本文選題：圖蘭扇頭蜱 + 線粒體基因組　； 參考：《石河子大學(xué)》2017年碩士論文

【摘要】：目的:(1)對圖蘭扇頭蜱線粒體全基因組序列進行測定并對其結(jié)構(gòu)特點進行分析,探討扇頭蜱屬系統(tǒng)發(fā)育關(guān)系。(2)設(shè)計3個新的圖蘭扇頭蜱種內(nèi)鑒定的基因標志物,對圖蘭扇頭蜱進行種內(nèi)分子鑒定。方法:(1)根據(jù)Gen Bank數(shù)據(jù)庫的扇頭蜱屬線粒體全基因組序列,設(shè)計出6對線粒體全基因組引物用于PCR擴增中國新疆圖蘭扇頭蜱線粒體全基因組序列。運用多個基因分析軟件進行線粒體全基因組序列的組裝、分析,并進行進一步的12S r DNA和cox1基因的最大似然法(Maximum Likelihood,ML)系統(tǒng)發(fā)生學(xué)分析。(2)采集中國新疆南北疆7個縣市等地區(qū)212只圖蘭扇頭蜱,阿爾巴尼亞4個縣市103只圖蘭扇頭蜱,結(jié)合蜱的形態(tài)學(xué)特征及蜱線粒體16S r RNA和cox1基因,對所采集的蜱進行蜱種的初步形態(tài)學(xué)及分子生物學(xué)鑒定并進行ML法系統(tǒng)發(fā)育分析。根據(jù)扇頭蜱屬線粒體全基因組中的保守區(qū)域,設(shè)計3個新的線粒體序列基因標志物—nad1-16S r RNA(簡稱為N1),nad2-cox1(簡稱為N2),cox1-t RNA-Lys(簡稱為C1),擴增和分析中國新疆和阿爾巴尼亞圖蘭扇頭蜱的N1、N2、C1片段并進行種內(nèi)基因差異分析和ML法系統(tǒng)發(fā)生學(xué)分析。結(jié)果:(1)在國際上首次測出圖蘭扇頭蜱全線粒體基因組長為14717 bp,其由13個蛋白質(zhì)編碼基因、22個t RNA基因、2個核糖體RNA基因和2個非編碼區(qū)組成,各基因排列順序與后溝類硬蜱相似。圖蘭扇頭蜱全線粒體基因組核苷酸組成為A=37.80%,T=40.00%,G=9.90%,C=12.30%,A+T含量為77.80%;ML系統(tǒng)進化樹顯示圖蘭扇頭蜱位于后溝類硬蜱分枝上,且與中國血紅扇頭蜱親緣關(guān)系較近,其和血紅扇頭蜱中國株一道,與血紅扇頭蜱美國株形成姊妹種關(guān)系。(2)本研究的3個新的基因標志物(N1、N2、C1)是有效的圖蘭扇頭蜱種內(nèi)鑒定的基因工具,這3個基因標志物在中阿兩國的圖蘭扇頭蜱的差異分別為3.57-4.07%、3.57-4.39%和3.18-4.69%,將中國和阿爾巴尼亞的圖蘭扇頭蜱分為兩個不同的亞種。結(jié)論:(1)本研究首次在國際上完成圖蘭扇頭蜱線粒體的全基因組測序;與大多數(shù)典型的后生動物線粒體基因組類似,圖蘭扇頭蜱線粒體基因組包含37個基因,包括13個蛋白質(zhì)編碼基因、2個r RNA基因和22個t RNA基因且僅有一個拷貝,具備2個非編碼控制區(qū),呈閉合環(huán)狀結(jié)構(gòu)。就目前的研究和數(shù)據(jù)而言,本研究的圖蘭扇頭蜱與血紅扇頭蜱中國株在系統(tǒng)發(fā)生關(guān)系上最近,但未來仍然需要擴增扇頭蜱屬的其他種的mt DNA來進行更廣闊的分析。(2)本研究設(shè)計的3個新的圖蘭扇頭蜱線粒體基因標志物—N1、N2、C1是有效的圖蘭扇頭蜱種內(nèi)鑒定基因標志物,將中國新疆和阿爾巴尼亞的圖蘭扇頭蜱成功分為2個不同的亞種進化枝。圖蘭扇頭蜱至少存在兩個不同的亞種,未來應(yīng)該大量采集世界其他地域的圖蘭扇頭蜱進行亞種的研究。
[Abstract]:Objective to determine the whole mitochondrial genome sequence of Tulan tick and analyze its structural characteristics, and to study the phylogenetic relationship. The intraspecific molecular identification of ticks was carried out. Methods according to the whole mitochondrial genome sequence of Gen Bank database, six pairs of mtDNA primers were designed to amplify the whole mitochondrial genome sequence of Tulan tick in Xinjiang, China. The whole mitochondrial genome sequence was assembled and analyzed by multiple gene analysis software. Further phylogenetic analysis of 12s r DNA and cox1 genes by Maximum Likelihoodtus (MLL) phylogenetic analysis was carried out. 212 Tulan head ticks were collected from 7 counties and cities in Xinjiang, China, and 103 from 4 counties in Albania. Combined with the morphological characteristics of ticks and the 16s r RNA and cox1 genes of ticks mitochondria, the preliminary morphological and molecular biological identification and phylogenetic analysis of the collected ticks were carried out by ML method. According to the conserved region of the mitochondrial genome of the genus Acarus, To design three new mitochondrial gene markers -nad1-16S r RNA-nad2-cox1 (abbreviated as N2C2Cox1-t RNA-Lys-Lys-Lys1), amplify and analyze the N _ 1N _ 2N _ 2N _ 2C _ 1 fragment of ticks from Xinjiang, China and Albania, and carry out intraspecific gene differential analysis and ML. Analysis of phylogeny of legal system. Results the length of the whole mitochondrial genome was 14717 BP for the first time in the world. It was composed of 13 protein coding genes, 22 t RNA genes, 2 ribosomal RNA genes and 2 non-coding regions. The sequence of genes was similar to that of Hypodes. The nucleotide composition of the whole mitochondrial genome of Trichophynchus thuringii is A37.800.0.0.The content of Cn12.30AT is 77.800.The phylogenetic tree shows that the tick is located on the branches of the posterior sulcus and is closely related to the Chinese Haematopsis, which is associated with the Chinese strain of Haemophorus. In this study, three new genetic markers, N _ (1) N _ (2) C _ (1), are effective genetic tools for intraspecific identification. The difference of the three gene markers between China and Albania was 3.57-4.073.57-4.39% and 3.18-4.69respectively. The Chinese and Albanian ticks were divided into two different subspecies. Conclusion the whole mitochondrial genome of Tulan head tick was sequenced in the world for the first time in this study, which was similar to that of most typical metazoa. The mitochondrial genome of Tulan head tick contained 37 genes. There are 13 protein coding genes, 2 r RNA genes and 22 t RNA genes with only one copy. There are 2 non-coding control regions with closed ring structure. As far as the current research and data are concerned, the phylogenetic relationship between the Chinese strains of Tulan tick and Hematoptera pyramidis is the most recent in this study. But in the future, mt DNA of other species of the genus Thalanopsis need to be amplified for a broader analysis.) the three new mitochondrial gene markers -N1N _ 2N _ 2C _ 1 designed in this study are effective markers for intraspecific identification of ticks. Two subspecies evolutionary branches were successfully divided into two subspecies from Xinjiang, China and Albania. There are at least two different subspecies in Tulan tick. In the future, a large number of species should be collected from other parts of the world.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R384.4

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