South China Sea (SCS) is the largest Western Pacific marginal sea. However, microbial studies have never been performed in the cold seep sediments in the SCS. In 2004, "SONNE" 177 cruise found two cold seep areas ...South China Sea (SCS) is the largest Western Pacific marginal sea. However, microbial studies have never been performed in the cold seep sediments in the SCS. In 2004, "SONNE" 177 cruise found two cold seep areas with different water depth in the northern SCS. Haiyang 4 area, where the water depth is around 3000 m, has already been confirmed for active seeping on the seafloor, such as microbial mats, authigenic carbonate crusts and bivalves. We investigated microbial abundance and diver- sity in a 5.55-m sediment core collected from this cold seep area. An integrated approach was employed including geochemistry and 16S rRNA gene phylogenetic analyses. Here, we show that microbial abun- dance and diversity along with geochemistry profiles of the sediment core revealed a coupled reaction between sulphate reduction and methane oxidation. Acridine orange direct count results showed that microbial abundance ranges from 105 to 106 cells/g sediment (wet weight). The depth-related variation of the abundance showed the same trend as the methane concentration profile. Phylogenetic analysis indicated the presence of sulphate-reducing bacteria and anaerobic methane-oxidizing archaea. The diver- sity was much higher at the surface, but decreased sharply with depth in response to changes in the geochemical conditions of the sediments, such as methane, sulphate concentration and total organic carbon. Marine Benthic Group B, Chloroflexi and JS1 were predominant phylotypes of the archaeal and bacterial libraries, respectively.展开更多
Diversity of bacteria was studied in deep-sea sediments from the Shikoku Basin in the Northwest Pacific Ocean by PCR, RFLP and sequence analysis of 16S rDNA and comparing with Genbank database. Based on the RFLP profi...Diversity of bacteria was studied in deep-sea sediments from the Shikoku Basin in the Northwest Pacific Ocean by PCR, RFLP and sequence analysis of 16S rDNA and comparing with Genbank database. Based on the RFLP profile generated, 77 clones from the 16S rDNA library were divided into 27 types. Phylogenetic analysis showed that the 27 independent clones fell into four groups: Proteobac-teria (62.96%), Chloroflexi (14.81%), Planctomycetes (14.81%) and Acidobacteria (7.41%). Among all sequenced clones, 6 were related to the sulfur or sulfate metabolism bacteria and the results also demonstrated that some bacteria in deep-sea sediments had relation to matter-energy circulation.展开更多
利用分子生物学技术分析南海北部神狐海域天然气水合物潜力区HS-373PC岩心表层沉积物中古菌多样性,从沉积物中提取总DNA并扩增古菌16SrRNA基因序列,对克隆文库进行系统发育分析。结果显示所有古菌序列均属于泉古菌(Crenarchaeota)和广古...利用分子生物学技术分析南海北部神狐海域天然气水合物潜力区HS-373PC岩心表层沉积物中古菌多样性,从沉积物中提取总DNA并扩增古菌16SrRNA基因序列,对克隆文库进行系统发育分析。结果显示所有古菌序列均属于泉古菌(Crenarchaeota)和广古菌(Euryarchaeota)。其中泉古菌以C3为主要类群,另有少量序列属于Marine Benthic Group(MBG)-B,MBG-C,Marine Crenarchaeotic Group I(MGI),Marine Hydrothermal VentGroup(MHVG)和Novel Group of Crenarchaea(NGC);广古菌以MBG-D为主,其它序列分别属于UnclassifiedEuryarchaeotic Clusters-1/2(UEC-1/2)。展开更多
利用提取且纯化的西北太平洋地区深海沉积物DNA为模板,利用细菌通用PCR引物扩增16S rDNA片段,构建其文库,建立阳性克隆子RFLP(Restriction Fragment Length Polymor-phism)酶切图谱.据酶切图谱选择部分克隆测序,并与数据库中的序列进行...利用提取且纯化的西北太平洋地区深海沉积物DNA为模板,利用细菌通用PCR引物扩增16S rDNA片段,构建其文库,建立阳性克隆子RFLP(Restriction Fragment Length Polymor-phism)酶切图谱.据酶切图谱选择部分克隆测序,并与数据库中的序列进行比对.结果表明,测序的27个序列分属于4个类群:变形细菌(Proteobacteria)、绿屈挠菌(Chloroflexi)、浮游霉菌(Planctomycetes)和酸杆菌(Acidobacteria);其中以变形细菌最多,占62.96%.展开更多
【目的】海洋沉积物中的古菌在全球生物地球化学循环中充当重要的角色,深入了解沉积物中古菌群落的结构及功能特征是探究海洋沉积物中古菌参与生物地球化学循环和生态学功能的基础。【方法】采用高通量测序技术,分别对南海北部陆坡不同...【目的】海洋沉积物中的古菌在全球生物地球化学循环中充当重要的角色,深入了解沉积物中古菌群落的结构及功能特征是探究海洋沉积物中古菌参与生物地球化学循环和生态学功能的基础。【方法】采用高通量测序技术,分别对南海北部陆坡不同海域(东部,西部和神狐海域的7个站位)沉积物中古菌16SrRNA基因进行Illumina Mi Seq测序。【结果】中国南海北部陆坡沉积物中古菌的主要门类是Bathyarchaeota、Thermoplasmata、Woesearchaeota(DHVEG-6)、Thaumarchaeota(Marine Group I)、Lokiarchaeota和Marine Hydrothermal Vent Group(MHVG),还存在少量的AK8、Marine Benthic Group A和Terrestrial Hot Spring Crenarchaeota Group(THSCG)等。在潜在水合物区沉积物中还发现了甲烷代谢相关古菌(Anaerobic methanotrophic archaea,ANME)类群,主要为ANME-1、ANME-2ab和ANME-2c等。甲烷代谢古菌的分布特征也从甲烷代谢保守功能基因mcr A(Methyl coenzyme-Mreductase A)的扩增中得到了验证。利用定量PCR对南海沉积物中的细菌、古菌的16SrRNA基因和mcrA基因进行了定量,发现细菌16SrRNA基因拷贝数为10~5-10~7 copies/g(湿重),古菌16SrRNA基因拷贝数为10~5-10~6 copies/g(湿重),潜在水合物区mcrA基因拷贝数为10~3-10~5 copies/g(湿重)。【结论】揭示了中国南海北部陆坡沉积物中具有丰富的微生物资源,其中古菌种类多样且丰度较高,同时发现冷泉特征古菌群落,为深入认识和理解南海沉积物中微生物丰度和古菌多样性,以及解析古菌地球化学功能奠定基础。展开更多
基金supported by the National Program on Key Basic Research Project(973 Program)(Grant No.2009CB219502)National Special Foundation(Grant No.GZH200200203-02-01)Non-profit Industry Financial Program of Ministry of Land and Resources of the PRC(Grant No.200811014-02)
文摘South China Sea (SCS) is the largest Western Pacific marginal sea. However, microbial studies have never been performed in the cold seep sediments in the SCS. In 2004, "SONNE" 177 cruise found two cold seep areas with different water depth in the northern SCS. Haiyang 4 area, where the water depth is around 3000 m, has already been confirmed for active seeping on the seafloor, such as microbial mats, authigenic carbonate crusts and bivalves. We investigated microbial abundance and diver- sity in a 5.55-m sediment core collected from this cold seep area. An integrated approach was employed including geochemistry and 16S rRNA gene phylogenetic analyses. Here, we show that microbial abun- dance and diversity along with geochemistry profiles of the sediment core revealed a coupled reaction between sulphate reduction and methane oxidation. Acridine orange direct count results showed that microbial abundance ranges from 105 to 106 cells/g sediment (wet weight). The depth-related variation of the abundance showed the same trend as the methane concentration profile. Phylogenetic analysis indicated the presence of sulphate-reducing bacteria and anaerobic methane-oxidizing archaea. The diver- sity was much higher at the surface, but decreased sharply with depth in response to changes in the geochemical conditions of the sediments, such as methane, sulphate concentration and total organic carbon. Marine Benthic Group B, Chloroflexi and JS1 were predominant phylotypes of the archaeal and bacterial libraries, respectively.
文摘Diversity of bacteria was studied in deep-sea sediments from the Shikoku Basin in the Northwest Pacific Ocean by PCR, RFLP and sequence analysis of 16S rDNA and comparing with Genbank database. Based on the RFLP profile generated, 77 clones from the 16S rDNA library were divided into 27 types. Phylogenetic analysis showed that the 27 independent clones fell into four groups: Proteobac-teria (62.96%), Chloroflexi (14.81%), Planctomycetes (14.81%) and Acidobacteria (7.41%). Among all sequenced clones, 6 were related to the sulfur or sulfate metabolism bacteria and the results also demonstrated that some bacteria in deep-sea sediments had relation to matter-energy circulation.
文摘利用分子生物学技术分析南海北部神狐海域天然气水合物潜力区HS-373PC岩心表层沉积物中古菌多样性,从沉积物中提取总DNA并扩增古菌16SrRNA基因序列,对克隆文库进行系统发育分析。结果显示所有古菌序列均属于泉古菌(Crenarchaeota)和广古菌(Euryarchaeota)。其中泉古菌以C3为主要类群,另有少量序列属于Marine Benthic Group(MBG)-B,MBG-C,Marine Crenarchaeotic Group I(MGI),Marine Hydrothermal VentGroup(MHVG)和Novel Group of Crenarchaea(NGC);广古菌以MBG-D为主,其它序列分别属于UnclassifiedEuryarchaeotic Clusters-1/2(UEC-1/2)。
文摘利用提取且纯化的西北太平洋地区深海沉积物DNA为模板,利用细菌通用PCR引物扩增16S rDNA片段,构建其文库,建立阳性克隆子RFLP(Restriction Fragment Length Polymor-phism)酶切图谱.据酶切图谱选择部分克隆测序,并与数据库中的序列进行比对.结果表明,测序的27个序列分属于4个类群:变形细菌(Proteobacteria)、绿屈挠菌(Chloroflexi)、浮游霉菌(Planctomycetes)和酸杆菌(Acidobacteria);其中以变形细菌最多,占62.96%.
文摘【目的】海洋沉积物中的古菌在全球生物地球化学循环中充当重要的角色,深入了解沉积物中古菌群落的结构及功能特征是探究海洋沉积物中古菌参与生物地球化学循环和生态学功能的基础。【方法】采用高通量测序技术,分别对南海北部陆坡不同海域(东部,西部和神狐海域的7个站位)沉积物中古菌16SrRNA基因进行Illumina Mi Seq测序。【结果】中国南海北部陆坡沉积物中古菌的主要门类是Bathyarchaeota、Thermoplasmata、Woesearchaeota(DHVEG-6)、Thaumarchaeota(Marine Group I)、Lokiarchaeota和Marine Hydrothermal Vent Group(MHVG),还存在少量的AK8、Marine Benthic Group A和Terrestrial Hot Spring Crenarchaeota Group(THSCG)等。在潜在水合物区沉积物中还发现了甲烷代谢相关古菌(Anaerobic methanotrophic archaea,ANME)类群,主要为ANME-1、ANME-2ab和ANME-2c等。甲烷代谢古菌的分布特征也从甲烷代谢保守功能基因mcr A(Methyl coenzyme-Mreductase A)的扩增中得到了验证。利用定量PCR对南海沉积物中的细菌、古菌的16SrRNA基因和mcrA基因进行了定量,发现细菌16SrRNA基因拷贝数为10~5-10~7 copies/g(湿重),古菌16SrRNA基因拷贝数为10~5-10~6 copies/g(湿重),潜在水合物区mcrA基因拷贝数为10~3-10~5 copies/g(湿重)。【结论】揭示了中国南海北部陆坡沉积物中具有丰富的微生物资源,其中古菌种类多样且丰度较高,同时发现冷泉特征古菌群落,为深入认识和理解南海沉积物中微生物丰度和古菌多样性,以及解析古菌地球化学功能奠定基础。