大足黑山羊与周边黑山羊品种(群体)mtDNA D-loop序列多态性研究

本实验旨在研究大足黑山羊与川、渝、滇三地的其他4个黑山羊品种(群体)遗传多样性及聚类关系,采用直接测序法分析了这些山羊品种(群体)的mtDNA D-loop序列多态性。结果表明:5个黑山羊群体的碱基组成没有明显差别;5个黑山羊品种mtDNA D-loop序列多态性较丰富;聚类分析显示,大足黑山羊与渝东黑山羊的遗传距离最近,大足黑山羊与川北黑山羊和乐至黑山羊遗传距离次之,与圭山黑山羊遗传距离最远,这与此5个黑山羊品种地理分布相符。     

5个黑山羊品种(群体)微卫星标记的遗传多样性研究

利用14个微卫星标记分析了攀枝花地方黑山羊、建昌黑山羊、云岭黑山羊、金堂黑山羊、乐至黑山羊5个品种(群体)的遗传多样性及亲缘关系。结果显示,14个微卫星座位多态信息含量(PIC)在0.5129~0.7646之间,均为高度多态位点;5个黑山羊品种(群体)的平均多态信息含量(PIC)为0.6405~0.6856,平均杂合度(H)在0.6985~0.7371之间,说明遗传多样性丰富;群体间平均遗传分化系数(Fst)为0.0296,说明群体间的变异仅为2.96%,群体间存在基因交流;攀枝花黑山羊和金堂黑山羊2个群体之间的Nei氏遗传距离最大(D=0.1286),金堂黑山羊和乐至黑山羊之间的遗传距离最小(D=0.0365);NJ聚类图中,攀枝花黑山羊、建昌黑山羊和云岭黑山羊聚为一类,金堂黑山羊和乐至黑山羊聚为一类,聚类结果与群体的地理分布较为一致。     

攀西黑山羊群体的AFLP遗传多样性研究

 研究采用随机片段长度多态性分析了攀西地区的5个黑山羊品种（类群）的遗传多样性。结果表明：15对引物共扩增出1003条多态条带,多态频率平均为99.01%,5个黑山羊群体的遗传多样性指数变异范围为0.0136~0.2131,其中建昌黑山羊平均遗传多样性指数最高（0.1346）,攀枝花黑山羊次之（0.1329）,乐至黑山羊最低（0.1101）。5个群体遗传距离范围为0.0062~0.0281,攀枝花黑山羊与建昌黑山羊之间的遗传距离最小（DR=0.0062）,云岭黑山羊与攀枝花黑山羊之间的遗传距离次之（DR=0.0128）,乐至黑山羊与金堂黑山羊间遗传距离最大（DR=0.0281）,金堂黑山羊与其它4个类群遗传距离较大（DR=0.0173-0.0281）,说明攀枝花黑山羊与建昌黑山羊的亲缘关系最近,与云岭黑山羊的亲缘关系较近,乐至黑山羊与金堂黑山羊的亲缘关系最远,金堂黑山羊与其它4个类群遗传距离均比较远。聚类结果与群体地理分布、生态条件差异情况基本一致。     

用5个微卫星标记分析四川7个地方山羊品种的遗传关系

选用5个微卫星标记对四川7个地方山羊品种(类群)进行了遗传关系分析。结果表明:5个微卫星标记检测到的等位基因数从1～7个不等,平均多态信息含量为0.537～0.716,其中ILSTS004、ILSTS030、McM038和OarFCB011这4个基因座位为高度多态性基因座,CSSM004为中度多态基因座,都能较好地用于山羊群体间的遗传多样性分析。7个山羊群体的平均多态信息含量为0.601～0.696,群体平均杂合度为0.530～0.655,群体间的遗传分化系数为0.1167,表明7个地方山羊群体的遗传多样性较为丰富。群体间的Nei氏遗传距离为0.0822～0.7240,以Nei氏遗传距离为基础构建的系统聚类树表明,藏山羊、建昌黑山羊、成都麻羊和北川白山羊聚为一大类,第2类为乐至黑山羊和金堂黑山羊,最后简阳大耳羊单独成一类。     

成都麻羊乳上皮黏蛋白MUC1的遗传多态性研究

采用SDS-PAGE研究成都麻羊乳上皮黏蛋白MUC1的遗传多态性,并以金堂黑山羊、乐至黑山羊、自贡黑山羊、藏山羊作对照进行比较分析。结果表明:成都麻羊与对照山羊乳MUC1的多态性丰富,成都麻羊有7种基因型、5个等位基因优势基因为C、D、E,金堂黑山羊7种基因型、6个等位基因优势基因为B、C、D,乐至黑山羊6种基因型、6个等位基因优势基因为B、C、D,自贡黑山羊5种基因型、5个等位基因优势基因为A、C、D,藏山羊3种基因型、3个等位基因优势基因为C、D。乳MUC1的基因型、等位基因的分布品种间有差异;根据乳MUC1等位基因频率对供试山羊品种进行聚类分析,成都麻羊与金堂黑山羊的遗传距离最小(D=0.2507)先聚为一类,再依次与乐至黑山羊(D=0.2831)、自贡黑山羊(D=0.3721)、藏山羊(D=0.4752)聚为一大类,结果较好的反映了成都麻羊与对照山羊品种间的亲缘关系。     

西南地区5个山羊遗传新资源的遗传多样性和亲缘关系分析

为研究新批准的西南地区5个山羊遗传资源的遗传多样性及这5个山羊遗传新资源与其他羊种的亲缘关系,研究扩增了山羊的mtDNA D-loop序列并测序,对序列进行了核苷酸多样性、单倍型和亲缘关系分析。结果表明:在西南地区5个新的山羊遗传资源中,大足黑山羊、黔北麻羊和酉州乌羊平均多态位点数比较接近,均高于渝东黑山羊和贵州黑山羊;大足黑山羊、黔北麻羊和酉州乌羊遗传多样性高于贵州黑山羊和渝东黑山羊,渝东黑山羊平均多态位点和遗传多样性在5个羊种中最低。从亲缘关系来看,酉州乌羊和渝东黑山羊与板角山羊亲缘关系近,贵州3个羊种(贵州白山羊、贵州黑山羊和黔北麻羊)遗传距离近,大足黑山羊与乐至黑山羊遗传距离较近。研究认为,渝东黑山羊和贵州黑山羊遗传多样性较低,需要加强遗传资源的保护和提纯。     

新批准的西南地区5个山羊遗传资源mtDNA D-loop遗传多样性与亲缘关系研究

为了研究新批准的西南地区5个山羊遗传资源的遗传多样性及其与其他羊种的亲缘关系,试验利用设计的引物对山羊的mtDNA D-loop序列进行了PCR扩增和测序,对序列数据进行了核苷酸多样性、单倍型和亲缘关系分析。结果表明:在西南地区新批准的5个山羊遗传资源中,大足黑山羊、黔北麻羊和酉州乌羊平均多态位点数比较接近,均高于渝东黑山羊和贵州黑山羊;大足黑山羊、黔北麻羊和酉州乌羊遗传多样性高于贵州黑山羊和渝东黑山羊;从亲缘关系来看,渝东黑山羊与板角山羊和南江黄羊亲缘关系较近,酉州乌羊与板角山羊亲缘关系最近,贵州3个羊种(贵州白山羊、贵州黑山羊和黔北麻羊)遗传距离近,大足黑山羊与乐至黑山羊遗传距离较近。说明渝东黑山羊和贵州黑山羊遗传多样性较低,需要加强遗传资源的保护和提纯;新批准的5个山羊遗传资源的遗传距离和亲缘关系与其所处的地理位置存在密切关系。     

基于SNP芯片的云上黑山羊遗传结构分析

为从分子水平对云上黑山羊遗传结构做出评价,本研究以云上黑山羊核心群母羊108只(来自7个育种核心场、10个群体)、育种素材云岭黑山羊和努比山羊各11和14只、对照组波尔山羊10只为试验动物,用Illumina公司Iselect Goat60k芯片技术在全基因组范围内进行单核苷酸多态性(SNP)分析,并运用GenAlEx 6.5、Cervus 3.0、SNPRelate、Plink 1.07、Mega 4.0进行遗传多样性参数统计计算、主成分分析和系统进化树构建。试验获得143个样本在48 116个SNPs位点上的分型结果,其中波尔山羊10个样本单独聚集,与云上黑山羊、努比山羊、云岭黑山羊存在较大的遗传距离,较小的遗传一致度和较小的基因流,在聚类进化树中显示为一个独立分支,这些结果与波尔山羊实际遗传背景完全一致,显示出它在本研究中的对照组作用,同时证明本研究方法可行、结果可靠;云上黑山羊108个样本具有高度一致性,在聚类系统树中这些样本聚为一个大簇,而努比山羊(14个)和云岭黑山羊(11个)的样本各自聚为一簇;UPGMA进化树显示,云上黑山羊与努比山羊关系较近,与云岭黑山羊关系稍远,这与云上黑山羊育种导血方案结果相一致。云上黑山羊新品种在基因组水平上可与其育种素材明显区分,在基因组层面已具备独立品种特点。     

利用微卫星标记对7个山羊品种的遗传多样性研究

为了研究小香羊与相邻产区山羊品种之间的亲缘关系,利用7个微卫星位点,以波尔山羊为对照,对中国6个山羊群体进行了遗传检测,计算了各品种群体的等位基因频率、平均杂合度(He)、平均多态信息含量(PIC)和平均有效等位基因数(Ne),同时分析了各品种群体内和群体间的遗传变异,并根据遗传距离进行了NJ聚类。结果表明,各群体的He、PIC、Ne分别为0.778~0.823、0.766~0.809、4.707~5.767;川东黑山羊和川东白山羊聚为一类,乌羊和南江黄羊聚为一类,其次是马头山羊、小香羊,最后是波尔山羊。微卫星标记分析结果与它们的地理分布及品种形成相一致。     

4个南方山羊品种BMP15和GDF9基因的RFLP分析

以已发现的绵羊高繁殖性能主基因FecXB、FecGH作为候选基因,以南方的4个山羊品种大足黑山羊类群、金堂黑山羊、南江黄羊、川东白山羊为研究对象,采用PCR-RFLP方法分析这两个候选基因在山羊中的多态性.结果发现.在这4个山羊品种(种群)中所有个体均没有检测到FecxB基因和FecGH基因的存在,所有个体均为野生纯合型,表明这两个候选基因位点对所研究的4个山羊品种(种群)的繁殖性能没有影响.     

四川5个山(绵)羊品种随机扩增多态DNA分析

本研究参阅文献挑选 80个随机引物 ,从中筛选出 3 0个重复性好的引物 ,对四川黑山羊、南江黄羊、北川白山羊、成都麻羊 4个山羊品种和藏绵羊进行RAPD分析 ,并进一步对 1 3只南江黄羊个体 ,1 8只黑山羊个体的基因组DNA进行PCR扩增。用Nei氏公式计算品种间的遗传距离 ,用UPGMA法构建树状聚类图。结果表明 :黑山羊与南江黄羊的遗传距离最小 ,亲缘关系较近 ;藏绵羊与各品种间的遗传距离都很大 ,亲缘关系远 ;RAPD技术可作为一种有效的分子标记用于山羊品种之间遗传亲缘关系的分析     

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.     

山羊生长分化因子9基因FecTT突变检测

本研究采用PCR-RFLP方法检测生长分化因子9(growth differentiation factor 9, GDF9)基因FecTT突变在济宁青山羊、贵州白山羊、大足黑山羊、陕南白山羊、古蔺马羊、马头山羊、波尔山羊、成都麻羊、萨能奶山羊、文登奶山羊、板角山羊、金堂黑山羊、关中奶山羊、川东白山羊、安哥拉山羊、辽宁绒山羊、内蒙古绒山羊等17个山羊品种中的分布。结果表明,在17个山羊品种中都未检测到FecTT突变,提示,GDF9基因影响Thoka绵羊高繁殖力的FecTT突变对以上17个山羊品种的繁殖力均没有显著影响。     

Mitochondrial DNA variation of indigenous goats in Narok and Isiolo counties of Kenya

Phylogenetic relationships among and genetic variability within 60 goats from two different indigenous breeds in Narok and Isiolo counties in Kenya and 22 published goat samples were analysed using mitochondrial control region sequences. The results showed that there were 54 polymorphic sites in a 481‐bp sequence and 29 haplotypes were determined. The mean haplotype diversity and nucleotide diversity were 0.981 ± 0.006 and 0.019 ± 0.001, respectively. The phylogenetic analysis in combination with goat haplogroup reference sequences from GenBank showed that all goat sequences were clustered into two haplogroups (A and G), of which haplogroup A was the commonest in the two populations. A very high percentage (99.90%) of the genetic variation was distributed within the regions, and a smaller percentage (0.10%) distributed among regions as revealed by the analysis of molecular variance (amova ). This amova results showed that the divergence between regions was not statistically significant. We concluded that the high levels of intrapopulation diversity in Isiolo and Narok goats and the weak phylogeographic structuring suggested that there existed strong gene flow among goat populations probably caused by extensive transportation of goats in history.     

Mitochondrial DNA diversity of Anatolian indigenous domestic goats

Anatolia has been an important region for civilizations and agricultural revolution as a major domestication centre for livestock species. Goats (Capra hircus) were among the earliest domesticated animals in this region. In this study, genetic diversity of Anatolian goat breeds was characterized by comparison of mitochondrial DNA hypervariable region 1. A total of 295 individuals, including 99 Anatolian Black goats, 96 Angora goats and 100 Kilis goats, were used. Haplogroup A was found to be the dominant haplogroup in all three breeds. The highest haplogroup diversity, including haplogroups A, B2, C and G, was observed in the Anatolian Black breed. Haplogroup D was only observed in Kilis and Angora goats. Haplogroup G was found in Angora and Anatolian Black breeds. The Anatolian goat breeds had high genetic diversity values and a weak phylogeographical structure. The nucleotide diversity values were found to be higher than those in previously studied goat breeds. The fact that Anatolia is a domestication centre and its geographical position as a junction of trade routes may have caused the higher genetic diversity of Anatolian goat breeds.     

Genetic diversity of Chinese domestic goat based on the mitochondrial DNA sequence variation

The aim of this study was to characterize the genetic diversity of domestic goat in China. For this purpose, we determined the sequence of the mitochondrial DNA (mtDNA) control region in 72 individuals of the Yangtze River delta white goat, and reanalysed 723 published samples from 31 breeds/populations across China. All goat haplotypes were classified into four haplogroups (A–D) previously described. The phylogenetic pattern that emerged from the mtDNA control region sequence was confirmed by the analysis of the entire cytochrome b sequence of eight goats representative of the four haplogroups. It appeared that in Chinese domestic goat, haplogroups A and B were dominant and distributed in nearly all breeds/populations, while haplogroups C and D were only found in seven breeds/populations. Four breeds/populations contained all four haplogroups. When grouping the breeds/populations into five geographic groups based on their geographic distributions and ecological conditions, the southern pasturing area had the highest diversity whereas the northern farming area had the lowest diversity. 84.29% and 11.37% of the genetic variation were distributed within breeds and among breeds within the ecologically geographical areas, respectively; only 4% of genetic variation was observed among the five geographic areas. We speculate that the traditional seasonal pastoralism, the annual long-distance migrations that occurred in the past, and the commercial trade would account for the observed pattern by having favoured gene flows.     

利用mtDNA细胞色素b分析贵州山羊的系统发育

试验选取贵州山羊4个群体(共55只),设计引物对其mtDNA细胞色素b基因片段进行测序,测序结果与GenBank报道的8个山羊品种mtDNA细胞色素b基因进行比对分析。结果发现,在55只贵州山羊中有11个变异位点,其中转换10次,T/G颠换1次;10种单倍型中有7种单倍型为1个群体独享,3种单倍型为多个群体共享。4个山羊类群间存在较近的亲缘关系,既是独立的类群,其间也存在基因交流,其中贵州黑山羊和贵州麻羊的亲缘关系最近,与贵州白山羊的亲缘关系次之,与贵州小香羊的亲缘关系最远。从山羊品种间遗传距离构建的邻接树可见,贵州4个山羊类群均起源于镰刀状角羊骨羊。