西太平洋菲律宾海海山区微生物原位富集及其物种多样性分析  

Deep-sea in situ cultivation and diversity analysis of bacteria in a seamount of Philippine Sea in western Pacific Ocean

作　　者：朱栩莹 夏静宇 李丁 曾令宇 董纯明 王丽萍 邵宗泽 ZHU Xuying;XIA Jingyu;LI Ding;ZENG Lingyu;DONG Chunming;WANG Liping;SHAO Zongze(Key Laboratory of Marine Genetic Resources,Third Institute of Oceanography,MNR,Xiamen 361005,China)

机构地区：[1]自然资源部第三海洋研究所、自然资源部海洋生物遗传资源重点实验室,福建厦门361005

出　　处：《应用海洋学学报》2024年第3期405-418,共14页Journal of Applied Oceanography

基　　金：国家自然科学基金(42030412);中国大洋矿产资源研究开发协会项目(DY-XZ-04,DY135B01)。

摘　　要：为了研究海山区微生物对外源有机物输入的响应情况,更好认识海山区微生物多样性以及它们在深海碳循环中的作用,本研究以硫粉烷烃混合物(CS)、活性污泥(HX)、几丁质(JDZ)、D型天冬氨酸(D-Asp)、二甲基砜(DMSO2)、聚-β-羟丁酸(PHB)等6种不同有机物为底物,通过深海水体原位定植培养系统,分别在西太平洋菲律宾海的海山山顶(2929 m)和山麓(4707 m)进行了为期10个月的微生物原位富集培养。结合高通量测序技术和分离培养方法,对深海原位富集样品进行了微生物多样性分析以及可培养微生物分离鉴定。16S rRNA基因高通量测序结果表明,6个样品共富集得到属于细菌域的15门27纲61目96科150属200种。不同底物富集样品的细菌物种组成差异较大,表明不同底物选择性富集了特定类群微生物。其中,硫粉烷烃混合物富集样品优势属包括Roseobacter_clade_NAC11-7_lineage属、摩替亚氏菌属(Moritella)等;活性污泥富集样品优势属为假交替单胞菌属(Pseudoalteromonas)、Fusibacter属等;几丁质富集样品优势属为弧菌属(Vibrio);二甲基砜富集样品优势属为假交替单胞菌属、鲁杰氏菌属(Ruegeria)等;D型天冬氨酸富集样品的优势属为假交替单胞菌属;聚-β-羟丁酸富集样品优势属为鲁杰氏菌属、Fusibacter属等。对6个原位富集样品进行平板分离培养,共分离鉴定得到277株可培养细菌,分属于5门8纲19目35科63属92种,其中潜在新种62株和潜在新科5株,表明菲律宾海海山环境蕴含了丰富多样的海洋微生物。它们在深海物质循环中的代谢机制和生态贡献还有待进一步研究。In order to research the response of the seamount bacteria to exogenous organic matters,and better understand the diversity of seamount bacteria and their roles in carbon cycles in the deep sea,a ten-month deep-sea in situ enrichment was developed at the top(2929 m)and bottom(4707 m)of the seamount of Philippine Sea in western Pacific Ocean.Different substrates including alkane-sulfur mixture(CS),activated sludge(HX),chitin(JDZ),D-Aspartic acid(D-Asp),Dimethyl sulfone(DMSO2)and poly-β-hydroxybutyric acid(PHB)were added into a circulation chamber of deep-sea water incubation system.The bacterial communities of enriched samples were analyzed with high-throughput sequencing technology and the cultivatable bacteria were isolated and identified with the spread plate method.Results of high-throughput sequencing of 16S rRNA genes showed that bacterial communities,including 200 species of 150 genera in 96 families and 61 orders in 27 classes of 15 phyla,were enriched in 6 samples.The bacterial composition of enriched samples with different substrates differed from each other significantly,indicating that different substrates selectively led to specific enriched bacterial communities.Results showed that Roseobacter_Clade_NAC11-7_Lineage and Moritella were dominated in alkane-sulfur mixture enrichments,Pseudoalteromonas and Fusibacter in activated sludge enrichments,Vibrio in chitin enrichments,Pseudoalteromonas in D-Asp enrichments,Pseudoalteromonas and Ruegeria in DMSO2 enrichments,Ruegeria and Fusibacter in PHB enrichments.Besides,a total of 277 bacterial strains were isolated and from these six different deep-sea in situ enriched samples and were identified to be 92 species of 63 genera in 35 families and 19 orders in 8 classes of 5 phyla.Among them 62 potential novel species and 5 potential novel families were obtained,indicating the diversity and novelty of marine microorganisms in the seamount of Philippine Sea.The potential ecological roles and metabolic mechanisms in element cycles of these cultural bacteria in d

关 键 词：海洋生物学 深海原位富集 微生物多样性 海山 菲律宾海 

分 类 号：P735[天文地球—海洋生物学]