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池塘养殖仿刺参(Apostichopus japonicus)肠含物、附着基和底泥中的菌群结构及其相关性
李 彬1, 廖梅杰1, 荣小军1, 张 正1, 王印庚1,2, 陈贵平1, 王 岚1, 范瑞用3
1.农业部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.青岛海洋科学与技术国家实验室 海洋渔业科学与食物产出过程功能实验室 青岛 266071;3.青岛瑞滋海珍品发展有限公司 青岛 266409
摘要:
2014年10月,利用PCR-DGGE技术分析了标准化养殖池塘中仿刺参(Apostichopus japonicus)肠含物及附着基和底泥中细菌的菌群结构及其相关性。结果显示, 仿刺参肠含物、附着基、底泥中细菌多样性较高,分别平均获得30.00±1.00、15.33±1.70、21.67±2.62条带,肠含物中细菌种类数显著高于附着基和底泥。聚类分析和戴斯系数表明,不同池塘的仿刺参肠含物样本单独聚为一支,相似性达到0.785,附着基和底泥样品聚为一支,平均相似性达到0.532。不同样品DGGE图谱中20条优势条带的切割、克隆、测序,共获得了20条细菌序列,表明肠含物细菌种类以聚球藻属(Synechococcus sp.)、脱硫杆菌属(Desulfobacterium)、脱硫叠球菌属(Desulfosarcina sp.)、极地杆菌属(Polaribacter)、Algibacter sp. 为主,附着基中主要以Robiginitalea sp. 和Silicibacter sp. 为主,底泥样品中主要以假单胞菌属(Proteobacterium)、噬胞菌属(Cytophaga)、Desulfosarcina sp.为主,肠含物中细菌特有种类的分离与鉴定为仿刺参养殖潜在益生菌的开发提供基础数据。
关键词:  PCR-DGGE  仿刺参  肠含物  环境  细菌
DOI:10.11758/yykxjz.20150727002
分类号:
基金项目:中央级科研院所基本科研业务费专项资金项目(20603022015015)、山东省自主创新成果转化专项(2013ZHZX2A0801)、青岛市海洋经济创新发展区域示范项目(海水健康养殖专用微生态制剂的开发应用与产业化项目)和海洋经济创新发展区域示范重点项目(山东省海水养殖重要疾病高效专用药物研发及产业化)共同资助
Correlation Analysis of Flora in the Habitat Environment and in the Gut Contents of Sea Cucumber (Apostichopus japonicus)
LI Bin1, LIAO Meijie1, RONG Xiaojun1, ZHANG Zheng1, WANG Yingeng1,2, CHEN Guiping1, WANG Lan1, FAN Ruiyong3
1.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;2.Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071;3.Qingdao Ruizi Precious Seafood Development Limited Company, Qingdao 266409
Abstract:
Here we studied the properties of flora in the gut content and attachments of sea cucumber (Apostichopus japonicus) as well as in the sediments of pond where sea cucumber were cultured. 16S rDNA PCR denaturing gradient gel electrophoresis (PCR-DGGE) was employed as the major method. The DGGE band numbers were 30.00±1.00, 15.33±1.70, and 21.67±2.62 for gut content, attachments, and pond sediments respectively. This indicated that the gut content had the highest diversity of bacterial species. UPGMA cluster analysis showed that flora from three gut content samples could be categorized into one branch with a similarity value of 0.785, and that flora from attachments and pond sediment samples were clustered into another branch with a similarity value of 0.532. Twenty specific DGGE bands from the three kinds of samples were cut and sequenced. The results showed that Synechococcus sp., Desulfobacterium, Desulfosarcina sp., Polaribacter, Algibacter sp. were the dominant bacterial species in the gut content; Robiginitalea sp., Silicibacter sp. were dominant in attachments; and Proteobacterium, Cytophaga, Desulfosarcina sp. were dominant in the sediment. The specific bacteria in the gut content could be candidates of probiotics in the sea cucumber culture.
Key words:  PCR-DGGE  Apostichopus japonicus  Gut content  Environment  Bacteria