Molecular Phylogeny Of Gekko Geckoy

As the second class national protected animals, Gekko gecko is classified as the animal of Gekkonidae.Since Gekko gecko named by Linnaeus in 1758, they have been considered as one species in animal classification . Since eighties ages of twenty century, population genetic differentiation between Gekko gecko has been disputed .Whether the differentiation can reach species or sub-species level or not need to be confirmed.In the study, normal PCR, Long-PCR amplification and primer walking sequencing methods were carried out for determing mitochondrial genome complete sequence of red-spot Gekko gecko.The mitochondrial genome of red-spot Gekko gecko is a circular molecule of 16,590 bp in length, containing 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and non-coding regions. In this mitochondrial genome, genes array compactly, no gene rearrangement happened, lighter strand replication orgin(OL) can form stem-loop structure and there are many tandem repeat sequences in the control region. The mito- chondrial genome of red-spot Gekko gecko is conservative relatively. Its component, gene order, low percentage of guanine and skews of base AT are idential with other vertebrates, except there are lots of (TA)n microsatellite repeats at the last 173bp of control region.In the study, we combined the red-spot Gekko gecko's mitochondrial DNA sequence data with those already published to assess phylogenetic relationship based on 12 protein- coding genes (except ND6 gene). PAUP4.0 was used for constructing NJ tree, MP tree and ML tree. The results are as follows: Lizards were divided into two branches, Gekkonidae animals as one branch, others as another branch. Gekkonids are monophyletic and Coleon yx variegatus might have a father relationship with the other four gekkonids. Amphisbaenia shoud not be separated from Lacertids, this view is different from the traditional morphologial analysis. Scincomorphas might be sister group to Iguanias. NJ tree and MLtree support these conclusions above by high bootstrap value. But still need more research to verify these conclusions because of complexity of the problem.This study selected different geographyical populations of Gekko gecko to discuss the genetic diversity based on Cytb gene and D-loop sequences. We constructed the phylogenetic trees with NJ tree, MP tree and UPGMA tree based on both Cytb gene and D-loop sequences by Mega 4.1 . The results are as follows: judging from the genetic distance , the genetic distance of four haploid types of the black-spot Gekko gecko was 0.09% - 0.36% and of four haploid types of the red-spot Gekko gecko was 1.01%-3.95%. But the genetic distance between the black-spot Gekko gecko and the red-spot Gekko gecko was up to 8.67% - 9.82%. NJ tree, MP tree and UPGMA tree showed that the black-spot Gekko gecko and red-spot Gekko gecko were divided into two branchs. The study support the view that genetic differentiation between Gekko gecko has reached sub-species level.