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。     

东北三省水稻遗传多样性和亲缘关系的SSR分析

以2006年辽宁、吉林、黑龙江三省水稻区域试验品种(系)为试材,采用SSR标记分析了东北三省35个水稻品种(系)的遗传多样性和亲缘关系,结果共检测出237个等位基因,分子量变异范围在95~320 bp。品种(系)间不同位点等位基因数目不等(2~8个),平均3.3857个,平均Nei基因多样性指数为0.4935,变幅为0.0328(PSM116)~0.8577(PSM363)。多态性位点的比率变化范围为71.25%~90.04%,多样性指数(H)介于0.3869~0.4903,信息指数(I)在0.6399~0.8489。吉林和黑龙江供试品种的SSR多样性低于辽宁水稻品系,而吉林和黑龙江基本相同。各稻区内品种的多样性均低于总体水平,显示了东北三省2006年水稻育种遗传基础的狭窄性。     

草原红牛微卫星标记的研究

选用草原红牛30头作为试验牛群体,经过牛血液基因组DNA的提取、8对微卫星引物的PCR扩增、扩增产物的聚丙烯酰胺电泳分型、计算机凝胶成像分析系统分析各位点等位基因及全部个体的标记基因型、PPAP3.0软件计算基因频率、多态信息含量(PIC)和杂合度等步骤从分子水平上分析了草原红牛在8个位点的遗传多态性。结果表明,IDVGA2、IDVGA46、TGLA44、BM1824、ETH225、BM2113、IDVGA44和IDVGA55等位基因数分别为6、6、6、4、4、2、6和4,多态信息含量分别为0.722 3、0.749 3、0.671 3、0.584 9、0.671 5、0.508 9、0.761 2和0.596 5,这8个位点均为高度多态位点。8个位点作为遗传标记应用于草原红牛遗传育种研究是可行的。     

虹鳟倍性鉴定SSR标记筛选与应用研究

虹鳟(Oncorhynchus mykiss)是我国冷水性鱼类的主要养殖品种。与二倍体相比,虹鳟三倍体具有生长速度快、肉质好和不育等优点。目前,鉴定虹鳟倍性主要是通过流式细胞术进行DNA含量分析,但这种方法需要采集鱼类血液或组织样本,会对其造成伤害。为了实现利用微创方法收集样本鉴定虹鳟倍性,本研究利用PCR扩增以及电泳分离技术对153个微卫星标记(SSR标记)进行分析,其中139个SSR标记能够在二倍体和三倍体中成功扩增,筛选出132个SSR标记具有多态性,最终鉴定出7个标记在三倍体中表现出较高的变异性。通过在52个已知倍性水平的参考样本和48个未知倍性的样本上进行验证,进一步从中筛选出3个SSR标记(SSR1054、SSR1056和SSR1468)足以区分倍性水平,并且根据3个标记序列重新设计了3对具有良好稳定性的引物序列进行倍性分析应用。本研究所筛选的SSR标记解决了现有虹鳟倍性鉴定中存在的技术流程复杂、耗材昂贵的问题,并有助于不同倍性虹鳟的遗传多样性研究。     

伏牛白山羊6个微卫星标记的遗传多样性分析

选取位于绵羊第6号染色体上与牛多胎基因FecB紧密连锁的2个微卫星基因座OarAE1 01、BM1329,牛第3号染色体上的微卫星基因座BMS1248,绵羊第4号染色体上的2个微卫星基因座MAF70、OarHH35及牛第8号染色体上的微卫星基因座BM1227,对伏牛白山羊遗传多样性进行检测。结果表明,6个微卫星基因座在伏牛白山羊中共检测到50个等位基因,其中平均多态信息含量(PIC)为0.789,且每个微卫星基因座的PIC>0.5,平均有效等位基因数(Ne)、平均杂合度(H)分别为5.165、0.903。由此表明,6个微卫星标记均具有高度多态性,可用于伏牛白山羊遗传多样性的分析。     

野生与家养鹌鹑种群杂交后代遗传变异机制初探

不同种群间的杂交是丰富遗传变异最常见的手段,也是保护野生濒危物种常用的方法之一,研究杂交后群体遗传变异水平和机制对动物遗传资源保护、评价具有重要的意义。运用微卫星DNA标记对中国境内分布的野生日本鸣鹑与家鹌鹑群体及其杂交后代(F1、F2代)的遗传变异机制进行分析,结果表明:F1代鹌鹑群体每个座位平均检测出4.14个等位基因,群体多态信息含量和平均杂合度分别为0.5834和0.6265,具有比杂交亲本更高的遗传多样性水平。F2代遗传多样性有所降低,平均检测出3.57个等位基因,有4个等位基因在杂交过程中因遗传漂变而丢失;同时发现F1代、F2代与杂交亲本的遗传分化趋势不一致,F2代鹌鹑与野生日本鸣鹑遗传分化系数最大,为0.0712;而F1与之最小,仅为0.0275,表明鹌鹑杂交后代的遗传分化受分散过程的作用,主要受遗传漂变的影响。表明杂交可能是造成群体无定向分化及遗传多样性丧失的主要机制之一。     

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.     

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.     

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.