云南省临沧市小管福寿螺COⅠ基因多态性分析

Genotyping and polymorphism analysis of cytochrome c oxidase subunit Ⅰ gene of Pomacea canaliculata from Lincang City in Yunnan Province

目的通过对云南省临沧市小管福寿螺(Pomacea canaliculata)COⅠ基因的遗传标记分析,了解当地小管福寿螺基因多态性,为云南省后续开展广州管圆线虫病监测提供科学依据。方法对采自云南省临沧市孟定镇38份小管福寿螺样本进行COⅠ基因扩增,并将PCR产物测序。运用MEGA 6.06软件Kimura-2参数模型,对来自于Gen Bank的福寿螺单倍型与本研究获得单倍型一起进行系统进化树构建和个体间遗传距离计算,分析其遗传多样性。结果共获得31条序列,分属3种单倍型(Haplotype1~Haplotype3),其中Haplotype1的频率较高,占整个样本的83.9%(26/31)。3种单倍型与小管福寿螺的遗传距离最小,为0~0.052;而与Pila conica的遗传距离最大,为0.021~0.239。进一步进化分析表明,3种单倍型均为小管福寿螺,与来自日本熊本(Gen Bank登录号:AB 433769)、中国香港(Gen Bank登录号:KT 313034)和美国夏威夷(Gen Bank登录号:EU 523129)的小管福寿螺序列聚成一大支,具有较近的亲缘关系;而与P.insularum(Gen Bank登录号:EF 514942)、P.camena(Gen Bank登录号:EF 515059)等序列的亲缘关系较远。结论云南省临沧市存在小管福寿螺,本研究获得的3种单倍型之间存在较大的遗传分化。

Objective To analyze the genetic diversity of Pomacea canaliculata based on the mitochondria DNA cytochrome c oxidase subunit Ⅰ（mt DNA COⅠ）gene as a molecular marker in Lincang City of Yunnan Province, so as to provide the scientific data for monitoring Angiostrongylus cantonensis in local areas.Methods The genotypes and polymorphisms of 38 specimens of P. canaliculata collected from Mengding Town of Lincang City were analyzed by sequencing COⅠ gene. The phylogenetic tree and genetic distances were produced based on the haplotypes from Gen Bank and the present study by using the neighbourjoining method with the software MEGA version 6.06.Results Totally 31 sequences were acquired in the present study and they produced 3 unique haplotypes. Haplotype 1 showed a higher frequency compared to the others and it accounted for 83.9 %（26/31）. The data showed that the least genetic distances ranged from 0 to 0.052 between P. canaliculata and 3 haplotypes,as well as the largest genetic distances ranged from 0.021 to 0.239 between Pila conica and 3 haplotypes. Otherwise,the analysis of the phylogenetic trees based on COⅠ gene sequences of P. canaliculata indicated that all of 3 haplotypes clustered into one big clade with that from Japan（Gen Bank accession number： AB433769）,China（Gen Bank accession number： KT313034）and USA（Gen Bank accession number： EU523129）,which owned the closet relationship amongst them. Their genetic relationships were distantly related to the Gen Bank＇s reference sequences of P. insularum（Gen Bank accession number： EF514942）,P. camena（Gen Bank accession number： EF515059）and so on.Conclusion There is a P. canaliculata species in Lincang City of Yunnan Province as well as a high genetic diversity amongst the acquired 3 haplotypes in this study.