Research Of The Complete Mitochondrial Genome Of Reed Parrotbill (Paradoxornis Heudei), Along With Isolation And Characterization Of Microsatellite Makers In The Paradoxornis

Mitochondrial genomes of all vertebrate animals analyzed to date have the same 37 genes,containing 13 protein-coding,2 ribosomal RNAs(rRNA),22 transfer RNAs (tRNAs)genes and the putative control region,whose arrangement in the circular DNA molecule varies only in the relative position of some genes.For the most avian orders, the mitochondrial organization has the following arrangement between Cytb and 12S rRNA genes:tRNA^Thr/tRNA^Pro/NADH6/tRNA^Glu/CR/tRNA^Phe.Another gene order between the Cyt b and 12S rRNA genes,tRNA^Thr/ CR/tRNA^Pro/NADH6/tRNA^Glu/nc/tRNA^Phe,was reported in orders Piciformes, Falconiformes,suboscine passeriforms and some oscines(Bensch and Harlid,2000; Singh et al.,2007).Using long-PCR amplification and primer-walking sequencing methods,the complete sequence of Paradoxornis heudei and gene segment between the Cyt b and 12S rRNA genes of Paradoxornis webbianus were obtained.The complete sequence of Paradoxornis heudei is 16928bp.The gene components,choice of coded chain,and low percentage of guanine in heavy chain and the content of A+T are more similar with other birds.The gene order of Paradoxornis heudei between the Cyt b and 12S rRNA genes is tRNA^Thr/CR/tRNA^Pro/NADH6/tRNA^Glu/nc/tRNA^Phe,Paradoxornis webbianus also has the same gene order.Obviously,the result is not identical to the conclusion of Mindell et al(1998)that the alternative mitochondrial gene orders in songbird only exists suboscines.The genus Phylloscopus is not the only oscine passeriformes group to have the derived arrangement.We suggest that the derived mitochondrial arrangement of Paradoxornis heudei and Paradoxornis webbianus is the result of a tandem duplication of the region tRNA^Pro/NADH6/tRNA^Glu/CR followed by the deletion of one of each of the two copies except for part of the duplicated CR:the NC region in the mitochondrial genome is the production of a tandem deletion of the CR copy.The derived mitochondrial arrangement of Paradoxornis heudei and Paradoxornis webbianus succeeded to a common ancestry;and the rearrangement of mitochondrial gene order in Paradoxornis origins independently.In order to evaluate the genetic variation and genetic structure of the isolated reed parrotbill populations,11 microsatellite loci were isolated using the fast isolation by AFLP of sequences containing repeats(FIASCO)method.Five to 11 alleles were identified in an analysis of 32 Paradoxornis heudei.with the expected and observed heterozygosity ranging from 0.6001-0.8647,0.037 to 0.80.Loci Parh05,Parh10, Parh12,Parh13,Parh21,Parh33 are significant deviations from the Hardy-Weinberg equilibrium and loci Parh05 and Parh13 showed linkage disequilibrium.We also tested the utility of these loci on 2 species of Timaliinae and Sylviidae.Although loci of Parh13,Parh25 failed to amplify in Paradoxornis webbianus,and Parh05,Parh33 failed to amplify in Acrocephalus orientalis,the other loci were successful at both of the species.