中国9个家驴品种mtDNA D-loop部分序列分析与系统进化研究

本研究利用Dnasp4.10软件对中国9个家驴品种162个个体的mtDNA D-loop区385bp进行遗传多样性分析,共检测到32种单倍型35个核苷酸多态位点,其单倍型多样度为0.8137～0.9722,核苷酸多样度为0.0182～0.0270,表明我国家驴的遗传多态性丰富;利用MEGA3.1软件采用邻接法与3个努比亚野驴、3个索马里野驴和6个亚洲野驴的序列构建NJ系统发育树,并进行系统进化分析。结果表明:我国家驴的母系起源是非洲野驴中的努比亚野驴和索马里野驴,亚洲野驴不是中国家驴的母系祖先。     

中国四个地方驴品种mtDNA D-Loop部分序列分析与系统进化研究

对中国4个家驴品种95个个体的mtDNAD—loop部分序列进行测序。用clustalX软件进行同源序列比对。Dnasp4-10软件用于遗传多样性分析，计算单倍型多样度、核苷酸多样度、平均核苷酸差异。MEGA3．1软件采用邻接法构建系统进化树并进行系统发生分析。以欧洲驴线粒体基因组为对照（Genbank登录号为X97337），中国家驴4个品种95个个体385bp序列共检测到核苷酸多态位点33个，其中31个位点为转换，1个位点为颠换，1个位点有插入现象。95个序列由31个单倍型组成，单倍型比例为32．6％。德州驴单倍型比例最高为100％，凉州驴单倍型比例最低为42．42％。31个单倍型与引用Genbank已登录的13条序列构建系统发育树，发现31个单倍型聚为两支，说明中国家驴可能有两个母系起源。以欧洲野驴线粒体D—loop为对照（登录号为AF403063、AF403063、AF403065），揭示本研究涉及的中国家驴品种与非洲野驴的进化亲缘关系较亚洲野驴近，并且从分子水平证明中国家驴可能起源于非洲野驴中的索马里野驴分支。     

非州野驴和亚州野驴

在马科动物中非洲野驴是最稀有的动物,这种野驴仅在埃塞俄比亚和索马里的东北部可见。这种有趣的野生动物分为两个亚种:努比亚亚种栖于埃塞俄比亚;索马里亚种可在索马里见到。学者们认为,努比亚亚种是家驴的前身,但其存活数极少(约20头)。 1937年曾经从非洲将1头公驴和3头努比亚亚种母驴运往西欧的慕尼黑。这些野     

六个家驴品种mtDNA D—Loop部分序列的遗传多样性分析

对中国的6个家驴品种(关中驴、德州驴、庆阳驴、泌阳驴、广灵驴、晋南驴)利用PCR-SSCP技术分析了其mtDNA D-Loop部分序列的遗传多样性。87个个体共检测到19个单倍型,单倍型比例为21.8%。单倍型多样度以关中驴、晋南驴较高,分别为0.834和0.904;其次为泌阳驴和广灵驴,分别为0.629、0.529;德州驴和庆阳驴较低,分别为0.467、0.599。文章从分子水平上揭示了家驴的遗传多样性,为我国家驴品种资源的评价和利用提供了一些依据。     

中国家驴生长激素基因内含子2的遗传多样性分析

为了从分子水平揭示家驴的遗传多样性,本研究以中国7个家驴品种(临县驴、关中驴、新疆驴、广灵驴、淮北驴、德州驴、晋南驴)174个个体为试验动物,利用PCR-SSCP技术研究生长激素(GH)基因第2内含子的遗传多样性并进行序列分析.结果显示:第2内含子表现多态性,174个个体检测到3个单倍型,单倍型比例为1.7%.单倍型多样度以临县驴、淮北驴较高,分别为0.678和0.542.广灵驴与关中驴的单倍型多样度(Haplotype diversity,H)比较接近,分别为0.409和0.462.其次为德州驴和晋南驴,分别为0.355和0.304,新疆驴最低为0.077.对该片段的纯合型分别测序发现,B单倍型在第735位碱基G→C.A单倍型在869位碱基G→T.上述结果首次证实驴GH基因内含子2存在多态性.     

家驴与藏野驴SRY基因多态性研究

研究旨在分析家驴与藏野驴SRY基因多态性,为藏野驴的种质资源保护提供理论依据。通过提取2头青海家驴和2头藏野驴的耳组织或毛发DNA,PCR扩增并测序得到1038bp的SRY基因序列,包含5′端调控区330bp和整个编码区708bp。序列比对分析表明,藏野驴与青海家驴SRY基因序列间存在4个核苷酸差异(-265GC,-131TC,493GA,629 AC),其中后2个核苷酸差异发生在编码区,导致相应的氨基酸发生变化(Thr165Ala和Thr210Lys)。构建的系统进化树结果显示,马属动物中家驴先与藏野驴并为一支,后与各种野生斑马相聚,最后与家马聚为一支,提示藏野驴与家驴亲缘关系较近。     

家鸭、媒鸭和野鸭mtDNA D-loop区的遗传变异

旨在从mtDNA水平探讨我国家鸭的遗传多样性及家鸭与野鸭间的亲缘关系.运用DNA测序方法,测定了不同地域、水域和表型性状的9种家鸭(北京鸭、高邮鸭、巢湖鸭、绍兴鸭、连城白鸭、吉安红毛鸭、文登黑鸭、云南麻鸭、建昌鸭)80个个体、4种媒鸭(西湖野鸭、钱江野鸭、枞阳媒鸭、媒头鸭)26个个体和斑嘴鸭8个个体,共计114个个体的mtDNA D-loop区667 bp序列.9种家鸭单倍型多样度和核苷酸多样度分别为0.250 00～0.607 14和0.000 38～0.001 18;9种家鸭、媒鸭、绿头鸭和斑嘴鸭之间的遗传距离为0.001～0.015;39种单倍型系统发生树和单倍型网络关系图表明,9种家鸭主要属于A1亚簇(绿头鸭单倍型簇).我国家鸭遗传多样性中等丰富,母系起源主要是绿头鸭,极少量个体含有斑嘴鸭血统.     

利用线粒体D-loop区分析家鸭品种遗传多态性与系统进化

通过线粒体DNA控制区的结构和多态性来研究我国家鸭的遗传多态性与系统进化。利用DNA测序技术测定了我国9个家鸭品种106个个体线粒体DNA控制区多变序列。序列分析结果显示:A、C、T、G碱基的平均含量分别为25.6%、33.3%、15.2%和25.9%。检测到34个变异位点,约占分析位点总数的5.1%,有转换、颠换、插入/缺失4种类型的变异。确定了31种单倍型,其中单倍型A7为家鸭的主体单倍型,品种之间有9种共享单倍型。9个家鸭品种单倍型多样度(Hd)平均为0.798,核苷酸多样度(Pi)平均为0.28%,单倍型多样度在荆江麻鸭中最高,其次是攸县麻鸭和恩施麻鸭,在文登黑鸭中最低。9个品种家鸭之间双参数距离范围为0.001 3~0.004 4。31个家鸭单倍型序列的系统发生分析表明,9个家鸭品种只有1个母系起源,没有发现东亚斑嘴鸭对9个家鸭品种起源有贡献的证据。     

青海省格尔木牦牛的母系遗传多样性及支系组成

[目的]从分子水平上探究青海省格尔木牦牛的母系遗传多样性、群体遗传结构及其遗传背景。[方法]对49头格尔木牦牛mtDNA D-loop区部分序列进行了测定,后使用DnaSP 5.10.01、Arlequin 3.11和MEGA 5.05等生物信息学软件确定其多态位点和单倍型数目,计算核苷酸多样度和单倍型多样度大小,并进行系统发育分析。[结果]在截取的618 bp格尔木牦牛D-loop区分析序列中,排除1处插入(缺失)后共检测到45处多态位点,包括10处单一多态位点和35处简约信息位点;根据序列间核苷酸变异共确定了18种单倍型,其中单倍型H4为优势单倍型,核苷酸多样度为0.015±0.008,单倍型多样度为0.911±0.022。与野牦牛及大通、天祝、金川等其他家牦牛品种相比,格尔木牦牛群体单倍型多样度和核苷酸多样度值均较高,表明该群体具有丰富的母系遗传多样性。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：格尔木牦牛群体18种单倍型分布在A、B、C、D和G 5种单倍型组中,且聚为3个大的分支,提示格尔木牦牛由3个母系支系组成,拥有3个母系起源。[结论]格尔木牦牛群体具有丰...     

我国部分地方鸭品种mtDNA D-loop区单倍型分析

为进一步研究鸭品种或遗传资源的遗传多样性,对22个地方鸭品种(遗传资源群体)、2个野鸭群体和2个番鸭群体,共计208个个体的mtDNA D-loop区部分DNA序列的多态性进行了检测,结果发现:单倍型多样度(Hd)为0.886±0.00033,平均核苷酸多样度(Pi)为0.02252,并且在所有群体中共发现了60种单倍型,表明我国地方鸭品种遗传多样性丰富,其中单倍型H8在2种野鸭中均有发现,为我国地方鸭品种起源判定提供了新的参考依据。     

The phylogenetic status of typical Chinese native pigs: analyzed by Asian and European pig mitochondrial genome sequences

China is one of the most diverse countries, which have developed 88 indigenous pig breeds. Several studies showed that pigs were independently domesticated in multiple regions of the world. The purpose of this study was to investigate the origin and evolution of Chinese pigs using complete mitochondrial genomic sequences (mtDNA) from Asian and European domestic pigs and wild boars. Thirty primer pairs were designed to determine the mtDNA sequences of Xiang pig, Large White, Lantang, Jinhua and Pietrain. The phylogenetic status of Chinese native pigs was investigated by comparing the mtDNA sequences of complete coding regions and D-loop regions respectively amongst Asian breeds, European breeds and wild boars. The analyzed results by two cluster methods contributed to the same conclusion that all pigs were classified into two major groups, European clade and Asian clade. It revealed that Chinese pigs were only recently diverged from each other and distinctly different from European pigs. Berkshire was clustered with Asian pigs and Chinese pigs were involved in the development of Berkshire breeding. The Malaysian wild boar had distant genetic relationship with European and Asian pigs. Jinhua and Lanyu pigs had more nucleotide diversity with Chinese pigs although they all belonged to the Asian major clade. Chinese domestic pigs were clustered with wild boars in Yangtze River region and South China.     

Mitochondrial diversity of native pigs in the mainland South and South-east Asian countries and its relationships between local wild boars

In this study, we analyzed DNA sequence of mitochondrial DNA (mtDNA) control regions on the 130 native domestic pigs and eight wild boars in the mainland South and South-east Asian countries including Bhutan, Cambodia, Laos, Myanmar, and Vietnam. Forty-four haplotypes were found in the 138 individuals, 41 were in the domestic and four were in wild boars. Only one haplotype was shared by domestic and wild population in Bhutan. In other cases, mtDNA of wild boars did not show close affinity to that of the domestic pigs in the same location, indicating that the native domestic pigs in these countries did not originate in the present habitat. Phylogenetic analyzes of mtDNA haplotypes recapitulated several major clusters identified in other studies, but 11 haplotypes were grouped in a new cluster we named MTSEA. In most cases, more than one lineage group were present in a sampling station, indicating that the present indigenous domestic pigs may have multiple origins. The MTSEA haplotypes were present in relatively high frequencies in domestic pigs in the mountainous area of mainland South-east Asia (Cambodia and Laos), with a few found in Myanmar and Bhutan. The distributions of MTSEA haplotypes are in great conformity with the distribution of present-day Mon-Khmer language and indicated the existence of yet another independent domestication. The D2 haplotypes that distribute high frequency (almost 100%) throughout the Chinese breeds were dominant in Bhutan, Myanmar, and Vietnam. These results suggest an existence of human-mediated dispersal of domestic pigs from north to the south during the historical expansion of Sino-Tibetan and Tai peoples. The D3 haplotypes previously reported in north India were found in sympatric domestic and wild pigs in Bhutan. The D3 haplotype is an important proof of independent domestication event and/or great gene flow between wild and domestic pigs in the foot of Himalaya.     

Haplotypic variability within the mitochondrial gene encoding for the cytochrome c oxidase 1 (cox1) of Cylicocyclus nassatus (Nematoda, Strongylida): evidence for an affiliation between parasitic populations and domestic and wild equid hosts

This study investigated the genetic variability within Cylicocyclus nassatus (Nematoda, Strongylida, Cyathostominae) collected from different domestic and wild hosts (i.e. horse, donkey, Przewalskii horse, tarpan and Turkmen kulan) and localities in Europe and/or USA. The ribosomal Internal Transcribed Spacer 2 (ITS2) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were PCR-amplified and sequences characterized from seventy individual parasitic specimens. While ITS2 displayed 0-0.6% variation rate among all individual adult specimens of C. nassatus examined, 22 different sequence variants (haplotypes) of cox1 were detected. Nucleotide variation was detected at 75 of the total 689 positions (overall 10.8% rate of intraspecific nucletidic difference) in the cox1, with the absence of invariable positions among specimens collected from each equid species or country. Conversely, two haplotypes were detected in horses from USA and in donkeys of Italy and Ukraine, respectively. The absence of haplotypes shared by the equid species suggests an affiliation of C. nassatus populations to their specific host. The results of the present study demonstrated that the characterization of mitochondrial regions may have important implications for studying the genetic structure and biology of equine cyathostomes, and to exploit taxonomic issues and practical implications related to the spread of anthelmintic resistance.     

D‐loop sequence mitochondrial DNA variability of Sarda goat and other goat breeds and populations reared in the Mediterranean area

To provide useful knowledge on goat breed origin and history, we studied the mitochondrial DNA (mtDNA) of 69 goats from five different breeds, Camosciata delle Alpi, Maltese, Nubian, Saanen and Sarda, and one population, the Tunisian. All goats analysed displayed a moderate haplotype and nucleotide diversity. The highest was in the Sarda – the autochthonous breed reared in Sardinia. On the basis of mtDNA control region sequences, animals showed a high genetic haplotype diversity, 35 haplotypes were each represented by a single sequence and only a few haplotypes were shared among the animals. New haplotypes of goats reared in the Mediterranean area were identified and the majority of Italian goats belonged to haplogroup A. This result confirmed worldwide distribution and diversity of haplogroup A.     

The novel polymorphism of the beta 3-adrenergic receptor gene and its distribution in domestic pigs and wild boars in Asia

The beta 3‐adrenergic receptor (ADRB3) is a G protein‐coupled receptor that is involved in regulating energy homeostasis. We have studied DNA sequences of porcine ADRB3 to find candidate genetic polymorphisms for economically important growth and performance traits in pigs. Five novel haplotypes derived from the three In/dels and 44 SNPs were identified among domestic pigs and wild boars. Three of them encode non‐synonymous amino acid sequences by five missense polymorphisms and a frameshift by a thymine insertion. The amino acid polymorphic sites were distributed as follows: one substitution was in extracellular loop 1, three substitutions were in intracellular loop3 and one substitution and the deletion of two amino acids were at the carboxyl‐terminal. There was no polymorphism in the transmembrane domains. In addition, we surveyed the allelic frequency of the thymine insertion that cause frameshift in South‐east Asian local pigs, including some commercial breeds and wild boars. This thymine insertion was distributed widely in the domestic pigs and wild boars. The frequencies of this allele were relatively low in Western improved breeds, while they were very common in Asian breeds and wild boars in Asia. This result indicates that this insertion originally occurred in ancient Asian wild boars and then circulated among Asian domestic pigs. This allele also spread over Western breeds, probably through the introgression of Asian pigs into European stocks during the 18th and 19th centuries.     

Genetic variations and sequences analysis of MTATP6 and MTATP8 genes among different Chinese pig breeds

To investigate the genetic variations of mtATP6 and mtATP8 genes among different Chinese pig breeds, two fragments of 425 and 743 bp containing the whole coding region of mtATP8 and mtATP6 genes were amplified with 805 individuals from 23 Chinese local pig breeds, three types of Chinese wild boars and three European pig breeds. Sequence comparison identified a total number of 17 substitutions including six variable sites in mtATP8 and eleven substitutions in mtATP6 gene. The restriction enzyme Fok I revealed four polymorphic sites (nt8086, 8176, 8514 and 7784), and four RFLP haplotype patterns (A, B, C and D) were identified in mtATP6 and mtATP8 genes among all tested samples. Our data showed AC combined haplotype originated from Asia and BD was regarded as European origin. The average frequency of Asian mtDNA haplotypes was 38.3% across the investigated European breeds but varied within each breeds (13.3～76.7%). Phylogenetic analyses were performed also considering some published sequences in the databases; the sequences were divided into three distinct groups, denoted A, E1, and E2. The Asian AC haplotype existed among the European domestic pigs was fully consistent with the results of previous molecular studies and well-documented history. This study will help us to better understand the genetic variations of mitochondrial genes among different Chinese pig breeds.