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。     

西藏部分山羊群体线粒体DNA D-loop区遗传多样性及系统进化分析

为了探究西藏不同山羊群体的遗传多样性和亲缘关系,提取4个山羊群体DNA,扩增其mtDNA D-loop区,并测序。结果显示:西藏山羊群体mtDNA D-loop区长度在1 200～1 212 bp,各群体山羊D-loop区富含A、T碱基,共发现106个多态位点,分离出21个单倍型。西藏山羊群体单倍型多样性(Hd)和核苷酸多样性(Pi)分别为0.085 7～1.000 0,0.007 04～0.019 14,4个群体山羊碱基突变率高,表明西藏山羊的遗传多样性非常丰富。核苷酸歧义度、NJ系统进化树表明西藏山羊群体间有共同母系血源,部分支系母系起源于镰刀型角野山羊(Capra aegagrus)和捻角山羊(Capra falconeri),同时存在其他的母系起源,支持山羊品种内的多起源说。     

中国部分地方水牛品种mtDNA D-loop区遗传多样性与起源研究

对我国10个地方水牛品种110个个体的mtDNA D-loop区序列(930 bp左右)进行分析,共检测到50种单倍型,107个核苷酸多态位点,其单倍型多样度(Haplotype diversity,Hd)为0.895 2±0.024 0,核苷酸多样度(Nucleotide diversity,π)为0.020 0±0.005 6,平均核苷酸差异(Average number of nucleotide differences,k)为18.445 0,表明我国水牛的遗传多态性丰富。构建的NJ进化树显示这10个品种的水牛主要有两个母系起源。     

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.     

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。     

蒙古家马和蒙古野马mtDNA D-loop区遗传多样性及母系起源分析

为了对马线粒体DNA(mtDNA)D-loop区序列的遗传变异进行分析,探讨蒙古家马、蒙古野马和驴之间的亲缘关系和遗传多样性,试验对各类马样本mtDNA D-loop区序列进行PCR扩增和测序,运用分子生物学软件进行数据处理。结果表明:共检测到5个单倍型,各类马单倍型多样性和核苷酸多样性分别介于0.400~1.000和0.009 25~0.028 90之间,说明蒙古家马有较高的遗传多样性,而蒙古野马单倍型多样性和核苷酸多样性最小,蒙古野马多态程度较低。根据D-loop区序列构建的NJ树发现,蒙古家马和蒙古野马有较近的亲缘关系,而且蒙古家马均匀地分散在各个分支中,说明蒙古家马有丰富的母系起源。     

溧阳鸡线粒体DNA D-loop区遗传多样性研究

为了研究溧阳鸡线粒体DNA(mtDNA)D-loop区遗传多样性,试验采用PCR产物直接测序法对30只溧阳鸡mtDNA的D-loop区序列进行分析。结果表明:溧阳鸡549 bp D-loop区序列的T、C、A、G平均含量分别为30.0%、29.9%、27.1%、13.0%,共检测到19个突变位点,其中单一多态位点3个,简约多态位点16个;序列的核苷酸多样性为0.007 63,单倍型多样性为0.662;共获得7种单倍型,其中Hap单倍型占56.7%,群体内遗传距离为0.008。结合NJ系统发生树发现,溧阳鸡存在3个分支,揭示溧阳鸡在遗传组成上具有3个母系来源。     

西林水牛和富钟水牛mtDNA D-loop序列的遗传多样性研究

该研究旨在探究西林水牛和富钟水牛mtDNA的遗传多样性和母系起源。采用PCR扩增、测序及生物信息学方法,测定了60头西林水牛和富钟水牛的mtDNA D-loop序列。结合GenBank数据库已公布的25条西林水牛和富钟水牛的mtDNA D-loop序列联合进行多态性分析,共检测到60个多态位点,定义了38种mtDNA单倍型。西林水牛和富钟水牛的单倍型多样度分别为0.909±0.028和0.913±0.035,核苷酸多样度分别为0.019±0.004和0.013±0.005,这一结果表明西林水牛和富钟水牛具有丰富的遗传多样性。NJ系统发育树显示,西林水牛和富钟水牛均属于沼泽型水牛,且有A支系和B支系两个母系起源。     

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。     

唐古拉山牦牛群体的母系遗传多样性及遗传结构分析

[目的]从分子水平上探究青海省唐古拉山牦牛群体的母系遗传多样性、群体遗传结构及其遗传背景。[方法] 对52头唐古拉山牦牛个体mtDNA D-loop区序列进行测定后,使用生物信息学软件分析确定其核苷酸变异位点和单倍型数目,计算单倍型多样度和核苷酸多样度大小,并进行系统发育分析。[结果] 在619 bp唐古拉山牦牛D-loop区序列分析中,排除2处插入（缺失）后共检测到31处多态位点,包括单一多态位点5处和简约信息位点26处。根据序列间核苷酸变异共确定了13种单倍型,单倍型多样度和核苷酸多样度分别为0.821±0.043和0.007±0.004。与我国其他18个家牦牛品种和野牦牛相比,唐古拉山牦牛群体单倍型多样度和核苷酸多样度值均较低,表明该群体遗传变异较为贫乏,母系遗传多样性水平较低。以美洲野牛为外群,邻接法（即NJ法）构建的系统发育树结果显示：唐古拉山牦牛群体13种单倍型分布在A、B、C、D和E五种单倍型组中,且聚为2个大的母系分支（即I和II）,支系Ⅰ占比为77%,提示唐古拉山牦牛由2个母系支系组成,拥有2个母系起源且以支系Ⅰ为主。 [结论] 唐古拉山牦牛母系遗传多样性水平较低,由2个母系支系组成,以支系Ⅰ为主,推测其有2个母系起源。     

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.     

Genetic Diversity of Mitochondrial DNA D-loop Sequences in Huili Black Goat

To explore the genetic diversity and origin for genetic resource protection of Huili Black goat, the mitochondrial DNA (mtDNA) D-loop was investigated. mtDNA D-loop sequences of 41 goats were analyzed by PCR, sequencing techniques, and biological information and the phylogenetic trees were constructed. The mtDNA sequences of the Huili Black goat ranged from 1211 to 1213 bp, and 2 sequences were 1211 bp, 29 sequences 1212 bp, and 10 sequences 1213 bp. The content of A+T (60.1%) was higher than one of G+C (39.9%). There were 9 haplotypes, and the haplotype diversity was 0.842+0.00368. The nucleotide diversity was 0.01542+0.00034. The phylogenetic analysis showed that Huili Black goat was distributed in a branch, and were closed to Jianchang Black goat, Chengdu Ma goat, Jintang Black goat, Guizhou White goat, Guizhou Black goat, but they were less related to Capra falconeri. Huili Black goats had rather abundant genetic diversity, and were greatly affected by other goat breeds in history.     

关中驴线粒体DNA D-loop多态性分析

本文对 6头关中驴线粒体DNAD loop区 399bp的核苷酸序列进行了分析。结果发现 ,关中驴的D loop区核苷酸变异只有转换 1种形式。 6头关中驴D loop区的核苷酸序列组成 3种单倍型 ,单倍型比例为 5 0 .0 0 % ,说明关中驴mtDNA遗传多样性正逐步丧失 ,需要加强驴种质资源保护。以欧洲驴D loop序列为对照 ,关中驴 3种单倍型的核苷酸变异率分别为 4 .2 1%、4 .5 1%和 0 .2 5 %。在获得的 399bpD loop碱基序列中 ,共检测出核苷酸多态位点18个 ,多态位点比例为 4 .5 1%。从D loop区核苷酸序列的 3种单倍型分析 ,发现关中驴可能有 2种不同的母系起源     

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.     

Phylogenetic relationships of German heavy draught horse breeds inferred from mitochondrial DNA D-loop variation

Summary We analysed a 610-bp mitochondrial (mt)DNA D-loop fragment in a sample of German draught horse breeds and compared the polymorphic sites with sequences from Arabian, Hanoverian, Exmoor, Icelandic, Sorraia and Przewalski's Horses as well as with Suffolk, Shire and Belgian horses. In a total of 65 horses, 70 polymorphic sites representing 47 haplotypes were observed. The average percentage of polymorphic sites was 11.5% for the mtDNA fragment analysed. In the nine different draught horse breeds including South German, Mecklenburg, Saxon Thuringa coldblood, Rhenisch German, Schleswig Draught Horse, Black Forest Horse, Shire, Suffolk and Belgian, 61 polymorphic sites and 24 haplotypes were found. The phylogenetic analysis failed to show monophyletic groups for the draught horses. The analysis indicated that the draught horse populations investigated consist of diverse genetic groups with respect to their maternal lineage.     

基于线粒体控制区的鸡不同杂交组合遗传多样性研究

旨在探讨鸡不同杂交组合线粒体控制区（mtDNA D-loop区）的遗传多样性和单倍型特性。选取固始鸡和隐性白羽鸡及其正、反交F1代、藏鸡以及F2代等6个群体共387个个体的mtDNA D-loop区进行测序,分析其遗传规律和单倍型特性,并与不同红色原鸡亚种进行聚类,分析其母系起源。结果显示,6个群体D-loop区全序列大小为1 231 bp,共检测到28个多态位点和1个C碱基缺失,共构成19种单倍型,分为A、B、C和E 4个单倍型群,其中,固始鸡和反交F1代主要为A、C单倍型,固始鸡A、C单倍型比例分别为53.42%和46.58%,反交F1代A、C单倍型比例分别为50.75%和49.25%;隐性白羽鸡、正交F1代和F2代优势单倍型均为E单倍型,占比分别为48.89%、48.84%和50.00%。6个鸡群体单倍型多样度（Hd）在0.496~0.729之间,核苷酸多样度（Pi）在0.003 40~0.005 41之间,Hd值和Pi值最大的均为正交F1代,其次为隐性白羽鸡和F2代,固始鸡和反交F1代群体遗传多样性接近。聚类分析显示,A、B单倍型群与滇南亚种交叉聚为一枝;E单倍型群与印度亚种交叉聚为一枝;C单倍型群与印度亚种、指名亚种、印尼亚种以及滇南亚种聚为一枝。结果提示,mtDNA D-loop区遵循严格的母系遗传,后代的遗传多样性和单倍型比例与其母本基本一致;我国家鸡群体具有多个红色原鸡母系起源,且主要起源于原鸡滇南亚种。     

A Genetic Diversity Analysis of Mitochondrial DNA D-loop Region in Four High Quality Chicken Breeds

This study was conducted to elucidate the genetic diversity of mitochondrial DNA (mtDNA) D-loop region in Qingyuan partridge chicken group 1,Qingyuan partridge chicken group 2,Yangshan chicken and Qingyuan Yellow feather black-bone chicken.The specific primers were designed according to mtDNA D-loop region of Gullus gullus spadiceus (accession No.:NC_007235.1) in GenBank.The sequence was analyzed after PCR amplification and sequencing,and the haplotype number,polymorphism number,haplotype diversity,nucleotide diversity and nucleotide mean difference were counted.The evolution divergence among breeds was calculated by Mega 5.10 software,and the phylogenetic tree was constructed.The results showed that the length of mtDNA D-loop region in four high quality chicken breeds was 591 bp,and 549 bp were used for subsequent analysis.The content of A,T,C and G were 27.2% to 27.3%,30.1% to 30.4%,29.5% to 29.8% and 12.8% to 12.9%,respectively,and the average content of G+C was 42.5%.There were 92 polymorphic sites which contained 14 singleton variable sites and 78 parsimony informative sites,and the percentage of transitions and transversions were 89.13% (82/92) and 10.87% (10/92),respectively.The haplotype diversity ranged from 0.682 to 0.835,and the nucleotide diversity ranged from 0.00849 to 0.01167.There were 32 haplotypes in all sequences,which could be divided into clades A,B,C and E,however,most of the individuals belonged to clades B (51.2%) and E (37.6%).The phylogenetic tree results showed that four high quality chicken breeds could be classified as 4 branches which were consistent with the haplotypes classification results.The results indicated that the four high quality chicken populations from Qingyuan had relatively high haplotype and nucleotide diversity and likely shared two or more common maternal lineages.     

Genetic diversity of Bangladeshi native chickens based on complete sequence of mitochondrial DNA D-loop region

ABSTRACT

1. The aim of this study was to explore genetic diversity and possible origin of Bangladeshi (BD) native chickens. The complete mtDNA D-loop region was sequenced in 60 chickens representing five populations; naked neck, full feathered, Aseel, Hilly and autosomal dwarf. The 61 reference sequences representing different domestic chicken clades in China, India, Laos, Indonesia, Myanmar, and other Eurasian regions were included. The mtDNA D-loop sequence polymorphism and maternal origin of five BD populations were analysed.

2. A total of 35 polymorphic sites, and 21 haplotypes were detected in 60 mtDNA D-loop sequences. The haplotype and nucleotide diversity of the five populations were 0.921 ± 0.018 and 0.0061 ± 0.0019, respectively. Both mtDNA network and phylogenetic analysis indicated four clades (four haplogroups) in BD populations (21 haplotypes) along with 61 reference haplotypes. Clade E contained the most individuals (20) and haplotypes (11) of BD chickens, followed by clade D (17, 6), clade C (12, 2) and clade F (11, 2), respectively.

3. The higher number of unique haplotypes found in Yunnan, China, suggested that the origin of BD chickens was in this region. The haplotypes from different haplogroups were introduced in Bangladeshi chickens from India, China and Myanmar. The phylogenetic tree showed a close relationship of BD chickens with the clusters from India, China, Myanmar and Laos, and indicated the dispersion of BD chickens from these sources. The phylogenetic information revealed high genetic diversity of BD chickens because of their origin from different lineages with high genetic variation and distance, which was determined from four cluster and neighbour-joining trees.

4. In conclusion, BD populations had high genetic diversity. The mtDNA network profiles and phylogenetic trees showed multiple maternal origins of BD chickens from India, China, Myanmar and Laos.