水生生物学报2011，Vol.35Issue(2)：203-209,7.DOI:10.3724/SP.J.1035.2011.00203
鲩肠袋虫18S rDNA的扩增、测序与分析
PCR AMPLIFICATION, SEQUENCING AND ANALYSIS OF 18S rDNA OF BALANTIDIUM CTENOPHARYNGODONI INHABITING GRASS CARP
摘要
Abstract
Balantidium ctenopharyngodoni, an obligatory intestinal symbiont and possibly opportunistic parasite of grass carp (Ctenopharyngodon idella), was first discovered and named by Chen (1955). It mainly inhabits the cream-like luminal contents and spaces between mucosal folds of the hindgut. In the present study, twenty two or three-summer grass carp were bought from Wuhan Fruit-Lake Market in March 2009 and transported alive to the laboratory for further examination. All fish samples were dissected and the intestines were cut to collect the luminal contents into Petri dishes for examination. Then 0.65％ saline solution was added to the contents and waited for a few minutes to allow B. ctenopharyngodoni swim free of luminal contents. The ciliates were collected with Pasteur micropipette and washed twice in distilled water. The parasite gDNA was isolated by proteinase K digestion overnight at 37℃, followed by phenol-chloroform protein extraction and ethanol precipitation. Its 18S rDNA was then sequenced, which contains 1638 base pairs. Phylogenetic trees were also constructed by the neighbour joining method, the maximum parsimony method and the Bayesian method, respectively. The results indicated that the subclass Trichostomatia was monophyletic and composed of the order Entodiniomorphida, Vestibuliferida and Australian clade (100％ Bay; 100％ MP; 100％ NJ). The oder Entodiniomorphida (100％ Bay; 98％ MP; 93％ NJ) and Australian clade (100％ Bay; 97％ MP; 99％ NJ) showed their monophyletic while the order Vestibuliferida showed its paraphyly. As to the phylogenetic position, B. ctenopharyngodoni first clustered with the Australian clade with higher bootstrap values (100％ Bay; 100％ MP; 100％ NJ).Then they formed a sister clade to “Entodiniomorphida+ Vestibuliferida” (100％ Bay; 85％ MP; 72％ NJ). B. coli, as an isolated taxon, was joined together with “Australian clade + B. ctenopharyngodoni” and “Entodiniomorphida + Vestibuliferida (part)” to form the subclass Trichostomatia (100％ Bay; 100％ MP; 100％ NJ). It also suggested that the genus Balantidium should be recognized as a higher taxon.关键词
鲩肠袋虫/18S rDNA/系统发育分类
生物科学引用本文复制引用
李明,王崇,汪建国,喻达辉,王文博,戈贤平,徐跑,谢骏,龚小宁..鲩肠袋虫18S rDNA的扩增、测序与分析[J].水生生物学报,2011,35(2):203-209,7.基金项目
国家自然科学基金(30870275)、(31000947) （30870275）
农业部海水养殖生态与质量控制重点实验室开放课题(2008C1201) （2008C1201）
淡水生态与生物技术国家重点实验室开放课题(2009FB08) （2009FB08）
农业部淡水鱼类遗传育种和养殖生物学重点实验室开放课题(BZ2009-02) （BZ2009-02）
淮安市科技支撑计划(SN1087),湖北省教育厅优秀中青年人才项目(Q20101702)资助 （SN1087）
