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

作　　者：高凤 伊珍珍 龚骏 Al-RASHEID Khaled A. S. 宋微波 

机构地区：[1]Laboratory of Protozoology, Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, College of Life Science, South China Normal University [2]Laboratory of Protozoology, Institute of Evolution and Marine Biodiversity, Ocean University of China [3]Zoology Department, King Saud University

出　　处：《Chinese Journal of Oceanology and Limnology》2010年第3期542-548,共7页中国海洋湖沼学报（英文版）

基　　金：Supported by the Natural Science Foundation of China (Nos. 30870264 and 40976099);the Center of Excellence in Biodiversity, King Saud University

摘　　要：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.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. pop1, Anteholosticha sp. pop2, A. manca, A. gracilis and Nothoholosticha fasciola, 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 (Hinf I, Hind III, Msp I, Taq I) yielded species-specific restriction patterns, and Hind III and Taq I produced different patterns 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.

关 键 词：CILIOPHORA phylogeny ARDRA SSrRNA gene ITS2 secondary structure 

分 类 号：Q951[生物学—动物学]