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。     

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。     

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。     

云南昭通黄牛mtDNA D-loop区遗传变异分析

为检测云南昭通黄牛的遗传多样性,测定并分析了云南昭通黄牛线粒体D-loop区段全序列.结果显示,D-loop序列四种碱基A、T、C、G的频率分别是:32.9%、28.6%、24.8%和13.6%;共检测到48个变异位点,平均核苷酸差异(k)为22.762,核苷酸多样度(π)为2.504%,存在6个单倍型,单倍型多样度(Hd)为0.952±0.096,显示出极高的遗传多样性.检索了与昭通黄牛地理分布相近的8个黄牛品种的D-loop序列资料,统计分析发现,9个黄牛品种间的遗传距离在0.0000～0.0376之间,昭通与其他黄牛品种的遗传距离最远,提示云南黄牛有着特殊的起源地位.     

黔西化屋苗寨黑土鸡mtDNA D-loop区遗传多样性分析

为了对黔西化屋苗寨黑土鸡线粒体DNA控制区进行分子遗传学研究,试验采用PCR扩增线粒体DNA控制区第Ⅰ高变区进行直接测序分析的方法。结果表明:在所分析的D-loop区序列(521 bp)中,A、G、C、T 4种碱基的平均含量分别为27.55%、12.60%、29.65%、30.20%;共发现16个核苷酸多态位点,核苷酸多样度(Pi)为0.008 26,单倍型变异度(Hd)为0.594,通过群体构建的NJ系统发育树发现,黔西化屋苗寨黑土鸡起源于红色原鸡,部分个体与乌蒙乌骨鸡、威宁鸡的亲缘关系较近。     

天水乌鸡线粒体DNA D-loop序列遗传多样性分析

试验旨在通过研究天水乌鸡线粒体DNA(mt DNA)D-loop序列的遗传多样性,确定天水乌鸡品种资源价值,从而进行保护和利用。随机抽取50只乌鸡血样,采用PCR和直接测序的方法测定天水乌鸡mt DNA D-loop序列。结果:在天水乌鸡mt DNA D-loop区中发现17个变异位点,10种单倍型;检测片段的G+C含量为47.60%,A+T含量为52.40%;单倍型多样度为0.876±0.023,核苷酸多样度为0.01654±0.01140;在NJ系统发生树上,天水乌鸡大致分为两大支,说明具有2个血统来源。研究表明:天水乌鸡的遗传多样性较为丰富,并且在其进化过程中应存在多个母系来源。     

拉伯高脚鸡线粒体DNA D-loop序列变异与起源分化研究

为从母系遗传角度探明云南拉伯高脚鸡的遗传多样性与起源分化,采用PCR产物直接测序技术对30只拉伯高脚鸡的线粒体DNA控制区(mtDNA D-loop)第一高变区序列进行了分析。结果表明:在所分析的mtDNA D-loop 527 bp序列中,共检测到17个变异位点,归结为5个单倍型。单倍型L1和L2属G世系,单倍型L3和L4属A世系,单倍型L5属B世系。G世系占整个群体的63.3%,A世系占33.3%。单倍型多样度为(0.763±0.049),群体内序列间核苷酸差异的平均数为7.205,核苷酸多样度为0.01315,平均遗传距离为0.016。由最大似然法构建的NJ系统进化树将拉伯高脚鸡聚为3大枝,分别与G、A、B 3个世系对应。与红原鸡及国内外鸡种聚类比较,拉伯高脚鸡在起源上具有其独特性。结果表明,拉伯高脚鸡存在较高的遗传多样性,起源上具有云南本地鸡特有的遗传特征。     

新疆野生盘羊mtDNA D-loop序列的多态性

根据GenBank中3条羊亚科动物线粒体DNA全序列设计并合成1对引物,成功扩增了新疆天山亚种野生盘羊线粒体(mtDNA)控制区(D-loop)全长序列片段,并进行序列分析。结果显示,1号和2号盘羊mtDNA D-loop序列全长分别为1 070,1 169 bp;在检测的2个个体上共发现73个突变位点,其中有7个插入,50个转换,11个颠换,5个缺失,表明碱基的替换有明显偏倚;1号和2号新疆天山亚种野生盘羊mtDNA D-loop区核苷酸突变位点分别为35个和38个,核苷酸变异率分别为3.27%和3.25%。这一结果表明,新疆天山亚种野生盘羊群体线粒体D-loop区存在着丰富的多态性,同时进一步说明了我国新疆野生盘羊遗传资源比较丰富。     

安宁果下马线粒体DNA D-loop序列的初步分析

为了解安宁果下马线粒体DNA(mtDNA)的序列特点,试验采用普通马mtDNA序列设计PCR引物,对4匹安宁果下马的mtDNA D-loop区进行扩增的方法,得到mtDNA D-loop区的全序列。结果表明:4匹马mtDNA D-loop区的平均核苷酸变异率为2.67%,核苷酸变异位点较多,变异类型包括颠换、转换、插入、缺失4种形式,其中转换最普遍;检测到重复序列(GTGCACCT)的数量为24个,不同于其他马种。4匹安宁果下马个体间mtDNA D-loop区差异较大,表明其mtDNA D-loop区丰富的多态性。     

寿光鸡遗传多样性及其起源进化的研究

为了更好地对寿光鸡种质资源进行保护和开发利用,以线粒体基因组(mitochondrial genome,mt DNA) D-loop区为分子标记,检测了58只寿光鸡的遗传多样性,同时探讨了寿光鸡在5个红色原鸡亚种的进化地位。结果显示:寿光鸡mt DNA D-loop区全长1 231～1 232 bp,核苷酸多样性(Pi)为(0. 005 3±0. 001 35),单倍型多样性(Hd)为(0. 864±0. 018),平均核苷酸差异(K)为6. 525;群体共发现23处单核苷酸变异位点,并由此界定了8种单倍型;系统发育分析发现,寿光鸡主要分布在A、B、C和E 4个进化分支,构成比分别为43. 1%、36. 2%、12. 1%和8. 6%。结果表明:寿光鸡在线粒体水平上具有很高的遗传多样性,有4个母系来源,本研究为寿光鸡种质资源遗传评估、有效保种和选育利用提供了理论依据。     

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.     

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 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.     

普洱毛脚乌鸡与南涧绿耳乌鸡线粒体DNA母系遗传分析

In order to elucidate the genetic differences between Puer Miaojiao and Nanjian Lver black bone chicken, the hypervariable region I sequence variations in mtDNA D loop of the respective 48 random samples from Puer Miaojiao and Nanjian Lver black bone chicken were examined by direct sequencing of PCR products in the present study, and an analysis of population genetics and phylogenetics were conducted on the data obtained. In Puer Miaojiao black bone chicken, a total of 24 polymorphic sites included 2 singleton polymorphic sites and 24 parsimony informative sites were detected, which defined 12 haplotypes and the unique haplotypes accounted for 50%. The haplotype diversity, nucleotide diversity and the average genetic distance within the population were 0.829±0.039,0.01559±0.00068 and 0.016±0.003, respectively. In Nanjian Lver black bone chicken, a total of 28 polymorphic sites included 0 singleton polymorphic site and 28 parsimony informative sites were detected, which defined 13 haplotypes and the unique haplotypes accounted for 53.85%. In this population, the haplotype diversity, nucleotide diversity and the average genetic distance within the population were 0.907±0.018,0.01590±0.00056 and 0.016±0.003, respectively. Phylogenetic and genetic divergence analysis revealed that there were A,B,E,F and G five matrilineages in Puer Miaojiao black bone chicken which accounted for 45.83%,2.08%,8.33%,16.67% and 27.09%, respectively. And there were A,B,C, E,F and G six matrilineages in Nanjian Lver black bone chicken which accounted for 22.92%,14.58%,4.17%,10.42%,16.67% and 31.25%, respectively. The results in this study indicated that the population genetic variation in Nanjian Lver black bone chicken are higher than that in Puer Miaojiao black bone chicken and the population genetic composition for the two chicken populations were significantly different. They are mainly for the native chicken descent by birth, but there may be a small amount of commercial chicken blood mixed.     

基于线粒体COⅠ基因的2个新发现鸡种资源DNA编码研究

以中国2个新发现鸡种资源金湖乌凤鸡、安义瓦灰鸡及地方鸡种狼山鸡为研究对象,利用DNA测序技术测定了线粒体细胞色素C氧化酶亚基Ⅰ（cytochrome c oxidase Ⅰ,COⅠ）基因序列,揭示2个新发现资源COⅠ基因的遗传多样性及其与地方鸡种的遗传关系。结果显示,选择的这段COⅠ基因序列在2个新发现资源和狼山鸡中有10个突变位点,为7个单倍型,其中有3个为特异性突变位点,5个特异性单倍型。金湖乌凤鸡、安义瓦灰鸡和狼山鸡群体内平均核苷酸多样度（Pi）分别为0.068%;0.051%和0.0369%,单倍型多样度（H）分别为0.4167、0.3030和0.9000。狼山鸡的遗传多样性大于2个新发现资源。3个地方鸡群体间的Kimura双参数遗传距离范围为－0.002～0.236;种间遗传距离小于种内遗传距离。结果表明,利用COⅠ这一特定基因的特定区段来做DNA条形编码的基础,进行不同鸡品种鉴定具有方便、快捷、经济、准确等优点,是可行和有效的。     

Genetic variation of major histocompatibility complex BLB2 gene exon 2 in Hebei domestic chicken

Genetic variation of MHC BLB2 gene exon 2 in Hebei domestic chicken was investigated, after PCR and sequencing of a 374bp fragment (containing entire exon 2 (270bp) of BLB2 gene) in 76 individuals. The results showed that along this fragment, there were 69 variable sites, of which 18 were novel variations, and 82 estimated haplotypes with the diversity of 0.960. In Hebei domestic chicken, the nucleotide diversity (π), the average number of nucleotide differences (k), the average number of nucleotide diversity of synonymous substitution (π(s)) and non-synonymous substitution (π(a)) in BLB2 gene exon 2 were 0.098, 24.688, 0.075, and 0.106, respectively; nine non-synonymous substitutions was exclusively found in the peptide-binding sites (PBS) region of BLB2 gene exon 2, inferring that these unique substitutions might be helpful to resist some special bacteria and pathogens. The higher genetic diversity of MHC BLB2 gene exon 2 in Hebei domestic chicken might be consistent with its more robust disease resistance.     

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.     

鸡MHC基因B-G区域的生物信息学分析

根据鸡主要组织相容性复合体（MHC）基因B-G区域外显子2的序列设计引物,采用直接测序的方法对该区域（207bp）进行序列测定,并对所获得核苷酸序列进行生物信息学分析。结果显示：该片段中碱基T、C、A、G的平均含量分别为22.0%、19.8%、24.8%、33.4%,C＋G含量为53.2%,T＋A含量为46.8%,C＋G含量高于T＋A含量;同时检测到27个核苷酸变异位点,导致14个氨基酸位点的变异;16条序列进行单倍型构建获得16种单倍型,单倍型多样度达到了高的多样水平。试验结果表明,鸡MHC-B-F基因外显子2的序列表现出了高度的多态性,而且其序列的多态性更多地表现在氨基酸水平上。     

坦布苏病毒鸡源分离株全基因组及遗传变异分析

为了解引起鸡产蛋骤降的坦布苏病毒全基因组分子特征,本试验运用RT-PCR技术克隆测定了2株坦布苏病毒鸡源分离株全基因组序列,其基因组为10 990nt,包含1个10 278nt的开放阅读框架,与GenBank数据库中登录的坦布苏病毒参考株一致,与2010年以来的坦布苏病毒毒株核苷酸及推导氨基酸同源性均在98%以上,与2010年前的坦布苏病毒核苷酸和氨基酸序列同源性仅分别为88%和96%左右。基于坦布苏病毒全基因组序列的分子系统进化分析,发现2株鸡源坦布苏病毒与2010年以来分离的各种禽源坦布苏病毒株均处于同一大的进化分支,且相互间的遗传距离均小于2%。以上研究结果表明,坦布苏病毒鸡源分离株与其他禽源病毒分离株的基因组差异不明显,各分离株亲缘关系非常近。