Investigating The Phylogenic Relationship Of Ribbed-Shelled Ang Smooth-Shelled Snails Of Oncomelania Hupesis Collected From Guangxi With These From Neighboring Provinces, China

Objective To investigate the morphology and genetic difference of cytochrome oxidase 1 (CO1) of smooth-shelled and ribbed-shelled snails of Oncomelania hupesis collected from Guangxi, and analyze the phylogenic relationship with these from neighboring provinces, China.Methods Ribbed-shelled(Hengxian county) and smooth-shelled (Jingxi and Luocheng counties) snails were collected from surviving snails areas in Guangxi zhuang autonomous region respectively by random sampling. Measuring and counting the whorl,height,width,length of penultimate whorl and thickness of outer lip of snails collected from 3 isolates by using magnifying glass and micrometer screw gauge. Genomic DNA was extracted from the snails. CO1 gene fragments were amplified by PCR, then purified and sequenced. Comparing the homology of CO1 between ribbed-snail and smooth-snail in Guangxi by using Clustal X and DNAMAN software. The data of CO1 gene sequences of Yunnan, Sichuan, Hunan, and Hubei isolates was obtained from the GenBank. Sequence of each isolate was edited by using Clustal X and DNAMAN software. The CO1 gene of Tricia was used as the reference of exogenous gene. The genetic distance was computed with Kimura method. The phylogenetic tree was constructed by the unweighted pair group method with arithmetic mean (UPGMA) as well as the methods of Neighbor-Joining (NJ) and Minimum evolution (ME) in the MEGA(3.1) software. Comparing the phylogenic relationship of two kinds of snails collected from Guangxi with these from neighboring provinces.Results NO. whorl of the smooth-snails in Luocheng and Hengxian and the ribbed-snails in Hengxian were 5.65±0.33, 5.49±0.40 and 5.58±0.48 respectively. The height of them were 6.75±0.37,6.59±0.26 and 6.75±0.38 respectively. The width of them were 3.38±0.52, 3.07±0.35 and 3.18±0.36. The length of penultimate whorl of them were 1.40±0.22,1.50±0.19 and 1.43±0.27. The thickness of outer lip were 1.40±0.22,1.50±0.19and 1.43±0.27. The genetic distance and homology of CO1 between Jingxi isolates(smooth-shelled) and Hengxian isolates (ribbed-shelled) was 0.0505 and 95%, respectively. The genetic distance and homology of CO1 between Jingxi isolates (smooth-shelled) and Luocheng isolates (smooth-shelled) was 0.0416 and 96%, respectively. The genetic distance and homology of CO1 between Jingxi isolates(smooth-shelled) and Hengxian isolates (ribbed-shelled) was 0.0154 and 98%, respectively. The genetic distance of Hengxian isolate(ribbed-shelled) with other isolates from Hunan, Hubei ,Yunnan and Sichuan province was 0.0102, 0.0171, 0.1401 and 0.1421, and the homology of that was 99%, 98%, 87% and 86%. The genetic distance of Luocheng isolate(smooth-shelled) with other isolates from Hunan, Hubei ,Yunnan and Sichuan province was 0.0068,0.0154,0.1362 and 0.1421, and the homology of that was 99%,98%,88% and 86%. Correspondingly, the genetic distance of Jingxi isolate (smooth-shelled)with isolates from Hunan, Hubei ,Yunnan and Sichuan province was 0.0451, 0.0505, 0.1382, and 0.1421, while the homology of that was 95%, 95%, 88% and 87%.Conclusion No significant genetic and morphological difference was found between the ribbed-shelled and smooth-shelled Oncomelania hupesis isolates from Hengxian county ,Luocheng county and Jingxi county of Guangxi zhuang autonomous region. They were phylogenetically closer, and more homologous to the isolates from Hunan and Hubei province. However, they showed longer genetic distance and less homologous to the isolates from Sichuan and Yunnan province.