Molecular Phylogenetics Analysis Of Haemaphysalis Longicornis And H.conicinna Based On COI, COII And ITS-2 Genes

Comparing with high speed development of molecular biology,molecular systematization has been used in all areas of species classified as a branch,included in parasitic species taxonomy.Ticks attached to the Arthropoda,Arachnida,Acari, Parasitiformes,Ixodoidea,is the terrestrial vertebrates,non-permanent parasites in vitro. They invasion of mammals,birds,reptiles,and amphibians,widely distributed around the world.Ticks harm for animals,not only as a parasite andmore importantly is as to spreading many human and animal diseases(including Fungi,Viruses,Rickettsiae's body, Proflagellata Bacteria and Protozoa) and seriously harm the animal and human health.In this paper,mtDNA gene included cytochrome coxidase subunitâ… (COâ… ) and cytochrome coxidase subunitâ…¡(COâ…¡),and ribosomal gene internal transcribed spacer 2 (ITS2) as targets were used to analyse phylogentic relation about Haemaphysalis Iongicornis and H.conicinna.Sequence alignments were used to analyse the difference of sequences by DNAStar and Clustal X program.Different evolutional approaches include Unweighted pair group with mathematical average(UPGMA),neighbour-joiner(N-J) and maximum parsimony(MP),from distance matrix and character state,were used to construct the phylogenetic tree by MEGA 3.1 program.The result of sequences alignment and phylogenetic tree were used to evaluate the evolutional relation between Haemaphysalis spp and other species included in the family Ixodidae.The experimental materials are Haemaphysalis longicornis and H.conicinna, combined with Genbank signed the relevant sequences.This text was designed ITS-2, COâ… ,COâ…¡three pairs of primers,through the PCR method successfully amplified the Haemaphysalis longicornis and H.conicinna ITS2 gene,COâ… gene and COâ…¡gene.The purpose of gene fragments,and through cloning and sequencing of the access to their respective nucleotide sequence.The three groups of nucleotide sequence to compare the results show that:Haemaphysalis longicornis and H.conicinna of ITS-2 gene nucleotide identity of 88.3%,Haemaphysalis longicornis and H.conicinna of COâ… gene nucleotide identity was 87.5%,Haemaphysalis longicornis and H.conicinna of the nucleotide COâ…¡gene nucleotide identity was 90.4%.Based on ITS2 gene sequence,Sarcoptes as outgroup.Based on the COâ… and COâ…¡gene sequence,Varroa destructor as outgroup.In the text three pairs genes were building three tree topology are similar,which makes the system more reliable.Based on ITS2 gene,Haemaphysalis longicornis and H.conicinna in each tree are gathered together, bootstrap values(100,99,100).Based on the COâ… gene,In N-J and the MP method Haemaphysalis longicornis and H.conicinna are gathered together,bootstrap values (99,96);UPGMA was taken from the change,Haemaphysalis longicornis and Rhipicephalus sanguineus gathered together,bootstrap values(69,78).Based on COâ…¡genes, Haemaphysalis longicornis,H.flava and H.conicinna are gathered together,the bootstrap values(90,89,60).The results show that Haemaphysalis longicornis and H.conicinna has a high homology and closely genetic distance.They are different species in the same genus.