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。     

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。     

运用模型分类法分析9个地方鸡品种间遗传结构和亲缘关系

以16个中度或高度多态性微卫星标记,运用基于模型分类法的Structure程序推断9个地方鸡品种群体遗传结构,构建聚类图分析相互间亲缘关系,并鉴定群体中迁移个体和具有复杂遗传基础个体的分布。结果表明：Structure程序在预先未标明品种起源的条件下准确推断出9个地方鸡种群体所属类别,揭示按照体型外貌、地理分布等为依据划分的9个地方鸡品种类型,也真实反映了其在遗传上的差异。Structure聚类图在K=2时将9个地方鸡种分为轻体型的鸡种（包括茶花鸡、藏鸡、仙居鸡、固始鸡和白耳鸡）和重体型的鸡种（包括大骨鸡、北京油鸡、鹿苑鸡和萧山鸡）两个类别;K=4、5、6时北京油鸡、固始鸡、大骨鸡分离出来形成独立类群;仙居鸡与白耳鸡在K=6、萧山鸡与鹿苑鸡在K=7、藏鸡与茶花鸡在K=8时仍聚在同一类别中,这与9个地方鸡种的地理分布及形成历史相吻合。9个地方鸡品种群体中均有具有复杂遗传基础的个体分布,迁移个体只分布在藏鸡、萧山鸡和鹿苑鸡群体中。     

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 isolation and cultured strain identification in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.     

SSR标记在小麦遗传育种中的应用研究进展

SSR(simple sequence repeat)标记是建立在PCR基础上的一种新型DNA分子标记。由于SSR在普通小麦中多态性丰富,随机分布于小麦的整个基因组中,多数表现为共显性,所以它是进行小麦遗传研究的理想工具。本文就SSR标记在构建小麦遗传连锁图谱、标记和定位目的基因、鉴定和标记染色体片段、鉴定品种、遗传多样性分析和标记辅助选择等方面的研究进展进行了综述。     

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.     

不同植物上桃蚜两种体色生物型的DNA多态性研究

用微卫星引物PCR方法分析了烟草、油菜、甘蓝3种不同寄主植物上桃蚜Myzus persica(Sulzer)红、 绿两种体色生物型的DNA多态性。结果表明:①来自不同寄主植物上的同一体色生物型个体之间的遗传关系 近于不同体色生物型个体之间的遗传关系。两种生物型内个体之间比较,红色生物型个体之间的遗传关系近 于绿色生物型个体之间的遗传关系。②来自3种不同寄主植物上的同一色型桃蚜个体中,其中同为十字花科 的甘蓝和油菜上的红色桃蚜或绿色桃蚜之间的遗传关系近于来自茄科寄主烟草上的红色桃蚜或绿色桃蚜。 ③3种寄毛上的红色桃蚜和绿色桃蚜的遗传关系相对较远,但并不明显,处于连续过渡状态。     

用微卫星标记研究南海四种石斑鱼的遗传变异与种间关系(英文)

[目的]从DNA分子水平揭示我国南海的四种重要经济海水鱼类点带石斑鱼(Epinephelusmalabaricus)、斜带石斑鱼(E.coioides)、鲑点石斑鱼(E.fario)、蜂巢石斑鱼(E.merra)的遗传群体变异水平与种间关系。[方法]本研究利用从GenBank中搜索到的21对微卫星引物,对点带石斑鱼、斜带石斑鱼、鲑点石斑鱼及蜂巢石斑鱼的等位基因数A、有效等位基因数Ne、多态信息含量PIC、观察遗传杂合度Ho、期望遗传杂合度He等遗传参数进行检测,并采用UPGMA类聚分析法探讨这四种石斑鱼的种间关系。[结果]点带石斑鱼群体的A、Ne、PIC、Ho和He等遗传参数的平均值分别为4.38±1.60、3.69±0.86、0.69±0.08、0.67±0.08、0.72±0.06,斜带石斑鱼群体的为3.88±1.13、3.55±1.04、0.66±0.10、0.68±0.21、0.70±0.08,鲑点石斑鱼群体的为6.00±1.07、4.68±0.65、0.78±0.03、0.73±0.25、0.79±0.03,蜂巢石斑鱼群体的为5.50±1.07、4.58±0.80、0.76±0.05、0.75±0.18、0.78±0.04。[结论]综合比较上述数值可知,四个石斑鱼群体的遗传变异程度依次为鲑点石斑鱼群体>蜂巢石斑鱼群体>点带石斑鱼>斜带石斑鱼群体;与其他海水鱼类的遗传变异水平比较,四种石斑鱼的群体遗传变异水平都较高,说明我国这些石斑鱼种类的遗传种质资源尚好。另外,遗传距离与类聚分析表明,点带石斑鱼与斜带石斑鱼遗传距离最近,而斜带石斑鱼与蜂巢石斑鱼的亲缘关系最远。