【摘要】： 【目的】研究湖北釘螺線粒體COl基因序列差異,建立分子系統(tǒng)發(fā)育樹:研究釘螺滇川亞種的遺傳多樣性,探討COI基因序列是否適合湖北釘螺分類學(xué)研究,從而為云南省血吸蟲病的防治,尤其是滅螺工作提供理論依據(jù)。 【方法】采集云南省血吸蟲病流行區(qū)釘螺,分別提取釘螺全基因組DNA,PCR擴(kuò)增線粒體DNA(mtDNA)的細(xì)胞色素C氧化酶I(COI)基因片段并測序。應(yīng)用DNAstar軟件對所測序列進(jìn)行編輯。從GenBank下載釘螺COI基因序列,應(yīng)用ClustalX1.83軟件對基因序列進(jìn)行排列比較。以雙臍螺Biomphalaria glabrata(AY380531)以及微小釘螺O.h.minima(DQ212795)作為外群參照,應(yīng)用MEGA4.0、PAUP*4.0、PHYML軟件,用鄰接法(NJ)、使用不加權(quán)算術(shù)平均組對法(UPGMA)、最大簡約法(MP)和最大似然法(ML)分別構(gòu)建系統(tǒng)發(fā)育樹進(jìn)行分析。應(yīng)用MEGA4.0軟件計算遺傳距離。 【結(jié)果】湖北釘螺COI基因序列的比較結(jié)果和所構(gòu)建系統(tǒng)發(fā)育樹顯示:所分析的湖北釘螺COI基因片段序列長度為638bp,釘螺滇川亞種的保守位點數(shù)目為533,變異位點數(shù)目為105,簡約信息位點數(shù)目為91,自裔位點為14。A、T、C、G平均含量分別為23.0%、37.6%、18.3%、21.1%,A+T含量為60.6%,而G+C含量為39.4%,A+T含量高于G+C含量。所分析釘螺滇川亞種的轉(zhuǎn)換與顛換的平均比值為6.2。堿基替換飽和性分析顯示湖北釘螺COI基因序列堿基替換基本未達(dá)到飽和。湖北釘螺系統(tǒng)發(fā)育樹分為三大支:滇川亞種支、夸氏亞種支和湖北釘螺其它各亞種支。三種方法構(gòu)建的釘螺滇川亞種系統(tǒng)發(fā)育樹拓?fù)浣Y(jié)構(gòu)基本一致,可分為兩大支:即云南支和四川支。釘螺滇川亞種與湖北釘螺其它各亞種間未校正P距離為0.1197-0.1313(0.1255±0.0058),Kimura二參數(shù)模型P距離為0.1485-0.1343(0.1414±0.0071);湖北釘螺其它各亞種間未校正P距離為0.0091-0.0252(0.01715±0.00805),Kimura二參數(shù)模型P距離為0.0093-0.0258(0.01755±0.00285);釘螺滇川亞種中云南支與四川支之間未校正P距離為0.0857,Kimura二參數(shù)模型P距離為0.0928。 【結(jié)論】①釘螺滇川亞種內(nèi)存在遺傳變異。②提示COI基因片段序列不適合近緣釘螺亞種的分類研究,但對于釘螺滇川亞種內(nèi)形成穩(wěn)定的分支具有很好的支持。③滇川亞種內(nèi)分兩個亞群,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 鹵0.0058), Kimura, P = 0.1485-0.1343, 0.1414 鹵0.0071). The uncorrected P distance between other subspecies of Oncomelania hupensis was 0.0091-0.0252 (0.01715 鹵0.00805), Kimura) and P distance was 0.0093-0.0258 (0.01755 鹵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.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R384

【引證文獻(xiàn)】

相關(guān)期刊論文 前2條

1 馬琳;趙玲;李石柱;尤平;周曉農(nóng);;景觀遺傳學(xué)及其在湖北釘螺研究中的應(yīng)用[J];中國血吸蟲病防治雜志;2010年06期

2 馬琳;李石柱;楊頻;尤平;周曉農(nóng);;湖北釘螺基因組5種抽提方法的比較[J];中國病原生物學(xué)雜志;2011年02期

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1 馬琳;中國大陸湖北釘螺不同地理景觀群體遺傳變異分析[D];陜西師范大學(xué);2011年

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