Studying genetic relationships among coconut varieties/populations using microsatellite markers

The extent of genetic diversity and the genetic relationships among 94 coconut varieties/populations (51 Talls and 43 Dwarfs) representing the entire geographic range of cultivation/distribution of the coconut was assessed using 12 pairs of coconut micro satellite primers. A high level of genetic diversity was observed in the collection with the mean gene diversity of 0.647±0.139, with that of the mean gene diversity of Talls 0.703±0.125 and 0.374±0.204 of Dwarfs. A phenetic tree based on D_AD genetic distances clustered all the 94 varieties/populations into two main groups, with one group composed of all the Talls from southeast Asia, the Pacific, west coast of Panama, and all Dwarfs and the other of all Talls from south Asia, Africa, and the Indian Ocean coast of Thailand. The allele distribution of Dwarfs highlighted a unique position of Dwarf palms from the Philippines exhibiting as much variation as that in the Tall group. The grouping of all Dwarfs representing the entire geographic distribution of the crop with Talls from southeast Asia and the Pacific and the allele distribution between the Tall and Dwarf suggest that the Dwarfs originated from the Tall forms and that too from the Talls of southeast Asia and the Pacific. Talls from Pacific Islands recorded the highest level of genetic diversity (0.6±0.26) with the highest number of alleles (51) among all the regions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Molecular Analysis of Local Potato Cultivars from Tenerife Island Using Microsatellite Markers

We have used 19 SSR markers to fingerprint 41 local potato cultivars from 10 locations of Tenerife Island. These varieties represent relicts of the early introductions originating from South America and have been characterised previously morphologically and ecophysiologically. The SSR primers generated a varying degree of polymorphisms. A total of 67 alleles were observed, 12 of them were present in all cultivars. Several accession and group specific alleles were detected. Similarity coefficients were computed from the molecular data and cluster analyses were performed. Generally, cultivar groups with identical or related common names showed the same SSR patterns or clustered closely together. According to the molecular patterns misleading or confounded names were evident for four accessions. The dendrogram clusters were generally in good agreement with previous classifications of the accessions as Solanum tuberosum subsp. andigena, S. tuberosum subsp. tuberosum and Solanum chaucha genotypes. However in four cases the molecular patterns showed discrepancies with previous species assignments suggesting the need for a more detailed and comparative study of these accessions.     

对家蚕多星纹基因ms的SSR标记定位分析

对家蚕幼虫斑纹突变多星纹基因ms进行分子标记定位,有益于对多星纹性状的分子标记辅助选择和对该基因的定位克隆研究。利用家蚕雌性染色体在减数分裂过程中不发生交换的特点,采用幼虫斑纹为素斑(p)的家蚕品系C108,幼虫斑纹为多星纹(ms)的家蚕品系g02,组配正反交群体(g02×C108)F1♀×g02♂和g02♀×(g02×C108)F1♂,分别记作BC1F和BC1M,利用已经构建的家蚕SSR分子标记连锁图谱中第12连锁群上的18对SSR标记引物在亲本间进行多态性筛选。BC1F群体中的普通斑个体均表现出与(g02×C108)F1相同的杂合型带型;而所有多星纹个体的带型与亲本g02一致,为纯合型。结果筛选出S1206、S1208、S1210、C5553S3共4个与家蚕ms连锁的SSR标记。利用BC1M群体构建家蚕ms及其连锁的SSR标记遗传连锁图,连锁图的图距为34.5 cM,4个SSR标记及ms的排列次序为S1206—S1208—S1210—C5553S3—ms,ms位于34.5 cM处,与ms最近的标记为C5553S3,遗传距离为12.4 cM。依据该SSR标记遗传连锁图谱可对ms进一步精确定位。     

茶树EST-SSRs信息分析

从NCBI公共数据库中公布的茶树表达序列标签中获得12378条茶树EST,通过CAP3软件进行拼接去冗余后再用Bathprimer3 vl.0(http://wheat.pw.usda.gov/demos/BatchPrimer3/)在线软件查找微卫星(Microsatellite,SSR)重复序列。结果表明:通过前处理得到全长为2 307.014kb的无冗余EST4779条,从中检索出27种重复基元类型共207个SSR,分布于156条EST中,出现频率是3.26%。其中包括二核苷酸重复72个(34.78%),三核苷酸重复88个(42.51%),四核苷酸重复31个(14.98%),五核苷酸重复16个(7.73%)。AG/CT和ACC/GGT分别是二、三核苷酸中的优势重复类型,分别占二、三核苷酸重复的86.11%和25.00%。     

苹果遗传连锁图谱构建及抗早期落叶病的基因定位

以“秦冠”ד富士”F1群体的158个株系为试材,得到最佳SSR- PCR反应体系:在20 μL总的反应体系中Taq DNA聚合酶0.5U、引物0.1μmol/L、Mg2+ 1.8 mmol/L、模板DNA浓度15 ng/μL、dNTPs 0.30 mmol/L.利用SSR分子标记,采用JoinMap 3.0软件构建“秦冠”ד富士”的遗传连锁图谱.结果表明:从340对SSR引物中筛选出49对具有多态性的引物,在群体中共获得75个SSR标记,其中17个不符合孟德尔遗传规律,其余27个位点可定位到10个连锁群上.对苹果杂交F1代早期落叶病进行抗性遗传分析,最后将抗早期落叶病基因定位到了遗传连锁图谱中的第10个连锁群上.与已经发表的苹果遗传连锁框架图对比结果表明,抗早期落叶病基因被定位在已发表图谱的第8连锁群上.     

苜蓿新品系Dy-2006亲缘关系的SSR分析

应用SSR分子标记对苜蓿新品系Dy-2006等6个苜蓿品种(品系)亲缘关系进行了分析.8对SSR引物共扩增出128个位点,106个位点显多态性,多态位点比率82.81%;不同苜蓿品种(品系)SSR多态性百分率存在一定差异,在32.81%～45.31%之间.遗传相似度(F)和遗传距离(D)的变化分别范围在0.9062～0.9689和0.0326～0.0762.聚类分析结果显示,新品系Dy-2006与龙牧801亲缘关系相对较近,与阿尔冈金亲缘关系较远,并与其它供试品种存在一定差异,为一个相对独立的品系.     

Exploitation of SSR,SRAP and CAPS-SNP markers for genetic diversity of Citrus germplasm collection

The present study was to assess informativeness and efficiency of three different molecular markers for genetic diversity among 24 Citrus and its relative species. Sixty one SSR, 33 SRAP and 24 CAPS-SNP markers were used to evaluate the level of polymorphism and discriminating capacity. A total of 596, 656 and 135 polymorphic amplicons were observed in SSR, SRAP and CAPS-SNP markers with average polymorphism information content (PIC) of 0.97, 0.98 and 0.89, respectively. High levels of polymorphism were recorded for SSR and SRAP compared with CAPS-SNP markers. The highest correlations (r = 0.930) were obtained between SSR and SRAP markers, whereas SSR and CAPS-SNP were poorly correlated (r = 0.833). Cluster analysis was performed to construct dendrograms using UPGMA. And the dendrogram from SSR data was most congruent with the general dendrogram. These findings provide basis for future efficient use of these molecular markers in the genetic analysis of Citrus and its relatives.     

Molecular characterization of chestnut plants selected for putative resistance to Phytophthora cinnamomi using SSR markers

A set of nine short sequence repeat (SSR) loci was used for the molecular characterization of 32 accessions of 15 chestnut trees selected in the field because of their putative resistance to the ink disease caused by Phytophthora spp. The goal of the present study was to determine the genetic identity of those selected European chestnut trees (Castanea sativa) or interspecific hybrids, considering that hybridization programs between European chestnut and Asiatic species (mainly Japanese chestnut, Castanea crenata) have been carried out in Galicia (Spain) since the early 20th century. The results showed that the analyzed SSR loci were useful to discriminate three Asiatic and the European species of Castanea. The joint information provided by a factorial correspondence analysis (FCA) and the presence of privative alleles allowed the putative molecular assignment of the selected plants to a certain identity. Most of them were determined as hybrids between C. crenata and C. sativa. The individuals coded C036 and C048 were assigned, with a high probability, to C. sativa due to their clustering with accessions of this species and because they had a number of privative alleles of this species. Only a few individuals could not be assigned to any particular genotype.     

甜瓜结实花初花节位QTL分析

 以WI998为母本（纯雌株、厚皮网纹甜瓜品系）,TopMark为父本（雄全同株纯合品系）,配置杂交组合,利用单粒传得到171个株系的重组自交系（F₂S₄）群体构建甜瓜SSR分子标记遗传连锁图谱。该连锁图谱包含19个连锁群,覆盖基因组长度为1 414.2 cM,标记间平均距离为10.2 cM。对结实花初花节位开展QTL分析,共检测到9个QTL,分别分布在第1、9、10、11连锁群上,各QTL的LOD值在2.89 ~ 9.42之间,4个QTL贡献率超过10%。位于第9连锁群的QTL Fp9.4贡献率最大,为18.57%（2010秋季LOD = 9.17）;位于第9连锁群的Fp9.3（2010春季8.64%,LOD = 6.44;2010秋季17.99%,LOD = 9.42）,位于第10连锁群的Fp10.1（2010春季3.59%,LOD = 2.89;2010秋季6.20%,LOD = 3.43）在两季的位置都很稳定。获得与结实花初花节位紧密连锁（< 10 cM）的8个特异标记（SSR01737、MU141991、TJ105、GCM206、SSR04910、MU173563、NR52、MU146331）,为进一步开展QTL精细定位提供参考。