| Schistosomiasis,the zoonotic infectious parasitic diseases,is one of the important public health problem in China.Oncomelania hupensis is the sole intermediate host of Schistosoma japonicum, the geographical distribution of schistosomiasis endemic area are same as Oncomelania hupensis. O. hupensis plays a key role in the transmission of Schistosoma japonicum. Snail mainly distribut in the southern areas, in China we can find it in the Yangtze River in 12 provinces:north of N 33°20', south of N 22°20', east of E 121°51', West of E 99°04'. Because O. hupensis is widespread and in the influence of the geographical isolation, there is very obvious variation between the different areas population in China's mainland, then evolving many different sub-species, and in the same condition, there's significant and different susceptivity to the S. japonicum. So hupensis species and population genetic differentiation are deeply researched, it provide important theoretical significance for the study of S. japonicum and O. hupensis, O. hupensis interaction diffusion mechanism of snail populations, epidemiology of schistosomiasis, schistosomiasis low prevalence of active monitoring.In this study, firstly, we compare five commonly used extraction method for extracting genome and amplificat mtDNA cytochrome C oxidase subunit II to compare the quality, in order to find a suitable extraction method to amplification mass samples, secondly, based on obtain hupensis mitochondrial genome sequence,we design 16 pairs of specific primers to amplify 12 mitochondrial genome sequence and analysis its sequence information, then 13 protein-coding genes in mitochondrial genome were selected and analysised to separate grade based on phylogenetic information molecular markers for population genetic study, and also studied the sampling strategy of population genetic analysis of O. hupensis, Finally, analysis population genetic structure of mtDNA cytochrome C oxidase subunit?and dcoxyribose oxidase subunit 3 at the landscape level in China's mainland to explore the specific characteristics at molecular level of O. hupensis in different geographical landscape, analysis the relationship between genetic diversity, migration and differentiation in different geographical landscape. The results are as follows: (1)Each method can extracte genomic DNA and amplified mitochondrial DNA of the CO?gene. The concentration of PCR product by Omega kit, TianGen kit and simpled method are higher. Omega kit and TianGen kit suitable for small sample size.(2) In this study we obtained 12 complete mitochondrial genome sequence, the length are 15183 kb to 15216 kb, the phylogenetic information efficiency between the mitochondrial protein coding genes are different, the gene can be divided into three grades, COI, ND2, ND5, ND3 are better, CO?, ATP6, ND1, Cytb, ND4 are middle, ATP8, ND6, COIII, ND4L are poor.(3)The ND3 gene in different geographic showed a different gene polymorphism, with the increase in sample size the gene polymorphism did not increase; It shown significant regional differences; therefore, in the genetic study by ND3 gene the sample size range should be between 10-30.(4)Based on mitochondrial ND3 gene and CO?gene sequences we divided O. hupensis in China's mainland into four main groups, the population representing the region of swamps and lakes along the Yangtze,the mountainous region population (from Sichuan and Yunnan provinces); the Karst region population (from Guangxi autonomous region), the littoral, hilly region population (from Fujian province). |
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