微卫星DNA在动物亲子鉴定中的应用及研究进展

本文概述了微卫星DNA进行亲子鉴定的基本原理与方法,将其用于亲子鉴定的优点以及研究进展,最后对未来的应用进行了展望。     

微卫星分子标记分析四川绵羊群体遗传多样性

为探究四川省6个绵羊群体的遗传多样性,实验应用12个微卫星标记计算基因频率、有效等位基因数、杂合度及多态信息含量来评估群体内遗传多样度,通过遗传距离聚类图、群体结构推测图、主成分分析及群体间分子方差分析来评估群体间遗传关系。结果表明:6个绵羊群体在12个微卫星位点的平均有效等位基因数为3.006~3.176,平均多态信息含量变化为0.559~0.612,平均期望遗传杂合度为0.610~0.670;6个绵羊群体间的遗传关系与地理分布情况及育成史实不完全一致,但遗传距离聚类图、群体结构推测图和主成分分析结果均显示,6个绵羊群体中布拖黑绵羊类群与贾洛绵羊类群遗传关系更近;6个绵羊群体间方差组分F统计量结果为0.112 39,处于中度分化水平。     

一个水稻三倍体×二倍体杂交F2群体的微卫星杂合性丧失

DB102为一田间发现的三倍体水稻,利用它做母本与二倍体水稻中作59杂交,得到一个早世代(F2)稳定群体,通过微卫星分析发现,该群体中有一些微卫星位点发生了杂合性丧失,且至少6条染色体上发生微卫星位点杂合性丧失(loss of heterzygoisity LOH)可能有DNA序列变化。而微卫星位点的杂合性丧失又可能与该杂交组合早世代稳定有关。     

Microsatellite Marker Based Assessment of Genetic Diversity among Cultivars, Landraces and Wild Relatives in Rice

India being the primary center of origin for rice had a very large treasure of local land races, most of which are out of cultivation today. The exact genetic potential and their differences from commercial varieties and the magnitude of heterogeneity still present in them are not well catalogued. Hence the need to characterize the available land races has become imminent in the modem day concept of crop improvement （Rezai and Frey, 1990）. The present study addresses the utility of SSR markers in divulging the genetic relationships at molecular level among the major component factions office gennplasm viz., local cultivars, land races and wild species collected from a wide range of agro-geographical regions of Indian subcontinent.     

Nomenclature and genetic relationships of apples and pears from Terceira Island

Heritage apple (Malus domestica Borkh. hybrids) and pear (Pyrus communis L. hybrid) trees grow in villages throughout Terceira Island, Azores, Portugal. Some of these pears have different names but similar morphology. The objective of this study was to determine synonymy, homology, and phylogeny of apples and pears from Terceira and to examine potential relationships of the island pears with standard apples and pears of Portuguese or American descent. Nine apple microsatellite markers were used to determine genetic relationships. Distance- and parsimony-based cluster analysis grouped these genotypes into separate apple and pear clades. The Terceira apples were divided into two clades: the maçā and the reineta-reinette. Among the 17 heritage apple genotypes, seven unique accessions were identified and four groups of synonyms, or possibly clones, were detected including: ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares; ‘Reineta Castanha’ and ‘Reineta Verde Miuda’; ‘Maçā Pêra,’ ‘Maçā Calhau’, ‘Pêro Branco’ from Salga and from Terra-Chā and ‘Maçā Marmelo’; and the five genotypes ‘Maçā Sao Joao’, ‘Malápio Rosa’, ‘Maçā Gaspar’, ‘Maçā Branca’ and ‘Maçā Pato’. In addition, two homonyms were detected. ‘Pêro Vermelho’ from Terra Chā was a separate genotype from a tree from Doze Ribeiras of the same name, but Pêro Branco from Terra Chā appears to be a clone that can be distinguished by an additional allele at CH1F07a from a tree with that name from Salga. One pair of apple clones, ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares appear to be derived from an unreduced gamete of ‘Golden Delicious.’ Another apple genotype ‘Maçā Acida’ could be a sibling of the ‘Maçā Pêra’ clonal group. Other tested standard apples from the US genebank were unrelated to Terceira genotypes. Of the seven heritage pears, five unique genotypes and one pair of synonyms were detected. ‘Pêra Papo Pintassilgo’ from Raminho and ‘Pêra Vermelha’ from the nursery of Serviço de Desenvolvimento Agario da Terceira (SDAT) were synonyms. ‘Passans du Portugal’ was related to ‘Pêra Cabaca’ but other standard pears from the US genebank were unrelated to Terceira genotypes. Future studies will include additional apple and pear cultivars from other Islands of the Azores and continental Portugal, and wild Asian species to further explore genetic relationships.     

Discrimination of ‘San Marzano’ accessions: A comparison of minisatellite, CAPS and SSR markers in relation to morphological traits

‘San Marzano’ (SM) is one of the most widely known tomato (Solanum lycopersicum L.) cultivars, and is a classic example of a local variety with a premium value. Unfortunately, the original cultivated form is underrepresented in the Protected Denomination of Origin (PDO) area because of the incidence of contaminant and phenotypically similar genotypes. Our aim was to examine the ability of three DNA marker systems (minisatellite, cleaved amplified polymorphic sequence (CAPS) and simple sequence repeat (SSR)) to reveal the genetic diversity of tomato accessions that were, based on a morphological analysis, very similar. The data indicate that both minisatellites and SSRs can be used to genetically distinguish the analysed materials. Furthermore, these two marker systems depict relationships consistent with the hierarchal pattern obtained by the morphological data. As locally cultivated tomato accessions are often characterised by some degree of genetic variability, our results will be valuable in facilitating the purification, management and breeding of tomato germplasms. The differences between the marker systems employed are also discussed in relation to their usefulness in the agro-food chain.     

Genetic diversity of apricot revealed by a set of SSR markers from linkage group G1

Eight polymorphic simple sequence repeat (SSR) markers located in the G1 linkage group of apricot (Prunus armeniaca L.) were previously developed and evaluated in a small set of cultivars. Those primers were used for studying variability in 77 apricot cultivars belonging to five different geographical groups, such as Chinese, Asian (Irano-Caucasian and Central Asian), North American, Mediterranean and Western European as well as Middle European cultivars. Six of the markers were polymorphic and revealed a total of 71 alleles ranging from 5 (aprigms11) to 20 (aprigms1) alleles per locus with a mean value of 11.83 alleles per locus. In conclusion, the SSR loci located in the G1 linkage group show a level of polymorphism which is similar to loci dispersed throughout the entire genome. The total number of alleles and the number of unique alleles were the highest in Chinese apricots and the lowest in Middle European cultivars. Heterozygosity also showed a decrease from Asia and China to Middle Europe. No association could have been observed between any SSR markers tested and plum pox virus (PPV) resistant phenotype of cultivars. PPV resistant cultivars did not form a separate clade on the dendrogram obtained by UPGMA cluster analysis. Middle European and Chinese cultivars formed separate clusters while other genotypes formed smaller multiple sub-groups or scattered among different clusters. Our results support previous hypotheses on the origin of PPV resistance in North American apricots. The allele data was also presented in a form that allowed the easy observation of allele frequencies in each geographical group at each locus. Using this data field, differences and similarities between cultivar groups can be easily assessed. The analysis demonstrated the links between the North American and Mediterranean apricot germplasm and confirmed that the Chinese and Eastern European cultivars are distantly related.     

微卫星技术对黄、渤海海域7个不同地理群体中国对虾的遗传结构和遗传分化研究

采用微卫星DNA技术对黄、渤海海域7个不同地理群体的中国对虾进行了遗传结构和遗传分化研究.7个地理群体分别来自辽东湾(LD)、渤海湾(BH)、海州湾 (HZ)、乳山湾 (RS)、海洋岛 (HYD)、朝鲜半岛西海岸(KW)以及朝鲜半岛南海岸(KS).7对多态性良好的微卫星引物共检测到109个等位基因,群体平均期望杂合度(He)范围为0.810～0.864.49个群体位点中,只有15个符合Hardy-Weinberg平衡.UPGMA聚类分析显示,各地理群体亲缘关系与地理位置关系密切.根据各地理群体间遗传距离及AMOVA遗传分化检验结果,可将中国对虾分为3个独立种群,分别为中国沿岸黄、渤海群体、朝鲜半岛西海岸群体及朝鲜半岛南海群体;其中中国沿岸黄、渤海群体内部也发生了一定程度的遗传分化,但是否达到种群水平还有待于进一步验证.     

Maintaining Genetic Resources of Peach Palm (<Emphasis Type="Italic">Bactris gasipaes</Emphasis> Kunth): The Role of Seed Migration and Swidden-fallow Management in Northeastern Peru

Knowledge of the effects of farmer practices on population genetic parameters of peach palm (Bactris gasipaes Kunth) is relevant to the improvement and conservation of the palm’s genetic resources. Microsatellite markers were used to assess genetic diversity and population structure of peach palm in swidden-fallow agroforestry systems in northeastern Peru. The study covered eight communities, comprising two study areas 160 km apart – one occupied by indigenous Amerindians and the other by mixed race campesinos. Simultaneous analysis of an ex situ peach palm germplasm collection provided a means to compare population genetic parameters. Farmers who were surveyed on seed selection practices for peach palm reported that an average of only four palms (4.3 for campesino and 1.5 for indigenous populations) were used to provide seed for the establishment of the forest gardens sampled. As expected, inbreeding coefficients observed within communities were relatively high (f = 0.105 − 0.210), however, observed heterozygosities within communities were also high (0.625–0.741). A metapopulation approach was used to describe migration within and among regions, implying a hierarchical structure of gene flow which maintains relatively high levels of genetic diversity. Seed migration was found to occur over longer distances (≤600 km) and at a higher frequency (46% of palms sampled) in the indigenous study area, and a proportionally greater number of alleles was found (49 vs. 43 over three loci) with twice as many private alleles occurring only in the indigenous populations. The farmers’ practice of preserving remnant palms through successive swidden generations may have contributed to the maintenance of alleles by reducing the severity of founder effects. Although the campesino study area exhibited a significant (20% of the variation; p < 0.01) isolation-by-distance relationship across 35 km distance, in general, both study populations had relatively limited genetic structure (θ = 0.012–0.03), which is believed to have resulted from the exchange of seeds over long distances and periods of time.     

中国蛋鸭品种资源的遗传变异

微卫星作为近年来应用较多的一种分子标记，可有效的进行基因鉴定和系谱分析，并可估算群体间的遗传距离。通过筛选的28个多态性较好的微卫星标记，检测了我国13个地方蛋鸭品种的遗传多样性。利用等位基因频率计算了各群体的遗传参数和群体间的遗传距离。采用邻近法构建了NJ聚类图。结果表明：我国所有蛋鸭品种全部群体的平均杂合度较低，其遗传多样性相对贫乏，各保种场（区）要加强多样性的保护；各品种的平均多态信息含量的高低与群体平均杂合度的顺序完全一致；遗传距离的计算说明了13个品种的遗传距离较远。分化时间较长；13个地方蛋鸭品种被聚类为3个类，微山麻鸭、攸县麻鸭、淮南麻鸭、广西小麻鸭、荆江麻鸭聚为一类；巢湖鸭和恩施麻鸭聚为一类；福建省的鸭种连城白鸭、莆田黑鸭、金定鸭、山麻鸭与三穗鸭、绍兴鸭聚为一类。聚类结果的分析表明了各蛋鸭品种的分子系统发生关系与其育成史和地理分布比较一致。