Molecular phylogeny and species separation of five morphologically similar Holosticha-complex ciliates （Protozoa, Ciliophora） using ARDRA riboprinting and multigene sequence data

• Published in: Chinese journal of oceanology and limnology Vol. 28; no. 3; pp. 542 - 548
• Main Author: 高凤 伊珍珍 龚骏 Al-RASHEID Khaled A. S. 宋微波
• Format: Journal Article
• Language: English
• Published: 2010
• Subjects: 分子系统发育; 原生动物; 基因序列; 数据分离; 核糖体DNA; 纤毛虫; 纤毛门
• ISSN: 0254-4059; 1993-5005

To separate and redefine the ambiguous Holosticha-complex, a confusing group of hypotrichous ciliates, six strains belonging to five morphospecies of three genera, Holosticha heterofoissneri, Anteholosticha sp. popl, Anteholosticha sp. pop2, A. manca, A. gracilis and Nothoholostichafasciola, were analyzed using 12 restriction enzymes on the basis of amplified ribosomal DNA restriction analysis. Nine of the 12 enzymes could digest the DNA products, four （HinfⅠ, Hind Ⅲ, Msp Ⅰ, Taq Ⅰ） yielded species-specific restriction patterns, and Hind Ⅲ and Taq Ⅰ produced different pattems for two Anteholosticha sp. populations. Distinctly different restriction digestion haplotypes and similarity indices can be used to separate the species. The secondary structures of the five species were predicted based on the ITS2 transcripts and there were several minor differences among species, while two Anteholosticha sp. populations were identical. In addition, phylogenies based on the SSrRNA gene sequences were reconstructed using multiple algorithms, which grouped them generally into four clades, and exhibited that the genus Anteholosticha should be a convergent assemblage. The fact that Holosticha species clustered with the oligotrichs and choreotrichs, though with very low support values, indicated that the topology may be very divergent and unreliable when the number of sequence data used in the analyses is too low.