利用微卫星技术分析中国部分地方鸡种的遗传结构

利用7个微卫星标记对鹿苑鸡、固始鸡、藏鸡、白耳鸡、仙居鸡、茶花鸡、大骨鸡、北京油鸡、狼山鸡、河南斗鸡、泰和乌骨鸡和萧山鸡等12个中国地方鸡种的等位基因频率、基因杂合度、平均基因杂合度、多态信息含量以及群体间的亲缘关系进行分析。研究结果表明,12个地方鸡种在7个微卫星座位上的基因频率存在一定的差异;鹿苑鸡的平均基因杂合度最高,为0．5929;茶花鸡的平均遗传杂合度最低,为0．3514。平均多态信息含量也出现了类似的结果,说明鹿苑鸡的遗传多样性最丰富。模糊聚类分析结果表明,12个地方鸡种间,泰和乌骨鸡与河南斗鸡的亲缘关系相对较近,而固始鸡与其他11个地方鸡种的亲缘关系相对较远。12个地方鸡种可以聚为3类：泰和乌骨鸡、河南斗鸡、狼山鸡、大骨鸡、萧山鸡、北京油鸡、鹿苑鸡聚为第1个类群;茶花鸡、藏鸡、仙居鸡、白耳鸡聚为第2类群;固始鸡为第3类群。     

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）-     

利用微卫星标记分析山东地方鸡品种的遗传多样性

微卫星是近几年来应用较多的一种分子标记,可有效地进行基因鉴定与系谱分析,并可估算群体间的遗传距离。通过选用5个微卫星标记,检测了山东省5个地方鸡种：日照麻鸡、寿光鸡、莱芜黑鸡、济宁百日鸡、鲁西斗鸡以及一个外来鸡种——安卡黄鸡和一个外省地方鸡种——广西黄鸡共7个鸡种的遗传多样性。根据测试结果计算了每个等位基因的频率,以基因频率为基础分析了品种内的遗传变异和品种间的DA遗传距离,并讨论了微卫星多态性在应用于群体遗传变异及亲缘关系等方面的意义。结果表明：共检测到40个等位基因,其中等位基因数最多的位点为。ADL0136(10个);等位基因数最低的位点为ADL0146(5个);而且每个位点的等位基因分布并不均匀,都有一种或几种优势基因存在。在7个品种中,杂合度最低的为寿光鸡,杂合度值为0．3327,因为此鸡种多年来一直由寿光市慈伦种鸡场进行纯繁保种,未与其他鸡种杂交,因此杂合度最小;其他鸡种杂合度也都低于0．4,据分析可能是由于日照麻鸡、济宁百日鸡群体较小;莱芜黑鸡是正在选育的一个品种,个体间遗传关系也不远;安卡黄鸡和广西黄鸡自从引人嘉明公司后,群体近交现象普遍,因此各鸡种杂合度都偏低。由此可见,通过对杂合度的计算,微卫星可以较好地反应群体内的变异。各品种PIC值的变动范围从0．6196(寿光鸡)到0．7027(莱芜黑鸡),PIC值的大小与杂合度的高低体现了较高的一致性。对DA遗传距离的计算表明：日照麻鸡与济宁百日鸡的距离最近,而鲁西斗鸡与其他鸡种距离均较远。用UPGMA法进行聚类分析,结果7个鸡种被聚为3类：山东的4个地方鸡种寿光鸡、日照麻鸡、莱芜黑鸡与济宁百日鸡聚为一类;安卡黄鸡和广西黄鸡聚在一起;鲁西斗鸡独自为一类。这与几个鸡种的分化与选育历史是一致的,因此聚类图能够比较正确地反映7个品种之间的亲缘关系。     

鸡生长激素基因5‘端部分调控区的克隆分析

利用ＰＣＲ技术扩增、克隆、测序了７个跨越不同生长速度的鸡品种（系）的生长激素（ＧＨ）基因的５＇端部分调控区。这７个品种（系）分别为：高生长速度的宝罗肉鸡父本;较高生长速度和一定的产蛋性能的宝罗肉鸡母本;肉蛋灵用型的芦花鸡;中等生长速度和中等体重的蛋鸡品种洛岛红和农大褐;生长速度较慢体重较轻的蛋鸡品种北京白鸡和生长速度很慢但又不是矮小型的地方品种丝毛乌骨鸡。所扩增的片段长度为７６０ｂｐ,包括了转录起     

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。     

家鸡和原鸡的线粒体DNA多态性比较

本文运用１１种限制性内切酶分析了家鸡（茶花鸡、尼西鸡、大理漾濞黄鸡）和原鸡共１０只个体的线粒体ＤＮＡ限制性片段长度多态性（ＲＦＬＰ）,平均每个个检测到的片段为４０条左右。但仅发现３种变异的限制性片制性格局,即ＳｔｕＩ－Ｂ,Ｅｃａ－Ｉ－ｂ和ＲＩ－Ｂ．其中ＳｔｕＩ－Ｂ和ＳｃａＩ－Ｂ为首次报道,而且均为原鸡所物有,ＥｃｏＲＩ－Ｂ则为大理漾濞黄鸡所特有。茶花鸡和尼西鸡拥有完全相同的限制性格局。经过计算,原     

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。     

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。     

藏鸡线粒体全基因组序列的测定和分析

通过PCR扩增,测序,拼接,获得藏鸡（Tibetan Chicken）线粒体全基因组序列并进行数据分析处理。藏鸡线粒体全基因组序列全长16783bp,共有13个蛋白质编码基因、2个rRNA基因、22个tRNA基因和1个D-loop区。模拟电子酶切结果显示,藏鸡DraI酶的酶切结果和先前报道的原鸡,茶花鸡,尼西鸡和大理漾濞黄鸡的酶切结果都不相同,为藏鸡特有。基于D-loop区全序列和13个蛋白质编码基因序列,采用N-J算法与原鸡属4个种,3个亚种和3个家鸡品系构建系统进化树：初步确定藏鸡起源于红原鸡,与家鸡中的来航鸡、白洛克鸡亲缘关系最近,但是藏鸡的进化与来航鸡、白洛克鸡这两个家鸡品系又显得相对独立。推测可能原因是藏鸡的祖先在进入高原以后处于相对封闭的环境,从而形成了独特群体遗传特性。     

中国红原鸡和泰国红原鸡遗传多样性分析

利用29个微卫星DNA标记对来自中国的红原鸡Gallus gallus spadiceus亚种和来自泰国的红原鸡Gallus gallus gallus亚种进行遗传多样性分析, 评估亚种内的遗传变异和亚种间的遗传分化, 结果表明: 共检测到168个等位基因, 每个位点的等位基因数从2到13不等, 所有位点平均的期望杂合度和PIC值分别为0.5780和0.53。中国和泰国红原鸡29个微卫星位点平均有效等位基因数分别为3.79和4.79, 平均基因杂合度为0.5379和0.6385, 两个红原鸡亚种均表现出较高的群体杂合度和丰富的遗传多样性。群体分化系数为19.4%(P<0.01), 两个红原鸡亚种间的Reynolds’遗传距离和Nm值分别为0.157和1.040。由此可见, Gallus gallus spadiceus亚种和Gallus gallus gallus亚种群体具有不同的群体遗传结构, 群体之间存在明显的遗传分化, 并不能将其认定为是同一亚种, 这也为中国家鸡具有独立的起源提供了一定的佐证。     

原鸡海南亚种的巢址特征

选择适宜的巢址对降低巢捕食风险,提高繁殖成效有重要意义。2008年3—7月,在海南省大田国家级自然保护区,采用随机样线法结合访问法在各种生境类型中系统地寻找原鸡海南亚种(Gallus gallus jabouillei)的巢,并在野外追踪观察基础上用样方法对原鸡的巢址选择进行研究。共记录到原鸡的巢16个,以巢为中心选取样方并测量反映巢址的13个特征生境参数,同时分析16个对照样方以进行比较,并通过主成分分析探讨影响巢址选择的主导因子。结果表明：（1) 原鸡的巢位于密集的丛生草本下或低矮的灌木丛基部。营巢背景多样,其中位于草本植物下10个(62.5%);灌草丛5个(31.3%);灌木下1个(6.25%)。（2) Mann–Whitney U检验的结果显示,原鸡偏好于比较开阔、地面落叶稀少而靠近的小道和林缘的位置营巢。（3) 主成分分析表明,植被盖度因子和空间位置因子贡献率最大,在降低巢捕食风险中有重要价值,是影响原鸡巢址选择的主要因素。     

红色原鸡群体遗传多样性

为了阐明红色原鸡的群体遗传结构,以对其有效保护提供遗传学依据,采用33个微卫星标记对其群体中56个个体进行了PCR-聚丙烯酰胺多态性电泳检测。33个微卫星座位共检测到140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2～8个之间,平均每个座位等位基因数4.24个,有效等位基因数3.30个。根据等位基因频率,计算出的群体表观杂合度、期望杂合度及多态信息含量分别为0.7980、0.6506和0.5948。结果表明,红色原鸡群体遗传多样性较丰富。     

A fourth allele in the plasma esterase-1 (Es-1) system of the domestic fowl

Plasma samples of fowl were analysed by horizontal polyacrylamide gel electrophoresis (pH 9.0). Evidence was presented for the subdivision of an earlier reported esterase-1 allele (Es-1A) into two alleles designated Es-1A1 and Es-1A2. Family data were consistent with the hypothesis that the Es-1 phenotypes were controlled by four codominant, autosomal alleles Es-1C, Es-1A1, Es-1A2 and Es-1B). The White Leghorn samples showed high frequency of Es-1A1 (about 0.7) and also had considerable frequency of Es-1A2 (0.2) and of Es-1B (0.1). The three meat-type breeds studied (White Plymouth Rock, Rhode Island Red and New Hampshire) showed a very high frequency of Es-1B (0.8-1.0).     

Study on the variation of precipitin reactions in normal sera of chickens

Breed distribution of precipitin reactants (of types A and B) was encountered in: Brahma and Columbian Plymouth Rock - high strength and frequencies; White Cornish, Dark Cornish, Minorca, Barred Plymouth Rock, New Hampshire, Rhode Island Red, Catalana and Cubalaya - low to medium strength and frequencies; Australorp, White Leghorn, Polish, Sexline - no reactions. Cocks exhibited only type A reactants. Both A and B substances were detected in egg yolks. Type A substances were encountered in sera of newly hatched chicks. Their decay was slow - the mean persistence time being about 15 days. Only two young chicks among 160 tested exhibited the reactants of type B. Hormonal treatments of White Leghorn and Rhode Island Red cocks with testosterone propionate, progesterone, oestradiol benzoate or oestradiol benzoate along with progesterone failed to induce the synthesis of precipitin reactants.     

Novel AluI-polymorphism in the fourth intron of chicken growth hormone gene

A novel AluI-polymorphism in the fourth intron of chicken growth hormone gene was shown. The cytosine to thymine transition in the restriction site for AluI was detected. Primers flanking the 460 bp fragment of the fourth intron containing a polymorphic restriction site for AluI were designed. The nucleotide sequence fragments amplified polymorphic variants were determined. Using designed primers, the genetic structure of populations of White Plymouth Rock, Poltava Clay, Rhode Island Red, and Borkovskaya Barvistaya chicken breeds were analyzed. It was found that growth hormone gene (by AluI-polymorphism in the fourth intron) was polymorphic in all experimental populations. Frequencies of alleles C and T in chicken population of White Plymouth Rock breed were 0.14 and 0.86, Rhode Island Red were 0.3 and 0.7, Poltava Clay were 0.04 and 0.96, and Borkovskaya Barvistaya were 0.08 and 0.92, respectively. The tendency to increase egg production and egg weight of chicken with C/C genotype and meat quality (live weight, carcass weight, weight of pectoral muscles) of chickens with genotype T/T of Rhode Island Red chicken breed was shown.     

原鸡海南亚种的巢址特征

选择适宜的巢址对降低巢捕食风险,提高繁殖成效有重要意义。2008年3—7月,在海南省大田国家级自然保护区,采用随机样线法结合访问法在各种生境类型中系统地寻找原鸡海南亚种(Gallus gallus jabouillei)的巢,并在野外追踪观察基础上用样方法对原鸡的巢址选择进行研究。共记录到原鸡的巢16个,以巢为中心选取样方并测量反映巢址的13个特征生境参数,同时分析16个对照样方以进行比较,并通过主成分分析探讨影响巢址选择的主导因子。结果表明:(1)原鸡的巢位于密集的丛生草本下或低矮的灌木丛基部。营巢背景多样,其中位于草本植物下10个(62.5%);灌草丛5个(31.3%);灌木下1个(6.25%)。(2)Mann–WhitneyU检验的结果显示,原鸡偏好于比较开阔、地面落叶稀少而靠近的小道和林缘的位置营巢。(3)主成分分析表明,植被盖度因子和空间位置因子贡献率最大,在降低巢捕食风险中有重要价值,是影响原鸡巢址选择的主要因素。     

Genetic diversity within chicken populations from different continents and management histories

Intra-population variation was assessed in 1970 chickens from 64 populations using 29 autosomal microsatellites. On average, 95% of the loci were polymorphic across populations. In 1456 ( c. 83%) of the 1763 combinations of populations and polymorphic loci, no departure from Hardy–Weinberg equilibrium was observed. On average, there were 11.4 alleles per locus and 3.6 alleles per population across loci. Within populations, the average observed heterozygote frequency was 0.46, with a range between 0.20 and 0.64. Dagu, a Chinese population, and the Red Jungle Fowl ( Gallus gallus gallus ) had the highest average heterozygote frequencies at 0.64 and 0.63 respectively. An inbred line used as a reference population for comparison showed the lowest average of observed heterozygote frequency (0.05), followed by the European population Hamburger Lackhuhn, whose average observed heterozygote frequency was 0.20. A total of 32 private alleles (alleles detected in only one population) for 20 loci were found in 18 populations. H'mong chickens, a Vietnamese population, carried the largest number of private alleles at five, followed by the Red Jungle Fowl with four private alleles. Genetic diversity within populations was low in the NW European fancy breeds and high in the non-commercial Asian populations, in agreement with population management history.     

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.