ABSTRACT
Objective:To identify 24 <italic>Rana</italic> species such as <italic>Rana dybowskii</italic> by mitochondrial cytochrome C oxidase subunit I （<italic>CO</italic>Ⅰ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of <italic>Rana</italic> species as well as the discovery of new species. Method:<italic>R. dybowskii</italic>， <italic>R. chensinensis</italic>， <italic>R. amurensis</italic>， <italic>R. culaiensi</italic>s， and <italic>R. huanrenesis</italic>， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification<italic> </italic>and sequencing. A total of 50 <italic>CO</italic>Ⅰ gene sequences were obtained. Then 163 <italic>CO</italic>Ⅰ gene sequences for 24 species of <italic>Rana</italic> and one <italic>CO</italic>Ⅰ gene sequence for <italic>Pelophylax</italic>，<italic> Odorrana</italic>， <italic>Nidirana</italic>， <italic>Hylarana</italic>， and <italic>Amolops</italic> were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of <italic>CO</italic>Ⅰ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis. Result:The <italic>CO</italic>Ⅰ gene sequences of 24<italic> Rana</italic> species including <italic>R. dybowskii</italic> were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> were clustered into one branch and some <italic>R. dybowskii</italic> and <italic>R. uenoi</italic> into one branch. There were two separate branches for <italic>R. chensinensis</italic> and the other species were all clustered independently. Conclusion:<italic>CO</italic>Ⅰ-based DNA barcoding enabled the identification of 24 species of <italic>Rana</italic> including <italic>R.dybowskii</italic>. The findings supported that <italic>R. sangzhiensis</italic>， <italic>R. zhengi</italic>， <italic>R. coreana</italic>， and <italic>R. kunyuensis</italic> were the same species. One branch of <italic>R. chensinensis </italic>might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that <italic>CO</italic>Ⅰ-based DNA barcoding allows not only the identification of 24 species of Rana including <italic>R. dybowskii </italic>but also the classification of ranidae species and the discovery of new species or subspecies.
ABSTRACT
Objective To apply the DNA barcoding technology for identification of rodent animals and to establish a rodent animal DNA barcode database in Gansu Province.Methods A total of 54 rodent animals were detected.DNA barcoding technology was used to analyze the DNA mitochondrial cytochrome C oxidase subunit Ⅰ (CO Ⅰ) gene sequence in Gansu Province.Results The intra-specific genetic distance was 0-2％ while the interspecific distance ranged from 18％ to 30％.Eight major clusters were apparently showed on a Neighbor joining tree.Conclusion DNA barcoding technology could overcome the shortcomings of the morphological identification,so it could be used to identify the rodent animals and has important implications for disease control and prevention in the natural focus of Gansu Province.
ABSTRACT
The mosquito Aedes albopictus is the primary vector for dengue virus transmission in Fujian Province.Despite that active dengue surveillance has been launched in several sites since 2005,the genetic diversity of A.albopictus from these sites remains exclusive.In this study,mosquito pools collected from dengue surveillance sites during 2015-2016 were randomly selected,the full-length mitochondrial cytochrome C oxidase subunit Ⅰ (mtDNA-COⅠ) were amplified and sequenced.Preliminary sequence alignment of 12 amplicons with the reference sequence indicated 99 ％ homology at nucleotide level,due to varia tions at 9 nucleotide sites.Three haplotypes,namely H02,H03 and H08,were determined based on phylogenetic analysis with 72 reference sequences of known haplotypes.These observations facilitate surveillance and control of arboviral diseases in Fujian.
ABSTRACT
Objective To apply the DNA barcoding technology for identification on host animal and to establish the host animal DNA bar code database on natural foci of plague in Shaanxi.Methods 139 host animals belonging to 3 orders,6 families and 12 genera and 62 residues belonging to 7 species from 8 different parts of the province,were detected.DNA barcoding technology was used to analyze the DNA CO Ⅰ gene sequence on the natural foci of plague in Dingbian county.Results The intra-specific genetic distance was less than 2％ while the inter-specific distance ranged from 8.9％ to 15.1％.Fourteen major clusters were apparently showed on a Neighbor-Joining tree.Residue samples could be detected regarding the objective gene.Alashan ground squirrel was previously noticed to carry 14 major clusters,which were previously mistakenly named as Citellus dauricus in Dingbian county.Conclusion DNA barcoding technology could overcome the shortcomings caused by the morphological identification so could be used to identify the host animal and residues in the natural focus of plague.
