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Alternative TitleTaxonomic and phylogenetic study of fish trypanosomes
Thesis Advisor汪建国
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword鱼类锥虫 分离 分类学 系统发育分析
Abstract本论文在建立鱼类锥虫分离方法的基础上,研究了我国一些锥虫种类的分类及系统发育。主要结果如下: 1.用Percoll试剂和DEAE-纤维素层析柱两步法分离纯化鳝锥虫及用重组气溶素蛋白分离纯化鱼类血液中锥虫,其中后者为首次运用。 2.选用13个形态特征对鳢锥虫进行了重新描述,发现鳢锥虫有三种形态和两个寄主。沙塘鳢是鳢锥虫新寄主。 3.用形态学和分子标记重新鉴定了鳜锥虫,确立了其种的有效性。通过形态学和分子数据分析、比较了寄生黄颡鱼的两种形态的锥虫和鲤的一种形态的锥虫。发现鲤中寄生的锥虫与鳜锥虫可能是同一种类,并发现了寄生于黄颡鱼的一种新的形态的锥虫,与报道的一株锥虫的18S rDNA序列相似性为99.8%,二者可能是同一种。在黄颡鱼中发现的另一种形态的锥虫为黄颡锥虫,表明同一种鱼类可以感染两种以上的锥虫。 4.利用透射电镜观察了黄颡锥虫的超微结构,发现锥虫表膜是一种三层结构的单位膜,外覆盖着一层糖蛋白的外套。鞭毛、鞭毛与身体黏附区及外周微管与其他锥虫的相似。在电镜图片中,动核、胞核、线粒体及空泡等细胞内含物均被观察到。但超微结构的异同在锥虫的进化关系研究是否具有重要意义并不确定。 5.以18S rRNA基因为分子标记进行了鱼类锥虫的系统发育分析,系统树中引入了7个我国鱼类锥虫的18S rDNA序列。所有的鱼类锥虫聚成一枝,海水鱼类锥虫与淡水鱼类锥虫形成姊妹关系。我国鱼类锥虫内分两个群。
Other AbstractThe methods of isolation of fish trypanosomes and the taxonomy and phylogenetic analyses were studied and discussed in the present dissertation. The main results and conclusions are summarized as follows: 1. The methods of isolation of fish trypanosomes using Percoll reagent and DEAE-cellulose column, and the recombinant aerolysin protein were discussed in this paper. The latter was first reported. 2. The trypanosomes from the blood of Ophiocephalus argus and Odontobutis obscura were used for light microscopy investigations and studied morphology using 13 morphometric characterizations. Infection with trypanosomes of Odontobutis obscura was recorded for the first time. 3. Trypanosoma siniperca was a valid species and redescribed using 13 morphometric characterizations and molecular data. The trypanosomes from the blood of Pseudobagras fulvidraco and Cyprinus carpio were compared with related species using morphological and molecular data. The results indicated the trypanosomes from Cyprinus carpio and T. siniperca probably were the same species; one form from Pseudobagras fulvidraco and Marv might be the same genotypes, the other might be tentatively identified as T. pseudobagri. 4. The fine structure of T. pseudobagri was observed with transmission electron microscopy. The pellicle of trypanosome was a unit membrane with characteristic 3-layered structure, which was covered by a well-developed glycoprotein surface coat. The structure and attachment of the flagellum and the subpellicular microtubules show the usual structural and organizational features. Kinetoplast, mitochondria, nucleus and vacuoles, which occur in trypanosomidae, were observed in electron micrographs. The significance of similarities and differences of these fine structures is uncertain in terms of the evolutionary relationship of trypanosomes. 5. Phylogenetic analysis was conducted by using 18S rRNA gene sequences, which contained 7 sequences of fish trypanosomes from China. The results indicated that all fish trypanosomes fell in a single clade, which freshwater fish trypanosomes was separate from trypanosomes of marine fish and formed sister group. Trypanosomes from China were nested within the fish clade and further subdivided into at least 2 subgroups.
Document Type学位论文
Recommended Citation
GB/T 7714
顾泽茂. 鱼类锥虫分类与系统发育研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2007.
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