Assessment of the uniformity of wheat and tomato varieties at DNA microsatellite loci

By analysing a number (20–38) of individuals from selected varieties of wheat and tomato, we have been able to assess intra-varietal uniformity at certain micro satellite (simple sequence repeat, SSR) loci. In total, 45 varieties of wheat were analysed at between 7–9 different SSR loci, and 10 varieties of tomato were analysed at six loci. The results showed that there was variation both between varieties and between microsatellites in the degree of non-uniformity observed, and it was possible to identify a number of different probable sources of non-uniformity. Twenty-four of the wheat varieties and nine of the tomato varieties were sufficiently uniform to meet the standards currently applied for distinctness, uniformity and stability (DUS) testing using phenotypic characteristics. The implications for the potential future use of SSRs in DUS testing are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

鲢三、四核苷酸重复微卫星标记的筛选及其特征分析

通过磁珠富集法筛选鲢(Hypophthalmichtys molitrix)的三、四核苷酸重复微卫星分子标记。提取鲢血液大片段基因组DNA,用限制性内切酶Sau3AI进行酶切,蔗糖密度梯度离心收集400~900 bp长度的片段,构建鲢全基因组PCR文库,用生物素标记的微卫星探针(TGA)8、(CAG)8以及(AGAT)6进行筛选,磁珠富集含有微卫星序列的DNA片段,获得含有微卫星的单链序列;将这些序列通过PCR扩增,连接pMD 18-T载体,转入感受态大肠杆菌(Escherichia coli)DH5α,获得微卫星文库;再用γ-32P标记的放射性同位素探针进行二次杂交筛选,获得1 758个阳性克隆,选择测序348个克隆序列中,共含有280个微卫星座位,其中完美型167个(59.64%),非完美型28个(10%),混合型85个(30.36%)。Primer 3.0软件设计60对引物,并用珠江水系的野生鲢群体对引物进行多态性位点检测。统计软件分析显示其中22对具有较高的筛选效率和多态性,可作为鲢种质评价等遗传分析的工具。研究亮点:首次利用多碱基(三、四核苷酸重复)探针筛选出微卫星分子标记结合同位素探针进行两次筛选...     

甜菜EST-SSR引物的开发与应用

利用NCBI公共数据库现有的甜菜(Beta vulgaris L.)表达序列标签(expressed sequence tags,EST)数据信息,开发了甜菜EST-SSR标记。在所有的29830条甜菜EST序列中共确认得到20109条非冗余EST序列,总长为11287.6kb。在含有微卫星重复的6951条EST序列中按照SSR引物设计要求,最终获得了2845个EST-SSR,平均每3.96 kb含有1个SSR。EST-SSR的分布频率和特征分析表明,A/T单碱基重复最多,其次是AAG/CTT三核苷酸重复,AG/CT二核苷酸重复,ACCTCC/AGGTGG等六核苷酸重复最少。随机合成了100对SSR引物,并分别选用6个甜菜品种进行多态性检验,将其按遗传相似性分为两组,多态信息含量(polymorphism information content,PIC)平均值为0.47。本研究证实这种全新的开发甜菜SSR标记的方法具有高效、多态性较高的特点,在甜菜遗传多样性分析、功能基因定位、遗传图谱构建以及比较基因组等研究方面有广阔的利用前景。     

Molecular markers as a method to evaluate the movement of Hypothenemus hampei (Ferrari)

The objective of this research was to develop a methodology to describe the movement of the coffee berry borer Hypothenemus hampei (Coleoptera: Curculionidae) in the field through: (i) the evaluation of allele variation of a microsatellite marker on polymorphic Colombian H. hampei populations; (ii) the invention of a device for releasing H. hampei adults; (iii) the standardization of a release-recapture technique for H. hampei populations; (iv) the estimation of the flight distance of the insect; and (v) the calculation of a mathematical expression that describes the movement of H. hampei in space over time. The results indicated that: (i) the microsatellite molecular marker HHK.1.6 was exclusively present in a population from Guapotá-Santander, was dominant and allows the evaluation of H. hampei movement for several generations; (ii) a device that released 88.8% of H. hampei adults in 2 s was designed; (iii) this device was used as H. hampei populations containing HHK.1.6 marker release strategy, and coffee seeds as recapture strategy; (iv) it was estimated that H. hampei adults flew as far as 65 m, however, 90% were recovered in a radius of <40 m. Finally, (v) the mathematical expression that described the movement of H. hampei in space over time was Y^=αβXi, being Y^ the average number of borer beetles recaptured per tree, and x the distance in meters. This method will allow to determine the movement of H. hampei from different environmental and ecological scenarios.     

文昌鸡群体遗传多样性的微卫星分析

利用10对来自鸡不同染色体的微卫星引物序列,对文昌鸡46个样本（雌雄各半）进行了遗传多样性检测。结果表明：该群体平均多态信息含量为0．808,平均观察杂合度为0．351,平均期望杂合度为0．841,平均等位基因14．6个,表明海南文昌鸡群体遗传变异较大,遗传多样性较为丰富。研究结果为文昌鸡品种资源合理保护与开发利用提供了科学依据。     

Genetic variation and the relationship between stream and lake ecotypes of a threatened desert Catostomid,the Warner sucker (Catostomus warnerensis)

Many fish species exhibit diverse life history strategies that help maintain population viability. An understanding of the relationships among these strategies is crucial for prioritising conservation actions. The Warner sucker, endemic to the Warner Lakes Basin in southern Oregon, USA, is one example of a taxon where a lack of information regarding relationships among life history strategies has hampered conservation efforts. Warner suckers have two distinct life history types: stream‐type fish that have a fluvial life history and lake‐type fish that have an adfluvial life history. There are advantages and disadvantages associated with each life history, and presently the relationship between life history types is not well understood. Our objectives were to determine the amount of genetic variation within and among tributary populations of Warner suckers and to determine the origins of suckers collected in the Warner Lakes. We collected individuals from four tributary populations, a refugial population, and Hart and Crump lakes and genotyped them at 16 microsatellite loci. Estimates of genetic variation among populations suggested low levels of gene flow (F_ST = 0.153) and genetic variation among populations seemed to be influenced by population and habitat characteristics. Nearly all of the individuals collected in Hart and Crump lakes originated in a single tributary, Deep Creek, which likely reflects reduced habitat connectivity between most other tributaries and the Warner Lakes. Data presented in this study are useful for evaluating the status of Warner sucker populations and for prioritising conservation actions such as the removal and modification of barriers.     

Genotyping of Anatolian doubled-haploid durum lines with SSR markers

In this study, doubled haploid lines generated from the durum wheat varieties, selections from Middle Anatolian landraces, ‘Çakmak-79’, ‘Berkmen-469’ and ‘Kunduru-1149’ (Savaskan et al., 1997), were analyzed using ten highly polymorphic microsatellite markers for genotyping and evaluation of genetic relationships between and within the doubled-haploid (DH) lines. The average PIC value was found to be 0.531. Populations of doubled-haploid lines of landrace selected cultivars ‘Çakmak-79’ and ‘Kunduru-1149’, were the two most distant populations with (δ μ)² = 1.42. ‘Berkmen-469’ x ‘Çakmak-79’ and ‘Berkmen-469’ x ‘Kunduru-1149’ yielded similar genetic distances, (δμ)² of 0.84 and 0.85, respectively. In addition, the genetic relationship between the progenitors of the DH lines together with other durum wheat varieties was analyzed. A meaningful relationship was obtained based on available pedigree information on the cultivars.     

DNA and morphological markers for a Russian wheat aphid resistance gene

The Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), is a significant insect pest of wheat worldwide. Morphological and molecular markers associated with RWA resistance could be used to increase the accuracy and efficiency of selection of resistant germplasm and facilitate transfer to desirable wheat genotypes. The objective of this work was to identify microsatellite (SSR) markers linked to the RWA resistance gene (Dn4) and glume-colour gene (Rg2) using a population of F₂-derived F₃ families originating from a cross between a susceptible line (synthetic hexaploid-11) and a resistance cultivar (Halt). Two microsatellite markers Xgwm106 and Xgwm337 flanked Dn4 on the short arm of chromosome 1D at 5.9 and 9.2 cM, respectively. Two other microsatellite markers, Xpsp2999 and Xpsp3000, at the distal part of this chromosome arm are linked to Dn4 and to Rg2. The accuracy and efficiency of marker-assisted selection were calculated for homozygous Dn4Dn4 genotypes in the F₂ generation. The gene Rg2 for red glume colour can also be used for marker-assisted selection of Dn4 gene individually and in combination with microsatellite markers. When used together, the closest markers Xgwm106 and Xgwm337, provide 100% accuracy and 75% efficiency. One hundred percent accuracy is also achieved when the morphological marker red glume is used in combination with either Xgwm106 or Xgwm337. Using these flanking markers, it may be possible to fix resistance to RWA in the first segregating generation of an F₂ population without infestation with aphids.     

Intrachromosomal mapping of genes for dwarfing (Rht12) and vernalization response (Vrn1) in wheat by using RFLP and microsatellite markers

For intrachromosomal mapping of the dominant GA-sensitive dwarfing gene Rht12 and the vernalization response gene Vrn1 on chromosome 5 A, an F₂ population was established using a wide (synthetic) wheat cross. In addition to restriction fragment length polymorphism (RFLP) probes four microsatellite markers were incorporated. Rht12 was mapped distally to four RFLP loci (Xmwg616, Xpsr164, Xwg114, Xpsr1201) and three microsatellite markers (Xgwm179, Xgwm410, Xgwm291), known to be located on the segment of chromosome SAL which was ancestrally translocated and is homoeologous to Triticeae 4 L. The map position of Rht12 suggests that it is homoeologous to the dominant GA-sensitive dwarfing gene Ddw1, present on chromosome 5RL. The vernalization response gene Vrn1 showed linkage to Xwg644, as might be expected from comparative maps.