百宜黑鸡与兴义矮脚鸡、长顺绿壳蛋鸡的遗传变异研究

测定了12只百宜黑鸡、13只贵州兴义矮脚鸡、8只长顺绿壳蛋鸡共33只鸡个体线粒体DNA控制区部分序列。结果:3个鸡群共33只个体线粒体DNA控制区部分序列为570 bp,共发现38个变异位点,占分析位点总数的6.67%,其中观测到转换23个,颠换15个。百宜黑鸡与其他2种鸡的遗传距离分别0.024 1和0.031 2,表明百宜黑鸡与兴义矮脚鸡、长顺绿壳蛋鸡是不同的种。     

百宜黑鸡mtDNA D-loop区遗传变异分析

为了研究百宜黑鸡线粒体DNA(mtDNA)D-loop区遗传变异情况,试验采用PCR和直接测序法测定了百宜黑鸡、兴义矮脚鸡、长顺绿壳蛋鸡共33个个体线粒体DNA控制区部分序列,长度约为570 bp,分析了百宜黑鸡与兴义矮脚鸡、长顺绿壳蛋鸡之间的遗传变异。结果表明:腺嘌呤(A)、鸟嘌呤(G)、胞嘧啶(C)、胸腺嘧啶(T)4种核苷酸的平均比例分别为26.44%、13.54%、30.75%、29.27%;其中C含量最高,表现出碱基组成的偏倚性。在系统发生树中,百宜黑鸡、兴义矮脚鸡和长顺绿壳蛋鸡各自聚成1支,说明百宜黑鸡与兴义矮脚鸡、长顺绿壳蛋鸡是不同品种。     

贵州兴义矮脚鸡遗传方式研究

本文对兴义矮脚鸡矮脚性状遗传方式进行了研究,结果表明:兴义矮脚鸡自交F1代中只出现矮脚和高脚两个性状,其分离比例接近2:1,经卡方检验.P>0.05,F1代观测数与期望数差异不显著,这是经典的孟德尔3:1比数的变异型.为了进一步确定其遗传方式,利用兴义鸡中高脚和矮脚进行正反交和高脚的自交.结果显示:正反交结果一致,矮脚、高脚的分离比接近于1:1.兴义矮脚鸡中的高脚自交F1代中全部为高脚.孵化过程中出现早期胚胎规律性死亡现象,我们对兴义矮脚鸡胚胎死亡时间和早期死亡率进行统计分析,结果显示:兴义矮脚鸡胚胎死亡时间主要集中在48～72h.早期胚胎死亡率平均达到24.3%,接近25%(P≥0.05).比较兴义鸡分离系矮脚鸡和高脚鸡0～12周龄胫长和体重变化,矮脚鸡胫长、体重显著低于高脚鸡(P<0.05).     

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。     

利用微卫星标记对两个世代百宜黑鸡遗传多样性的研究

采用微卫星标记对百宜黑鸡第4、第5世代的遗传多样性进行研究，结果表明：10个微卫星座位在百宜黑鸡群体中的等位基因频率变化范围较大，为0．0124-0．4876。百宜黑鸡第4世代和第5世代的多态信息含量（PIC）分别为0．6649和0．6418，呈高度多态（PIC〉0．5），说明百宜黑鸡第4世代和第5世代均有丰富的遗传多样性。     

矮脚黄鸡、兴义矮脚鸡血清蛋白多态性的比较研究

利用聚丙烯酰胺凝胶梯度电泳对贵州矮脚黄鸡、兴义矮脚鸡的血清前白蛋白(Pa)、白蛋白(Alb)、后白蛋白(Po)、运铁蛋白(Tf—1,Tf—2)5个位点多态性进行检测。结果表明:除Pa位点在矮脚黄鸡表现为单态,在兴义矮脚鸡表现为多态外,其它4个血清蛋白位点在两鸡种均具多态性,诸位点分别受2～3个(复)等位基因控制。Hardy-Weinberg平衡状态检测结果表明,两群体的Tf—1、Tf—2位点基因型频率都偏离了Hardy-Weinberg平衡状态。兴义矮脚鸡的有效等位基因数和期望杂合度(1.8792,0.4539)较大,矮脚黄鸡(1.7538,0.3772)较小,表明兴义矮脚鸡较矮脚黄鸡的遗传变异度大。     

贵州兴义矮脚鸡与矮脚黄鸡生产性能研究

试验对贵州兴义接脚鸡与矮脚黄鸡生产性能进行了对比研究,结果:兴义矮脚鸡0～45日龄时体重与矮脚黄鸡接近,而在45～90日龄显著高于矮脚黄鸡(P<0.05);胫长在0～75日龄时极显著低于矮脚黄鸡(P<0.01),而在75日龄后低于矮脚黄鸡,表明对兴义矮脚鸡的优势性能值得更进一步开发和利用.兴义矮脚鸡和矮脚黄鸡从初产至12月龄平均产蛋率为42.42%与52.38%,试验结果表明矮脚黄鸡产蛋率高于兴义矮脚鸡.     

略阳乌鸡线粒体DNA控制区(D-loop)的遗传多样性分析

为了研究略阳乌鸡线粒体DNA控制区（mtDNA D-loop）的遗传多样性和起源,本研究对30只略阳乌鸡样品的mtDNA D-loop全序列进行了PCR扩增和测序,结合GenBank中公布的其他鸡的D-loop区序列,分析略阳乌鸡线粒体多态性及其起源。结果表明,略阳乌鸡mtDNA D-loop区全序列中,A、C、G、T平均含量分别为26.6%、26.6%、13.4%和33.4%,26个核苷酸多态位点均为转换位点,核苷酸多样度（Pi）为0.00705,单倍型变异度（Hd）为1.000,中性检验Tajima's D值为-0.47272。通过群体构建的系统进化树发现,略阳乌鸡样品在系统进化树上聚为4大分支。研究结果表明,略阳乌鸡群体内个体序列变异程度较大,遗传多样性丰富,揭示略阳乌鸡在遗传组成上具有4个母系来源。     

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。     

五指山猪线粒体控制区序列分析及遗传多样性研究

为了研究五指山猪遗传多样性和系统地位,本研究采用PCR产物直接测序法得到55条五指山猪线粒体控制区(mtDNA D-Loop)全序列,并结合GenBank中发表的1条五指山猪mtDNA D-Loop全序列进行结构分析、遗传多样性研究及系统发育分析。结果发现,56条序列中出现21次保守区变异,即"TTATAAAACAC"序列重复,共定义13个单倍型,发现14个变异位点,单倍型多样度(Hd)和核苷酸多样度(Pi)分别为0.838和0.00334,表明五指山猪遗传多样性较丰富。五指山猪线粒体控制区序列构建的系统发育树和Network网络分析呈现两大分支,推测有两个母系起源,且与GenBank中其他23个猪种的聚类结果表明,五指山猪与长江流域以南地区的猪种有着较近的亲缘关系。     

中国家鸡和红色原鸡mtDNA控制区遗传多态性及系统进化分析

通过线粒体DNA控制区的结构和多态性来研究中国家鸡和红色原鸡的遗传多态性与系统进化。测定14个中国地方鸡种和红色原鸡2个亚种的256个个体线粒体DNA控制区部分序列约560bp,结果表明,A、C、G、T这4种核苷酸的平均比例分别为25．∞％、37．40％、4．40％和33．20％。共发现44个变异位点,约占分析位点总数的7．86％,没有观测到插入／缺失,颠换和转换之比为0．13;共具有32种单倍型,9种为共享单倍型;16个群体内单倍型多样度从0到0．964,单倍型变异度总体为0．909&#177;0．014,整体的平均核苷酸差异数为7．276,核苷酸多样度为1．851％。群体间核苷酸分歧度（Dxy）在0．747％～3．125％之间变化,核苷酸净遗传距离（Da）为0．015％～2．633％。16个群体表现出较高水平的遗传多态性,群体间表现出显著的遗传分化。群体遗传多态性和亲缘关系分析表明,一些中国家鸡的群体（如固始鸡和仙居鸡）起源于泰国红色原鸡Gallus gallu sgallus亚种,一些中国家鸡的群体（如茶花鸡和藏鸡等）起源于中国红色原鸡Gallus gallus spadiceus亚种,在一些中国地方鸡种还同时具有这2种红色原鸡的遗传贡献;认为中国家鸡起源于泰国或单纯起源于中国的观点都是不全面的。     

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。     

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.     

基于线粒体DNA D-loop区全序列分析儋州鸡遗传多样性及其起源进化关系

为了研究儋州鸡的遗传多样性及其起源进化关系,本研究对36只儋州鸡样品的线粒体DNA(mtDNA) D-loop区全序列进行PCR扩增和测序,结合GenBank中公布的部分品种鸡的mtDNA D-loop区全序列,利用生物信息学方法进行数据处理,分析儋州鸡的遗传多样性及其起源进化关系。结果显示,儋州鸡mtDNA D-loop区扩增片段长度为1 210 bp,A+T含量为59.9%,C+G含量为40.1%,变异区在167～1 215 bp之间,高变区主要集中在167～367 bp之间,存在6种单倍型,共有20个变异位点,单倍型变异度(Hd)为0.571,平均核苷酸差异(k)为6.449,核苷酸多样度(Pi)为0.00537,中性检验的Tajima’s D值为1.61643,6种单倍型可分为A、B、C 3个世系,以B世系为主。研究结果表明,儋州鸡群体遗传多样性和单倍型多样性相对偏低,结合群体构建的系统进化树发现,儋州鸡的遗传组成来自3个母系祖先,缅甸红原鸡、爪哇红原鸡及红原鸡海南亚种均是其潜在的祖先,受外来鸡种影响较小,是一个较为封闭的原始鸡种。     

Study on Genetic Diversity and System Evolution of mtDNA D-Loop Region in Xianglushan Chicken

This study was aimed to analyze the genetic diversity and phylogenetic evolution in Xianglushan chicken.The full-length sequences of mitochondrial DNA D-Loop (mtDNA D-Loop) region of 121 Xianglushan chickens were analyzed by PCR amplification combined with bidirectional sequencing,and the composition,variation and maternal origin of mtDNA D-Loop region were discussed.The results showed that the total length of mtDNA D-Loop region in Xianglushan chicken was 1 231-1 233 bp.The contents of A,T,C and G were 26.62%,33.55%,26.49% and 13.34%,respectively.The content of T was the highest,the content of G was the lowest,and the content of A+T was significantly higher than that of G+C,it indicated that region might have certain base hobbies.Analysis of 121 full-length sequences were found to coexist in 11 haplotypes and 26 mutation sites,of which 2 were single polymorphic information sites,24 were simple information sites,4 bases were inserted and 2 bases were missing.The genetic diversity analysis results showed that the haplotype diversity was 0.814,the nucleotide diversity was 0.00447,the average nucleic acid difference was 5.494,which indicated that the genetic diversity of Xianglushan chicken was relatively rich,the effect of preservation was better,which had a certain breeding space.Tajima's D was 0.39378 and the test results were not significant (P>0.10),in line with neutral mutations.The cluster analysis results showed that Xianglushan chicken and Gallus gallus gathered as one,indicating that Xianglushan chicken originated from Gallus gallus, and there were 4 branches inside the branch,indicating that Xianglushan chicken had many matrilineal origins.The results could provide some reference data for the protection and exploitation of pheasant germplasm resources in Xianglushan chicken.     

Genetic Diversity and Evolution of Danzhou Chicken Assessed with Mitochondrial Complete DNA D-loop Region Sequencing

The objective of this study was to determine the genetic diversity and evolution of Danzhou chicken.The complete mitochondrial DNA (mtDNA) D-loop regions of 36 Danzhou chickens were amplified,sequenced and analyzed.The sequencing reads were compared with the complete mtDNA D-loop sequence of several relative strains of chicken annotated in GenBank,and analyzed by bioinformatics methods.The genetic diversity and its evolutionary relationship in Danzhou chicken were analyzed.The results showed that the lengths of PCR products at the D-loop region were 1 210 bp,with 59.9% being A+T and 40.1% as C+G.The variable regions were 167-1 215 bp,and the high variable regions were mainly 167-367 bp.A total of 20 variable sites that defined 6 haplotypes were identified.The average haplotype diversity (Hd) and average number of nucleotide difference (k) were 0.571 and 6.449,respectively,the nucleotide diversity (Pi) was 0.00537,and the Tajima's D value of neutrality test was 1.61643.6 haplotypes could be grouped to 3 haplogroups (A,B and C) as determined by phylogenic analysis,with B clade,as the most abundant population.It concluded that the genetic diversity and haplotype diversity of Danzhou chicken were relatively low.Phylogenetic tree showed that the genetic composition of Danzhou chicken came from 3 maternal ancestors,Gallus gallus spadiceus,Gallus gallus bankiva and Gallus gallus jabouillei were potential ancestors.There was few influence of exotic lineage detected,which indicated that Danzhou chicken was a relatively conserved breed.     

Genetic Diversity Analysis of Three Newly Discovered Native Chicken Breeds in Yunnan Using mtDNA D-loop Region Sequences

This study was aimed to evaluate the information of their genetic background of Frizzle chicken (FM),Naked-neck chicken (CB) and YN chicken (YN) that were three newly discovered native chicken genetic resources with excellent characteristics,which had been found in Nujiang prefecture and mountainous area of Yunnan province.The variation in a total of 168 individuals sampled from the three chicken populations was assessed using the mitochondrial DNA (mtDNA) D-loop region sequences as genetic marker.The results showed that there were a total of 27 haplotypes were defined in the three native chicken as well as the haplotype diversity of these three chicken were 0.947,0.938 and 0.596,respectively.The nucleotide diversity of these three chicken were 0.01268,0.01434 and 0.00239,respectively.Phylogenetic tree displayed that all 168 individuals distributed in maternal lineage A,B,C,E,F and G.Frizzle chicken contained lineages E,F and G,and Naked-neck chicken contained all 6 lineages,of which the main lineages were E,F and G.YN chicken included lineages E,F and G,and lineage E was the highest percentage in this population.Also,it found that YN chicken was closely related to Gallus gallus murghi,White Plymouth Rock chicken,White Leghorn chicken and New Hampshires chicken,while Naked-neck chicken was closely related to Gallus gallus spadiceus,Gallus gallus jabouillei,Gallus gallus gallus Linnaeus,Indonesian cockfight and Laos chicken.The Frizzle chickens shared more haplotypes with Naked-neck chicken,and the evolutionary relationship between the two species was closer.This study provided a basis for the origin and genetic assessment of three newly discovered Yunnan local chicken breeds.     

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1200 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。