DNA Barcodes Information And Phylogenetic Analysis Of Partial South China Fisheries Animals

There are a lot river systems in South China,with abundant fishery animal resources and the important germplasm resource library in our country.Due to the defects of the morphological identification and combined with the traditional taxonomists is decreasing,The development of fishery biology Identification and classification become more and more difficult.So it is badly in need of a fast,cheap and specification of species identification method to solve.DNA barcode is the DNA sequences which can be used to represent the species.And the DNA barcode technology can make up for the traditional morphological identification of defects.In this research,1653 fishery animal species in South China was collected and prepared as specimens,research for DNA barcode number 837(COI gene).The Marbled Goby(Oxyeleotris marmoratus)and Sleepy Cod(Oxyeletris lineolatus)are difficult to identify in seedling stage,while they were collected in our research.And their prices have huge differences,so some mistakes often exist in seedlings market.The complete mitochondrial genome of Oxyeletris lineolatus was determined and compared with the complete mitochondrial genome of Oxyeleotris marmoratus.We found some sites existing differences in COI,so some species-specific primers was designed on the different sites.This method can be used to identify two species easily and quickly.Based on the DNA barcode(COI gene),the genetic distance and phylogenetic analysis of th Cyprinidae,the Eleotridae,the Siluriformes,the Decapoda and the Chelonia were analysed.The analysis results enrich the achievements of traditional taxonomy,also providing some supporting data for the problems of species affiliation.The research contents of this thesis areas follows:1.Determination of the Oxyeletris lineolatus mitochondrial genomeOxyeletris lineolatus is native to Queensland in northeastern of Australia,and was introduced to the pearl river region in south of China in 1996.Oxyeletris lineolatus is people's love because of its delicious taste.Based on the complete mitochondrial genome sequence of Oxyeleotris marmoratus in the Genbank,PCR primers were designed using Primer 5.0 and Oligo6.0 softwares to cover the complete mitochondrial genome sequence.The PCR products were further isolated and sequenced,then they were aligned and analyzed.The complete mitochondrial genome sequence of Oxyeletris lineolatus was firstly determined.As other bony fishes,the mitochondrial DNA of Oxyeletris lineolatus is 16519 bp circular DNA,37 genes(22 tRNA genes,2 rRNA genes,13 protein-coding genes)with three termination codon of protein-coding genes(T,TA,TAA).The mitochondrial DNA of Oxyeletris lineolatus can provide the data to identify species and research the genetic evolution.2.The molecular mark for species identification of the Oxyeleotris marmoratus and Oxyeletris lineolatusVia the alignment and analysis of COI sequences of the Oxyeleotris marmoratus and Oxyeletris lineolatus,many primers were designed basing on the differences sites and one specific primers(XW1)were selected.Amplified with XW1,there was the 170 bp band in the Oxyeletris lineolatus,while nothing in the Oxyeleotris marmoratus.Then 20 individuals per fish were randomized to verify the primers,the results showed that the specific primers have specificity for species.The specific primers can distinguish two species simply and rapidly.This method can provide a molecular marker identification method for identifying.3.The aquatic animal DNA barcode and genetic evolution analysisThis study collected aquatic animal 1653 samples,involving 81 kinds,837 total DNA barcode number(COI gene)in south China area.Through tissue samples of DNA extraction and universal primer PCR amplification,COI gene sequences were aligned and confirmed.Based on the COI genes of the Cyprinidae,the Eleotridae,the Siluriformes,the Decapoda and the Chelonia,genetic distance and system evolution were analysed.The Mauremys genus is still controversial,so the COI gene analysis and research can provide a reference for the chelonia classification.The analysis results show that the interspecific genetic distance of Megalobrama hoffinanni and Parabramis pekinensis,Erythroculter pseudobrevicauda and Culter alburnus,were relatively close and rare differences.The Bostrychus sinensis,Oxyeleotris marmoratus and Oxyeletris lineolatus gathered for a sister branch,Eleotris fusca and Eleotris oxycephala gathered for a sister branch.The endemic species of Pearl River cranoglanis bouderius and Pelteobagrus fulvidraco?Siniperca scherzeri?Silurus soldatovi meridionalis Chen gathered for a sister branch,and it has a close relationship with yellow Sang fish.Euryhaline Exopalaemon carinicauda and Macrobrachium nipponense?Macrobrachium rosenbergii gathered for a sister branch.Genetic distance between Mauremys reevesii and other species of Mauremys are 0.03 to 0.08,and the phylogeny gathered for a sister branch.Our results suggested that tortoise should divide into Mauremys.