Some Poultry MtDNA Sequence Determination And Application In Different Sources Identification Of Poultry

Meat mixture and shoddy phenomenon has emerged on the market repeatedly, Simple and fast testing technique becomes necessary for it. Mitochondrial genome (mtDNA) has advantages in rapid evolution, rich polymorphism and maternally inheritance without gene recombinations, which has become an ideal molecular markers of population genetics, phylogenetics, molecular ecology and taxonomy. This paper Selected Galloanserae common poultry as the the research object and choosed mitochondrial gene as target gene. Mitochondrial genome sequences of Anser cygnoides (Anas poecilorhyncha, Chinese goose ancestry) and Anas platyrhynchos (Anas poecilorhyncha, one of the duck ancestors) were amplified, after analyzing we targeted the cytochrome oxidase I gene, then we designed primers to amplified six kinds of typical poultry mitochondrial DNA barcode region. Through sequence alignment et al methods we established a rapid identificate poultry molecular biology method. The main results were as follows:We amplified and obtained mitochondrial genome sequence of Spotbill(Anas poecilorhyncha), and upload it to Genbank (AccessionNo.KF751616). The results showed whole mitochondrial genome sequence of Anas poecilorhyncha was 16606 bp, including Twenty-two tRNA, two rRNA genes, thirteen protein-coding genes and D-loop. Base composition of T, C, A and G was 22.2%,32.8%,29.2% and 15.8%, respectively. Besides, the base preference of AT was not determined. Twenty-two kinds of tRNA were typically cloverleaf structures. Compared to 12S rRNA of brown head crow(Larus brunnicephalus) and black-tailed ground crow(Podoces hendersoni), we found the secondary structure of 12S rRNA contains four domains, thirty-seven stem-loop. The D-loop control region was analyzed and found goose hairpin, E-box, F-box, D-box, Bird similarity-box and CSB1-box. Finally, we taked the original chicken as an out group, the phylogenetic tree was constructed based on mitochondrial genome sequences and D-loop region(ducks were four kinds,three kinds of ducks, Gallus gallus) using NJ algorithm and ML algorithm. The results showed that three mallard ducks breed and Anas platyrhynchos had closer relationship.We amplified and obtained mitochondrial genome sequence mitochondrial genome sequence of Swan goose (Anser cygnoides), and upload it to Genbank (GenBank accession No. KJ124555). In this study. The results showed that whole mitochondrial genome sequence of Swan goose was 16 739 bp, including Twenty-two tRNA, two rRNA genes, thirteen protein-coding genes and D-loop. Base composition of T, C, A and G were 22.49%,32.24%, 30.21% and 15.06%, respectively. Besides, the base preference of AT was not determined. Twenty-two kinds of tRNA were all typically cloverleaf structures. Compared to 12S rRNA of Red Junglefowl (Gallus gallus) and Mongolian Ground Jay (Podoces hendersoni), we found the secondary structure of 12S rRNA included four domains, thirty-seven stemloops and thirteen salients in the Swan goose, and LSP/HSP, ETAS1-2, goose hairpin, E-box, F-box, D-box, Cbox, Bird similarity-box, CSB1-box, CSB-like and OH in the D-loop control region. Finally, we taken Red Junglefowl as an outgroup, the phylogenetic tree was constructed based on mitochondrial genome sequences using Neighbor-joining (NJ) algorithm, Maximum-likelihood (ML) algorithm and Bayesian model. The results showed that Swan goose, Graylag goose, Bean goose, white-fronted goose and Canada goose had closely genetic relationship. The findings enrich the ducks mitochondrial genome sequences and provide a theoretical basis for the study of geese phylogeny.By means of Galloanserae mitochondrial genome sequence analysis, the upstream of the gene COI barcode region was targeted and designed poultry universal primers(P1-5), with a bird COI common primers(P8) for comparison, which choose chicken, Turkey, duck, goose, quail and pigeon six kinds of poultry meat as experimental material. Experiment results showed six kinds of poultry in P4 general primer amplificate and digest by enzyme resulted in product in glue presented different band combination, and P8 universal primers amplification out different band combination, thus P4 and P8 both COI universal primers can accurate identify six different of poultry.