安集延鸡mtDNA D-loop遗传多样性及系统进化研究

本研究通过分析mtDNA,对安集延鸡的母系遗传多样性和母系来源进行调查。随机采集了29个安集延鸡DNA样品,测定了D-loop区1231bp序列。结果显示,共检测到30个多态位点和10个单倍体型,在859bp位置发现一个插入/缺失变异,此变异在安集延鸡中的分布频率为51.72%;单倍体型多样度和核苷酸多样度适中,分别为0.776和0.00614;Tajima′D检验和Fuli′D检验值分别为-0.0041和-0.0192;遗传距离与红色原鸡最近,为0.007;Network结构图显示安集延鸡群体分为五大世系,可划分到家鸡世系的A、B、C、D和E分支中;世系D所占比例最高为45%,据以往报道,D型世系中分布的大部分为红色原鸡和斗鸡,结果表明安集延鸡母系大部分来源于红色原鸡和斗鸡,并且也掺入一些家鸡母系,为多母系构成的群体。     

中国西南地区5个地方绵羊群体mtDNA遗传多样性及系统进化研究

采用PCR—SSCP技术及mtDNA D-loop序列分析相结合的方法，对我国云南昭通绵羊、腾冲绵羊、宁蒗绵羊及西藏的多玛绵羊、江孜绵羊5个地方绵羊群体共232个个体进行遗传多样性及系统进化分析。PCR-SSCP分析显示，在西藏的多玛绵羊和江孜绵羊中均检测到线粒体编码区的Cytb和ND2基因的3种单倍型A、B和C，且在西藏的多玛绵羊、江孜绵羊中C单倍型比例高于B型；而在云南的昭通绵羊、腾冲绵羊和宁蒗绵羊中只检测到单倍型A和B。根据不同的单倍型从5个群体中筛选出39个样品进行mtDNAD-loop区克隆测序，经过系统进化分析揭示西藏绵羊存在A、B、C3种mtDNA单倍型；而云南绵羊只存在A、B2种mtDNA单倍型。以上基于PCR-SSCP和D-loop区序列的分析结果一致提示西藏绵羊有3个母系来源，云南绵羊有2个母系来源。基于mtDNAD-loop序列的多态性分析结果显示西藏多玛绵羊和江孜绵羊的单倍型多样度（Hd）、核苷酸多样度（Pi）及平均核苷酸差异数（k）均高于云南3个地方绵羊品种，提示西藏绵羊遗传多样性较丰富，云南绵羊遗传多样性相对贫乏。     

郏县红牛mtDNA D-loop区遗传多样性与母系起源研究

［目的］探究郏县红牛的mtDNA D-loop遗传多样性与母系起源。［方法］采用生物信息学方法。［结果］在46头郏县红牛mtDNA D-loop区全序列共检测到60个变异位点,定义20种mtDNA D-loop单倍型,平均单倍型多样度(Hd)为0.8530,平均核苷酸多样度(Pi)为0.0254,表明郏县红牛有丰富的母系遗传多样性。构建的IQ系统发育树表明郏县红牛具有瘤牛和普通牛两个母系支系。［结论］郏县红牛具有丰富的母系遗传多样性,有普通牛和瘤牛两个母系起源。     

建昌马mtDNA D-loop区遗传多样性及系统进化分析

试验旨在以线粒体DNA(mitochondrial DNA,mtDNA)为切入点,研究建昌马的母系遗传多样性与系统进化。从建昌马(n=39)血液中提取基因组DNA,用PCR方法扩增mtDNA D-loop区并直接测序,分析其高变区247 bp序列信息,统计mtDNA D-loop区的单倍型及变异位点,计算单倍型多样性(haplotype diversity,Hd)、核苷酸多样性(nucleotide diversity,Pi)和平均核苷酸变异数(average number of nucleotide differences,K)。构建包括建昌马在内的19个品种马的NJ系统进化树,计算各品种间的遗传距离。结果显示,试验获得了清晰的PCR扩增产物,并通过直接测序方法获得了约1 200 bp的序列。39匹建昌马mtDNA D-loop区247 bp序列(其中1个样品缺失1 bp)的AT碱基含量为61.45%,属AT碱基对富集区,检测到33个多态性位点,共显示26种单倍型,其中4种为共享单倍型,且Hap7和Hap1为优势单倍型,单倍型多样性为0.947,核苷酸多样性为0.02399,平均核苷酸变异数为5.901,显示丰富的母系遗传多样性;NJ系统进化树显示,建昌马分布在A、C、D、E、F、G共6个支系中,约50%的样品分布在A支系,显示出复杂的母系起源;建昌马与关中马的遗传距离最小(0.021),其次是三河马、文山马、韩国车巨马(0.024),与韩国济州岛马遗传距离最大(0.032)。本研究结果表明,建昌马的mtDNA D-loop高变区遗传多样性丰富,具有多个母系起源,且A支系占有明显优势,与关中马、文山马可能有共同的母系起源。     

西藏牦牛mtDNA ND6遗传多样性及系统进化分析

为从分子水平上探究西藏牦牛的序列多态性、遗传多态性及其系统进化关系,对西藏15个牦牛类群150个个体的mtDNA ND6基因全序列进行了测定,确定了多态位点和单倍型数目,计算了单倍型多样性(Hd)和核苷酸多样性(Pi),并构建了分子聚类关系图和单倍型系统发育树。结果表明:西藏牦牛mtDNA ND6基因全序列长度均为528bp,T、C、A和G 4种碱基的平均比例分别为42.2%、7.6%、20.9%、29.3%,存在一定的碱基组成偏倚性。序列变异中存在转换、颠换2种核苷酸变异类型,其中转换37次,颠换2次,表现出较强的转换偏倚性。根据序列间核苷酸变异共确定7种单倍型,其中Hap_1为西藏牦牛类群的主流单倍型,其余6种单倍型为部分类群所特有。平均单倍型多样性(Hd)、平均核苷酸多样性(Pi)分别为0.2978、0.00191,揭示西藏牦牛mtDNA ND6遗传多样性较贫乏。根据遗传距离构建了分子聚类关系图,表明西藏15个牦牛类群可分为2大类;根据7种单倍型序列构建了单倍型系统发育树,表明西藏牦牛存在2个母系起源。     

青海省门源白牦牛mtDNA D-loop区遗传多样性及母系起源

为从分子水平上探究青海省门源白牦牛的母系遗传多样性及遗传背景,本研究对31头门源白牦牛线粒体DNA(mtDNA)D-loop区序列进行测定和比对分析,确定其多态位点和单倍型数目,计算核苷酸多样度、单倍型多样度及平均核苷酸差异数大小,并构建系统发育树。结果表明,门源白牦牛mtDNA D-loop区序列长度为892～896bp,排除5处插入/缺失后共发现38处多态位点,包括24处单一变异位点和14处简约信息位点;依据序列间核苷酸变异共确定了13种单倍型,单倍型多样度为0.901±0.030,核苷酸多样度为0.006±0.004,平均核苷酸差异数为5.17,提示门源白牦牛具有较丰富的母系遗传多样性。以普通牛为外群,邻接法(NJ法)构建的系统发育树结果显示,13种单倍型分为明显的2个分支,表明门源白牦牛有2个母系起源。综上所述,本研究结果表明门源白牦牛具有较丰富的母系遗传多样性,由2个母系遗传分支组成,具有2个母系起源。     

山东猪种mtDNA D-loop区部分序列遗传多样性及系统进化研究

为了解山东省猪种的遗传资源状况,阐明猪种间的亲缘关系,对山东省猪种(莱芜黑猪、大蒲莲黑猪、里岔黑猪、烟台黑猪、沂蒙黑猪、五莲黑猪、昌潍白猪和鲁烟白猪)及引进猪种(杜洛克、大约克和长白猪)共165个样本的mtDNA D-loop区部分序列(1 102 bp)进行PCR扩增和序列测定,并采用现代生物信息学软件进行了数据处理分析,结果表明:山东各猪种之间遗传距离较小(0.000 5～0.004 6),远小于国外猪种杜洛克、长白与山东各猪种间的遗传距离(>0.14),但大约克与山东各猪种的平均遗传距离较小(0.005).构建的NJ分子系统进化树将所测样本分为2个独立的支系,第一支由山东各猪种和部分大约克构成,另一支全部为引进猪种.莱芜黑猪和大蒲莲黑猪在NJ系统树上分布集中,各自聚为一小类,其它6个猪种都很分散,说明莱茺黑猪和大蒲莲黑猪母源血统单一,其它6个猪种母源血统遗传复杂,彼此之间基因交流多.     

庆阳驴mtDNA D-loop区遗传多样性及起源分析

【目的】研究庆阳驴养殖群体的遗传多样性与母系起源,了解其遗传信息,为保护庆阳驴种质资源、选育和遗传改良工作提供理论依据。【方法】随机选取133头庆阳驴,对其线粒体DNA(mitochondrial DNA,mtDNA)D-loop区序列进行PCR扩增、测序及比对,并探讨庆阳驴的遗传多样性与母系起源。【结果】在获得的520 bp D-loop碱基序列中,AT含量(57.3%)高于GC含量(42.8%),表现出碱基的偏倚性;检测到38个变异位点,包含8个碱基对的转换;其核苷酸多样性(Pi)、单倍型多样性(Hd)、平均核苷酸差异(K)分别为0.01591、0.895和8.274,与欧洲家驴和中国家驴研究的平均值相比较低,说明该驴品种核苷酸变异较为贫乏。庆阳驴mtDNA D-loop区存在35个单倍型,单倍型之间的遗传距离为0.002～0.042。系统进化结果显示,庆阳驴存在2个线粒体支系,表明其具有2个母系起源,且遗传距离表明,庆阳驴与克罗地亚家驴之间的遗传距离较近。【结论】本研究从分子水平初步揭示庆阳驴核苷酸变异比较贫乏,杂交程度高,mtDNA遗传多态性正逐步丧失,应加强庆阳驴品种的遗传资...     

mtDNA D-loop在家畜遗传多样性和系统分化中的应用

本文简要综述了mtDNA D-loop区结构的研究现状,着重概述了D-loop区研究在山羊、绵羊以及牛、猪遗传多样性和系统分化中的应用,并对该领域的深入研究进行了讨论。     

淮南猪线粒体DNA D-loop遗传多样性及母系起源研究

为探究淮南猪的遗传多样性和母系起源,本实验对32头淮南猪线粒体DNA D-loop区序列进行PCR扩增和测序,并结合GenBank中国内外猪种及黄河流域古家猪D-loop序列,分析淮南猪线粒体多态性及其起源。结果显示:淮南猪1 170 bp的D-loop序列核苷酸多样性(Pi)及单倍型多样性(Hd)分别为0.004 25和0.968;32个测序结果共检测到27个多态位点,界定了20种单倍型;中性检验表明淮南猪在驯化进程中曾发生过种群扩张;邻接法(NJ)构建分子系统发育树明显将群体分为国内和国外2个类群,淮南猪20个单倍型都聚集在国内分支上;进一步Network分析显示,淮南猪单倍型在国内猪种中呈广泛分布,但与华中、华北型猪种基因交流较多,且与黄河流域古家猪遗传关系较近,表明淮南猪为多母系起源,主要来自于黄河流域古家猪。     

云南水牛mtDNA D-loop遗传多样性研究

[目的] 探究云南3个水牛品种(德宏水牛、滇东南水牛、盐津水牛)的mtDNA D-loop区的遗传多样性与母系起源。[方法]采用PCR扩增、测序及生物信息学方法。[结果] 本研究共分析了215条云南水牛mtDNA D-loop全序列,检测到107个多态位点,定义了86种单倍型。结果表明,云南3个水牛品种的mtDNA遗传多样性非常丰富,其中,滇东南水牛的遗传多样性最高(Hd: 0.946±0.017,Pi: 0.0162±0.0018),盐津水牛的遗传多样性最低(Hd: 0.805±0.063,Pi: 0.0141±0.0031)。系统发育分析结果表明,在3个云南水牛品种中检测到2个主要支系(A和B支系及其亚支系)及一个稀有支系C,其中A支系为主要支系(79.07%),且经历了强烈的群体扩张。[结论] 云南3个水牛品种具有丰富的mtDNA遗传多样性,主要有A与B两个母系起源。     

广西的尼里—拉菲水牛和摩拉水牛mtDNA D-loop区遗传多样性研究

[目的]探究引入广西南宁的尼里-拉菲水牛和摩拉水牛的mtDNA D-loop区遗传多样性与母系起源。[方法]采用PCR扩增、测序及生物信息学方法。[结果]对从广西南宁采集的52个江河型水牛（尼里-拉菲水牛25个,摩拉水牛27个）mtDNA D-loop序列与GenBank下载的20条尼里-拉菲水牛和23条摩拉水牛序列进行联合分析,共检测到112个变异位点,定义42个单倍型,发现摩拉水牛（Hd: 0.934±0.027）与尼里-拉菲水牛的遗传多样性（Hd: 0.929±0.017）都很丰富。NJ系统发育树显示尼里-拉菲水牛和摩拉水牛含有江河型和沼泽型水牛mtDNA支系,表明尼里-拉菲水牛和摩拉水牛引入中国后均与沼泽型水牛进行了杂交,在外貌上很难区分。[结论]引入广西南宁的尼里-拉菲水牛和摩拉水牛与沼泽型水牛存在广泛的血缘混杂现象。     

Genetic Diversity Analysis of Shaanxi Moschus berezovskii Based on mtDNA Cytb Gene and D-loop Region Sequence

【Objective】 This study was aimed to explore the genetic diversity of Shaanxi Moschus berezovskii population,and understand the genetic information of Moschus berezovskii.【Method】 The hair of Moschus berezovskii was collected to extract DNA,the mitochondrial DNA(mtDNA) cytochrome b(Cytb) gene and D-loop sequences of 43 Moschus berezovskii individuals were determined,and the base composition was counted.All sequences were integrated and compared using ClustalX 2.0 software to obtain nucleotide polymorphic sites (SNPs) in the population.The nucleotide diversity (Pi),number of haplotype (H),haplotype diversity (Hd) and average number of nucleotide differences (K) were calculated by DNASP 5.10 software.The genetic distance among different haplotypes of Cytb gene and D-loop sequences was calculated by Mega 7.0 software,and Neighbor-Joining (NJ) phylogenetic tree was constructed.【Result】 The AT content of Cytb gene and D-loop region were higher than GC content,indicating there was bias in base composition.There were 241 and 383 SNPs of Cytb gene and D-loop region,respectively.The nucleotide diversity of Cytb gene and D-loop region were 0.28343 and 0.07707,and the haplotype diversity was 0.983 and 0.975,respectively,indicating that the population genetic diversity was rich.The genetic distances of 35 haplotypes of Cytb gene ranged from 0.002 to 0.831,and 29 haplotypes of D-loop region ranged from 0.006 to 1.342.The phylogenetic tree showed that there were two mitochondrial lineages,indicating that there were two mitochondrial maternal origins.The evolutionary analysis of D-loop region also supported this conclusion.【Conclusion】 The nucleotide diversity and haplotype diversity of Moschus berezovskii population were high,and the genetic diversity was rich.At the same time further supported the view of Moschus berezovskii and Moshus moschiferus belonged to a branch of the view.     

中国水牛mtDNA D-loop区遗传多样性与母系起源

[目的]检测中国水牛21个群体232条线粒体DNA D-loop 915 bp全序列的遗传多样性及系统进化关系.[方法]PCR扩增、测序和生物信息学方法.[结果]发现232条序列中共有87种单倍型,其核苷酸多态位点88个,其中有79个转换,6个颠换,3个颠换与转换共存.中国水牛mtDNA D-loop区核苷酸多样度(π值) 为0.01403±0.00178, 单倍型多样度(H)为0.8460±0.0240,表明中国水牛mtDNA 遗传多样性丰富.根据单倍型构建了中国水牛的NJ分子系统树,发现中国水牛变异类型主要为两个大支系A和B,表明中国水牛有两个主要母系起源.进一步分析发现支系B具有较大的差异,可以细分为两个亚支B1 和B2.[结论]中国水牛mtDNA 遗传多样性丰富,有2个母系起源.     

基于线粒体DNA控制区(mtDNA D-loop)序列分析贵妃鸡的遗传多样性

采用PCR产物直接测序技术对30只贵妃鸡样品的线粒体DNA控制区(mtDNA D-loop)第Ⅰ高变区序列进行了分析。结果表明,在所分析的D-loop区部分序列中(520 bp),A、G、C、T平均含量分别为26.8%、13.0%、31.1%和29.1%;共发现13个核苷酸多态位点,均为转换位点,未检测到插入/缺失和颠换,核苷酸多样度(Pi)为0.0059,单倍型变异度(Hd)为0.538,中性检验Tajima’s D值为-0.67065。通过群体构建的NJ聚类图分子系统树发现,贵妃鸡起源于红原鸡。     

Analyses of Genetic Diversity and Phylogeny of mtDNA D-loop from Yak in Karakoram-Pamir Area

This experiment was conducted to clarify the genetic diversity,genetic differentiation and phylogenetic status of yak in Karakoram-Pamir area.The mtDNA D-loop region sequence was selected as a molecular marker,and the sequence and genetic diversity of the mtDNA D-loop region of yak in Karakoram-Pamir area were analyzed by PCR direct sequencing and bioinformatics methods.The yak sequence in GenBank was used.The maximum likelihood method was used to construct the phylogenetic tree and the intermediary network relationship.The results showed that the mtDNA D-loop sequence of yak in Karakoram-Pamir area was rich in A and T bases,with AT content of 61.2%,and there were 63 polymorphic loci,accounting for 7.04% of the total number of nucleotides.The results indicated that A and T bases were rich in the mtDNA D-loop sequences at 61.2%.There were 63 mutation sites,accounting for 7.04% of all nucleotides,The average haplotype diversity (Hd) was 0.806,the average nucleotide diversity (π) was 0.01528,and the average nucleotide difference (K) was 13.509,indicating that the yak was rich in genetic diversity in Karakoram-Pamir area;Through phylogenetic analysis,there were two branches in yak in China,forming two branches and six small clades.The yak in Karakoram-Pamir area involved in this study had two different maternal origins.Additionally,yak in the Karakoram-Pamir area was less shared with other breeds of yak haplotypes.In the branch C,the yak group in the Karakoram-Pamir area accounts for a large proportion and was shared with wild yak.The yak population in Karakoram-Pamir area had a unique genetic background,which might be the result of early domestication of wild yaks.It was suggested to increase the identification of yak breeds and the formulation of breed standards in this area,and strengthen the protection of yak genetic resources in this area.According to the current situation of the population,wild blood yaks were introduced for purification and rejuvenation to prevent breed degeneration and decrease of genetic diversity.The introduction of foreign yak breeds and disorderly hybridization were reduced to ensure the characteristics of this breed of high-quality yak breed resources.     

陆川猪mtDNA D-loop序列遗传多样性分析

本研究测定了陆川猪17个个体mtDNA D-loop高变区序列,共检测到5个单倍型,单倍型多样度和核苷酸多样度分别为0.640和0.0074,表明陆川猪有较高的遗传多样性。结合已报道的中国地方猪种的mtDNA D-loop序列进行分析,结果发现,陆川猪和属于华中型的宁乡猪、大花白猪有很近的遗传关系,但与传统上同属华南型的香猪和滇南小耳猪的遗传关系较远,这与传统的分类方法不一致。     

The Genetic Diversity and Phylogenetic Relationships of Five Guangxi Pig Breeds and Three Exotic Pig Breeds by Analysis of mtDNA D-loop Sequences

In order to compare the genetic diversity and phylogenetic relationships between Guangxi pig breeds and three exotic pig breeds,one pair of primers was used to amplify 85 mtDNA D-loop sequences of five Guangxi pig breeds (Bama Xiang pig,Dongshan pig,Guizhong spotted pig,Luchuan pig,Huanjiang Xiang pig),and we downloaded 45 mtDNA D-loop sequences of three exotic pig breeds (Duroc,Yorkshire,Landrace) from GenBank.Then the haplotype,genetic diversity and phylogenetics analysis were carried out.The results showed that 38 haplotypes were identified in 130 sequences,and the mean distances among Guangxi pig breeds were smaller than that among Duroc,Landrace and Guangxi pig breeds.The NJ phylogenetic tree classified all samples into two clusters.One contained all of Guangxi pig breeds and partial Yorkshire,the other was composed wholly of exotic pig breeds,which mean there were two distinct maternal origins.In the cluster of Guangxi pig breeds,Luchuan pig were clustered,and the others were rather dispersive.It indicated that Luchuan pig had simple maternal bloods,and the other breeds had complex maternal bloods,and they had gene flow with each other.The maternal pig resources in Guangxi were not affected by the introduction of exotic pig breeds,and they were still suitable for use as a female parent in their development and utilization.     

Genetic Diversity of Lueyang Black-bone Chicken Based on Mitochondrial DNA D-loop Region Sequence

This study was aimed to assess mitochondrial DNA D-loop sequence diversity and origin of Lueyang Black-bone chicken.The mtDNA D-loop sequence of 30 individuals from Lueyang Black-bone chicken were amplified by PCR and subsequently sequenced.The mtDNA D-loop sequence of other chicken were collected from GenBank and used as reference sequences to analyze the diversity and origin of Lueyang Black-bone chicken.The results revealed that the average values of base composition of A,C,G and T in mtDNA D-loop the sequence of Lueyang Black-bone chicken were 26.6%,26.6%,13.4% and 33.4%,respectively.26 nucleotide polymorphic sites were transition.The average nucleotide diversity (Pi) of the sites and haplotype diversity (Hd) were 0.00705 and 1.000,and the value of Tajima's D was -0.47272.Phylogenetic tree showed that samples were clusted in 4 clades. In the research, it could be concluded that the genetic diversity was relatively rich and wealthy and there were 4 maternal origins to Lueyang Black-bone chicken population.