云南省血吸虫病流行区湖北钉螺COI基因序列分析
发布时间：2018-11-17 12:38
【摘要】： 【目的】研究湖北钉螺线粒体COl基因序列差异,建立分子系统发育树:研究钉螺滇川亚种的遗传多样性,探讨COI基因序列是否适合湖北钉螺分类学研究,从而为云南省血吸虫病的防治,尤其是灭螺工作提供理论依据。 【方法】采集云南省血吸虫病流行区钉螺,分别提取钉螺全基因组DNA,PCR扩增线粒体DNA(mtDNA)的细胞色素C氧化酶I(COI)基因片段并测序。应用DNAstar软件对所测序列进行编辑。从GenBank下载钉螺COI基因序列,应用ClustalX1.83软件对基因序列进行排列比较。以双脐螺Biomphalaria glabrata(AY380531)以及微小钉螺O.h.minima(DQ212795)作为外群参照,应用MEGA4.0、PAUP*4.0、PHYML软件,用邻接法(NJ)、使用不加权算术平均组对法(UPGMA)、最大简约法(MP)和最大似然法(ML)分别构建系统发育树进行分析。应用MEGA4.0软件计算遗传距离。 【结果】湖北钉螺COI基因序列的比较结果和所构建系统发育树显示:所分析的湖北钉螺COI基因片段序列长度为638bp,钉螺滇川亚种的保守位点数目为533,变异位点数目为105,简约信息位点数目为91,自裔位点为14。A、T、C、G平均含量分别为23.0%、37.6%、18.3%、21.1%,A+T含量为60.6%,而G+C含量为39.4%,A+T含量高于G+C含量。所分析钉螺滇川亚种的转换与颠换的平均比值为6.2。碱基替换饱和性分析显示湖北钉螺COI基因序列碱基替换基本未达到饱和。湖北钉螺系统发育树分为三大支:滇川亚种支、夸氏亚种支和湖北钉螺其它各亚种支。三种方法构建的钉螺滇川亚种系统发育树拓扑结构基本一致,可分为两大支:即云南支和四川支。钉螺滇川亚种与湖北钉螺其它各亚种间未校正P距离为0.1197-0.1313(0.1255±0.0058),Kimura二参数模型P距离为0.1485-0.1343(0.1414±0.0071);湖北钉螺其它各亚种间未校正P距离为0.0091-0.0252(0.01715±0.00805),Kimura二参数模型P距离为0.0093-0.0258(0.01755±0.00285);钉螺滇川亚种中云南支与四川支之间未校正P距离为0.0857,Kimura二参数模型P距离为0.0928。 【结论】①钉螺滇川亚种内存在遗传变异。②提示COI基因片段序列不适合近缘钉螺亚种的分类研究,但对于钉螺滇川亚种内形成稳定的分支具有很好的支持。③滇川亚种内分两个亚群,即云南支和四川支。
[Abstract]:[objective] to study the difference of mitochondrial COl gene sequence in Oncomelania hupensis and to establish a phylogenetic tree. To study the genetic diversity of Oncomelania hupensis subspecies, and to explore whether the COI gene sequence is suitable for the taxonomic study of Oncomelania hupensis. It provides theoretical basis for the prevention and control of schistosomiasis in Yunnan Province, especially for snail control. [methods] Oncomelania hupensis was collected from schistosomiasis endemic area in Yunnan Province, and the whole genome DNA,PCR of snail was extracted to amplify the I (COI) gene fragment of cytochrome C oxidase (I (COI) of mitochondrial DNA (mtDNA) and sequenced. The DNAstar software was used to edit the measured sequence. The COI gene sequence of Oncomelania hupensis was downloaded from GenBank and compared with the sequence by ClustalX1.83 software. Biomphalaria glabrata (AY380531 and O.h.minima (DQ212795) were used as outer group reference, MEGA4.0,PAUP*4.0,PHYML software was used, (NJ), adjacent method was used to use unweighted arithmetic average group pair (UPGMA),. The phylogenetic tree was constructed by maximum minimization method (MP) and maximum likelihood method (ML) respectively. Genetic distance was calculated by MEGA4.0 software. [results] the results of comparison of COI gene sequence and phylogenetic tree showed that the length of COI gene fragment was 638 BP, the number of conserved loci was 533 and the number of mutation sites was 105 in the subspecies of Oncomelania hupensis. The number of parsimonious information sites was 91, and the average content of C was 23.037. 6% and 60.6%, respectively, while the content of G C was 39.4%, respectively, with the number of simple information sites being 91, and the average content of C being 14. 0% and 37. 6%, respectively, and 18. 3% and 21. 1%, respectively. The content of A T is higher than that of G C. The average ratio of transformation to transversion of subspecies of Oncomelania hupensis was 6.2. Base substitution saturation analysis showed that the base substitution of COI gene sequence of Oncomelania hupensis was basically not saturated. The phylogenetic tree of Oncomelania hupensis is divided into three branches: branch of Yunnan-Sichuan subspecies, branch of Kua's subspecies and branches of other subspecies of Oncomelania hupensis. The phylogenetic tree of Oncomelania hupensis subspecies in Yunnan and Sichuan has the same topological structure and can be divided into two branches: Yunnan branch and Sichuan branch. The uncorrected P distance between Yunnan and Sichuan subspecies of Oncomelania hupensis and other subspecies of Oncomelania hupensis was 0.1197-0.1313 (0.1255 卤0.0058), Kimura, P = 0.1485-0.1343, 0.1414 卤0.0071). The uncorrected P distance between other subspecies of Oncomelania hupensis was 0.0091-0.0252 (0.01715 卤0.00805), Kimura) and P distance was 0.0093-0.0258 (0.01755 卤0.00285). The uncorrected P distance between Yunnan branch and Sichuan branch in subspecies of Oncomelania hupensis was 0.0857 and the P distance of Kimura two-parameter model was 0.0928. [conclusion] 1 there is genetic variation in subspecies of Oncomelania hupensis, 2 it is suggested that the sequence of COI gene fragment is not suitable for the classification of related subspecies of Oncomelania hupensis. But it has good support for the formation of stable branches within the subspecies of Oncomelania hupensis. 3 the subspecies of Yunnan and Sichuan are divided into two subgroups: the branch of Yunnan and the branch of Sichuan.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R384
本文编号：2337808
[Abstract]:[objective] to study the difference of mitochondrial COl gene sequence in Oncomelania hupensis and to establish a phylogenetic tree. To study the genetic diversity of Oncomelania hupensis subspecies, and to explore whether the COI gene sequence is suitable for the taxonomic study of Oncomelania hupensis. It provides theoretical basis for the prevention and control of schistosomiasis in Yunnan Province, especially for snail control. [methods] Oncomelania hupensis was collected from schistosomiasis endemic area in Yunnan Province, and the whole genome DNA,PCR of snail was extracted to amplify the I (COI) gene fragment of cytochrome C oxidase (I (COI) of mitochondrial DNA (mtDNA) and sequenced. The DNAstar software was used to edit the measured sequence. The COI gene sequence of Oncomelania hupensis was downloaded from GenBank and compared with the sequence by ClustalX1.83 software. Biomphalaria glabrata (AY380531 and O.h.minima (DQ212795) were used as outer group reference, MEGA4.0,PAUP*4.0,PHYML software was used, (NJ), adjacent method was used to use unweighted arithmetic average group pair (UPGMA),. The phylogenetic tree was constructed by maximum minimization method (MP) and maximum likelihood method (ML) respectively. Genetic distance was calculated by MEGA4.0 software. [results] the results of comparison of COI gene sequence and phylogenetic tree showed that the length of COI gene fragment was 638 BP, the number of conserved loci was 533 and the number of mutation sites was 105 in the subspecies of Oncomelania hupensis. The number of parsimonious information sites was 91, and the average content of C was 23.037. 6% and 60.6%, respectively, while the content of G C was 39.4%, respectively, with the number of simple information sites being 91, and the average content of C being 14. 0% and 37. 6%, respectively, and 18. 3% and 21. 1%, respectively. The content of A T is higher than that of G C. The average ratio of transformation to transversion of subspecies of Oncomelania hupensis was 6.2. Base substitution saturation analysis showed that the base substitution of COI gene sequence of Oncomelania hupensis was basically not saturated. The phylogenetic tree of Oncomelania hupensis is divided into three branches: branch of Yunnan-Sichuan subspecies, branch of Kua's subspecies and branches of other subspecies of Oncomelania hupensis. The phylogenetic tree of Oncomelania hupensis subspecies in Yunnan and Sichuan has the same topological structure and can be divided into two branches: Yunnan branch and Sichuan branch. The uncorrected P distance between Yunnan and Sichuan subspecies of Oncomelania hupensis and other subspecies of Oncomelania hupensis was 0.1197-0.1313 (0.1255 卤0.0058), Kimura, P = 0.1485-0.1343, 0.1414 卤0.0071). The uncorrected P distance between other subspecies of Oncomelania hupensis was 0.0091-0.0252 (0.01715 卤0.00805), Kimura) and P distance was 0.0093-0.0258 (0.01755 卤0.00285). The uncorrected P distance between Yunnan branch and Sichuan branch in subspecies of Oncomelania hupensis was 0.0857 and the P distance of Kimura two-parameter model was 0.0928. [conclusion] 1 there is genetic variation in subspecies of Oncomelania hupensis, 2 it is suggested that the sequence of COI gene fragment is not suitable for the classification of related subspecies of Oncomelania hupensis. But it has good support for the formation of stable branches within the subspecies of Oncomelania hupensis. 3 the subspecies of Yunnan and Sichuan are divided into two subgroups: the branch of Yunnan and the branch of Sichuan.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R384
【引证文献】
相关期刊论文 前2条
1 马琳;赵玲;李石柱;尤平;周晓农;;景观遗传学及其在湖北钉螺研究中的应用[J];中国血吸虫病防治杂志;2010年06期
2 马琳;李石柱;杨频;尤平;周晓农;;湖北钉螺基因组5种抽提方法的比较[J];中国病原生物学杂志;2011年02期
相关硕士学位论文 前1条
1 马琳;中国大陆湖北钉螺不同地理景观群体遗传变异分析[D];陕西师范大学;2011年
，本文编号：2337808
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2337808.html
最近更新
教材专著
