水牛、牦牛及马Y染色体分子遗传多样性研究

发布时间：2019-06-19 12:10

【摘要】：水牛是我国重要的役用家畜。目前,对于水牛起源进化的研究在考古学、形态学与分子遗传学方面均有涉及,大量线粒体DNA的研究虽能提供水牛母系起源的证据,但仍需Y染色体分子遗传多样性方面的研究来提供水牛父系起源的遗传学证据。本研究以10个中国沼泽型水牛品种共196头公牛为研究对象,利用已发表的42对黄牛Y染色体微卫星(Y-STR)标记,采用常规的银染Y-STR分型技术,筛选出更多的水牛Y-STR多态标记,并用国际先进的荧光Y-STR分型技术加以验证,以获得可靠的水牛Y-STR多态标记。牦牛是一个我国古老的牛种,在青藏高原地区畜牧业经济中有着有不可替代的生态经济地位,是重要的自然资源、生态资源和经济资源,同时也是牛科家畜基因库的重要组成部分。对不同牦牛品种进行遗传多样性研究从Y染色体角度对不同牦牛品种分子遗传多样性研究具有重大意义。本研究以2个青海高原牦牛类群和大通牦牛共108头公牛为研究对象,对8对黄牛的Y-STR标记进行筛选,以获得可靠的牦牛Y-STR多态标记。中国拥有世界上最丰富的马品种遗传资源,居世界第一位,但从父系遗传(即Y染色体)角度对马品种分子遗传多样性的研究报道较少,中国马Y-SNP研究更不多,还没有发现Y-SNP。揭示马品种的Y染色体分子遗传多样性状况,将为马品种分子遗传多样性数据库的建立、育种改良提供理论依据。本研究对13个中国马品种在4对Y染色体SNP标记中的分子遗传多样性进行了分析。本研究对水牛、牦牛及马Y染色体分子遗传多样性分析,获得以下主要结果:1.水牛Y-STRs筛选与遗传多样性研究(1)选择黄牛42对Y-STR标记,对水牛群体进行Y-STR标记特异性与多态性筛选,初步发现35对Y-STR标记为水牛Y染色体特异性标记,其中12对(UMN0108、UMN2008、UMN0705、UMN0907、UMN1113、INRA126、BC1.2、UMN2713、UMN2908、PBR1F1、UMN2001、INRA189)在水牛中有多态性。另外7对Y-STR标记(UMN2303、UMN2405、UMN0929、UMN0803、UMN0301、UMN1307、UMN1514)不是水牛Y染色体特异的微卫星标记。(2)荧光Y-STR分型结果表明,3对标记(INRA189、PBR1F1和UMN2001)在水牛不同品种中存在多态性,表明采用常规银染方法所筛选的Y-STR多态性标记有一定的可靠性。(3)中国10个水牛品种具有较高的Y-STR多态性,聚类分析表明,中国水牛有2个父系起源。2.牦牛Y-STRs筛选与遗传多样性研究(1)采用常规银染方法检测了青海高原牦牛2个类群和大通牦牛8个Y-STR标记的特异性,其中5个Y-STR标记(BM861,BYM-1,INRA126,UMN0307和UMN0103)不是牦牛Y染色体特异微卫星标记;1个标记(INRA124)在牦牛类群中无扩增产物;2个标记(INRA189和UMN2404)是牦牛Y染色体特异微卫星标记,且具有多态性。(2)荧光Y-STR分型结果表明,2个标记(INRA189和UMN2404)是牦牛Y染色体特异微卫星标记,且具有多态性。首次证明UMN2404标记能用于牦牛Y染色体遗传多样性研究。(3)青海高原牦牛2个类群和大通牦牛之间Y-STR多态性较低,2个类群和大通牦牛没有明显的遗传分化。3.马Y-SNP筛选及遗传多样性研究(1)首次在中国马中发现3个Y-SNPs(Y-45701/997,G→T突变;Y-50869,T→A突变;Y-45288,T碱基缺失)。(2)在这3个Y-SNPs位点中,中国马的Y-SNP多态性比欧洲及美洲马低。中国西北地区的马具有较高的Y-SNP多态性。
[Abstract]:The buffalo is an important domestic animal in our country. At present, research on the origin of buffalo has involved in the aspects of archaeology, morphology and molecular genetics. But it is still necessary to study the genetic diversity of the Y-chromosome molecule to provide the genetic evidence of the origin of the father of the buffalo. In this study, a total of 196 bulls were used as the research object in 10 Chinese swamp buffaloes, and the published 42 pairs of Y-STR markers were used to screen the Y-STR polymorphic markers of buffalo. And a reliable buffalo Y-STR polymorphic marker is obtained by using the international advanced fluorescent Y-STR typing technology. Yak is an ancient Chinese species, and has an irreplaceable ecological economic status in the animal husbandry economy of the Qinghai-Tibet Plateau. It is an important natural resource, ecological resource and economic resources, and is also an important part of the cattle gene bank of the cattle family. It is of great significance to study the genetic diversity of different yak breeds from the Y chromosome angle. In this study, the Y-STR markers of 8 yellow cattle were selected to obtain a reliable Y-STR polymorphic marker. China has the most abundant genetic resources in the world, ranking the first in the world, but the study of the genetic diversity of the species of the horse from the father's genetic (i.e., Y chromosome) angle is less, and the Chinese horse Y-SNP study is much less, and the Y-SNP is not found. The genetic diversity of the Y chromosome of the horse variety is revealed, which will provide a theoretical basis for the establishment of the genetic diversity database of the horse variety and the improvement of the breeding. The molecular genetic diversity of 13 Chinese horse breeds in 4 pairs of Y-chromosome SNP markers was analyzed. In this study, the molecular genetic diversity of buffalo, yak and horse Y was analyzed, and the following main results were obtained:1. The selection of Y-STRs and genetic diversity of water buffalo (1) selection of yellow cattle 42 to the Y-STR markers and the selection of Y-STR markers for buffalo groups, and the preliminary finding of 35 pairs of Y-STR markers for buffalo Y chromosome-specific markers,12 pairs (UMN0108, UMN2008, UMN0705, UMN0907, UMN1113, INRA126, BC1.2, UMN2713, UNU2908, PBR1F1, UMN2001, INRA189) were polymorphic in buffaloes. In addition,7 pairs of Y-STR markers (UMN2303, UMN2405, UMN0929, UMN0803, UMN0301, UMN1307, UMN1514) are not microsatellite markers specific to the Y chromosome of the buffalo. (2) The results of fluorescent Y-STR typing show that 3 pairs of markers (INRA189, PB1F1 and UMN2001) have a polymorphism in different varieties of buffalo, indicating that the Y-STR polymorphic marker screened by the conventional silver staining method has certain reliability. (3) Chinese buffalo had a high Y-STR polymorphism, and the cluster analysis showed that the Chinese buffalo had 2 paternal origins. The selection and genetic diversity of Yak Y-STRs (1) were used to test the specificity of 2 taxa and 8 Y-STR markers of Yak in Qinghai Plateau. The five Y-STR markers (BM861, BYM-1, INRA126, UMN0307 and UMN0103) were not specific microsatellite markers of Yak Y chromosome. One marker (INRA124) has no amplification product in yak group, and 2 markers (INRA189 and UMN2404) are specific microsatellite markers of Yak Y chromosome and have polymorphism. (2) The results of fluorescent Y-STR typing show that 2 markers (INRA189 and UMN2404) are specific microsatellite markers of Yak Y chromosome and have polymorphism. The first time that the UMN2404 marker can be used to study the genetic diversity of Y chromosome in Yak. (3) The Y-STR polymorphism between the two groups of Yak and the Yak in Qinghai plateau was lower, and there were no obvious genetic differentiation between the two groups. Three Y-SNPs (Y-45701/997, G-T mutation, Y-50869, T-A mutation, Y-45288, T-base deletion) were found in Chinese horses for the first time. (2) In these three Y-SNPs sites, the Y-SNP polymorphism of Chinese horse is lower than that in Europe and America. The horse in the northwest of China has a higher Y-SNP polymorphism.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S813.1

【相似文献】