Molecular Phylogenetic Study Of Chinese Common Wolf Spiders (Lycosidea) Inferred From 16S RRNA Sequences

It was the first time reported that analyzed a sequence dataset of a portion of mitochondrial 16S rRNA gene of the ribosomal large subunit for 26 species of the wolf spider (Lycosidae) and the funnel-web spider Agelena limbata (Agelenidae) from China applying DNA sequence in this paper. The wolf spider of 26 species are belonged to six genera (Hippasa, Pirata, Atctosa, Trochosa, Pardosa, Wadicosa) and four subfamilies(Hippasinae, Lycosinae, Pardosinae, Wadicosinae). Reference was making 16S rRNA sequence of two North American spider (Pardosa milvina, Pardosa takahashii) and the Agelena limbata, which were regarded as outside groups. Based on the obtained data, a consensus phylogenetic tree of the Lycosidae was constructed using NJ and MP and ML methods of Phylip software and UPGMA method of Mega software. The experimental results as follows:1. PCR-amplification and DNA-sequencing were used by oneself designing primers: (5' -AGCCGCTTTTGTGCTAAGGTA-3' ) and (5' -TAGAAGATAGAAACCGACCTG-3' ). We can get the band between 400 and 450bp after the electrophoresis.2.After sequencing for all individuals, the length of a portion of 16S rRNA gene from 340bp to 360bp. Its average base is made up: T: 36.1%; C: 12.2%; A: 39.0%; G: 12.8%; the analyzed sequences of 26 species had an average T/A higher than C/G. content. And base content of the sequence was similar with North America psrdosa' s. 3.After multiple sequence aligning, of these 133 variable and 90 were also phylogentically informative in parsimony analysis. It was proved that 16S rRNA sequence is a conservative sequence, and it was suitable for the study of systematic development in the species. 4.Datas were analyzed by applying the four methods in parallel, using Mega and Phylip. This was done to test for the inference of the methods used on the resulting phylogentic hypothesis. Since four methods are based on different approaches and algorithms. Congruent...