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作 者:沈辉[1,2] 万夕和[1] 何培民[2] 黎慧[1] 乔毅[2] 蒋葛
机构地区:[1]江苏省海洋水产研究所,江苏南通226007 [2]上海海洋大学水产与生命学院,上海201306
出 处:《微生物学通报》2015年第10期1922-1928,共7页Microbiology China
基 金:国家科技支撑计划课题(No.2012BAC07B03);江苏省农业科技自主创新资金项目(No.CX(13)2040);江苏省水产三项工程项目(No.PJ2014-51);南通市科技项目(No.HL2014014)
摘 要:【目的】研究海水池塘养殖条件下的脊尾白虾肠道微生物菌群组成及多样性。【方法】采用PCR-RFLP技术,以直接提取的脊尾白虾肠道细菌总DNA为模板进行16S rRNA基因扩增,产物与T载体连接建立质粒文库,从RFLP建立的文库中筛选出不同细菌来源的克隆子,将测定的差异克隆子16S rRNA片段序列与Gen Bank数据库进行比对。【结果】从脊尾白虾肠道的菌群文库中共获得114个克隆子,HaeⅢ和Msp I双酶切得到11种差异克隆子;对差异克隆子测序后,其中8个差异序列与已知细菌具有较高的同源性,分别是假单胞菌属(Pseudomonas)、肠杆菌属(Enterobater)、冰冻小杆菌属(Frigoribacterium)、褐杆菌属(Phaeobacter)、弧菌属(Vibrio)、赤杆菌属(Erythrobacter)、气单胞菌属(Aeromonas)、葡萄球菌属(Staphylococcus)和其他未培养细菌。【结论】初步揭示了脊尾白虾肠道微生物菌群结构的组成,为开发脊尾白虾专用微生态制剂提供基础资料。[Objective] To study bacterial communities and diversity in shrimp (Exopalaemon carinicauda Holehuis) intestine. [Methods] 16S rRNA gene was amplified and a library was constructed by genomic DNA that was extracted from the bacteria in shrimp intestine. [Results] Different RFLP patterns of the clones were obtained from analysis with Hae Ⅲ and Msp I. By comparing the obtained sequence with the published sequence in the GenBank database, 114 clones were obtained in the 16S rRNA clone library, and 11 different RFLP patterns of the clone were affirmed by Hae Ⅲ and Msp I. There were 8 clone sequences in high identity with known bacteria sequence, respectively belonging to Pseudomonas (17.5%), Enterobater (21.1%), Frigoribacterium (8.8%), Phaeobacter (5.3%), Fibrio (10.5%), Erythrobacter (9.6%), Aeromona (4.4%), and Staphylococcus (2.6%). In addition, uncultured bacteria accounted for 19.3%. [Conclusion] This study discovered the bacterial communities in the intestine of Exopalaemon carinicauda Holehuis, and the results would enhance our understanding about the bacterial community composition in Exopalaemon carinicauda Holehuis intestine and provide valuable data for the development of disease prevention mechanisms for shrimp cultivation.
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