塔里木马鹿微卫星遗传多样性与产茸量的相关性研究

利用7个微卫星遗传标记,采用PCR扩增,12%非变性聚丙烯酰胺凝胶电泳、Sanguinetti银染法显色,对新疆塔里木马鹿亚种遗传多样性进行检测,统计各群体的等位基因组成、平均有效等位基因数(E)和平均基因纯合率(Rh),根据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,利用分子进化遗传分析软件(MEGA),采用邻结法(NJ)重建系统发生树。根据等位基因频率,利用PHYLIP(3.6)分析软件,采用最大似然法(ML)构建系统发生树,应用自举检验(bootstrap test)估计系统树中节点的自引导值(bootstrap value),并进行了系统发生分析。同时对部分马鹿群体个体基因型与产茸量之间进行了相关分析。结果表明:7个微卫星位点在3个塔里木马鹿中的多态信息含量除BMS2508和Celjp0023没有多态性,以及BM5004为中度多态外,其他4个微卫星均为高度多态,可作为有效的遗传标记用于3个塔里木马鹿群体遗传多样性和系统发生关系的分析。所有马鹿群体的平均PIC为(0.5196)、h(0.5552)和E(2.45),其基因多态性和遗传多样性相对丰富。塔里木马鹿35团群体与沙雅群体的血缘关系及遗传距离近于阿拉尔群体,3个塔里木马鹿群体的系统发育关系基本符合其地理分布和育成史。微卫星位点BM4208的166bp/185bp基因型和微卫星BM888的208bp/208bp基因型可以对塔里木马鹿35团群体高产茸量进行分子遗传标记。     

Genetic variation within and relationships among five subpopulations of Slovak Thoroughbred

Genetic variation at six microsatellite loci was analysed for five Thoroughbred subpopulations to determine the magnitude of genetic differentiation and the genetic relationships among the subpopulations. Significant deviations from Hardy-Weinberg equilibrium were shown for a number of locus-population combinations, with all subpopulations. The genetic diversities and relationships of five Thoroughbred subpopulations were evaluated using six microsatellites recommended by the International Society of Animal Genetics (ISAG). The allele frequencies, the effective numbers of alleles, and the observed and expected heterozygosities were calculated. POPGENE v. 1.31 (Yeh et al., 1997) was used to test for deviations from the Hardy-Weinberg (H-W) equilibrium and to assign F(IS) estimates (Weir, 1990). The utility of microsatellites for evaluating genetic diversity of horses is discussed.     

Analysis of genetic diversity and phylogenetic relationship of red deer subspecies in XinJiang,China

Polymorphisms for seven microsatellite loci in three red deer subspecies (9 populations) found in XinJiang were detected by polymerase chain reaction (PCR), 12% nondenaturation polyacrylamide gel electrophoresis and the Sanguinetti silver staining method. Numbers of alleles, average effective numbers of alleles (E) and the average rate of homozygosity, allelic frequencies of seven microsatellite loci, polymorphism information content (PIC), mean heterozygosity (H) and genetic distances among the populations were calculated for each population. Dendrograms were constructed based on genetic distances by the neighbor‐joining method (NJ), utilizing molecular evolutionary genetics analysis software PHYLIP (3.6). The phylogenetic tree was constructed based on allelic frequencies using maximum likelihood (ML); the bootstrap value was estimated by bootstrap test in the tree. Lastly, phylogenesis was analyzed. The results showed that four of the seven microsatellite loci were highly polymorphic, but BMS2508 and Celjp0023 showed no polymorphism and BM5004 was a neutral polymorphism. It is our conclusion that the four microsatellite loci are effective DNA markers for the analysis of genetic diversity and phylogenetic relationships among the three red deer subspecies. The mean PIC, H and E‐values across the microsatellite loci were 0.5393, 0.5736 and 2.64, which showed that these microsatellite loci are effective DNA markers for the genetic analysis of red deer. C.e. songaricus populations from Regiment 104, 151 and Hami are clustered together. C.e. yarkandensis populations from Regiment 35, Xaya and Alaer are clustered together. These two clusters also cluster together. Lastly, C.e. sibiricus populations from Burqin, Regiment 188 and the first two clusters were clustered together. The phylogenetic relationship among different red deer populations is consistent with the known origin, history of breeding and geographic distributions of populations.     

新疆塔里木马鹿遗传多样性的微卫星分析

利用5个微卫星标记对新疆塔里木马鹿遗传多样性进行了检测。统计了塔里木马鹿3个群体的等位基因组成、平均有效等位基因数(E)和平均基因纯合率(Rh),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)。结果表明5个微卫星位点在库尔勒、阿拉尔和沙雅县塔里木马鹿3个群体的平均多态信息含量分别为0.5884、0.5754、0.5344,除微卫星位点BM5004外均为高度多态,可作为有效的遗传标记用于塔里木马鹿遗传多样性分析;塔里木马鹿三个群体总平均PIC、h、Rh和E分别为:0.5661、0.5995、0.4474和2.7。分析认为塔里木马鹿遗传变异度较高,遗传多样性相对丰富,具有较大的遗传潜力。     

Screening the Polymorphic Tri-nucleotide Repeat Microsatellites from the Genome of Maiwa Yak and Their Genetic Diversity Analysis

In order to study the genetic diversity and differentiation characteristics of the existing population of Maiwa yak and provide a theoretical basis for utilization in the future, a total of 9 466 tri-nucleotide repeats microsatellites loci were screened based on the release of genome of yaks by bioinformatics method. The tri-nucleotide repeats microsatellites were amplified by PCR using the genomic DNA of 191 Maiwa yaks as templates,and the polymorphic microsatellite loci were genotyped through polyacrylamide electrophoresis experiment. After the Hardy-Weinberg equilibrium test, 5 tri-nucleotide microsatellite loci were deviated from the Hardy-Weinberg equilibrium (P <0.05). 15 microsatellite loci were highly polymorphic (PIC >0.5), the mean observed heterozygosity (Ho),mean expected heterozygosity(He) and mean polymorphic information content(PIC) were 0.5041,0.8152 and 0.7831,respectively. Maiwa yak population exhibited no characteristics of differentiation by Structure 3.0 software analysis. Therefore, Maiwa yak had plentiful genetic polymorphism while no signs of genetic differentiation were observed within the population.     

麦洼牦牛基因组三碱基重复微卫星的挖掘及其遗传多态性分析

为了解麦洼牦牛现有群体的遗传多样性及种群遗传分化特征,试验通过生物信息学方法从已公布的牦牛基因组中筛选到9 466个三碱基重复微卫星位点,以191个麦洼牦牛基因组DNA样品为模板对各个位点进行PCR扩增和聚丙烯酰胺凝胶电泳基因分型。经过哈迪-温伯格平衡检验,在15个三碱基微卫星位点中,有5个位点偏离哈迪-温伯格平衡（P<0.05）。15个微卫星位点均为高度多态性（PIC>0.5）,群体平均观测杂合度（Ho）、平均期望杂合度（He）和平均多态信息含量（PIC）分别达到0.5041、0.8152和0.7831。进一步通过Structure 3.0软件分析发现,麦洼牦牛群体结构单一,无分化特征,表明麦洼牦牛品种虽然具有较高的遗传多态性但是无分化迹象。     

基于微卫星标记的中国斑翅山鹑遗传多样性及遗传结构分析

斑翅山鹑(Perdix dauuricae)是具有很高经济价值的猎用禽.为了获得其群体遗传多样性及遗传变异信息,为我国斑翅山鹑进一步有效保护和合理利用提供科学依据,本研究共采用了8个微卫星标记位点对分布于我国北方的斑翅山鹑23个地理种群285个个体进行了群体遗传多样性及遗传结构分析.结果表明,我国斑翅山鹑为遗传多样性较丰富的群体,且每一群体均表现出较高的遗传多样性,其中柴达木盆地地区的种群拥有最高的遗传多样性.遗传分化分析显示组间及组内种群间均表现为遗传分化显著,遗传分化系数(FST)值表明大部分种群间表现出显著的遗传分化.系统树构建和贝叶斯聚类分析结果均显示斑翅山鹑23个地理种群被分化为明显的两个组群.另外,通过BOTTLENECK对各地理种群的分析显示斑翅山鹁种群曾经历过近期瓶颈效应.本研究结果表明,柴达木盆地地区种群、华北平原地区种群、静宁种群、张家川种群以及六盘山地区种群应予以相应关注,以保证有足够的种群大小来保持这些种群的遗传多样性.     

An evaluation of genetic diversity indices of the Red Bororo and White Fulani cattle breeds with different molecular markers and their implications for current and future improvement options

The genetic diversity of the Red Bororo and White Fulani cattle breeds of Cameroon and Nigeria was assessed with a panel of 32 markers. Estimates for the various indices of genetic diversity, total number of alleles (TNA), mean observed number of alleles (MNA), mean effective number of alleles (MNE), observed heterozygosity (H _ob) and expected heterozygosity (H _ex), were higher at microsatellite loci than at protein loci. Mean H _ex values were above 71% at microsatellite loci in all the breeds and ranged from 37% to 41.6% at milk protein loci and from 40.9% to 45.6% at blood protein loci. The highest TNA and MNA of microsatellites were recorded for the Nigerian White Fulani. MNE of milk protein loci was highest in the Cameroonian Red Bororo, while TNA of blood protein loci was highest in the Cameroonian White Fulani. The high genetic diversity levels indicate the presence of the necessary ingredients for improvement breeding and conservation. Multi-locus estimates of within-population inbreeding (f), total inbreeding (F) and population differentiation (θ) of the breeds were significantly different from zero, except for θ of blood proteins. A high level of gene flow was found between the breeds (5.829). The phylogenetic relationship existing among the four breeds is greatly influenced by location. The high gene flow between the breeds may lead to a loss of genetic diversity through genetic uniformity and a reduction in opportunities for future breed development. We propose an improvement scheme with aims to prevent loss of genetic diversity, improve productivity and reduce uncontrolled genetic exchanges between breeds.     

Use of microsatellites for genetic variation and inbreeding analysis in Sarda sheep flocks of central Italy

A set of 11 polymorphic microsatellites has been used to assess the distribution of genetic variability in 17 flocks of the Sarda sheep breed in the Viterbo province, located in central Italy. The suitability of samples size and number of loci analysed were tested using a bootstrap procedure. The data obtained were used to estimate the genetic diversity within and among flocks, and to test the presence of inbreeding within flocks. To study the genetic relationship among flocks, a principal component analysis based on Nei standard distances was performed. The results of genetic analysis show a significant excess of homozygotes in some of the flocks. The use of genetic data for maximum likelihood tests of assignment is investigated in order to establish suitable breeding policies. This research represents a case study for the development of mating strategies suitable for incrementing genetic variability within flocks that can be applied in other domestic species showing similar breeding problems (goat, buffalo and small populations of cattle).     

利用微卫星分析中国家兔的遗传多样性

利用10个微卫星DNA标记对七个家兔品种进行了遗传检测。利用变性聚丙烯酰胺凝胶电泳检测微卫星的PCR扩增产物,计算了七个家兔品种（9个家兔群体）样本在10个微卫星基因座上的等位基因频率、微卫星的多态信息含量（PIC）、各群体遗传杂合度、群体间的遗传距离、并根据遗传距离对这几个群体进行了聚类分析。结果表明：所选择的10个微卫星位点在所检测群体中均表现了较好的多态性,平均多态信息含量为0．569905。平均杂舍度范围在0．5885～0．6826。表明所研究的家兔品种的遗传多样性较丰富,还可以做进一步的选择利用。根据遗传距离所做的聚类分析图表明吉戎-Ⅰ系和吉戎-Ⅱ系、哈尔滨大白兔、塞北兔的亲缘关系依次较近,与它们的培育历史相吻合。     

Genetic diversity and differentiation of five Cuban cattle breeds using 30 microsatellite loci

Conservation and improvement strategies in farm animals should be based on a combination of genetic and phenotypic characteristics. Genotype data from 30 microsatellites were used to assess the genetic diversity and relationships among five Cuban cattle breeds (Siboney de Cuba, Criollo Cubano, Cebú Cubano, Mambí de Cuba and Taíno de Cuba). All microsatellite markers were highly polymorphic in all the breeds. The expected heterozygosity ranged from 0.67 ± 0.02 in the Taíno de Cuba breed to 0.75 ± 0.02 in the Mambí de Cuba breed, and the observed heterozygosity ranged from 0.66 ± 0.03 in the Cebú Cubano breed to 0.73 ± 0.02 in the Siboney de Cuba breed. The genetic differentiation between the breeds was significant (p < 0.01) based on the infinitesimal model (F_ST). The exact test for Hardy‐Weinberg equilibrium within breeds showed a significant deviation in each breed (p < 0.0003) for one or more loci. The genetic distance and structure analysis showed that a significant amount of genetic variation is maintained in the local cattle population and that all breeds studied could be considered genetically distinct. The Siboney de Cuba and Mambí de Cuba breeds seem to be the most genetically related among the studied five breeds.     

Genetic diversity in Egyptian and Italian goat breeds measured with microsatellite polymorphism.

Seven microsatellite markers were used to study genetic diversity of three Egyptian (Egyptian Baladi, Barki and Zaraibi) and two Italian (Maltese and Montefalcone) goat breeds. The microsatellites showed a high polymorphic information content (PIC) of more than 0.5 in most of the locus-breed combinations and indicated that the loci were useful in assessing within- and between-breed variability of domestic goat (Capra hircus). The expected heterozygosity of the breeds varied from 0.670 to 0.792. In the geographically wider distributed Egyptian Baladi breed there were indications for deviations from random breeding. Analysis of genetic distances and population structure grouped the three Egyptian goat breeds together, and separated them from the two Italian breeds. The studied Mediterranean breeds sampled from African and European populations seem to have differentiated from each other with only little genetic exchange between the geographically isolated populations.     

Detection of occurrence of a recent genetic bottleneck event in Indian hill cattle breed Bargur using microsatellite markers

The effective number of breedable individuals is a crucial determinant for maintaining genetic variability within a population. The population of Bargur, the hill cattle of South India, has gone down drastically by more than 93 % in the past three decades, and only a few thousand animals are available at present. The present study was undertaken to evaluate Bargur cattle for mutation drift equilibrium and to detect the occurrence of recent genetic bottleneck event, if any, in this population. About 50 unrelated animals, true to the type, were sampled and genotyped at 25 microsatellite loci. The mean observed heterozygosity (0.808?±?0.023) was higher than the mean expected heterozygosity (0.762?±?0.008) with 15 out of 25 microsatellite loci exhibiting heterozygosity excess when assumed under Hardy–Weinberg equilibrium. To evaluate Bargur cattle for mutation drift equilibrium, three tests were performed under three different mutation models, viz., infinite allele model (IAM), stepwise mutation model (SMM) and two-phase model (TPM). The observed gene diversity (H _e) and expected equilibrium gene diversity (H _eq) were estimated under different models of microsatellite evolution. All the 25 loci were found to exhibit gene diversity excess under IAM and TPM, while 22 loci were having gene diversity excess under SMM. All the three statistical tests, viz., sign test, standardized differences test, and Wilcoxon sign rank test, revealed significant (P?<?0.01) deviation of Bargur cattle population from mutation-drift equilibrium under all the three models of mutation. Furthermore, the qualitative test of allele frequency distribution in Bargur cattle population revealed a strong mode shift from the normal L-shaped form suggesting that the population had experienced genetic bottleneck in the recent past. The occurrence of genetic bottleneck might have led to the loss of several rare alleles in the population, which point towards the need for efforts to conserve this important cattle germplasm. The present study is the first report in demonstrating the genetic basis of demographic bottleneck in an Indian cattle population.     

长江中下游以及东南沿海的7个山羊群体的遗传多样性分析

利用23个微卫星标记对宜昌白山羊、马头山羊、湘东黑山羊、福清山羊、戴云山羊、黄淮山羊、长江三角洲白山羊等7个山羊群体进行了遗传多样性检测,结果显示:23个基因座中21个座位在所有群体中都具有多态性,基因座BM0203在各个群体中分别为纯合基因座,但存在群体间变异,基因座BM6444在湘东黑山羊、宜昌白山羊群体中为单态,但在其他群体都存在多个等位基因。总群体基因杂合度为0.819 0,7个群体PIC平均值在0.630 5~0.691 9之间。根据Nei氏遗传距离用UPGMA方法对7个群体进行亲缘关系聚类,马头山羊和湘东黑山羊首先聚为一类,福清山羊、戴云山羊、长江三角洲白山羊、黄淮山羊再和前者依次聚类,最后,宜昌白山羊和它们整个聚为一类。     

Genetic diversity among some cattle breeds in the Alpine area

Seven autochthonous Italian cattle breeds bred in the alpine area (Aosta Black Pied, Aosta Red Pied, Aosta Chestnut, Oropa Red Pied, Grey Alpine, Rendena and Burlina) were investigated in order to characterise their genetic structure and to study their phylogenetic origin. Two cattle breeds from Germany (Original German Brown and Holstein) and four from Switzerland (Simmental, Herens, Evolene and Brown Swiss) were included in the study in order to determine the genetic diversity existing among Italian local breeds, similar breeds bred on the other side of the Alps and in the well known Holstein.
Seventeen microsatellites, of the internationally accepted panel for the study of cattle biodiversity, were used for the analysis.
Microsatellites were highly polymorphic with a mean number of 5,5 alleles (ranging from 2 to 12 per locus). For each locus, allelic frequencies, heterozygosity (H) and the Polymorphism Information Content (PIC) were computed. The genetic equilibrium according to Hardy–Weinberg was calculated for each population and for each locus. Allele frequencies were used to estimate genetic distances and to draw a phylogenetic tree. The two closest breeds were Aosta Red Pied and Aosta Black Pied, while the two genetically most different were Holstein and Aosta Chestnut. Aosta valley breeds, Evolene and Herens constituted a tight cluster in the phylogenetic consensus tree. Principal component analysis showed a similar pattern for all the alpine breeds, while Holstein and Original German Brown were far away. The genetic differences among breeds were in accordance with their geographical and historical origins.     

中国美利奴（新疆军垦型）种羊场超细品系核心群群体遗传结构分析

利用10个微卫星标记对新疆紫泥泉国家二级种羊场中国美利奴（新疆军垦型）羊超细品系核心群基因组DNA的遗传多样性进行了检测。统计了各位点的多态信息含量(PIC) 、遗传杂合度(H) 、有效等位基因数(E) 和基因纯合率(Rh)等遗传指标。结果表明：除BMS772（PIC=0.2567），BM6506（PIC=0.4697）为中度多态外，其余微卫星位点均为高度多态；群体的平均PIC、H、E、Rh分别为0.5848、0.6333、3.0315和0.4841，该品系核心群的基因纯合率处于中等水平，具有较为丰富的多样性；10个微卫星标记中只有OARAE57位点处于哈代 温伯格平衡(P>0.05)。     

Genetic variability of Trypanosoma evansi isolates detected by inter-simple sequence repeat anchored-PCR and microsatellite

Studies on genetic variability in Trypanosoma evansi have been limited by a lack of high-resolution techniques. In this study, we have investigated the use of inter-simple sequence repeats (ISSR) and microsatellites in revealing polymorphism among T. evansi isolates. Twelve ISSR primers and five microsatellite loci were used to generate polymorphic bands and alleles, respectively, to investigate the genetic variability among T. evansi isolates from Africa and Asia. Seven of the twelve ISSR primers showed variability between isolates with a total of 71 fragments of which 49(69%) were polymorphic. Microsatellite analysis revealed a total of 60 alleles. On average the ISSR markers revealed a higher genetic diversity (23%) than microsatellites (21.1%). The two techniques showed a strong agreement of r=0.95 for Dice and r=0.91 for Jaccard indices in estimating the genetic distances between isolates. The distance UPGMA tree revealed two major clusters of T. evansi which correlate with the minicircle classification of subtype A and B. The cophenetic correlation coefficient between Dice and Jaccard based matrices were r=0.79 for microsatellites and r=0.73 for ISSR indicating a strong agreement between dendrograms. The results suggest that both ISSR and microsatellites markers are useful in detecting genetic variability within T. evansi.     

Biodiversity in Mediterranean buffalo using two microsatellite multiplexes

The present study was carried out to investigate genetic diversity in Nile-Delta and Southern-Egypt buffalo populations in comparison with the Italian buffalo utilizing two microsatellite multiplexes. A total of 104 animals classified into three groups were used, 28, 38 and 38 representing the Nile-Delta, Southern-Egypt and Italian buffalo, respectively. The 15 studied microsatellites were CSSM38, CSSM70, CYP21, CSSM42, CSSM60, MAF65, BM0922, CSSM19, INRA006, ETH02, BM1706, BMC1013, CSSM47, INRA026 and CA004. All studied microsatellites showed allelic polymorphism. Number of polymorphic alleles ranged between 4 alleles (CSSM38, CSSM70 and CYP21) and 11 alleles (CA004). Pairwise Chi-square test for Nile Delta and Southern Egypt showed significant differences in allelic distribution at five loci, CSSM70, CSSM38, BM0922, ETH02 and BM1706. Italian buffalo showed the lowest percentage of observed heterozygotes (65%), while the Southern Egypt showed the highest (71%). Both the Italian and the Delta populations deviated significantly (P < 0.05) from HW equilibrium. Italian buffalo is relatively the most inbred population while the Southern-Egypt buffalo is the only outbred population. High level of genetic differentiation (F_ST estimates) between the Italian group and each of the Delta and Southern-Egypt group (0.083 and 0.076, respectively) was observed while Southern-Egypt group showed a lower level of genetic differentiation with the Delta group (0.014). Italian buffalo had the greatest genetic distance values with the two Egyptian groups (0.25 and 0.23) while much lower values between the Southern-Egypt and the Delta groups (0.06) was observed. Genetic variation between the Italian buffalo and the Egyptian buffalo was detected in 14 (out of 15) microsatellite loci. While a lower level (than that between Egyptian and Italian) of genetic variation between Southern-Egypt and Delta buffalo populations was expressed by 5 loci. It was concluded that the Southern-Egypt buffalo could be considered as distinct population from the Delta buffalo. In addition, Southern-Egypt group, being the only group with non-significant global deviation from HW equilibrium, the most heterozygous, and the only outbred population as well, is expected to respond more favorably to selection.     

The Genetic Polymorphism Analysis in Tree Shrew(T.belangeri chinensis) based on Fluorescent Microsatellite

A total of 10 fluorescent labeling microsatellite markers detected by capillary electrophoresis were selected to analyze the genetic diversity of tree shrew(T.belangeri chinensis).Microsatellite Toolkit and PopGene softwares were used to calculate the genetic correlation indexes to assess the genetic diversity of the population.64 alleles with 6.4 alleles per locus were identified among 10 microsatellite loci.The average effective number of alleles, observed heterozygosity, expected heterozygosity and polymorphic information content were 3.7892, 0.6156, 0.6963, and 0.6367, respectively.Seven of all loci were high polymorphic, and three were moderate polymorphic.Five loci extremely significantly deviated from Hardy-Weinberg equilibrium(P<0.01).These novel polymorphic microsatellite markers indicated high genetic diversity, which will be used to evaluate genetic diversity in tree shrew.     

Microsatellite analysis of genetic diversity in the Catalonian donkey breed

A total of 111 individuals (79 females and 32 males) of the endangered Catalonian donkey breed were analysed by using a commercial equine paternity polymerase chain reaction typing kit. Eleven of the 12 horse microsatellites were amplified when using donkey's DNA. One locus, ASB2, did not amplify in any sample. The allele range for HTG4 overlapped with HMS7 and their results could not be interpreted. The mean number of alleles detected per locus was 7.7 (±1.0), being the average of genetic diversity detected in the population, and measured as the unbiased average expected heterozygosity ( H _e), of 0.712 ± 0.038. All the analysed loci showed disagreement with Hardy–Weinberg proportions except HTG7 (from permutation tests). The within-population-inbreeding estimate was highly significant (p < 0.001) and equal to 15.4% (as measured by F _IS-statistic). The cumulative exclusion probability was 0.999. The deficit of heterozygotes can be partly explained by a population subdivision effect ( F _ST=3.5%), although the main factor that has provoked this lack of heterozygotes can be attributed to consanguinity.