Genetic diversity studies of Kherigarh cattle based on microsatellite markers

We report a genetic diversity study of Kherigarh cattle, a utility draught-purpose breed of India, currently declining at a startling rate, by use of microsatellite markers recommended by the Food and Agriculture Organization. Microsatellite genotypes were derived, and allelic and genotypic frequencies, heterozygosities and gene diversity were estimated. A total of 131 alleles were distinguished by the 21 microsatellite markers used. All the microsatellites were highly polymorphic, with mean (±s.e.) allelic number of 6.24 ±1.7, ranging 4–10 per locus. The observed heterozygosity in the population ranged between 0.261 and 0.809, with mean (±s.e.) of 0.574 ±0.131, indicating considerable genetic variation in this population. Genetic bottleneck hypotheses were also explored. Our data suggest that the Kherigarh breed has not experienced a genetic bottleneck in the recent past.     

Genetic diversity in Indian cucumber based on microsatellite and morphological markers

Genetic variation among 44 cucumber accessions was assessed using morphological and SSR markers. High genetic variability was observed for days to 50% female flowering (37–46 days from sowing), number of fruits per plant (1.4–6.0), individual fruit weight (0.04–0.552 kg) and root length (14.25–32.8 cm). The pair-wise Jaccard similarity coefficient ranged between 0.25 and 0.85 indicating that the accessions represent genetically diverse populations. The allelic diversity of polymorphic markers ranged from 0.001 to 0.9396 with an average of 0.31 based on polymorphic information content. The clustering pattern of SSR markers was not in consonance with the groupings based on quantitative traits. The accession of Indian state i.e.; Madhya Pradesh and Uttar Pradesh were diverged from the accessions of other parts of India. The study provides information for future exploration and collection of cucumber germplasm in India and utilization of diverse germplasm for developing cultivars/hybrids for specific traits.     

Genetic diversity in Capsicum germplasm based on microsatellite and random amplified microsatellite polymorphism markers

A sound knowledge of the genetic diversity among germplasm is vital for strategic germplasm collection, maintenance, conservation and utilisation. Genomic simple sequence repeats (SSRs) and random amplified microsatellite polymorphism (RAMPO) markers were used to analyse diversity and relationships among 48 pepper (Capsicum spp.) genotypes originating from nine countries. These genotypes covered 4 species including 13 germplasm accessions, 30 improved lines of 4 domesticated species and 5 landraces derived from natural interspecific crosses. Out of 106 SSR markers, 25 polymorphic SSR markers (24 %) detected a total of 76 alleles (average, 3.04; range, 2–5). The average polymorphic information content (PIC) was 0.69 (range, 0.29–0.92). Seventeen RAMPO markers produced 87 polymorphic fragments with average PIC of 0.63 (range, 0.44–0.81). Dendrograms based on SSRs and RAMPOs generated two clusters. All 38 Capsicum annuum genotypes and an interspecific landrace clustered together, whereas nine non-annuum (three Capsicum frutescens, one Capsicum chinense, one Capsicum baccatum and four interspecific landraces) genotypes clustered separately. Genetic variation within non-annuum genotypes was greater than the C. annuum genotypes. Distinctness of interspecific derivative landraces grown in northeast India was validated; natural crossing between sympatric Capsicum species has been proposed as the mechanism of their origin.     

Genetic diversity of Triticum turgidum L. based on microsatellite markers

Using microsatellite (SSR) markers, the genetic diversity and genetic relationships among 48 Triticum turgidum L. accessions, including 30 Triticum turgidum L. ssp. turgidum, 7 Triticum turgidum L. ssp. durum, 4 Triticum turgidum L. ssp. carthlicum, 3 Triticum turgidum L. ssp. paleocolchicum, 2 Triticum turgidum L. ssp. turanicum and 2 Triticum turgidum L. ssp. polonicum accessions, were investigated. A total of 97 alleles were detected on 16 SSR loci. At each locus, the number of alleles ranged from 2 to 14, with an average of 6.1. The Genetic similarity (GS) value ranged from 0.20 to 0.92, with the mean of 0.59. In cluster analysis, it was found the 48 Triticum turgidum L. accessions could be distinguished easily by SSR markers, whereas the 6 subspecies taxonomic entities of T. turgidum L. could not differentiate with each other, indicating that the morphological differences present among the 6 subspecies could not be reflected by the SSR markers. These results suggested that SSR markers had the superiority in detecting the genetic diversity of T. turgidum L., while it was not good for the studies of the phylogenic relationships among the subspecies of T. turgidum L.     

Genetic diversity analysis of BMY cattle based on microsatellite DNA markers

cattle (1/2 Brahman, 1/4 Murray Grey and 1/4 Yunnan Yellow cattle) has been inter se breeding since 1980s. Genetic diversity of BMY cattle was extensively investigated using 16 microsatellite markers. A total of 130 microsatellite alleles and high allele size variance were detected. All loci displayed high genetic diversity with overall mean of N _a = 8.13, PIC = 0.7224 and H _e = 0.7666, which were higher than those of many other beef breeds. The allele-sharing neighbour-joining tree clearly displayed the new genotypic combinations and the minglement from both BMY cattle and Brahman. The results provided the genetic information to match the standards of new beef breed in South China.     

Genetic diversity of Hungarian indigenous chicken breeds based on microsatellite markers

Six local chicken breeds are registered in Hungary and are regarded as Hungarian national treasures: Hungarian White, Yellow and Speckled, and Transylvanian Naked Neck White, Black and Speckled. Three Hungarian academic institutes have maintained these genetic resources for more than 30 years. The Hungarian Yellow, the Hungarian Speckled and the Transylvanian Naked Neck Speckled breeds were kept as duplicates in two separate subpopulations since time of formation of conservation flocks at different institutes. In this study, we investigated genetic diversity of these nine Hungarian chicken populations using 29 microsatellite markers. We assessed degree of polymorphism and relationships within and between Hungarian breeds on the basis of molecular markers, and compared the Hungarian chicken populations with commercial lines and European local breeds. In total, 168 alleles were observed in the nine Hungarian populations. The F _ST estimate indicated that about 22% of the total variation originated from variation between the Hungarian breeds. Clustering using structure software showed clear separation between the Hungarian populations. The most frequent solutions were found at K  = 5 and K  = 6, respectively, classifying the Transylvanian Naked Neck breeds as a separate group of populations. To identify genetic resources unique to Hungary, marker estimated kinships were estimated and a safe set analysis was performed. We show that the contribution of all Hungarian breeds together to the total diversity of a given set of populations was lower when added to the commercial lines than when added to the European set of breeds.     

Genetic diversity of Triticum turgidum L. based on microsatellite markers

Using microsatellite (SSR) markers, the genetic diversity and genetic relationships among 48 Triticum turgidum L. accessions, including 30 Triticum turgidum L. ssp. turgidum, 7 Triticum turgidum L. ssp. durum, 4 Triticum turgidum L. ssp. carthlicum, 3 Triticum turgidum L. ssp. paleocolchicum, 2 Triticum turgidum L. ssp. turanicum, and 2 Triticum turgidum L. ssp. polonicum accessions, were investigated. A total of 97 alleles were detected at 16 SSR loci. At each locus, the number of alleles ranged from two to fourteen, with an average of 6.1. The genetic similarity (GS) value ranged from 0.20 to 0.92, with the mean of 0.59. In cluster analysis, it was found the 48 Triticum turgidum L. accessions could be distinguished easily by SSR markers, whereas the six subspecies taxonomic entities of T. turgidum L. could not differentiate with each other, indicating that the morphological differences present among the six subspecies could not be reflected by the SSR markers. These results suggested that SSR markers had superiority in detecting the genetic diversity of T. turgidum L., while they were not good for studies of the phylogenic relationships among the subspecies of T. turgidum L. The text was submitted by the authors in English.     

Genetic diversity analysis of BMY cattle based on microsatellite DNA markers

BMY cattle (1/2 Brahman, 1/4 Murray Grey and 1/4 Yunnan Yellow cattle) has been inter se breeding since 1980s. Genetic diversity of BMY cattle was extensively investigated using 16 microsatellite markers. A total of 130 microsatellite alleles and high allele size variance were detected. All loci displayed high genetic diversity with overall mean of N(a) = 8.13, PIC = 0.7224 and H(e) = 0.7666, which were higher than those of many other beef breeds. The allele-sharing neighbour-joining tree clearly displayed the new genotypic combinations and the minglement from both BMY cattle and Brahman. The results provided the genetic information to match the standards of new beef breed in South China.     

Genetic diversity and molecular differentiation of Chinese toad based on microsatellite markers

Genetic diversity and population structure of 9 populations of Bufo gargarizans with total 111 samples in China were assessed using seven microsatellite loci. The analysed microsatellite markers produced 161 alleles, varied from 9 to 38 alleles each locus. The number of alleles per population per locus ranged from 4.43 to 10.29. Polymorphic information content showed that all seven loci were highly informative (mean = 0.810 ± 0.071). The average observed heterozygosity was less than the expected (0.353 ± 0.051 and 0.828 ± 0.067, respectively). All tested populations gave significant departures from Hardy–Weinberg equilibrium. Genetic differentiation among the populations was considerably high with the overall and pairwise F _ST values (mean = 0.160 ± 0.039), and showed fairly high level of inbreeding (indicated by a mean F _IS value of 0.504 ± 0.051) and global heterozygote deficit. In comparison to other amphibian studies; however, our results suggested that the level of genetic structuring in B. gargarizans was relatively low in the geographical scale of the study area. Interestingly, the speculated population bottleneck was found to be absent and the analyses provide only weak evidence for a recent contraction in size even though there was severe inbreeding (indicated by the F _IS value) in the Chinese toad populations.     

基于微卫星标记的5个尼罗罗非鱼品系的遗传多样性分析

为了对生产上应用的5个尼罗罗非鱼(Oreochromis niloticus)品系的遗传背景进行本底调查,以期为育种工作提供基础数据, 本文用19对微卫星引物对5个尼罗罗非鱼品系(苏丹品系、台湾品系、吉富品系、超雄品系Ⅰ和超雄品系Ⅱ)进行了遗传多样性分析,计算并统计了等位基因数、多态信息含量( PIC)、杂合度、遗传相似性系数、遗传距离等参数。结果表明19个微卫星位点在5个罗非鱼品系中共检测到113个等位基因,平均期望杂合度在0.578–0.692之间,平均多态信息含量在0.473–0.628之间。5个尼罗罗非鱼品系有较丰富的遗传多样性,而超雄品系Ⅰ的遗传多样性相对较为贫乏。     

雪峰乌骨鸡自然群体遗传多样性的微卫星分析

雪峰乌骨鸡是具有独特遗传特性的优质肉药兼用地方家禽品种.为了评价雪峰乌骨鸡现有品种资源的遗传背景、群体遗传结构等,提出合理的保种方法和措施,本研究筛选了家鸡基因组23条染色体上的23个微卫星标记,对采自湖南怀化的50只纯种乌骨鸡个体进行了多态性检测.结果表明:23个微卫星座位共检测到79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2-6个之间,平均为3.435个.群体表现出较高的平均杂合度和平均多态信息含量,分别为0.6285和0.5496,说明雪峰乌骨鸡自然群体具有较丰富的遗传多样性.研究结果将为雪峰乌骨鸡种的保护和利用提供重要的理论依据.     

Genetic diversity and differentiation of Chinese domestic buffalo based on 30 microsatellite markers

To determine genetic diversity and evolutionary relationships among Chinese buffalo populations, 18 indigenous swamp buffalo populations and two introduced river buffalo breeds were genotyped for 30 microsatellite loci. The mean number of alleles across the 30 loci was 8.13, and the expected heterozygosity ranged from 0.517 (Yanjin) to 0.609 (Dehong). Although there was abundant genetic variation, genetic differentiation between Chinese buffalo populations was low, with only 2.8% of the total genetic variance among populations. The genetic differentiation pattern and genetic relationships among Chinese buffalo populations were consistent with their geographic distribution. The Dehong population was discerned as a distinct indigenous population, but suffered genetic admixture with river buffalo. The remaining populations were divided into four major clusters, i.e. the Upper and Middle Reaches of Yangtze Valley cluster (Guizhou, Guizhoubai, Yanjin, Fuling, Enshi and Jianghan), the Lower Reaches of Yangtze Valley cluster (Haizi, Shanqu and Dongliu), the South China cluster (Fuan and Xinfeng) and the Southwest China cluster (Xinglong, Xilin, Diandongnan and Dechang).     

家兔遗传多样性及系统发生关系的微卫星分析

为了揭示我国家兔资源的遗传现状,本文从30个微卫星位点中筛选出10个位点,对22个家兔品种(系)的遗传变异进行了分析.结果表明:22个家兔品种(系)在10个微卫星座位上的平均有效等位基因数为3.O-4.6;平均多态信息含量在0.4997-0.7009之间,多为高度多态,说明家兔有丰富的遗传多样性.各品种之间的遗传距离(Ds)较近,在0.0087-1.2712之间,其中以美系獭兔和德系獭兔间距离最近,为0.0087.德国大白兔与长毛兔之间距离最远,为1.2712.根据遗传距离做聚类图,可以看出中国22个家兔品种被分为三大类:吉戎-Ⅰ系和吉戎-Ⅱ系成一类,塞北兔、哈白兔聚为一类,其他品种聚为一大类.该研究结果将为中国家兔品种的保护和利用提供重要的科学依据.     

Genetic diversity and relationship among southern Indian goat breeds based on microsatellite markers

Genetic diversity and relationship among goat breeds of southern India were investigated based on microsatellite markers. All five breeds of south India, namely Attappady, Osmanabadi, Sangamneri, Malabari and Kanniadu, along with Ganjam of eastern India were considered for the investigation. In total, 190 alleles were observed from 288 DNA samples analysed with 25 microsatellite loci across six breeds. The most diverse breed was Kanniadu and the least was Osmanabadi. Gene diversity for each breed ranged from 0.73 in Kanniadu to 0.61 in Osmanabadi. The genetic distance tended to be least (0.22) between Ganjam and Malabari and the widest (0.83) between Kanniadu and Malabari. The genetic differentiation between different pairs of the breeds was significantly different from zero. Majority of the loci in almost all the breeds were heterozygote deficit. The overall F_is value (0.20) was moderate and significantly different from zero. Principal-component analysis showed the clustering of the goat breeds according to their geographical origin. Therefore, the geographical origin of the breeds should be taken into consideration while deciding conservation and improvement options for these breeds.     

中国5个地方牦牛品种遗传多样性的微卫星分析

本文以大额牛（Bos frontalis）为外群,应用16对微卫星DNA标记结合荧光-多重PCR技术,评估了5个中国地方牦牛（Bos grunniens）（帕里牦牛、斯布牦牛、西藏高山牦牛、麦洼牦牛和九龙牦牛）品种内遗传变异和品种间遗传关系。6个群体的16个微卫星座位上共检测到159个等位基因,其中有33个等位基因为5个牦牛品种所特有。6个群体的有效等位基因数（Ne）在2.2043-3.2754之间,平均杂合度（H）在0.4858-0.6153之间,平均多态信息含量（PIC）在0.4230-0.5711之间。5个牦牛品种的微卫星座位有丰富的遗传多样性;而大额牛的遗传多样性相对较贫乏。5个牦牛群体间的遗传分化系数（Gst）为0.0527,表明牦牛亚群体间遗传分化水平很低。采用邻近结合法构建聚类图和模糊聚类分析表明,5个牦牛品种分为两大类,其中斯布牦牛、西藏高山牦牛、帕里牦牛和麦洼牦牛为一大类,九龙牦牛为一类。研究结果将为中国地方牦牛品种的保护和利用提供重要的理论依据。     

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。     

利用微卫星标记分析我国13个地方灰羽鹅品种的遗传多样性

我国地方鹅品种具有很多优良性状,但长期以来由于没有形成科学的选育制度和培育方法,品种的遗传多样性没有得到完整保存。为了深入了解我国地方鹅品种的遗传结构,使之得到更好的保护和利用,作者选用31个多态性较高的微卫星标记(其中19个是首次用磁珠富集法从AFLP片段中分离),检测了我国13个地方灰羽鹅(An-sercygnnoides)品种的遗传多样性。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)和群体间的遗传距离(DA)。结果表明:13个地方灰羽鹅品种中,平均多态信息含量为0.323–0.398,平均杂合度为0.4985–0.6727,各品种的杂合度都较高,最高的是狮头鹅(0.6727),最低的是雁鹅(0.4985)。用UPGMA法进行聚类分析的结果显示,13个品种被聚为4类:丰城灰鹅、武冈铜鹅、兴国灰鹅、狮头鹅、乌棕鹅、阳江鹅、马冈鹅、钢鹅、雁鹅聚为第1类;伊犁鹅自聚为第2类;长乐鹅、右江鹅聚为第3类;永康灰鹅自聚为第4类。本研究为鹅遗传育种提供了参考资料。     

Genetic diversity estimation and core collection construction of Sinojackia huangmeiensis based on novel microsatellite markers

Sinojackia huangmeiensis is a critically endangered tree species in the Styracaceae endemic to China. To create tools to better evaluate the genetic diversity of this species, we used a modified genomic library enrichment method, the PETUER (Probe Extension and TEACl Universal EnRichment) method, to develop genetic markers. The resulting 18 microsatellite loci had high polymorphism with 3–12 alleles per locus and showed negligible stutter. Observed (H_O) heterozygosities were 0.057–0.724, and polymorphism information content (PIC) was 0.109–0.794. No significant linkage disequilibrium between pairs of loci was found. All 123 individuals found in the National Natural Reserve of Longgan Lake was genotyped at the 18 loci and used to estimate a UPGMA dendrogram. Using an iterative clustering method, a set of 18 individuals was established as a core collection to represent genetic diversity and would be useful to facilitate the conservation for S. huangmeiensis. In addition, we tested the 18 loci for cross-species amplification in three other Sinojackia species and the closely related Changiostyrax dolichocarpa. These polymorphic loci will be valuable for future population genetic and phylogenetic studies of S. huangmeiensis and congeneric species, as well as in closely related Styracaceae.     

Genetic diversity and parentage assignment in Dojo loach,Misgurnus anguillicaudatus based on microsatellite markers

In this study, genetic diversity and structure of three Misgurnus anguillicaudatus populations from three different geographical locations in China (Hunan, Hubei and Henan province) were investigated using microsatellite markers. High level of genetic diversity of all three populations was revealed by expected heterozygosity (H_e), observed heterozygosity (H_o) and allele number. Significant genetic differentiations were found between all pairs of populations. The efficiency of eight microsatellite markers in parentage assignment of 540 progeny from twenty full-sib families was evaluated. Simulation based on allele frequency data demonstrated that probabilities of exclusion per locus range from 0.313 to 0.825 when no parent information is available and 0.504 to 0.904 when one parent is known. The assignment success rate based on the real data using eight markers was 96.85%. This study indicates that these M. anguillicaudatus resources are valuable genetic and breeding material for aquaculture and the microsatellite markers will be useful for investigation of genetic background and molecular marker-assisted selective breeding in this species.