东山湾软珊瑚共附生可培养细菌的多样性和产酶活性鉴定  

Phylogenetic diversity analysis and enzyme activity identification of cultured symbiotic bacteria of soft coral in Dongshan Bay

作　　者：刘阳[1,2] 赖其良[2] 陈华[2] 王建宁[2] 余志伟[2] 王丽萍[2] 邵宗泽[2] 

机构地区：[1]华中农业大学生命科学技术学院,湖北武汉430070 [2]国家海洋局第三海洋研究所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》2013年第3期396-403,共8页Journal of Applied Oceanography

基　　金：福建省自然科学基金资助项目(2009J01177);国家海洋局海洋公益性行业科研专项资助项目(201005032)

摘　　要：利用稀释10倍的2216E培养基,采用涂布平板法分离软珊瑚共附生可培养细菌,然后提取各菌株基因组DNA,扩增其16S rRNA基因并测序,通过比对分析确定菌株的分类地位,进而研究东山湾软珊瑚共附生可培养细菌的多样性;同时测定各菌株产淀粉酶、蛋白酶和酯酶能力,进而获得具有产酶活性的细菌资源.结果表明:一共分离到244株细菌,剔除重复后共获得128株不同的细菌.128株细菌包括变形菌纲(86株)、放线菌纲(高G+C革兰氏阳性菌,17株)、厚壁菌门(低G+C革兰氏阳性菌,22株)和拟杆菌纲(3株)4大类群,共包含44个属,72个种,多样性十分丰富,其中有9株细菌的16S rRNA基因相似度低于98.0%,为分离到的潜在新种.同时,128株细菌3种酶活检测的结果表明:16株细菌具有淀粉酶活性,73株细菌具有酯酶活性,23株细菌具有蛋白酶活性,其中15株细菌同时具有3种酶活性.本研究首次报道了东山湾软珊瑚共附生可培养细菌丰富的多样性,同时获得了具有良好的产酶特性菌株资源,具有进一步研究和应用价值.The objective of this study was to study the diversity of cultured symbiotic bacteria of soft coral in Dongs- han Bay, and to obtain more microbial resources. Cultured symbiotic bacteria of soft coral were isolated and puri- fied with 2216E media diluted 10 times and spread plate method, and then the genomic DNA was extracted, ampli- fied for 16S rRNA gene and sequenced, and determined their taxonomic status for diversity analysis. Meanwhile, the amylase, protease and esterase were identificated on special plates for strain resources with the enzyme activity. Totally, 244 strains were isolated and finally 128 strains unique in 16S rRNA gene sequence were obtained. They belonged to Proteobaeteria（86 strains）, Actinomycetes （high G ＋ C Gram-positive bacteria） （ 17 strains）, Firmi- cutes（low G ＋ C Gram-positive bacteria） （22 strains） and Bacteroidetes （3 strains）. They covered 72 species of 44 genera among which 9 strains had the similarities of 16S rRNA gene less than 98.0% with type strains, referring to a potential novel species. From these isolates, 16, 73 and 23 strains had extracellular activities of amylase, es- terase and protease, respectively. What is more, 15 strains of them had 3 enzyme activities. This study is the first report on the diversity of cultured symbiotic bacteria of soft coral in Dongshan Bay and Some strains with enzyme ac- tivities were obtained and can be used for further application.

关 键 词：海洋生物学 软珊瑚 共附生细菌 多样性 酶活鉴定 

分 类 号：P735[天文地球—海洋生物学]