我国地方鸡品种保种群微卫星多态性分析与分子标记档案的建立

利用20个微卫星标记对国家家禽品种资源基因库中保存的11个地方鸡品种保种群进行了遗传检测,计算各群体的等位基因频率、平均基因杂合度、平均多态信息含量及各群体间的遗传距离,并用类平均法进行聚类分析。研究结果表明：20个微卫星标记在11个地方鸡品种保种群共检测到176个等位基因,平均为8.8个,基因频率分布在0.013～0.838之间。检测到等位基因中,有45个等位基因为11个地方鸡品种所共有;11个地方鸡品种平均杂合度在0.6800～0.7432之间。其中藏鸡最高,为0.7432;狼山鸡最低,为0.6800;平均多态信息含量在0.6329～0.7023之间,均大于0.5,表现为高度多态性;11 个鸡品种聚为4类。丝羽乌骨鸡、茶花鸡、仙居鸡、藏鸡、萧山鸡聚为一类,鹿苑鸡、狼山鸡聚为一类,固始鸡、北京油鸡、大骨鸡聚为一类,河南斗鸡单独聚为一类;通过利用20个微卫星基因座检测不同世代群体中等位基因及其频率、群体基因平均杂合度和多态信息含量,建立地方鸡品种保种群微卫星标记档案,并分析世代间的差异,预期可以达到监测保种效果的目的。     

红色原鸡和中国家鸡遗传多样性及亲缘关系

对红色原鸡（Gallus gallus）和中国家鸡（Gallus domesticus）之间的遗传多样性和亲缘关系进行系统分析,可以为中国家鸡的起源进化、种质资源保护以及科学开发利用等研究提供理论依据,但迄今尚未见用微卫星标记进行分析比较的公开报道．本研究利用29对微卫星引物对红色原鸡Gallus gallus spadiceus亚种和Gallus gallus gallus亚种以及14个中国家鸡品种的568只个体进行扫描,共检测到286个等位基因,平均值为9．86±6．36;所有群体的期望杂合度为0．6708±0．0251,皖南三黄鸡的期望杂合度最高（0．6442）,固始鸡的最低（0．4532）．单个位点偏离Hardy-Weinberg平衡的群体数从0-7不等．整个群体平均遗传分化为17．9％（P〈0．001）,约有16．7％的遗传变异源自群体间的差异,所有的位点都极显著地贡献于这一结果（P〈0．001）;杂合子缺失的水平很高,为0．015（P〈0．01）,有8个位点显示显著的杂合子缺失．群体间的Reynolds’遗传距离从0．036（萧山鸡-鹿苑鸡）～0．371（泰国红色原鸡-河南斗鸡）不等,而Nm值变异范围为从0．583（泰国红色原鸡-河南斗鸡）～5．833（萧山鸡鹿苑鸡）．NJ系统发生树中,鹿苑鸡、萧山鸡、北京油鸡、河南斗鸡、淮南麻黄鸡、狼山鸡等重体型的鸡种首先形成一大类;鹿苑鸡与萧山鸡以及茶花鸡与藏鸡有着较近的遗传关系;茶花鸡和藏鸡与两个红色原鸡亚种亲缘关系较近,中国红色原鸡Gallus gallus spadiceus亚种与所有中国家鸡品种的亲缘关系比泰国红色原鸡Gallus gallus gallus亚种要近．结果表明：29个微卫星位点均具有较高的多态性,红色原鸡和中国家鸡群体间存在着极显著的遗传分化;中国家鸡和红色原鸡两个亚种的亲缘关系从近到远的排序是：进化型品种-原始型品种（茶花鸡与藏鸡）-中国红色原鸡亚种（Gallus gallus spadiceus）-     

我国部分地方鸡种肤色伴性遗传初步观察

对泰和鸡、仙居鸡、固始鸡、北京油鸡、萧山鸡、狼山鸡（Ｎ系）进行肤色伴性遗传观察,结果表明：泰和鸡常染色体上含有１对黑色素基因ＰＰ,仙居鸡、萧山鸡、北京油鸡性染色体上含有Ｉｄ抑制色素基因,狼山鸡、固始鸡、泰和鸡性染色体上含有ｉｄ基因,泰和鸡♂与含有Ｉｄ基因鸡种（仙居鸡、萧山鸡、北京油鸡）♀杂交,Ｆ１代肤色能自别雌雄,公鸡为黄皮肤,母鸡为黑皮肤。     

家鸡腺苷琥珀酸裂解酶基因多态性与肌苷酸含量的关联及其与红原鸡的亲缘关系

根据腺苷琥珀酸裂解酶(adenylosuccinate lyase, ADSL)基因外显子2的序列设计引物, 用PCR-SSCP的方法对隐性白羽鸡、丝羽乌骨鸡、白耳鸡、藏鸡以及红色原鸡两个亚种进行了单核苷酸多态性分析, 并检测到了多态性, 表现为3种基因型, 对两种纯合子进行直接测序, 结果发现3484位碱基处发生C→T突变。对3种基因型的肌肉肌苷酸含量的最小二乘分析结果显示TT型(突变型)个体的肌肉肌苷酸含量极显著地高于CT型、显著地高于CC型个体, CT型个体也稍高于CC型, 但差异不显著, 初步推测该位点可能与肌肉肌苷酸含量有关。根据该多态位点的基因频率, 基于Nei氏的遗传距离运用NJ聚类法构建系统发生树, 进行家鸡与原鸡的亲缘关系分析, 结果发现, 丝羽乌骨鸡与白耳鸡的亲缘关系最近, 藏鸡和中国红原鸡亚种的亲缘关系也较近, 中国地方家鸡品种与中国红原鸡亚种的亲缘关系较近,而与泰国红色原鸡的亲缘关系较远,隐性白羽鸡与原鸡亲缘关系最远, 初步得出中国家鸡有自己独自的血缘来源的结论。     

利用微卫星标记分析四川8个地方鸡品种遗传多样性

通过选用30个多态性较好的微卫星标记,检测了四川省8个地方鸡品种：峨嵋黑鸡、泸宁鸡、旧院黑鸡、米易鸡、石棉草科鸡、凉山崖鹰鸡、兴文乌骨鸡、沐川乌骨鸡的遗传多样性。利用等位基因频率计算出各群体的平均遗传杂合度（H）、多态信息含量（PIC）和群体间的DA遗传距离。结果表明：30个微卫星位点中有24个微卫星位点在8个鸡群体中的多态信息含量均为高度多态,可作为有效的遗传标记用于各鸡品种的遗传多样性和系统发生关系的分析。各鸡种的杂合度都较高,平均杂合度范围是0.629~0.681,最高的是泸宁鸡（0.681）,最低的是旧院黑鸡（0.629）。据分析可能是由于交通闭塞,形成了家禽品种的多种多型,而且杂合度的高低与PIC值的大小体现了较高的一致性。对DA遗传距离的计算表明：用UPGMA法进行聚类分析,结果8个鸡品种被聚为3类：Ⅰ类：峨嵋黑鸡、米易鸡、泸宁鸡、旧院黑鸡聚为一类。米易鸡、泸宁鸡先聚在一起,然后又与峨嵋黑鸡在较近的距离聚在一起,然后再与旧院黑鸡聚在一起;Ⅱ类：石棉草科鸡、兴文乌骨鸡、沐川乌骨鸡聚为一类。兴文乌骨鸡、沐川乌骨鸡在较近的距离聚在一起,然后又与石棉草科鸡在较近的距离聚在一起;Ⅲ类：凉山崖鹰鸡独自聚为一类。这与几个鸡品种的来源、分化、选育历史及地理分布是一致的。     

利用微卫星技术分析不同类群鸡的遗传多样性

利用8个微卫星标记分析了6个生产类群鸡的遗传多样性。计算了各群体在各位点上的等位基因频率,并据此计算出各群体的平均遗传杂合度(h)、多态信息含量(PIC)和有效等位基因数(Ne)。结果表明:6个鸡群在8个微卫星座位上的基因频率存在明显的差异,其平均基因杂合度为0.7317,平均多态信息含量为0.6815。其中,群体平均杂合度最高的是安卡红鸡,为0.7716;平均杂合度最低的是新罗曼鸡,为0.7073。所选的8个微卫星座位均为高度多态,可作为有效的遗传标记用于鸡群体遗传多样性的分析。     

茶花鸡群体遗传多样性

茶花鸡是我国具有独特遗传特性的地方家禽品种,为了进一步阐明其群体遗传变异和遗传结构状况,采用了33个家鸡特异性的微卫星标记对该鸡种自然群体中30个个体进行了多态性电泳检测。33个微卫星座位共检测到105个等位基因,所有座位都呈现出多态性,每个座位的等位基因数在2~5个之间,平均每个座位等位基因数3.20个。群体平均杂合度和平均多态信息含量分别为0.612 9和0.527 6。结果表明,茶花鸡自然群体遗传多样性较丰富。     

红原鸡与家鸡的亲缘关系研究

对中国红原鸡滇地亚种和海南亚种与我国茶花鸡,泰和鸡和寿光鸡等地方鸡种以及芦花鸡,洛岛红等外国鸡种进行了血型（３个位点,１３个等位基因）,蛋白质（酶）多态（５个位点,１１个等位基因）和ＤＮＡ指纹分析,结果表明,红原鸡与茶花鸡（原始型品种）的亲缘关系较近;与泰和鸡,寿光鸡,芦花鸡,洛岛红（进化型品种）的亲缘关系较远,呈红原鸡－茶花鸡－泰和鸡,寿光鸡或芦花鸡,洛岛红这样一个进化阶梯,以上结果与国外资料（     

应用微卫星标记分析中国地方鸡种的遗传变异

利用 8个微卫星位点对中国 9个地方鸡种和 1个引进品种进行了遗传检测。计算出了各品种的平均杂合度、平均多态信息含量 (PIC)及品种间的遗传距离 ,并进行了系统聚类。结果表明 :8个微卫星位点上共检测到了5 4个等位基因 ,每个位点上平均为 6 .75个。各位点平均多态信息含量为 0 .5 0 71～ 0 .74 34,均表现出了高度多态性。各群体平均杂合度较高 ,为 0 .5 5 6 4～ 0 .7135 ,说明我国地方鸡种有着较丰富的遗传多样性。地方鸡种间的遗传距离相对较远 ,10个鸡种共分为三大类。研究结果对我国鸡种资源的评估、保存和预测杂种优势具有一定的指导意义     

湖北省三品种猪27个微卫星座位的遗传变异

采用国际动物遗传学会（ISAG）和联合国粮农组织（FAO）共同推荐的27个微卫星标记,对湖北省3个主要地方猪种（通城猪、清平猪和阳新猪）的遗传变异进行了检测。计算出各个品种的基因杂合度、各个座位的多态信息含量及品种间的遗传距离。结果表明,3个地方猪种的平均基因杂合度分别为0.7489、0.6987和0.6273,遗传多样性比较丰富;通城猪和清平猪亲缘关系较近,而两者与阳新猪亲缘关系略远。     

Analysis of genetic diversity and phylogenetic relationships among red jungle fowls and Chinese domestic fowls

Genetic diversity and phylogenetic relationships among 568 individuals of two red jungle fowl subspe- cies (Gallus gallus spadiceus in China and Gallus gallus gallus in Thailand) and 14 Chinese domestic chicken breeds were evaluated with 29 microstaellite loci, the genetic variability within population and genetic differentiation among population were estimated, and then genetic diversity and phylogenetic relationships were analyzed among red jungle fowls and Chinese domestic fowls. A total of 286 alleles were detected in 16 population with 29 microsatellite markers and the average number of the alleles observed in 29 microsatellite loci was 9.86±6.36. The overall expected heterozygosity of all population was 0.6708±0.0251, and the number of population deviated from Hardy-Weinberg equilibrium per locus ranged from 0 to 7. In the whole population, the average of genetic differentiation among population, measured as FST value, was 16.7% (P<0.001), and all loci contributed significantly (P<0.001) to this differentiation. It can also be seen that the deficit of heterozygotes was very high (0.015) (P<0.01). Reynolds' distance values varied between 0.036 (Xiaoshan chicken-Luyuan chicken pair) and 0.330 (G. gallus gallus-Gushi chicken pair). The Nm value ranged from 0.533 (between G. gallus gallus and Gushi chicken) to 5.833 (between Xiaoshan chicken and Luyuan chicken). An unrooted consensus tree was constructed using the neighbour-joining method and the Reynolds' genetic distance. The heavy-body sized chicken breeds, Luyuan chicken, Xiaoshan chicken, Beijing Fatty chicken, Henan Game chicken, Huainan Partridge and Langshan chicken formed one branch, and it had a close genetic relationship between Xiaoshan chicken-Luyuan chicken pair and Chahua chicken-Tibetan chicken pair. Chahua chicken and Tibetan chicken had closer genetic relationship with these two subspecies of red jungle fowl than other domestic chicken breeds. G. gallus spadiceus showed closer phylogenetic relationship with Chinese domestic chicken breeds than G. gallus gallus. All 29 microstaellite loci in this study showed high levels of polymorphism and significant genetic differentiation was observed among two subspecies of red jungle fowl and 14 Chinese domestic chicken breeds. The evolutional dendrogram is as follows: evolutional breeds→primitive breeds (Chahua chicken and Tibetan)→red jungle fowl in China (G. gallus spadiceus)→red jungle fowl in Thailand (G. gallus gallus). The results supported the theory that the domestic fowls might originate from different subspecies of red jungle fowl and Chinese domestic fowls had independent origin.     

Analysis of genetic structure and relationship among nine indigenous Chinese chicken populations by the <Emphasis Type="Italic">Structure</Emphasis> program

The multi-locus model-based clustering method Structure program was used to infer the genetic structure of nine indigenous Chinese chicken (Gallus gallus) populations based on 16 microsatellite markers. Twenty runs were carried out at each chosen value of predefined cluster numbers (K) under admixture model. The Structure program properly inferred the presence of genetic structure with 0.999 probabilities. The genetic structure not only indicated that the nine kinds of chicken populations were defined actually by their locations, phenotypes or culture, but also reflected the underlying genetic variations. At K = 2, nine chicken populations were divided into two main clusters, one light-body type, including Chahua chicken (CHA), Tibet chicken (TIB), Xianju chicken (XIA), Gushi chicken (GUS) and Baier chicken (BAI); and the other heavy-body type, including Beijing You chicken (YOU), Xiaoshan chicken (XIA), Luyuan chicken (LUY) and Dagu chicken (DAG). GUS and DAG were divided into independent clusters respectively when K equaled 4, 5, or 6. XIA and BIA chicken, XIA and LUY chicken, TIB and CHA chicken still clustered together when K equaled 6, 7, and 8, respectively. These clustering results were consistent with the breeding directions of the nine chicken populations. The Structure program also identified migrants or admixed individuals. The admixed individuals were distributed in all the nine chicken populations, while migrants were only distributed in TIB, XIA and LUY populations. These results indicated that the clustering analysis using the Structure program might provide an accurate representation of the genetic relationship among the breeds.     

采用微卫星DNA标记分析部分地方鸡种保种场的保种效果

采用28对微卫星引物分析了我国两个地方鸡种大骨鸡和北京油鸡不同保种场的保种效果。检测了大骨鸡和北京油鸡共计125个个体的基因型,通过计算等位基因数、等位基因频率、遗传杂合度(H)、多态信息含量(P,G)、F统计量、Nei氏遗传距离,并采用UPGMA聚类法分析了大骨鸡和北京油鸡群体内与群体间的遗传变异,比较了两个鸡种不同保种场的保种效果。在所检测的4个群体中,各群体均有较高的多态性,其杂合度都超过了0．55,各位点等位基因的数目为2～22。除LE110194和MCW0032外,其余26个微卫星位点都处于基因平衡状态。结果表明,4个保种场均较好的保存了这些品种的遗传多样性,但同一品种保种场间保种群体已经产生了差异。     

New variations in intron 4 of growth hormone gene in Chinese native chickens

Polymorphism in intron 4 of chicken growth hormone (cGH) gene was studied in 20 Chinese native chicken populations and broiler or layer populations. A total of eight restriction digestion profiles were identified in intron 4 and confirmed by sequencing. Among 20 populations, there were distinctively different allele numbers and frequencies of intron 4 restriction fragment length polymorphisms (RFLPs) between Chinese native chickens and broilers or layers. Two new alleles, allele D and allele E, were identified in Taihe Silkies. Allele D was also identified in other Chinese native breeds and a 50 bp fragment deletion was identified in allele E.     

鸡的血型研究 Ⅹ.我国11个地方鸡种的血型和血浆蛋白质多态分析

对我国11个地方鸡种1087羽鸡群进行了血型和血浆蛋白质多态7个位点22个等位基因的群体遗传学分析。结果表明,血型A、C位点,基因频率各鸡种存在显著差异,B位点则没有显著差异,血型和血浆蛋白质多态位点基因纯合系数普遍较低,血型因子分布在群体中较为分散,表明这些地方品种存在较大的选择潜力。血浆蛋白质多态位点,中国与外国的某些鸡种比较,存在着较大的相同或相异,可能意味着我国和外国鸡种起源上的同缘或品种起源、进化上的多样性。聚类分析表明,这11个地方鸡种可系统聚类成4个类群,其结果与目前人们对这些鸡种的表征分类不尽一致。     

浙江省地方鸡种的遗传多样性研究

测定了浙江省5个地方鸡种及来航鸡（对照）线粒体D－环区的部分序列（539bp）,构建了鸡种的分子系统树。在所测序列中共有24个变异位点,变异率为4．45％。结果显示,浙江省地方鸡种分为两支,有两个母系来源,一支为仙居鸡,它和外来品种来航鸡关系较近,有共同的母系祖先。另一支为灵昆鸡、白银耳鸡、乌骨鸡和萧山鸡,它们有着共同的母系祖先,其中灵昆鸡,白银耳鸡、乌骨鸡较近,萧山鸡与此3种鸡关系较远。