著高于七角井,嗜盐细菌分布于37门590属,优势类群包括拟杆菌门(Bacteroidota,33.41%)、芽孢杆菌门(Bacillota,24.71%)、放线菌门(Actinomycetota,14.64%)和假单胞菌门(Pseudomonadota,10.58%)。碳酸盐型南湖以拟杆菌门(Bacteroidota,35.05%)为主,硫酸盐型七角井则以Bacillota (44.66%)为主。南湖碱湖嗜盐细菌丰富度显著高于七角井盐湖,Pontibacter为两盐湖的优势类群。嗜盐菌分离培养结果显示,两盐湖共获得1 130株,隶属于4门7科9属,Bacillota占40.53%、Actinomycetota占36.81%、Pseudomonadota占21.15%。在7种分离培养基中,F6培养基对嗜盐微生物的选择性最佳。两盐湖的优势物种组成相似,多为群落中的低丰度类群,以拟诺卡氏菌属(Nocardiopsis)和芽孢杆菌属(Bacillus)为主。酶活筛选结果显示,46.81%的嗜盐细菌产酯酶,44.07%的菌产纤维素酶,20.88%的菌产淀粉酶,其中Bacillus的综合产酶能力最强。【结论】 新疆硫酸盐型(七角井)与碳酸盐型(南湖碱湖)盐湖的嗜盐微生物种类存在显著差异,碳酸盐型(南湖碱湖)的嗜盐细菌群落多样性更高,并展现出广泛的酶活性。本研究为盐湖微生物资源的开发利用和生态保护提供了科学依据。

, correspAuthors=王芸, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=wYjr5v1Yix4qb2Yu+uPaNw==, magXml=4hhO8upK+kwUWr+5vx5G8A==, pdfUrl=null, pdf=zYrQlb79hT1rpBf3gd4bVA==, pdfFileSize=1946533, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=hlsxOcD6ktz2tfpd+nby5Q==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=olxv+rmiWV3vDOaRNJVz9Q==, mapNumber=null, authorCompany=null, fund=null, authors=

作者贡献声明

高琳:利用传统微生物学方法对两盐湖进行菌种分离,以及初稿的撰写;谢卓斌:高通量测序数据分析,以及全文撰写与修改;王芸:全文指导与修改;蒋刚强:盐湖土壤样品的采集;韩燕燕:土壤样品理化性质的测定;陈雪莹:数据收集和处理;孙鹏:提供技术支持。

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Applied Microbiology and Biotechnology, 2023, 107(10): 3183-3190., articleTitle=Halomonas elongata: a microbial source of highly stable enzymes for applied biotechnology, refAbstract=null)], funds=[Fund(id=1227680965120688727, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, awardId=2024D01A25, language=EN, fundingSource=Natural Science Foundation of Xinjiang Uygur Autonomous Region(2024D01A25), fundOrder=null, country=null), Fund(id=1227680965292655203, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, awardId=2024D01A25, language=CN, fundingSource=新疆维吾尔自治区自然科学基金(2024D01A25), fundOrder=null, country=null), Fund(id=1227680965401707113, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, awardId=31400438, language=EN, fundingSource=National Natural Science Foundation of China(31400438), fundOrder=null, country=null), Fund(id=1227680965535924853, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, awardId=31400438, language=CN, fundingSource=国家自然科学基金(31400438), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1227680955893219405, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, xref=1., ext=[AuthorCompanyExt(id=1227680955901608014, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, companyId=1227680955893219405, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.School of Science and Technology, Lingnan Normal University, Zhanjiang, Guangdong, China), AuthorCompanyExt(id=1227680955905802319, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, companyId=1227680955893219405, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.岭南师范学院 生命科学与技术学院,广东 湛江)]), AuthorCompany(id=1227680956010659924, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, xref=2., ext=[AuthorCompanyExt(id=1227680956019048534, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, companyId=1227680956010659924, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.Urumqi Customs Technology Center, Urumqi, Xinjiang, China), AuthorCompanyExt(id=1227680956027437143, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, companyId=1227680956010659924, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.乌鲁木齐海关技术中心,新疆 乌鲁木齐)]), AuthorCompany(id=1227680956140683358, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, xref=3., ext=[AuthorCompanyExt(id=1227680956161654880, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, companyId=1227680956140683358, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.Xinjiang Forestry School, Urumqi, Xinjiang, China), AuthorCompanyExt(id=1227680956174237792, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, companyId=1227680956140683358, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.新疆林业学校,新疆 乌鲁木齐)])], figs=[ArticleFig(id=1227680961555530157, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Figure 1, caption=Microbial species diversity in the two salt lakes. A: Phylum level; B: Genus level; C: The difference in the abundance of the main bacterial flora (phylum level); D: Principal coordinate analysis generated using Bray Curtis distance; E: Venn maps of microbial differences obtained by two methods (genus level). N: Nanhu Alkaline Lake; Q: Qijiaojing; a: Illumina MiSeq sequencing; b: Pure culture; *: P<0.05; ***: P<0.001; ns: No significant., figureFileSmall=c5NhX24zUi8II00PxFxhoQ==, figureFileBig=py7IOHQzHWTEN8dNOVp+Jg==, tableContent=null), ArticleFig(id=1227680961664582069, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=图1, caption=两盐湖微生物物种多样性。A:门水平;B:属水平;C:主要细菌群落的丰度差异(门水平);D:基于Bray Curtis距离的主坐标分析;E:Illumina MiSeq测序与培养法的微生物差异韦恩图(属水平)。N:南湖碱湖;Q:七角井;a:Illumina MiSeq测序;b:可培养;*:P<0.05;***:P<0.001;ns:无显著性。, figureFileSmall=c5NhX24zUi8II00PxFxhoQ==, figureFileBig=py7IOHQzHWTEN8dNOVp+Jg==, tableContent=null), ArticleFig(id=1227680961794605508, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Figure 2, caption=High abundance halophiles bacteria diversity in the two salt lake. A: Halophiles bacteria diversity; B: Moderate halophiles bacteria diversity. S: Slight halophiles bacteria; M: Moderately halophiles bacteria; E: Extreme halophiles bacteria; Un: Unclassified; N: Nanhu Alkaline Lake; Q: Qijiaojing; Relative abundance>0.5%., figureFileSmall=xYCJUy2W1xU/LuiIvssenw==, figureFileBig=iRda8ixepAx8/AffTmnwBA==, tableContent=null), ArticleFig(id=1227680961916240339, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=图2, caption=两盐湖嗜盐细菌多样性。A:嗜盐细菌多样性;B:中度嗜盐细菌多样性。S:轻度嗜盐细菌;M:中度嗜盐细菌;E:极端嗜盐细菌;Un:未分类;N:南湖碱湖;Q:七角井;相对丰度>0.5%。, figureFileSmall=xYCJUy2W1xU/LuiIvssenw==, figureFileBig=iRda8ixepAx8/AffTmnwBA==, tableContent=null), ArticleFig(id=1227680962029486557, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Figure 3, caption=Comparison of halophilic bacterial diversity obtained from different culture media. A: Diversity of culturable halophiles in two salt lakes (N: Nanhu Alkaline Lake; Q: Qijiaojing); B: Diversity composition of halophiles in seven media., figureFileSmall=iShSmskY8GZnknL6yfGOSg==, figureFileBig=a1eD7PwllekjfF3cZT4Ifg==, tableContent=null), ArticleFig(id=1227680963451355628, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=图3, caption=不同培养基获得的嗜盐细菌多样性比较。A:两盐湖可培养菌群多样性(N:南湖碱湖;Q:七角井);B:7种培养基嗜盐细菌的多样性。, figureFileSmall=iShSmskY8GZnknL6yfGOSg==, figureFileBig=a1eD7PwllekjfF3cZT4Ifg==, tableContent=null), ArticleFig(id=1227680963577184755, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Figure 4, caption=Functional diversity of halophiles in two salt lakes. A: Functional diversity of moderate halophiles bacteria at genus level; B: Enzyme activity in two salt lakes (N: Nanhu Alkaline Lake; Q: Qijiaojing)., figureFileSmall=Tax2cilA/7/41smkHTv1MA==, figureFileBig=ki8XC1hgUtYoyU8La27aMQ==, tableContent=null), ArticleFig(id=1227680963711402493, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=图4, caption=两盐湖功能酶分析。A:嗜盐细菌的功能酶活多样性;B:两盐湖的酶活性分布(N:南湖碱湖;Q:七角井)。, figureFileSmall=Tax2cilA/7/41smkHTv1MA==, figureFileBig=ki8XC1hgUtYoyU8La27aMQ==, tableContent=null), ArticleFig(id=1227680963791094278, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Table 1, caption=

Physicochemical properties of two salt lake soil samples

, figureFileSmall=null, figureFileBig=null, tableContent=
SamplepH

Conductivity

(ms/cm)

Physical and chemical factors (g/kg)
Total saltCO32-HCO3-Cl-SO42-Ca2+Mg2+Na+K+
Nanhu Alkaline Lake8.51±0.52**35.84±21.21158.61±95.750.05±0.040.39±0.23**22.66±17.9175.12±54.844.46±1.980.49±0.3543.26±26.162.64±7.43*
Qijiaojing8.44±0.2459.28±30.41227.15±121.300.03±0.030.17±0.0790.82±74.77*48.45±43.876.81±4.112.23±1.61***69.94±41.950.30±0.25
), ArticleFig(id=1227680963895951886, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=表1, caption=

两盐湖土壤样品的理化性质

, figureFileSmall=null, figureFileBig=null, tableContent=
SamplepH

Conductivity

(ms/cm)

Physical and chemical factors (g/kg)
Total saltCO32-HCO3-Cl-SO42-Ca2+Mg2+Na+K+
Nanhu Alkaline Lake8.51±0.52**35.84±21.21158.61±95.750.05±0.040.39±0.23**22.66±17.9175.12±54.844.46±1.980.49±0.3543.26±26.162.64±7.43*
Qijiaojing8.44±0.2459.28±30.41227.15±121.300.03±0.030.17±0.0790.82±74.77*48.45±43.876.81±4.112.23±1.61***69.94±41.950.30±0.25
), ArticleFig(id=1227680964009198103, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Table 2, caption=

Alpha diversity index of microbial community in two salt lakes

, figureFileSmall=null, figureFileBig=null, tableContent=
Diversity indexIllumina MiSeq sequencingPure culture
Nanhu Alkaline LakeQijiaojingNanhu Alkaline LakeQijiaojing
Simpson0.94±0.04***0.74±0.0040.06±0.110.18±0.21
Shannon4.06±0.58***2.83±0.320.13±0.230.28±0.28
), ArticleFig(id=1227680964109861405, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=表2, caption=

两盐湖微生物群落α多样性指数

, figureFileSmall=null, figureFileBig=null, tableContent=
Diversity indexIllumina MiSeq sequencingPure culture
Nanhu Alkaline LakeQijiaojingNanhu Alkaline LakeQijiaojing
Simpson0.94±0.04***0.74±0.0040.06±0.110.18±0.21
Shannon4.06±0.58***2.83±0.320.13±0.230.28±0.28
), ArticleFig(id=1227680964231496228, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Table 3, caption=

Correlation analysis between soil physical and chemical properties and main bacteria (genus level)

, figureFileSmall=null, figureFileBig=null, tableContent=
PhylumGenuspHConductivityTotal saltCO32-HCO3-Cl-SO42-Ca2+Mg2+Na+K+
ActinomycetotaBifidobacterium-0.340.260.18-0.23-0.470.52*-0.330.070.59*0.22-0.20
Kocuria0.75***0.130.200.75***0.86***-0.210.77***0.10-0.330.180.34
Nitriliruptor0.38-0.34-0.230.160.59*-0.53*0.39-0.19-0.40-0.220.01
BacteroidotaBacteroides-0.240.360.31-0.07-0.380.60*-0.300.250.53*0.29-0.12
Parabacteroides-0.120.410.360.02-0.320.62**-0.260.320.51*0.35-0.02
Pontibacter0.54*0.180.330.460.66**-0.250.75***0.150.020.290.28
BalneolotaAliifodinibius-0.24-0.56*-0.49-0.31-0.24-0.49-0.25-0.33-0.33-0.54*0.04
BacillotaClostridium-0.34**0.190.11-0.29*-0.53***0.56-0.52***0.150.480.15-0.11
Dialister-0.220.290.21-0.12-0.390.62*-0.330.150.50*0.230.04
Faecalibacterium-0.160.390.34-0.01-0.360.65**-0.290.320.55*0.34-0.00
Fusicatenibacter-0.240.350.26-0.09-0.410.67**-0.350.210.52*0.28-0.01
Planococcus0.55*-0.030.100.300.72**-0.400.67**0.08-0.120.090.25
Roseburia-0.140.410.360.00-0.340.65**-0.270.310.53*0.35-0.01
Ruminococcus-0.150.410.360.00-0.350.65**-0.270.300.54*0.35-0.01
MethanobacteriotaHalalkalicoccus0.53*0.220.250.450.67**0.050.55*0.42-0.040.210.62*
PseudomonadotaSphingomonas0.00-0.56*-0.46-0.100.06-0.66**-0.01-0.61*-0.56*-0.44-0.20
RhodothermaeotaSalinibacter-0.53*-0.09-0.13-0.31-0.53*0.180-0.41-0.200.15-0.17-0.01
), ArticleFig(id=1227680964369908268, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=表3, caption=

土壤理化性质与主要细菌的相关性(属水平)

, figureFileSmall=null, figureFileBig=null, tableContent=
PhylumGenuspHConductivityTotal saltCO32-HCO3-Cl-SO42-Ca2+Mg2+Na+K+
ActinomycetotaBifidobacterium-0.340.260.18-0.23-0.470.52*-0.330.070.59*0.22-0.20
Kocuria0.75***0.130.200.75***0.86***-0.210.77***0.10-0.330.180.34
Nitriliruptor0.38-0.34-0.230.160.59*-0.53*0.39-0.19-0.40-0.220.01
BacteroidotaBacteroides-0.240.360.31-0.07-0.380.60*-0.300.250.53*0.29-0.12
Parabacteroides-0.120.410.360.02-0.320.62**-0.260.320.51*0.35-0.02
Pontibacter0.54*0.180.330.460.66**-0.250.75***0.150.020.290.28
BalneolotaAliifodinibius-0.24-0.56*-0.49-0.31-0.24-0.49-0.25-0.33-0.33-0.54*0.04
BacillotaClostridium-0.34**0.190.11-0.29*-0.53***0.56-0.52***0.150.480.15-0.11
Dialister-0.220.290.21-0.12-0.390.62*-0.330.150.50*0.230.04
Faecalibacterium-0.160.390.34-0.01-0.360.65**-0.290.320.55*0.34-0.00
Fusicatenibacter-0.240.350.26-0.09-0.410.67**-0.350.210.52*0.28-0.01
Planococcus0.55*-0.030.100.300.72**-0.400.67**0.08-0.120.090.25
Roseburia-0.140.410.360.00-0.340.65**-0.270.310.53*0.35-0.01
Ruminococcus-0.150.410.360.00-0.350.65**-0.270.300.54*0.35-0.01
MethanobacteriotaHalalkalicoccus0.53*0.220.250.450.67**0.050.55*0.42-0.040.210.62*
PseudomonadotaSphingomonas0.00-0.56*-0.46-0.100.06-0.66**-0.01-0.61*-0.56*-0.44-0.20
RhodothermaeotaSalinibacter-0.53*-0.09-0.13-0.31-0.53*0.180-0.41-0.200.15-0.17-0.01
), ArticleFig(id=1227680964516708915, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Table 4, caption=

Taxonomic position and functional enzyme activity of moderate halophiles isolated from Xinjiang salt lakes

, figureFileSmall=null, figureFileBig=null, tableContent=
PhylumGenusStrainCountThe closest species

Similarity

(%)

GenBank

NaCl

optimum (%)

AmylaseCellulaseEsterase
ActinomycetotaMyceligenerans4 3646Myceligenerans salitolerans XHU 503199.68PP4119485-++++
Nocardiopsis4 004235Nocardiopsis aegyptia DSM 44442T100.00PP4119545--+++
4 07420Nocardiopsis aegyptia DSM 44442T100.00PP4119475--+++
4 0761Nocardiopsis akebiae HDS12T100.00PP4119655NDNDND
4 13013Nocardiopsis akebiae HDS12T98.95PP4119555++++++
4 2101Nocardiopsis akebiae HDS12T99.05PP4119535-+++++
4 32592Nocardiopsis alba DSM 43377T98.80PP4119685NDNDND
4 38223Nocardiopsis alba DSM 43377T98.80PP41196910--+++
4 0533Nocardiopsis chromatogenes YIM 90109T99.68PP41194610--+++
4 4021Nocardiopsis eucommiae HDS5T98.63PP4119705++++++
4 1481Nocardiopsis halophila KCTC 9825T99.14PP41196610-+++++
4 3932Nocardiopsis synnemataformans DSM 44143T98.75PP4119455+++++++
Streptomyces4 2011Streptomyces mangrovicola GY199.83PP4119595+++-
4 2871Streptomyces ochraceiscleroticus NRRL ISP-559499.14PP41196710+-+++
4 3871Streptomyces ochraceiscleroticus NRRL ISP-559499.14PP4119565-++++
4 1783Streptomyces pini PL19T99.38PP4119525+++++++
4 08712Streptomyces sparsus YIM 90018T99.83PP41196310-+++
BalneolotaFodinibius4 15913Aliifodinibius salipaludis WN023T97.78PP4119725NDNDND
Bacillota4 1641Aliifodinibius salipaludis WN023T97.7PP41197310++++-
4 2403Aliifodinibius salipaludis WN023T98.23PP41197510NDNDND
Alteribacillus4 1264Alteribacillus bidgolensis IBRC-M10614T99.37PP41197110++++-
Bacillus4 057186Bacillus vallismortis DV1-F-3T99.84PP41195010+++++++++
4 25915Bacillus vallismortis DV1-F-3T99.84PP41194910+++++++++
4 3083Bacillus vallismortis DV1-F-3T99.83PP4119645++++++++
4 368244Bacillus vallismortis DV1-F-3T99.84PP4119515-++-
Gracilibacillus4 0415Gracilibacillus saliphilus YIM 91119T99.66PP4119625+++--
4 3951Gracilibacillus salitolerans SCU50T99.83PP4119575-++-
PseudomonadotaHalomonas4 169235Halomonas elongata DSM 2581T100.00PP41197415NDNDND
4 3901Halomonas xinjiangensis TRM 0175T98.15PP4119605+--
4 3991Halomonas xinjiangensis TRM 0175T98.15PP4119615-+++++
Microbulbifer4 1742Microbulbifer halophilus YIM 91118T99.68PP4119585-++++
), ArticleFig(id=1227680964638343739, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=表4, caption=

新疆两盐湖嗜盐微生物代表菌株分类地位及产酶活性

, figureFileSmall=null, figureFileBig=null, tableContent=
PhylumGenusStrainCountThe closest species

Similarity

(%)

GenBank

NaCl

optimum (%)

AmylaseCellulaseEsterase
ActinomycetotaMyceligenerans4 3646Myceligenerans salitolerans XHU 503199.68PP4119485-++++
Nocardiopsis4 004235Nocardiopsis aegyptia DSM 44442T100.00PP4119545--+++
4 07420Nocardiopsis aegyptia DSM 44442T100.00PP4119475--+++
4 0761Nocardiopsis akebiae HDS12T100.00PP4119655NDNDND
4 13013Nocardiopsis akebiae HDS12T98.95PP4119555++++++
4 2101Nocardiopsis akebiae HDS12T99.05PP4119535-+++++
4 32592Nocardiopsis alba DSM 43377T98.80PP4119685NDNDND
4 38223Nocardiopsis alba DSM 43377T98.80PP41196910--+++
4 0533Nocardiopsis chromatogenes YIM 90109T99.68PP41194610--+++
4 4021Nocardiopsis eucommiae HDS5T98.63PP4119705++++++
4 1481Nocardiopsis halophila KCTC 9825T99.14PP41196610-+++++
4 3932Nocardiopsis synnemataformans DSM 44143T98.75PP4119455+++++++
Streptomyces4 2011Streptomyces mangrovicola GY199.83PP4119595+++-
4 2871Streptomyces ochraceiscleroticus NRRL ISP-559499.14PP41196710+-+++
4 3871Streptomyces ochraceiscleroticus NRRL ISP-559499.14PP4119565-++++
4 1783Streptomyces pini PL19T99.38PP4119525+++++++
4 08712Streptomyces sparsus YIM 90018T99.83PP41196310-+++
BalneolotaFodinibius4 15913Aliifodinibius salipaludis WN023T97.78PP4119725NDNDND
Bacillota4 1641Aliifodinibius salipaludis WN023T97.7PP41197310++++-
4 2403Aliifodinibius salipaludis WN023T98.23PP41197510NDNDND
Alteribacillus4 1264Alteribacillus bidgolensis IBRC-M10614T99.37PP41197110++++-
Bacillus4 057186Bacillus vallismortis DV1-F-3T99.84PP41195010+++++++++
4 25915Bacillus vallismortis DV1-F-3T99.84PP41194910+++++++++
4 3083Bacillus vallismortis DV1-F-3T99.83PP4119645++++++++
4 368244Bacillus vallismortis DV1-F-3T99.84PP4119515-++-
Gracilibacillus4 0415Gracilibacillus saliphilus YIM 91119T99.66PP4119625+++--
4 3951Gracilibacillus salitolerans SCU50T99.83PP4119575-++-
PseudomonadotaHalomonas4 169235Halomonas elongata DSM 2581T100.00PP41197415NDNDND
4 3901Halomonas xinjiangensis TRM 0175T98.15PP4119605+--
4 3991Halomonas xinjiangensis TRM 0175T98.15PP4119615-+++++
Microbulbifer4 1742Microbulbifer halophilus YIM 91118T99.68PP4119585-++++
), ArticleFig(id=1227680964789338693, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=EN, label=Table 5, caption=

Diversity index of culturable halophiles bacteria

, figureFileSmall=null, figureFileBig=null, tableContent=
Culture mediaDiversity index
SimpsonShannon
F10.540.83
F20.670.80
F30.080.20
F40.480.67
F50.020.06
F60.520.81
F70.380.85
), ArticleFig(id=1227680964910973516, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226855189949367181, language=CN, label=表5, caption=

可培养嗜盐细菌物种多样性指数

, figureFileSmall=null, figureFileBig=null, tableContent=
Culture mediaDiversity index
SimpsonShannon
F10.540.83
F20.670.80
F30.080.20
F40.480.67
F50.020.06
F60.520.81
F70.380.85
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新疆典型硫酸盐和碳酸盐型盐湖嗜盐细菌群落特征及酶活特性
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高琳 1 , 谢卓斌 1 , 王芸 1, * , 蒋刚强 2 , 韩燕燕 3 , 陈雪莹 1 , 孙鹏 1
微生物学报 | 研究报告 2025,65(6): 2499-2513
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微生物学报 | 研究报告 2025, 65(6): 2499-2513
新疆典型硫酸盐和碳酸盐型盐湖嗜盐细菌群落特征及酶活特性
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高琳1, 谢卓斌1, 王芸1, * , 蒋刚强2, 韩燕燕3, 陈雪莹1, 孙鹏1
作者信息
  • 1.岭南师范学院 生命科学与技术学院,广东 湛江
  • 2.乌鲁木齐海关技术中心,新疆 乌鲁木齐
  • 3.新疆林业学校,新疆 乌鲁木齐
Community structure and enzyme activity characteristics of halophilic bacteria in sulfate- and carbonate-type salt lakes in Xinjiang, China
Lin GAO1, Zhuobin XIE1, Yun WANG1, * , Gangqiang JIANG2, Yanyan HAN3, Xueying CHEN1, Peng SUN1
Affiliations
  • 1.School of Science and Technology, Lingnan Normal University, Zhanjiang, Guangdong, China
  • 2.Urumqi Customs Technology Center, Urumqi, Xinjiang, China
  • 3.Xinjiang Forestry School, Urumqi, Xinjiang, China
出版时间: 2025-06-04 doi: 10.13343/j.cnki.wsxb.20240836
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【目的】 探究嗜盐细菌在中国新疆2种不同类型盐湖中的分布特征及其酶学特性。【方法】 采集代表性硫酸盐型(七角井)和碳酸盐型(南湖碱湖)盐湖土壤样本,分析其理化性质,并结合培养实验比较2个盐湖的微生物多样性、嗜盐细菌优势类群及其酶活特性。【结果】 2种类型盐湖的理化性质存在显著差异,七角井盐湖盐度高达227.15 g/kg,高于南湖碱湖的158.61 g/kg,其余pH、HCO3-、Cl-、Mg2+和K+含量也均有显著差异。Spearman相关性分析发现,Cl-和Mg2+含量与多数优势菌属丰度呈正相关,如海洋杆菌属(Pontibacter)和拟杆菌属(Bacteroides)。细菌16S rRNA基因Illumina MiSeq测序结果显示,南湖碱湖的Simpson和Shannon指数均显著高于七角井,嗜盐细菌分布于37门590属,优势类群包括拟杆菌门(Bacteroidota,33.41%)、芽孢杆菌门(Bacillota,24.71%)、放线菌门(Actinomycetota,14.64%)和假单胞菌门(Pseudomonadota,10.58%)。碳酸盐型南湖以拟杆菌门(Bacteroidota,35.05%)为主,硫酸盐型七角井则以Bacillota (44.66%)为主。南湖碱湖嗜盐细菌丰富度显著高于七角井盐湖,Pontibacter为两盐湖的优势类群。嗜盐菌分离培养结果显示,两盐湖共获得1 130株,隶属于4门7科9属,Bacillota占40.53%、Actinomycetota占36.81%、Pseudomonadota占21.15%。在7种分离培养基中,F6培养基对嗜盐微生物的选择性最佳。两盐湖的优势物种组成相似,多为群落中的低丰度类群,以拟诺卡氏菌属(Nocardiopsis)和芽孢杆菌属(Bacillus)为主。酶活筛选结果显示,46.81%的嗜盐细菌产酯酶,44.07%的菌产纤维素酶,20.88%的菌产淀粉酶,其中Bacillus的综合产酶能力最强。【结论】 新疆硫酸盐型(七角井)与碳酸盐型(南湖碱湖)盐湖的嗜盐微生物种类存在显著差异,碳酸盐型(南湖碱湖)的嗜盐细菌群落多样性更高,并展现出广泛的酶活性。本研究为盐湖微生物资源的开发利用和生态保护提供了科学依据。

嗜盐细菌  /  盐湖  /  Illumina MiSeq测序  /  可培养微生物  /  功能酶

[Objective] To study the distribution characteristics and enzyme potential of halophilic bacteria in two distinct types of salt lakes located in Xinjiang, China. [Methods] Soil samples were collected from sulfate-type (Qijiaojing) and carbonate-type (Nanhu Alkaline Lake) salt lakes, and their physicochemical properties were analyzed. The diversity, dominant taxa, and enzyme activities of halophilic bacteria were compared between the two salt lakes by Illumina MiSeq and culture experiments. [Results] The physicochemical properties of soil differed significantly between the two salt lakes, and the soil salinity of Qijiaojing salt lake (227.15 mg/g) was higher than that of Nanhu Alkaline Lake (158.61 mg/g). Significant differences were also observed in pH, HCO3-, Cl-, Mg2+, and K+ content. Spearman correlation analysis revealed positive correlations between Cl- and Mg2+ content and the relative abundance of dominant bacterial genera such as Pontibacter and Bacteroides. Illumina MiSeq results of bacterial 16S rRNA genes indicated that the Simpson and Shannon indexes of Nanhu Alkaline Lake were significantly higher than those of Qijiaojing. Halophilic bacteria belonging to 590 genera of 37 phyla were identified, including Bacteroidota (33.41%), Bacillota (24.71%), Actinomycetota (14.64%), and Pseudomonadota (10.58%). The dominant phylum was Bacteroidota (35.05%) in Nanhu Alkaline Lake, while it was Bacillota (44.66%) in Qijiaojing. The richness of halophilic bacteria in Nanhu Alkaline Lake exceeded that in Qijiaojing, with Pontibacter identified as the dominant genus in both lakes. A total of 1 130 strains were obtained from two salt lakes, belonging to 9 genera, 7 families of 4 phyla, among which Bacillota, Actinomycetota, and Pseudomonadota accounted for 40.53%, 36.81%, and 21.15%, respectively. The results of culture experiments with seven different media indicated that the F6 medium exhibited the highest selectivity towards halophilic microorganisms. Culture experiments demonstrated similar dominant species in both lakes, primarily comprising low-abundant bacteria, such as Nocardiopsis and Bacillus. Enzyme activity screening results revealed that 46.81%, 44.07%, and 20.88% of halophilic bacteria produced esterase, cellulase, and amylase, respectively, with Bacillus exhibiting the highest overall enzyme production capability. [Conclusion] There are significant differences in the halophilic bacterial diversity between sulfate- and carbonate-type salt lakes in Xinjiang. The halophilic bacteria in the carbonate-type Nanhu Alkaline Lake salt lake have higher diversity and exhibit stronger enzyme activities. This investigation contributes valuable insights for the advancement and sustainable utilization of microbial resources and the ecological preservation in salt lakes.

halophilic bacteria  /  salt lake  /  Illumina MiSeq  /  culturable microorganism  /  functional enzyme
高琳, 谢卓斌, 王芸, 蒋刚强, 韩燕燕, 陈雪莹, 孙鹏. 新疆典型硫酸盐和碳酸盐型盐湖嗜盐细菌群落特征及酶活特性. 微生物学报, 2025 , 65 (6) : 2499 -2513 . DOI: 10.13343/j.cnki.wsxb.20240836
Lin GAO, Zhuobin XIE, Yun WANG, Gangqiang JIANG, Yanyan HAN, Xueying CHEN, Peng SUN. Community structure and enzyme activity characteristics of halophilic bacteria in sulfate- and carbonate-type salt lakes in Xinjiang, China[J]. Acta Microbiologica Sinica, 2025 , 65 (6) : 2499 -2513 . DOI: 10.13343/j.cnki.wsxb.20240836
盐湖是地球上独特而重要的生态系统,具有高盐度、高pH值和高矿化度等极端环境特征,是重要的矿产资源和生物资源宝库[1]。作为拥有众多盐湖的国家,中国分布有1 000个大小不等的盐湖[2]。一般而言,盐浓度大于50 g/L的湖泊即可称为盐湖[3]。盐湖卤水的化学性质也是湖泊类型划分的重要依据,根据瓦里亚什科水化学分类方案,盐湖可分为碳酸盐型、硫酸盐型和氯化物型盐湖[4]。在盐湖的演化进程中,随着水分蒸发,饱和矿物离子析出沉淀,盐湖卤水内的主要离子成分也随之发生变化,表现为从碳酸盐型向硫酸盐型直至氯化物型的演化历程[3,5]
极端环境条件孕育了盐湖中独特的微生物群落,其中嗜盐微生物为主要组成。根据嗜盐菌最适生长NaCl浓度的不同,Kushner[6]将其划分为轻度嗜盐菌(0.2-0.5 mol/L NaCl)、中度嗜盐菌(0.5-2.0 mol/L NaCl)和极端嗜盐菌(2.0-5.2 mol/L NaCl)。其中,中度嗜盐菌分布广泛,涵盖了现有已知微生物的主要类群。嗜盐菌能够在高盐环境中生存,得益于它们进化出的一系列适应机制[7-8],如相容溶质的积累、离子泵的调节等,使其能够产生一些具有特殊性质的酶和代谢产物,如嗜盐淀粉酶、蛋白酶、脂肪酶等[9]。这些特殊的酶和代谢产物在食品、制药、环境治理等领域具有广阔的应用前景[9]。因此,研究盐湖嗜盐细菌的多样性及其功能特性,对于了解极端环境微生物的适应机制,并为开发新型生物资源提供重要理论基础。
我国新疆地处欧亚大陆腹地,远离海洋,气候干旱,蒸发强烈,是我国盐湖资源最为丰富的地区之一[3]。新疆盐湖类型多样,其卤水水化学类型以硫酸盐型居多,氯化物型和碳酸盐型盐湖极少,与青海、内蒙古和西藏盐湖片区相比,新疆盐湖有着自身独特性[3]。不同类型盐湖的理化性质存在显著差异,可能会导致其微生物群落结构和功能的差异。因此,对比研究不同类型盐湖的微生物多样性具有重要的科学意义。
当前对新疆盐湖嗜盐菌的研究相对有限,尤其是缺乏不同类型盐湖嗜盐细菌多样性的比较研究,以及对其功能特性的深入分析。本研究选择新疆典型的硫酸盐型(七角井)和碳酸盐型(南湖碱湖)盐湖作为研究对象,采用Illumina MiSeq测序技术与传统培养方法相结合的方法,揭示新疆典型硫酸盐型和碳酸盐型盐湖嗜盐细菌的多样性特征,比较两类盐湖嗜盐细菌群落组成的异同,分析土壤理化性质与微生物群落多样性的关系,评估不同培养基的分离效果,筛选具有潜在应用价值的嗜盐细菌株。本研究结果将有助于深入了解新疆盐湖微生物资源,为盐湖生态系统的保护和可持续利用提供科学依据。
土壤样品采集自新疆七角井盐湖和南湖碱湖,两盐湖位于我国新疆维吾尔自治区哈密地区吐哈盆地,气候终年干旱,从自20世纪50年代起两盐湖已无地表水,属于典型的干盐湖,七角井盐湖常有人开采制盐。根据郑喜玉[3]对两盐湖矿物质的测定,七角井盐湖属于硫酸盐型盐湖,南湖碱湖属于碳酸盐型盐湖。在两盐湖核心区域采集不同地貌植被类型的土壤样本,使用无菌袋分装后带回实验室,于4 ℃保存。南湖碱湖采样区位于42°34′-42°31′N,93°36′-93°43′E区域内,七角井采样区位于43°26′N,91°29′E区域内。
HS Taq酶,TaKaRa公司;PowerSoil® DNA Isolation Kit,深圳市安必胜科技有限公司;DNA marker,生工生物工程(上海)股份有限公司。
PCR仪,ThermoFisher Scientific公司;pH计,上海理达仪器厂;电导率仪,上海仪电科学仪器股份有限公司;火焰光度计,上海精密科学仪器有限公司。
F1培养基(g/L)[10]:酵母膏0.5,l-天门冬酰胺0.1,甘油1 mL,K2HPO4 0.1,KNO3 0.5,琼脂20,pH 7.2-7.5,去离子水1 000 mL;F2培养基:在F1基础上改良,将去离子水改为海水,其余成分不变;F3培养基(g/L)[11]:酵母提取物0.02,胰蛋白胨0.02,甘氨酸0.01,MgSO4·7H2O 5.0,Na2CO3 1.0,NaHCO3 0.5,山梨醇0.02,琼脂20.0,pH 7.2-7.5;F4培养基[11]:将F3中的山梨醇改为海藻糖,其余成分不变;F5培养基(g/L)[12]:淀粉10.0,酸水解酪蛋白0.3,KNO3 2.0,MgSO4·7H2O 0.05,K2HPO4 2.0,CaCO3 0.02,FeSO4 0.001,琼脂20.0,蒸馏水1 000 mL,pH 7.2-7.5;F6培养基(R2A) (g/L):蛋白胨0.5,酵母浸出粉0.5,葡萄糖0.5,酸水解酪蛋白0.5,可溶性淀粉0.5,KH2PO4 0.3,MgSO4 0.024,丙酮酸钠0.3,琼脂20.0,海水1 000 mL,pH 7.2;F7培养基(高氏) (g/L):可溶性淀粉20.0,KNO3 1.0,K2HPO4 0.5,MgSO4·7H2O 0.5,FeSO4·7H2O 0.01,琼脂20.0,蒸馏水1 000 mL,pH 7.4-7.6。
采集南湖碱湖土壤样品8份,七角井盐湖土壤样品7份。盐湖土壤理化性质测定方法具体参照《土壤农业化学分析方法》[13]:土壤水溶性盐总量的测定采用残渣烘干—质量法;pH采用电位测定法;土壤导电率采用导电法;土壤CO32-和HCO3-采用双指示剂中和法;Ca2+和Mg2+采用EDTA络合滴定法;Cl-采用AgNO3滴定法;K+和Na+采用火焰光度法;SO42-采用EDTA间接滴定法。
根据PowerSoil® DNA Isolation Kit的操作步骤提取盐湖土壤微生物基因组DNA。将提取满足要求的基因组DNA交由北京诺禾致源科技股份有限公司利用Illumina MiSeq平台进行高通量测序。引物选取细菌的16S rRNA基因的V3-V4区,引物为341F (5′- CCTACGGGNGGCW GCAG-3′)和785R (5′-GG ACTACHVGGGTWT CTAATCC-3′)[14]。数据分析采用QIIME 2软件包,根据97%的相似度进行可操作分类单元(operational taxonomic unit, OTU)划分、嵌合体移除和分类地位鉴定等分析[15]
为了评估当前常用嗜盐细菌分离培养基的分离效果,本研究通过文献调研,选取7种培养基,在5%、10%和15% NaCl浓度条件下进行嗜盐菌株分离。首先取出冰箱中保存的合并好的2份盐湖土样,各称取2 g置于锥形瓶中,用无菌水按1:10比例制作土壤稀释液,将其振荡混合充分后吸取100 μL稀释液,采用涂布平板法接种到各分离培养基,使其均匀分布,倒置于37 ℃恒温箱培养7-14 d。根据菌株的形态特征、颜色和大小进行筛选,选择形态存在明显差异的菌株并记录菌株生长信息,挑选每种培养基中的单菌落在F1培养基平板进行菌株纯化,去除重复菌株。
基因组提取与扩增参考文献[16]完成,使用Chelex-100法提取菌株基因组DNA,使用细菌通用引物27F (5′-AGAGTTTGATCMTGGCT CAG-3′)和1492R (5′-TACGGYTACCTTGTTA GGACTT-3′)进行细菌16S rRNA基因扩增。PCR反应体系(50 μL):DNA 1 μL,2×Taq PCR MasterMix溶液25 μL,上、下游引物(20 μmol/L)各1 μL,ddH2O 22 μL。PCR反应程序:95 ℃预变性5 min;94 ℃变性1 min,55 ℃复性5 min,72 ℃延伸1.5 min,共29个循环;72 ℃终延伸10 min。将扩增产物送至生工生物工程(上海)股份有限公司进行测序。测序结果通过EzBioCloud数据库(https://www.ezbiocloud.net/)[17]进行相似性比对,确定其分类地位。
按照李泉泉等[18]的研究方法,分别配制淀粉酶、纤维素酶和酯酶筛选培养基对代表菌株进行酶活功能筛选。
应用SPSS (v17.0)对土壤理化性质环境因子进行独立样本t检验(检验显著性水平为P<0.05)和Spearman相关性分析。利用PAST (v4.09)软件对分离微生物进行α多样性分析,采用GraphPad Prism (v8.0)绘图。
南湖碱湖和七角井两盐湖土壤的主要理化性质存在明显差异。如表1所示,两盐湖土壤的pH值均为8.50左右,属于盐碱土。七角井土壤的电导率和总盐含量分别为59.28 ms/cm和227.15 g/kg,高于南湖碱湖土壤。此外,两盐湖土壤的pH、HCO3-、Cl-、Mg2+和K+含量差异显著:南湖碱湖土壤的HCO3-和K+含量明显高于七角井,而七角井土壤的Cl-和Mg2+含量则高于南湖碱湖土壤4倍以上。
Illumina MiSeq测序数据分析显示(图1A),新疆两盐湖微生物多样性丰富,嗜盐细菌分布于37门82纲150目286科590属。门级优势菌群主要为拟杆菌门(Bacteroidota,33.41%)、芽孢杆菌门(Bacillota,24.71%)、放线菌门(Actinomycetota,14.64%)和假单胞菌门(Pseudomonadota,10.58%)。南湖碱湖以Bacteroidota (35.05%)和Bacillota (19.83%)为优势门,七角井以Bacillota (44.66%)和拟杆菌门(27.43%)为优势门。两盐湖放线菌门和芽孢杆菌门的丰度差异显著(图1B)。
在属水平(图1C),两盐湖微生物群落结构显著不同。南湖碱湖以海洋杆菌属(Pontibacter)为优势属(7.22%),而在七角井中仅为2.29%,七角井则以拟杆菌属(Bacteroides)为主(13.68%)。七角井中栖粪杆菌属(Faecalibacterium)、瘤胃球菌属(Ruminococcus)、巨球形菌属(Megasphaera)和双歧杆菌属(Bifidobacterium)等类群丰度接近4.64%-6.96%,与Bacteroides共同组成主要群落。与之相比,南湖碱湖解腈杆菌属(Nitriliruptor)、别样矿生菌属(Aliifodinibius)、盐坑微菌属(Salinimicrobium)和鞘氨醇单胞菌属(Sphingomonas)等类群丰度为2.32%-2.77%,与Pontibacter共同构成优势群落。两盐湖未知属级类群分别占57.79%和24.36%,显示出丰富的微生物资源。
通过对比2个盐湖的微生物α多样性指数(表2),发现南湖碱湖的Simpson指数明显高于七角井。这表明碳酸盐型的南湖碱湖微生物群落多样性远高于硫酸盐型的七角井。同时,南湖碱湖的Shannon指数与七角井存在显著差异,表明南湖碱湖具有更高的物种丰富度和均匀度。基于Bray Curtis距离的主坐标分析(PCoA)结果(图1D)显示,新疆碳酸盐型和硫酸盐型干盐湖的土壤微生物群落组成差异显著。
通过Spearman相关性分析(表3),本研究探讨了2个盐湖中高丰度土壤微生物类群(>1%)与相关土壤理化因子之间的关系。结果表明,Cl-和Mg2+是关键土壤理化因子,与大部分细菌属丰度呈显著正相关;pH与考克氏菌属(Kocuria)正相关,与梭菌属(Clostridium)负相关;电导率与AliifodinibiusSphingomonas呈显著负相关;CO32-Kocuria正相关;HCO3-KocuriaPontibacter、游动球菌属(Planococcus)和盐碱球菌属(Halalkalicoccus)正相关,与Clostridium负相关;SO42-KocuriaPontibacterPlanococcus正相关,与Clostridium负相关;Ca2+Sphingomonas负相关;Na+Aliifodinibius负相关;K+Halalkalicoccus正相关。总盐含量与菌属无显著相关性,可能是由于样品间差异较大所致,尤其是七角井盐湖,部分样品含有结晶盐矿,总盐含量高达488.5 g/kg。
通过7种培养基、3种NaCl浓度以及平板稀释涂布,共获得1 130株菌株(南湖碱湖391株,七角井739株)。在5%和10% NaCl浓度的培养基中主要分离到的是嗜盐细菌和放线菌。大部分菌落呈白色或乳白色,对这些菌株进行形态学特征分析后进行纯化和去重复。选取50株代表性嗜盐菌株进行16S rRNA基因序列相似性分析,登录号为PP411945-PP411975 (表4)。菌株隶属于4门7科9属,Bacillota占40.53%、Actinomycetota占36.81%、Pseudomonadota占21.15%、Balneolota占1.50% (图1A)。南湖碱湖以Bacillota为主(67.26%),七角井以Actinomycetota (39.24%)和Pseudomonadota (32.07%)为优势。Illumina MiSeq测序显示Bacteroidota为绝对优势门,但可培养实验未获得更多该门菌株,这表明目前常用的嗜盐菌分离培养基的配方设计还有待进一步完善。
在属水平(图1C),南湖碱湖的优势菌群为Bacillus (67.01%)和Nocardiopsis (30.18%)。七角井的优势菌群为Nocardiopsis (37.08%)、盐单胞菌属(Halomonas,31.80%)和Bacillus (25.17%)。南湖碱湖特有的菌属为生菌丝菌属(Myceligenerans),七角井特有的菌属为另类芽孢杆菌属(Alteribacillus)、矿生菌属(Fodinibius)和微泡菌属(Microbulbifer)。尽管南湖碱湖和七角井的优势物种组成相似,但菌属类群的丰度存在显著差异。通过韦恩图比较发现(图1E),可培养获得的9个属中,绝大部分并非Illumina MiSeq测序中的高丰度类群,仅BacillusHalomonas在南湖碱湖中丰度较高。
表2所示,2个盐湖中微生物物种分布及其α多样性指数存在差异。七角井所获的嗜盐微生物数量(739株)远多于南湖碱湖(391株)。七角井的Simpson指数和Shannon指数高于南湖碱湖,但差异未达到统计学显著性水平。
Illumina MiSeq测序结果无法区分轻度、中度和极端嗜盐菌的组成。为进一步了解盐湖中不同嗜盐细菌的分布情况,本研究选取丰度大于0.5%的49个属级类群进行分析。通过查阅相关文献并结合盐度实验数据,确定了这些菌属的嗜盐性特征[原始数据储存在国家微生物科学数据中心(http://nmdc.cn),编号为NMDCX0002079]。研究发现,18个属为轻度嗜盐菌,包括BacteroidesNitriliruptorRuminococcus等;19个属为中度嗜盐菌,包括AliifodinibiusBacillusHalomonas等;以及1个极端嗜盐菌属盐场杆菌属(Salinibacter)。此外,还有11个属尚未见嗜盐性相关报道。
Illumina MiSeq测序显示(图2A),南湖碱湖和七角井盐湖中丰度超过0.5%的属级类群均为22个,但嗜盐细菌分布差异显著。南湖碱湖中度嗜盐细菌属有11个,占群落丰度的12.13%;七角井仅有2个,占1.59%。在两盐湖的高丰度类群中,轻度嗜盐细菌占有较高比例;而极端嗜盐细菌仅在七角井中发现有较高丰度的属。两盐湖高丰度中度嗜盐细菌属存在明显差异(图2A),例如Aliifodinibius在南湖碱湖中的丰度为2.72%,而在七角井中仅为0.22%;乳杆菌属(Lactobacillus)在七角井中的丰度为0.74%,而在南湖碱湖中仅为0.007%。
总体而言,两盐湖间嗜盐细菌群落结构差异大,以轻度嗜盐细菌为主,南湖碱湖中的中度嗜盐细菌相较于七角井更为丰富。
本研究采用7种不同的分离培养基从2个盐湖土壤样本中分离获得1 130株嗜盐细菌(图3A)。结果表明,培养基F6能最有效地分离嗜盐细菌,共分离到490株,涵盖了5个菌属;其次是培养基F5和F3,分别分离得到344株和194株。不同培养基的多样性指数分析结果也表明,F1、F2、F4、F6和F7培养基分离的可培养微生物多样性明显高于F3和F5培养基(表5)。进一步分析种属发现(图3B),Nocardiopsis在F4、F5和F7培养基中占绝对优势,尤以F5培养基最高达98.84%。Bacillus在F1、F3和F6培养基中为优势类群,以F3培养基最高达93.88%。此外,Alteribacillus仅在F5培养基中分离获得,而MyceligeneransMicrobulbifer属则仅在F6培养基中分离获得。综上所述,合理选择培养基对于获得目标菌属至关重要。本研究发现,F5培养基对放线菌Nocardiopsis的分离效果最佳,而F6培养基则对BacillusHalomonas的分离效果最优。
通过可培养嗜盐细菌功能酶筛选发现(图4A),产酯酶和纤维素酶的菌株数量最多,分别占菌株总数的46.81%和44.07%,产淀粉酶的菌株占20.88%。从菌群类别来看,Bacillus在产淀粉酶和纤维素酶方面的能力最强,占产淀粉酶和纤维素酶菌数的86.44%和89.96%。相比之下,Nocardiopsis在产酯酶方面的能力最强,占56.52%,Bacillus次之(38.56%)。从菌株分布来看(图4B),产淀粉酶和酯酶的菌株主要分布在七角井,产纤维素酶的菌株在南湖碱湖和七角井的分布量相近。综合分析可知,两盐湖土壤可培养菌株中,有69.56%的菌株具有产胞外酶的能力,Bacillus在3种功能酶的综合产酶能力方面最为突出。
嗜盐微生物是盐湖微生态系统中的主要类群,它们通过积累胞内小分子相容溶质以抵御渗透压,其多样性的代谢特征和产多功能酶的特性展现出巨大的工业应用潜力[9]。七角井盐湖和南湖碱湖虽然地理位置接近,且均为干盐湖,但水化学性质存在差异,是研究盐湖演化类型对嗜盐微生物群落组成和结构影响的理想样地。
根据盐湖演化进程,硫酸盐型盐湖的成盐年代比碳酸盐型盐湖更早[3],其含盐量通常高于碳酸盐型盐湖,主要理化性质也存在显著差异,因此推测其微生物群落可能存在明显差异。本研究中两盐湖土壤理化性质测定结果表明,硫酸盐型(七角井)盐湖的总盐含量高于碳酸盐型(南湖碱湖),其余因子如pH、HCO3-、Cl-、Mg2+、K+含量也存在显著差异,这些理化因子在一定程度上对两盐湖土壤微生物群落组成和数量具有显著影响[9]。来自111项研究的统计结果表明,盐度是影响微生物群落结构的一项决定性因素[19],通常认为随着盐度的升高,会导致环境中细菌多样性下降[20-21]。对两盐湖Illumina MiSeq测序结果进行α多样性分析发现,总盐度更高的七角井盐湖,其嗜盐细菌群落多样性低于南湖碱湖,现有研究表明地理位置、盐度、温度和季节等环境因素都会显著影响不同盐湖中微生物群落的结构和功能[22]。本研究的相关性分析结果揭示,Cl-和Mg2+浓度是关键的土壤理化因子,对盐湖土壤微生物群落结构产生重要影响。这与先前研究发现Cl-含量是影响盐碱土土壤细菌群落组成的重要因素一致[23]。Wallis等[24]的研究也发现,Mg2+含量与土壤细菌群落结构和组成显著相关。综上所述,在分离嗜盐微生物时,不仅需要考虑NaCl浓度,还可以加入适量浓度的MgCl2
通过Illumina MiSeq测序分析,发现两盐湖均蕴含丰富的嗜盐细菌微生物资源,共有37门82纲150目286科590属。BacteroidotaBacillotaActinomycetotaPseudomonadota为两盐湖普遍存在的核心类群。对比其他盐湖研究结果发现,盐湖中优势的嗜盐微生物类群在门级分类水平上存在一定的共性,主要优势类群包括假单胞菌门、芽孢杆菌门、拟杆菌门[25];然而,由于不同盐湖的水化学特性或其他因素存在差异,这些优势类群在各盐湖中的相对丰度也会有所不同。韩晶等[26]对硫酸盐型艾比湖研究发现Pseudomonadota为第一优势群,朱德锐等[27]发现青藏高原盐湖微生物以Bacillota为优势群,且Bacillota的丰度比例与盐湖总盐度相关,这与七角井的结果一致。嗜盐微生物会根据不同的环境变化进行适应性调整,例如Bacteroidota可能是逐渐适应干旱区的一类微生物[28],这与盐湖水域面积减少、逐步趋向干旱化密切相关[29]Pseudomonadota在环境中广泛存在,在大部分有水盐湖中通常为优势门或次优势门[25-26,29],而在两盐湖中为低丰度类群,可能与两盐湖均为干盐湖有关。
两个盐湖在属级类群分布上存在显著差异,除Pontibacter为两盐湖共有的高丰度类群外,其余丰度>0.5%的类群在另一湖中的丰度普遍低于0.1%。七角井中丰度较高的FaecalibacteriumRuminococcusMegasphaeras均属于Bacillota,其内生孢子的能力使其能在极端条件下正常繁殖和生存[30]。值得注意的是,七角井富含高丰度的Bifidobacterium (6.97%),文献报道Bifidobacterium是维持人和动物肠道健康的重要益生菌,最适生存pH在6.0-7.0之间,当NaCl浓度超过2%时,其生长会受到显著抑制[31-32]。目前尚未有报道在盐湖中分离获得Bifidobacterium,极端环境下的双歧杆菌的生理特性有待进一步探索。
尽管通过可培养方法可以探究盐湖嗜盐细菌的多样性,但该方法存在明显局限性。大部分分离菌株属于低丰度类群,如MyceligeneransNocardiopsisStreptomycesFodinibiusAlteribacillusGracilibacillusMicrobulbifer,其丰度均低于0.5%,并非盐湖中主要的嗜盐细菌群。本研究主要从5%盐浓度培养基中分离获得了中度嗜盐细菌,其中南湖碱湖的优势类群为Bacillus,七角井的优势类群为放线菌NocardiopsisBacillusNocardiopsisHalomonasStreptomyces等属被认为是代表性的中度嗜盐菌,广泛分布于盐湖[33-34]、海洋沉积物[35]盐场[36]等盐环境,是目前研究较为深入的嗜盐细菌。值得注意的是,在碳酸盐型南湖碱湖中,Pontibacter作为第一优势类群,在硫酸盐型七角井中的丰度也达2.29%,但本次分离未获得该菌属。根据文献报道,Pontibacter可在0-8%盐浓度下生长,最佳生长盐浓度为2%-3%[37],培养基盐度设置过高可能是本研究未获得该属的主要原因。Pontibacter已在新疆沙漠、胡杨根际土壤等干旱环境中有报道[38-39]。本研究还发现Pontibacter的丰度与盐湖HCO3-和SO42-含量呈显著正相关,盐湖HCO3-和SO42-含量是划分碳酸盐型和硫酸盐型的重要理化因子。因此,在未来的可培养实验中,应充分考虑微生物生长所需的环境条件。此外,盐湖中还有19.82%的未分类菌属类群,表明盐湖中丰富的新物种资源有待进一步开发。
不同培养基分离结果显示,F6 (R2A)培养基获得的嗜盐细菌株数和菌株种类最为丰富,是一款优秀的分离培养嗜盐细菌的培养基。Oren[40]曾指出,丙酮酸钠对嗜盐细菌的生长非常重要,我们在此前分离极端嗜盐菌时也发现了这一规律[18]。由此可见,在分离微生物菌群时,尤其是获取某一特定菌群,应当结合其原始生存环境特性和对不同离子成分的偏好等特征设计培养基,以简化菌属分离培养工作,从而获得目标菌群。
本次酶活筛选结果显示,可培养嗜盐细菌产功能酶的能力较强,其中产酯酶和纤维素酶的嗜盐细菌数量最多,而产淀粉酶菌株较少。Bacillus是两盐湖中主要的产功能酶类群,芽孢杆菌被广泛应用于工业生产多种功能酶和有机代谢物,据估计,总酶市场中近50%的酶由枯草芽孢杆菌生产[41]。如Nocardiopsis通过分泌纤维素、淀粉、蛋白质等水解酶从环境中吸收营养,使其能够适应多种环境。天然高盐碱生拟诺卡氏菌具有丰富的物种多样性、代谢产物多样性和产酶能力,是拟诺卡氏菌资源研究的热点,且在食品、环境等领域展现出巨大的应用潜力[42]。同时,结果进一步说明高氏培养基也适用于嗜盐放线菌的分离。研究表明,嗜盐酶和耐盐酶通常对高盐环境具有高耐受性,在生物能源、生物修复和生物医学等领域具有潜在的应用价值[43]。综上所述,嗜盐细菌具有强大的生物技术应用和发展潜力,深入研究嗜盐细菌群的功能及其酶系统有助于我们更好地将其转化为功能性生产资源。
本研究联合Illumina MiSeq测序和传统培养方法,探究了新疆碳酸盐型(南湖碱湖)和硫酸盐型(七角井) 2种类型盐湖中嗜盐细菌的生物多样性。研究发现,2个盐湖的微生物群落结构存在显著差异,碳酸盐型南湖碱湖更有利于嗜盐细菌的生存。不同培养基分离获得的嗜盐微生物类群为后续特定菌株的获取提供了理论参考。此外,两盐湖微生物的功能特性研究对开发利用盐湖嗜盐微生物在生物资源库、酶制剂和生物新功能等领域具有重要的理论意义。
作者声明不存在任何可能会影响本文所报告工作的已知经济利益或个人关系。
  • 新疆维吾尔自治区自然科学基金(2024D01A25)
  • 国家自然科学基金(31400438)
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2025年第65卷第6期
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doi: 10.13343/j.cnki.wsxb.20240836
  • 接收时间:2024-12-22
  • 首发时间:2026-02-07
  • 出版时间:2025-06-04
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  • 收稿日期:2024-12-22
  • 录用日期:2025-03-02
基金
Natural Science Foundation of Xinjiang Uygur Autonomous Region(2024D01A25)
新疆维吾尔自治区自然科学基金(2024D01A25)
National Natural Science Foundation of China(31400438)
国家自然科学基金(31400438)
作者信息
    1.岭南师范学院 生命科学与技术学院,广东 湛江
    2.乌鲁木齐海关技术中心,新疆 乌鲁木齐
    3.新疆林业学校,新疆 乌鲁木齐

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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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