柴达木山羊群体遗传结构及系统地位的研究

采用简单随机抽样法对柴达木山羊群体进行了遗传检测,从毛色、外型特征、体尺、血液蛋白与DNA多态性等5个方面进行了群体遗传结构分析,并探索了其系统地位。结果表明：(1)毛色和形态特征的平均表型异质度分别为0.3419、0.5207;(2)血液蛋白在6个基因座上具有多态性,平均基因座杂合度为0.2584;(3)DNA-RAPD标记表现丰富的多样性,基因多样度为0.4085～0.5318。通过对柴达木山羊与其他国内山羊的系统聚类,初步探索了该山羊群体的形成及所属,这些都表明柴达木山羊是一个比较古老的未经较高强度选育的地方山羊品种,其基因资源是我国山羊遗传资源中宝贵的一部分。Abstract: Genetic structure and character of Chaidamu Goats were studied through simple random sampling. Genetic structure was analysed from five aspects, and phylogeny status was also investigated. The results indicated that: (1) the average phenotypic heterogeneity degree of coat color and morphological character were 0.3419 and 0.5207,respectively; (2) polymorphous blood albumen existed in 6 loci and the average loci heterozygosity was 0.2584; and (3) polymorphism existed in marked genes by DND-RAPD with diversity of 0.4085～0.5318. Phylogeny status was investigated through clustering by Ward’s method on Chaidamu Goats and other domestic goats. All these indicated that Chaidamu Goats was a domestic goat with less intensively selected breed.     

中国地方黄牛的遗传多样性*

中国黄牛起源复杂,我国地方黄牛群体不同品种在毛色、形态外貌、细胞遗传学、血液蛋白座位分析均表现出多样性。计算我国黄牛群体6个毛色座位平均杂合度和6个血液蛋白座位平均杂合度分别为0.3144和0.4873,表明我国地方黄牛群体的遗传多样性非常丰富。计算我国黄牛群体的6个毛色座位和6个血液蛋白座位的基因分化系数分别为0.3404和0.095,表明我国黄牛群体毛色差异中有34.04％是由品种间的差异造成的。血液蛋白的多态性有9.5％是由品种间的差异造成的。我国黄牛群体的遗传多样性主要来自品种内的遗传多样性。保存我国黄牛品种资源多样性不仅要从整个中国黄牛群体上考虑,而且要针对每个品种（或类群）进行保种。     

7个地方山羊品种遗传多样性及遗传结构分析

检测了7个地方山羊群体在15个微卫星位点的遗传多样性和遗传结构,旨为地方山羊群体的保护利用奠定基础。采集燕山绒山羊、辽宁绒山羊、承德无角山羊、济宁青山羊、太行山羊、武安山羊血液样品及内蒙古绒山羊的耳组织,利用微卫星方法分析遗传多样性及遗传结构。结果表明,7个山羊群体的平均有效等位基因数为4.235 8、平均期望杂合度为0.718 8、平均多态信息含量为0.706 8,均具有高度的遗传多样性;总群体平均近交系数为0.088 3,平均遗传分化系数为0.544 2,平均基因流为0.209 4。武安山羊和承德无角山羊的遗传距离最近,太行山羊和燕山绒山羊的遗传距离最远。系统进化树聚类分析表明,燕山绒山羊与辽宁绒山羊聚为一类,济宁青山羊、承德无角山羊、武安山羊、内蒙古绒山羊和太行山羊聚为一类。综上,7个地方山羊群体遗传多样性丰富,群体间遗传分化程度大,基因交流少,受近交程度影响小,具有较高的利用价值和潜力。     

中国绒山羊遗传多样性现状和系统发生关系的微卫星分析

为了调查中国绒山羊遗传资源现状, 作者应用联合国粮农组织和国际家畜研究所推荐的19对微卫星引物并结合荧光PCR技术, 对9个中国地方产绒山羊群体和1个西非山羊品种进行了遗传多样性检测。14个微卫星座位在10个山羊群体中显示为高度多态, 可作为山羊遗传多样性分析的有效标记。多态信息含量和遗传杂合度等数据表明: 目前中国地方产绒山羊群体的遗传多样性较为丰富, 并且大部分保种场较好地保存了这些地方资源。采用非加权配对算术平均法构建的聚类图和采用主成分分析法得到的散点图均显示, 中国山羊与西非山羊为不同的2类; 中国产绒山羊中河谷山羊、河西绒山羊与其他山羊的遗传距离较远; 其他山羊又大致分为2类: 一类由辽宁绒山羊、新疆山羊、柴达木山羊、陕北山羊组成, 另一类由内蒙古绒山羊组成。此研究结果为开展我国地方绒山羊种质特性研究及资源保护和利用提供了科学依据。     

群体遗传不平衡条件下的结构基因遗传共适应特性

本研究以柴达木山羊、柴达木绒山羊和辽宁绒山羊三个群体共147只山羊为材料,利用聚丙烯酰胺凝胶电泳（PAGE）技术检测了5种血液蛋白质（酶）基因座的遗传多态性,并进行了结构基因遗传共适应的研究,结果发现：45个基因座组合中有10个基因座组合处于遗传不平衡状态,并且这些遗传不平衡皆单纯由遗传共适应差异造成;除辽宁绒山羊Tf-PA-3组合的遗传不平衡包含非等位基因间的遗传共适应差异外,其他基因座组合的遗传不平衡皆由等位基因间的共适应差异,即单基因座的遗传不平衡造成;LAP-EsD组合的共适应差异在群体间有遗传传递现象。 Abstract:With the technology of PAGE,the genetic polymorphism of blood protein and enzyme was investigated,and genetic co-adaptability among structural genes was studied in three goat populations(147 goats) including Chaidamu goat(CS),Chaidamu Cashmere goat(CRS) and Liaoning Cashmere goat(LRS) in Qinghai Province,China.The results were showed that the genetic disequilibrium of 10 locus combinations was found among 45 locus combinations in the three goat populations,and these genetic disequilibria were caused only by the difference of genetic co-adaptability among genes,because there didn′t exist the linkage disequilibrium among non-allelic genes.The genetic disequilibrium including the difference of genetic co-adaptability between non-allelic genes was only found at Tf-PA-3 locus combinations in LRS population,the other ones were all caused by the genetic disequilibrium at a single locus.The difference of genetic co-adaptability of LAP-EsD locus combinations could be messaged among different populations.     

山羊MyoD基因家族多态性及与体尺性状的相关性

用PCR-SSCP技术研究了波尔山羊和徐淮山羊2个群体共147个个体MyoD基因家族中3个基因座位的单核苷酸多态性(single nucleotide polymorphism, SNP)。结果表明, 在徐淮白山羊群体中, myf-5基因座发现有3种基因型AA、AB和BB, 波尔山羊均为AA型。在myf-6基因座和myoD 5′侧翼区基因座, 两个山羊群体均检测到了AA和AB型个体。对山羊myf-5、myf-6基因座, myoD 5′侧翼区基因座不同基因型与两品种山羊体尺性状相关分析表明, myf-5基因座对管围和管围指数的效应差异显著(P<0.05)。myf-6基因座对徐淮白山羊体尺性状的效应均不显著(P>0.05), 而对波尔山羊的体高和管围指数效应差异显著(P<0.05)。两个山羊品种myoD 5′侧翼区不同基因型个体的体尺性状差异均不显著。     

黄河下游家绵羊与家山羊遗传关系的微卫星分析

为了探讨家养绵羊与山羊的属间遗传关系,我们利用13个定位于绵羊染色体上的微卫星基因座,分析了黄河下游4个地方绵羊品种、4个地方山羊品种和1个杂交绵羊类群的遗传结构及其系统发生关系.经Hardy-Weinberg平衡检验和中性测试,发现地方绵羊和山羊种群均处于不平衡状态(P＜0.01),61.53%的基因座属于中性位点,说明所研究种群属于非随机交配,可能受到选择、迁移等进化因素的影响.对有效等位基因数、多态信息含量、Shannon信息指数、观察杂合度和期望杂合度等遗传多样性参数进行比较,发现绵羊种群的遗传变异程度明显(P＜0.01或P＜0.05)高于山羊种群,但不同基因座上的差异效应不一致;结合F统计量和亲缘关系等参数,可推测绵羊和山羊虽然均存在不同程度的近交现象(He＞Ho,FIS＞0),但分别属于杂交和近交繁殖.通过群体遗传分化和系统发育拓扑结构分析,证明绵羊属由共同祖先分化而来的时间晚于山羊属,两属间的遗传距离为1.0708-1.5927,遗传分化时间约为19,807-28,955年;绵羊属内品种间的遗传分化程度(FST＜0.05)均低于山羊属内品种间的分化(FST＞0.15).本研究揭示了人工选择对同域家养绵羊与山羊交配系统的形成及群体遗传分化具有深刻的影响.     

用微卫星标记技术对国内BALB/c小鼠遗传质量的分析

为了解和掌握国内BALB/c小鼠遗传质量状况,验证微卫星标记技术在近交系小鼠遗传检测中应用的可靠性,应用所筛选的小鼠不同染色体上的14个微卫星基因座,通过PCR扩增对北京、上海、沈阳、广州、长春、重庆和哈尔滨7个地区11个厂家提供的BALB/c小鼠进行遗传质量分析.结果北京、上海、哈尔滨及广州地区7家BALB/c小鼠在14个基因座均呈现一条清晰条带,且群体间呈单态性.沈阳、广州、长春和重庆4个群体有8个基因座在群体内表现杂合或呈多态性;其中沈阳和长春分别在1个基因座上表现多态性和杂合;广州另一群体有4个基因座出现杂合或多态性;重庆群体有7个基因座表现为杂合或多态性,在D10Mit180基因座与上海群体比较呈现多态性.     

北方汉族人群8个多拷贝Y-STR基因座的遗传多态性

为分析8个多拷贝Y-STR基因座在北方汉族群体中的遗传多态性和突变率,并同其他群体进行群体间差异分析,探讨其法医学和群体遗传学研究的应用价值。采用复合扩增体系对508名北方汉族无关男性个体和156对父子的8个多拷贝Y-STR基因座进行扩增,应用ABI3730分析仪进行基因分型,采用相应软件计算基因(单倍型)频率、基因多态性和突变率并对群体间差异进行分析;北方汉族群体的8个多拷贝Y-STR基因座的基因多态性普遍较高,基因多态性在0.673 8(DYS389I)~0.968 8(DYS464)之间,共检出484个单倍型,单倍型多态性为0.9998,家系调查中共发现5次基因突变,其中,各个基因座的每代突变率在0~9.62×10~(-3)(DYS535)之间,总平均突变率为1.60×10~(-3)(95%CI 0.5-3.7×10~(-3));群体间比较显示:8个基因座的遗传多态性在群体间存在差异,遗传距离在0.001 64~0.098 02之间。北方汉族群体的8个多拷贝Y-STR基因座的基因多态性普遍较高,适合应用于法医学个体识别和亲权鉴定中。     

柚品种的等位酶变异研究

研究了柚的48个品种的等位酶变异,利用等位酶分析技术对柚的酯酶（EST）,6－磷酸葡萄糖异的酶（PGI）,6－磷酸葡萄糖变位酶（PGM）,莽草酸脱氢酶（SKD）,超氧化物歧化酶（SOD）共5个酶系统的10个等位酶基因座进行了分析,除PGI－1,PGI－2两个基因座外,其它8个均为多态性基因座;10个等位酶基因座共观察到的等位基因25个,平均每个基因座的有效等位基因数目为1．55,基因多样度0．2805,柚的品种间具有较为丰富的等位酶标记遗传多样性,但柚类种质资源群体总的遗传多样性水平偏低。柚的较低的有效等位基因数目与基因多样度可能由于人工选择及资源流失造成。     

Ear mites from domestic goats in Florida

Ear canals of 284 domestic goats from Florida were examined via swabbing for the presence of ear mites; 57% were infested withPsoroptes cuniculi. Raillietia caprae was recovered from 2 goats in one herd. Breeds examined were Nubian, Alpine, Saanen, Toggenburg, La Mancha, Spanish/ Woods and Pygmy. Goats with dependent ear types were infested more commonly than those with erect ears; no goats with vestigial ears were found to harbor mites. Goats less than 1 year old had a significantly higher mite prevalence than did goats over 1 year of age. Gross lesions were noted in the ear canals of only three individuals.This paper was previously accepted for publication in the journal Goat and Sheep Research prior to that journal's demise.     

Genome-wide association study for antibody response to Mycobacterium avium ssp. paratubeculosis in goats

Mycobacterium avium ssp. paratuberculosis (MAP), causes Johne's disease (JD), or paratuberculosis, a chronic enteritis of ruminants, which in goats is characterized by ileal lesions. The work described here is a case–control association study using the Illumina Caprine SNP50 BeadChip to unravel the genes involved in susceptibility of goats to JD. Goats in herds with a high occurrence of Johne's disease were classified as healthy or infected based on the level of serum antibodies against MAP, and 331 animals were selected for the association study. Goats belonged to the Jonica (157) and Siriana breeds (174). Whole-genome association analysis identified one region suggestive of significance associated with an antibody response to MAP on chromosome 7 (p-value = 1.23 × 10⁻⁵). These results provide evidence for genetic loci involved in the antibody response to MAP in goats.     

Genetic characterisation of a domestic dog<Emphasis Type="Italic">Canis familiaris</Emphasis> breed endemic to South African rural areas

Allozyme electrophoresis (horizontal starch gel and PAGE) and histochemical staining techniques were used to study the genetic composition of an endemic southern African domestic dogCanis familiaris Linnaeus, 1758, the Africanis breed. Genetic differentiation was analysed at 21 protein-coding loci. The results were compared to those for three other populations/breeds: representatives of established Western breeds, crossbred dogs of Western descent from rural areas in South Africa, and indigenous Saluki dogs from the Middle East. Nine polymorphic loci were found (Ak-1,-2, Ck, Per, Hb, Po-A-1 to-3 andPo-Tf). Two unique alleles at theCk andPo-A-2 loci separated the Africanis breed from the other groups. There were also significant differences between Africanis and the other breeds in pair-wise comparisons of allelic frequencies at polymorphic loci. An assignment test, fixation index values, gene flow and genetic distance values indicated a closer genetic association between the Africanis and Saluki breeds than with dogs of Western origin. This finding supports archaeological evidence that the endemic Africanis breed was introduced from the Middle East into Africa thousands of years ago, and not through later western influences. The average heterozygosity ranged from 0.106–0.15, with least heterozygosity in the Africanis and most in the rural crossbred group. The percentage of polymorphic loci, the mean number of alleles per locus (biologically more significant than heterozygosity), and conformation of genotypes to Hardy-Weinberg proportions showed no evidence of recent loss of genetic diversity in Africanis. Genetic differentiation and support of archaeological evidence by genetics indicate that the endemic southern African domestic dog breed is unique.     

中国部分家养山羊mtDNA D环区遗传多样性与进化

对我国10个家养山羊品种140只个体的mtDNA D-loop区进行测序分析, 测得结果: 整个D-loop区为 1 211~1 213 bp, 共检测到84种单倍型, 171个多态位点。其中核苷酸多样性(Nucleotide diversity, Pi)为: 0.02063 ± 0.00225, 单倍型多样性(Haplotype gene diversity, Hd)为: 0.988 ± 0.003, 平均核苷酸差异数(Average number of nucleotide differences) k为: 24.896。表明我国家养山羊品种遗传多样性丰富。通过构建NJ网络进化树, 得出中国家养山羊主要分为两大支系, 并且其中一支与角骨羊(Capra aegagrus)聚在一起, 而旋角野山羊(Capra fal-coneri)单独聚为一支, 说明角骨羊对中国家养山羊贡献较大。     

Mitochondrial diversity and phylogeographic structure of Chinese domestic goats

China has numerous native domestic goat breeds, but so far there has been no extensive study on genetic diversity, population demographic history, and origin of Chinese goats. Here, we examined the genetic diversity and phylogeographic structure of Chinese domestic goats by determining a 481-bp fragment of the first hypervariable region of mitochondrial DNA (mtDNA) control region from 368 individuals representing 18 indigenous breeds. Phylogenetic analyses revealed that there were four mtDNA lineages (A-D) identified in Chinese goats, in which lineage A was predominant, lineage B was moderate, and lineages C and D were at low frequency. These results further support the multiple maternal origins of domestic goats. The pattern of genetic variation in goat mtDNA sequences indicated that the two larger lineages A and B had undergone population expansion events. In a combined analysis of previously reported sequences and our sequences belonging to lineage B, we detected two subclades, in which one was unique to eastern Asia and another was shared between eastern and southern Asia. A larger genetic variation in eastern Asia than southern Asia and the pattern of phylogeographic variation in lineage B suggest that at least one subclade of lineage B originated from eastern Asia. There was no significant geographical structuring in Chinese goat populations, which suggested that there existed strong gene flow among goat populations caused by extensive transportation of goats in history.     

Cross-species amplification of Bovidae microsatellites and low diversity of the endangered Korean goral

The Korean goral (Nemorhaedus caudatus) is an endangered species of wild goat. The conservation and management of this species could benefit from a better understanding of its genetic diversity and structure. Cross-species amplification of 34 Bovidae microsatellite loci was tested on a panel of 6 Korean gorals and 10 domestic goats. After polymerase chain reaction (PCR) optimization, 29 (85.3%) microsatellite loci amplified successfully for the Korean gorals and 27 (79.4%) for the domestic goats. Of the amplified products, 16 (55.2%) were polymorphic in the Korean goral and 22 (81.5%) in domestic goats. Nei's unbiased mean heterozygosity and mean allele number per locus were, respectively, 0.356 and 2.6 in the Korean goral and 0.636 and 4.8 in domestic goats. Low genetic diversity in the Korean gorals observed in this preliminary microsatellite survey suggests an urgent need for further detailed study of genetic diversity in Korean goral populations and a population management strategy based on these studies. Current results of cross-species amplification of domestic Bovidae microsatellites could be employed for the necessary population genetic studies on the Korean goral and other endangered Caprinae species.     

Power of 22 microsatellite markers in fluorescent multiplexes for parentage testing in goats (Capra hircus)

Two multiplex systems, each containing 11 microsatellite loci, were developed for semiautomated parentage testing in goats. Eight of the loci originate from goats, nine from cattle and five from sheep. Eighteen of the loci have been mapped to 16 different autosomes (in goats and cattle). Parentage exclusion probabilities were computed from allele frequencies in approximately 30 unrelated individuals from each of four economically important breeds: Mongolian Native Cashmere, Turkish Angora, Swiss Saanen, and Spanish Murciana-Grenadina. In cases where genotypes are known for one parent and an offspring, the 22 markers will exclude an (erroneously) alleged parent with a probability of > 0.999999 in the cashmere breed, > 0.99999 in Angora and Murciana-Grenadina, and > 0.9999 in Saanen. The multiplexes provide very high power for individual identification as the probability of finding two identical genotypes for the 22 loci is < 1 in 1.10(15) in each of the four breeds. The multiplexes will also be useful for studies of population structure, history, and diversity in goats and also in wild Capra species that represent important resources for genetic improvement of domestic breeds.     

Genetic relationship among Mongolian native goat populations estimated by blood protein polymorphism

Several local strains and populations of goats distinguished by morphogenetic and performance characteristics are kept by goat breeders in different natural climatic regions of Mongolia, namely Bayandelger, Ulgii Red, Erchim Black, Dorgon and Zavkhan Buural. The genetic relationships among eight native goat populations in Mongolia at 33 biochemical genetic loci was assessed. A total of 440 animals in eight regional zones were studied. Twelve loci, i.e. the serum transferrin, serum amylase, serum alkaline phosphatase, serum prealbumin-3, cell esterase-D, hemoglobin (Hb) β, hemoglobin (Hb) α-II, cell peptidase-B, cell tetrazolium oxidase, cell esterase-1, cell esterase-2 and cell catalase loci, were found to be polymorphic. The data indicated that Mongolian native goats are not highly differentiated (D=0.0002–0.0038) genetically. To set Mongolian native goats in a larger context, the present data were compared with those on other goat breeds and populations in east and southeast Asia that were previously reported. The average heterozygosity in the Mongolian native goats did not significantly differ from those in other Asian goat populations and breeds. A phylogenetic tree of the gene constitution of the Mongolian native goats and other Asian goat breeds and populations was constructed and revealed that genetically the Mongolian native goats had diverged slightly from the group consisting of Chinese, Japanese, Korean and Indonesian native goats, but markedly from the Indian goat group.     

Microsatellite analysis of genetic diversity in the Chinese alligator (<Emphasis Type="Italic">Alligator sinensis</Emphasis>) Changxing captive population

Chinese alligator (Alligator sinensis) is a critically endangered species endemic to China. In this study, the extent of genetic variation in the captive alligators of the Changxing Reserve Center was investigated using microsatellite markers derived from American alligators. Out of 22 loci employed, 21 were successfully amplified in the Chinese alligator. Sequence analysis showed loci in American alligators had a bigger average size than that of the Chinese alligators and the longest allele of an individual locus almost always existed in the species with longer stretch of repeat units. Eight of the 22 loci were found to be polymorphic with a total of 26 alleles present among 32 animals scored, yielding an average of 3.25 alleles per polymorphic locus. The expected heterozygosity (H _E) ranged at a moderate level from 0.4385 to 0.7163 in this population. Compared to that in the American alligators, a lower level of microsatellite diversity existed in the Changxing population as revealed by about 46% fewer alleles per locus and smaller H _E at the homologous loci. The average exclusion power and the ability to detect shared genotypes and multiple paternity were evaluated for those markers. Results suggested that when the polymorphic loci were combined, they could be sensitive markers in genetic diversity study and relatedness inference within the Chinese alligator populations. The level of genetic diversity present in the current Changxing population indicated an important resource to complement reintroductions based on the individuals from the other population. In addition, the microsatellite markers and their associated diversity characterized in this population could be utilized to further investigate the genetic status of this species.