Tracking Phytophthora infestans with SSR markers within and between seasons – a field study in Sweden

The dynamics of a late blight epidemic and sexual reproduction in Phytophthora infestans were studied in an experimental field in mid‐Sweden. The field was inoculated with six isolates of P. infestans taken from another potato field where sexual reproduction of the pathogen was suspected. Three weeks after inoculation single‐lesion leaflets were sampled and the resulting isolates characterized using microsatellites (SSRs) and mating type as markers. Among the 151 isolates analysed, the inoculum genotypes constituted more than 80% of the genotypes found, with three other genotypes making up the remainder. The following year, P. infestans obtained from soil samples taken from this field were analysed, and six novel genotypes were identified. Genotypes from the previous summer’s population were not detected. Analysis of the genotypes recovered was consistent with them being recombinants, with the previous summer’s population acting as parents. These findings are consistent with the hypothesis that oospores produced during a summer epidemic in Sweden can overwinter and cause infection the next year.     

Genealogical Analysis of Diversity of Russian Winter Wheat Cultivars (Triticum aestivum L.)

With the aid of genealogical analysis the genetic diversity of Russian winter Triticum aestivum L. wheat cultivars was studied. The change of diversity in time from 1929 to 2002 shows an increase in diversity of Russian cultivars due to the use of foreign material in the breeding programs. At the same time, the genetic erosion of the released diversity occurred. In the 1950s and the 1960s about 50% of the Russian local and old varieties had dropped out of the pool of landrace ancestors. The set of the modern cultivars included in the Russian Official List 2002 has a cluster structure. The overwhelming majority (96%) are the descendants of cultivars Bezostaya 1 and/or Mironovskaya 808. The low diversity of cultivars recommended for cultivation in the Central and Volga-Vyatka regions is apparent. This situation can result in losses of a yield due to a uniform susceptibility to pathogens.     

梅花杂种鉴定中SSR分子标记引物的筛选

利用SSR分子标记技术对以‘淡丰后’(或‘丰后’)为母本,分别与其他梅花品种及近缘种杂交的F1代及其亲本进行杂种鉴定。用47对从梅花近缘种(包括桃、李、杏、酸樱桃、甜樱桃)中已开发出的SSR引物和3对梅花EST-SSR引物对亲本进行SSR-PCR扩增,其中8对引物(aprigms18、BPPCT001、BPPCT002、BPPCT004、BPPCT034、UDP96005、UDP98409、PES16)对23个父本中的21个有稳定、特异扩增(相对于母本),包括12个梅花品种和9个近缘种,有效扩增率为91.3%。再用以上8对引物对10个杂交组合进行扩增,有6对引物(BPPCT001、BPPCT004、BPPCT034、UDP96005、UDP98409、PES16)能够初步鉴定出其中7个杂交组合中21个杂种F1代的真实性(父本分别为江梅、‘变绿萼’、‘白须朱砂’、‘小红朱砂’、‘辽梅’山杏、毛樱桃、‘寿红’桃),鉴别率为77.8%。证明梅花近缘种基因组SSR引物及梅花EST-SSR引物在梅花杂种鉴定中的适用性,拓宽了SSR分子标记技术在梅花杂交育种中的应用。     

利用微卫星DNA技术进行绵羊亲子鉴定

利用9个微卫星DNA对9头陶赛特羊DNA进行扩增,每3对微卫星DNA扩增产物混合后经聚丙烯酰胺凝胶电泳和银染检测.采用已确定母女关系的2头陶赛特羊和7个嫌疑父亲的耳组织为样本,对1头母本已知的陶赛特羊在嫌疑父本中找出父本,并计算父权概率为99.997%.     

青藏高原青稞育成品种的亲缘系数分析

【目的】分析青藏高原青稞育成品种的遗传多样性,评估其遗传基础,探索拓宽其遗传基础的对策,推动青稞新品种选育工作。【方法】以1950s后,影响力高、推广面积大的105个藏区青稞育成品种和3个引进品种为研究对象,采用亲缘系数分析法分析参试品种的遗传基础,进而以亲缘系数为基础,采用兰氏距离、类平均法对参试品种进行聚类分析;采用累积直系亲缘系数,筛选藏区主要骨干亲本。【结果】18.01%的品种组合间存在亲缘关系,各组合亲缘系数的变异范围为0.000—0.750,组合平均亲缘系数为0.053,显著高于中国大麦主栽品种以及北美大麦品种。根据参试品种的育成时间,以十年为期,将参试品种划分为7个时期,1970s和1980s 2个时期,参试育成品种的组合平均亲缘系数较高,分别达到0.115和0.107;1990s和最近十年,参试育成品种的组合平均亲缘系数较低,分别为0.040和0.032。西藏自治区、青海省、四川省、甘肃省的参试育成品种数分别为48、27、17和12个;甘肃省的参试育成品种组合平均亲缘系数最高,达到0.220,西藏自治区、青海省、四川省的参试育成品种组合平均亲缘系数较为接近,分别为0.094、0.122和0.138。聚类分析的结果显示,参试品种聚为7类,Ⅱ、Ⅲ类群共包含25个参试品种,各类群的组合平均亲缘系数较低;其他5个类群共包含83个参试品种,各类群的组合平均亲缘系数较高。40个参试品种的累积直系亲缘系数大于0,其中9个育成品种是藏区青稞的重要骨干亲本,其衍生了67个参试品种。9个骨干亲本及其67个衍生种占参试育成品种总量的72.38%。1960s,骨干亲本及其衍生种的数量占该时期参试品种总量的41.76%,之后的4个时期,骨干亲本及其衍生种的数量占各时期参试育成品种总量的比例分别达到80.00%、80.77%、76.47%和72.22%;最近十年,该指标虽有所下降,但仍然高达68.75%。对于西藏自治区、青海省、四川省和甘肃省,骨干亲本及其衍生品种数占各地区参试育成品种总量的比例分别达到79.17%、62.96%、70.59%和75.00%。参试育成品种所使用的亲本合计118个,其中本地亲本97个,占比82.20%,外引亲本15个,占比12.71%;剩余6个亲本的来源信息缺失。【结论】藏区青稞的9个骨干亲本及其67个衍生品种构成了育成品种的主体,外引大麦种质利用率低,从而导致青藏高原青稞育成品种的遗传多样性较低,遗传基础相对狭窄、脆弱。     

微卫星多重PCR分析食蚊鱼系统发育关系

食蚊鱼是全球入侵性物种之一。探讨和研究食蚊鱼快速入侵的原因对进一步的生态学研究有重要意义。实验选取11对多态性微卫星位点,检测了来自欧洲境内3条河流流经的10个不同城市53个家系的755个幼鱼样本。利用微卫星多重PCR技术及Colony统计学软件,分析食蚊鱼系谱关系。食蚊鱼的繁殖能力强,繁殖期的雌性食蚊鱼可一胎产下大量雌雄幼鱼,平均9尾,最多可达72尾;更重要的是实验中发现食蚊鱼存在多重授精现象。结果显示,来自同一父本的幼鱼胚胎最多可达41尾;多重授精加快了食蚊鱼的基因流;系统发育树证实,食蚊鱼来源于遥远的海洋,实验采集的10个群体在系统发育树中主要聚为3支。食蚊鱼入侵性强的关键原因是多重授精,强壮精子具有更强的竞争受精能力,增加了物种的多样性,但受精频率与雌鱼大小可能不成比例。在长距离的迁移中,基因流也在快速变化。     

Genotyping,pedigree reconstruction and endocrinological characterization of Acipenser naccarii (Bonaparte, 1836) using microsatellite markers and plasma steroid levels

This study aimed to set up a method for the long‐term management of Adriatic sturgeon, through the analysis of sex steroids and the genetic profiling of individuals in order to maximize the already reduced genetic variability of this species. Ten A. naccarii adults (nine of captive origin, one captured in the Ticino river and then moved into captivity) and eight subadults located in a semi‐natural land‐locked pond in Abbiategrasso (Milan, Italy) were analysed. Plasma testosterone differed significantly in the eight analysed subadults allowing their differentiation into two groups: the first group with an average testosterone concentration of 5.42 ± 1.31 ng/ml (probably female) and the second group with an average of 423.14 ± 75.97 ng/ml (probably male), as subsequently confirmed by artificial stripping. The plasma testosterone level was also significantly different between adult males and females (371.37 ± 43.58 vs. 95.34 ± 51.10 ng/ml), while the E2 levels showed no significant differences. Animals were genotyped on the basis of 10 microsatellite loci and their parental relationships were defined: four adults, two females and two males, generated the eight subadults. On the basis of pedigree analyses and genetic distances, 15 unrelated couples were identified for the future breeding seasons. Finally, the adult female captured in the Ticino River showed an interesting genetic profile, widely different from all of the other 17 specimens analysed, and represents a valuable source of genetic diversity.     

Parentage assignment in Haliotis midae L.: a precursor to future genetic enhancement programmes for South African abalone

The establishment and evaluation of family lines using pedigree information provides an advanced understanding of the variability that exists for complex, economically valuable traits and is a necessary step in the execution of an effective breeding programme. The aim of this study was to assign parentage to mass‐spawned Haliotis midae juveniles using species‐specific microsatellite markers. Screening of wild abalone individuals revealed that the nine loci selected complied with the minimum requirements for parentage analyses: a null allele frequency <5% as well as a high number and frequency of alleles per locus. A total of 598 individuals were genotyped (198 breeding individuals and 400 F1 progeny) from two farms, with parentage results yielding 91% and 90% successful assignment for Farms A and B respectively. This study, therefore, provided the necessary pedigree information required for controlled breeding of individual adult abalone and indicated the usefulness of the panel of microsatellite markers selected for parentage assignment.     

Development of two microsatellite multiplex PCR systems for high throughput genotyping in Populus euphratica

Eighteen microsatellite primer pairs previously developed at Oak Ridge National Laboratory for Populus tremuloides Michx. and Populus trichocarpa Torr. & Gray were screened for amplification in Euphrates poplar, Populus euphratica Oliv. Thirteen loci were found to express polymorphisms ranging from two to 17 alleles. The eight most variable loci were selected to set up and optimize two multiplex polymerase chain reaction (PCR) assays. Three populations containing altogether 436 trees were used to characterize the selected loci and ascertain their applicability for parentage analysis and genotyping studies. Through cross-checking of clonal identity against sex of the genotyped trees we estimated the maximum error rate for merging genotypes to be less than 0.045. Foundation project: This study was supported by the Deutsche Forschungsgemeinschaft DFG (grant number SCHN 1080/1-1)