Molecular characterization and genetic relationship among almond cultivars assessed by RAPD and SSR markers

RAPDs and SSRs were used to study the genetic diversity of Iranian almond cultivars and their relationship to important foreign cultivars and three related species. Eight unidentified almond Shahrodi cultivars and three wild almonds (Prunus communis, Prunus orientalis and Prunus scoparia) were also included. Of the primers tested, 42 (out of 80) RAPD and 18 (out of 26) SSR primers were selected for their reproducibility and high polymorphism. A total of 664 polymorphic RAPD bands were detected out of 729 bands. The number of presumed alleles revealed by the SSR analysis ranged from 3 to 10 alleles per locus with a mean value of 6.64 alleles per locus. Both techniques discriminated the genotypes very effectively, but only RAPDs were able to discriminate the cultivars Monagha and Sefied. Results demonstrated an extensive genetic variability within the tested cultivars as well as the value of SSR markers developed in peach for characterization of almond and related species of Prunus. Dice similarity coefficient was calculated for all pair wise comparisons and was used to construct a UPGMA dendrogram. For both markers a high similarity in dendrogram topologies was obtained although some differences were observed. All dendrograms, including that obtained by the combined use of both the marker data, depicted the phenetic relationships among the cultivars and species, depending upon their geographic region and/or pedigree information. Almond cultivars clustered with accession of P. communis showing their close relationship. P. orientalis and P. scoparia were clustered out of the rest of P. dulcis.     

Detecting<Emphasis Type="Italic">Orobanche</Emphasis> species by using cpDNA diagnostic markers

Some species of the genusOrobanche are among the most devastating parasitic weeds, causing extensive damage in agricultural fields. Considering the difficult control due to seed longevity in the soil, small seed size, high fecundity and a subterranean phase that allows them to parasitize the host before they emerge and become evident, the development of diagnostic markers is highly recommended. In our study we identified potential molecular diagnostic markers from the plastid genome in order to distinguish among the most importantOrobanche species attacking crops in Andalusia, the southern region of the Iberian Peninsula. The study has consideredO. crenata, O ramosa andO. cumana causing serious losses in legumes, solanaceous crops and sunflower fields, respectively, andO. minor that, although abundant in Andalusia, has to our knowledge not yet been found parasitizing agricultural hosts. We amplified a non-coding region from the plastid genome, studied sequence differences among the amplified fragments and digested those of the same length with selected restriction enzymes. Here, we propose a molecular protocol to distinguish the main parasitic plants in crop fields of southern Spain. Different applications such as identification ofOrobanche seeds in soil or crop seed lots are discussed in order to offer right crop recommendations or to prevent new infestation of parasite-free fields. Recommendations for further development of these diagnostic markers are also considered. http://www.phytoparasitica.org posting Jan. 15, 2007.     

晋大62×诱处4号杂交后代群体遗传多样性分析

选用晋大62×诱处4号杂交亲本和后代群体为试验材料,通过微卫星(SSR)标记分析,利用SPSS软件对其进行了聚类分析,探讨了亲本及后代群体间的遗传特性和亲缘关系,研究了大豆群体的遗传多样性,为大豆杂交育种亲本的选配、杂交后代的筛选、种质创新及大豆遗传多样性研究提供了理论和实践依据。结果表明,以亲本的基因组DNA为模板,选用40对不同的引物进行电泳共筛选出23对位于多条染色体上的差异引物,其多态性比例为55.0%。通过聚类分析所有材料可聚类为以下几类:母本晋大62(CK1)和后代6~13号材料聚为第一大类,父本诱处4号(CK2)和后代25~31号材料聚为第二大类,而1~5、14~20聚为第一小类,21~24、32~35聚为第二小类,这两小类材料与亲本的亲缘关系较远。     

黄瓜叶面积遗传及QTL定位分析

选用2个叶面积大的黄瓜品种（D9320、D2005）和2个叶面积小的黄瓜品种（D0455、D0401）配制杂交组合，筛选叶面积差异较大的亲本组合D0401×D9320构建F2分离群体，采用加性－显性（AD）模型进行遗传分析。结果表明：黄瓜叶面积是由多基因控制的数量性状，以显性效应为主，存在一定的加性效应，狭义遗传力和广义遗传力较高。采用SSR分子标记构建遗传连锁图谱，利用复合区间法对黄瓜叶面积进行QTL定位。结果表明：CSWACC03、CSWCT05BZ和CSCTTT15a位于同一个连锁群上，连锁群总长度为68.3 cM；检测出1个与黄瓜叶面积有关的QTL，与最近标记的距离为3.6 cM，且变异贡献率为8.65 %。     

Bulked Segregant Analysis to Detect QTL Related to Heat Tolerance in Rice （Oryza sativa L.） Using SSR Markers

The study was undertaken to assess the genetic effect of quantitative trait loci （QTLs） conferring heat tolerance at flowering stage in rice. A population consisting of 279 F2 individuals from the cross between 996, a heat tolerant cultivar and 4628, a heat-sensitive cultivar, was analyzed for their segregation pattern of the difference of seed set rate under optimal temperature condition and high temperature condition. The difference of seed set rate under optimal temperature condition and high temperature condition showed normal distribution, indicating the polygenic control over the trait. To identify main effect of QTL for heat tolerance, the parents were surveyed with 200 primer pairs of simple sequence repeats （SSR）. The parental survey revealed 30% polymorphism between parents. In order to detect the main QTL association with heat tolerance, a strategy of combining the DNA pooling from selected segregants and genotyping was adopted. The association of putative markers identified based on DNA pooling from selected segregants was established by single marker analysis （SMA）. The results of SMA revealed that SSR markers, RM3735 on chromosome 4 and RM3586 on chromosome 3 showed significant association with heat tolerance respectively, accounted for 17 and 3% of the total variation respectively. The heat tolerance during flowering stage in rice was controlled by multiple gene. The SSR markers, RM3735 on chromosome 4 and RM3586 on chromosome 3 showed significant association with heat tolerance respectively, accounted for 17 and 3% of the total variation respectively. The two genetic loci, especially for RM3735 on chromosome 4, can be used in marker-assistant-selected method in heat tolerance breeding in rice.     

用基于PCR的分子标记分析甘蓝型油菜抗感菌核病品系间的遗传距离

用离体叶接种法鉴定了甘蓝型油菜选系对菌核病的抗感性，结果表明绝大多数抗性品系与感病品种间的差异达到显著或极显著水平。以两种基于ＰＣＲ的标记估计所选１０ 份材料间的遗传距离。１９ 个随机引物扩增出了２６ 条重复性很好的ＲＡＰＤ 标记，１ 对依据植物Ｒ基因的ＮＢＳ区段设计的简并引物扩增出了３ 条多态性的带。依据２９ 个标记进行聚类分析，结果表明抗、感性材料在遗传距离为０．１２０６ 处被明显地区分开。５份抗性材料中，宁ＲＳ—１ 与中油８２１、中Ｒ８８８ 等其它抗性品系的遗传距离较远     

利用RIL群体和自然群体检测与花生含油量相关的SSR标记

以远杂9102×中花5号杂交后代衍生的重组近交系F₈代家系为材料，在含油量测试的基础上，选用10份低油材料(平均含油量52.91%)、12份高油材料(平均含油量58.85%)以及亲本进行SSR引物筛选，通过631对SSR引物扩增，筛选出来源于7对引物的13个有显著差异的片段可以有效区分低油材料和高油材料。以这7对差异引物在F₈ RIL群体中扩增，对20份低油家系材料(含油量<55%)和45份高油家系材料(含油量>56%)进行统计分析，获得1个与花生含油量相关的分子标记2A5-250/240，其中，标记2A5-250为低油材料(含油量<55%)所拥有，相符率为95.0%，标记2A5-240为高油材料(含油量>56%)所拥有，相符率为88.9%。用SSR标记2A5-250/240检测11份高油(平均含油量为55.93%)栽培种花生和11份低油(平均含油量为48.41%)栽培种花生，结果表明，标记2A5-240与高油栽培种花生的符合率为63.6%，2A5-250与低油栽培种花生的符合率为90.9%。在19份高油(平均含油量为58.60%)野生花生中，10份野生花生能检测到标记2A5-240。综合分析RIL群体和自然群体的研究结果表明，标记2A5-250/240可用于花生含油量分子标记辅助选择。     

利用SSR技术对黄瓜新品种津优35号进行种子纯度鉴定

利用天津科润农业科技股份有限公司建立并优化的黄瓜种子纯度鉴定技术体系,筛选黄瓜新品种津优35号特异分子标记。所试验的80多对SSR引物中,有2对引物(85号和212号)在杂交种和亲本间表现多态性,杂交种条带为父母本的互补型,且特异性强,适合做杂交种纯度鉴定。用该2对引物对津优35号50粒种子进行SSR鉴定。结果表明:该2对引物鉴定结果相同,且与田间鉴定结果吻合率高达96%以上,表明所筛选到的2个SSR标记可以作为津优35的特异分子标记用于其种子纯度的鉴定。     

利用SSR技术对黄瓜新品种津优35号进行种子纯度鉴定

利用天津科润公司建立并优化了的黄瓜种子纯度鉴定技术体系,筛选黄瓜新品种津优35号特异分子标记。试验选用80多对SSR引物中,有2对引物(85号和212号)在杂交种和亲本间表现多态性,杂交种条带为父母本的互补型,且特异性强,适合做杂交种纯度鉴定。用这2对引物对津优35号50粒种子进行了SSR鉴定。结果表明:这2对引物鉴定结果相同,且与田间鉴定结果吻合率高达96%以上,表明所筛选到的2个SSR标记可以作为津优35的特异分子标记用于其种子纯度的鉴定。     

快速大量提取甜菜DNA的新方法

以甜菜幼嫩叶片为材料,通过冷冻真空干燥将材料变成干粉,利用CTAB法对DNA提取程序进行了研究,建立了一种简单、快速的甜菜DNA提取程序,既节省时间,又可获得高质量的DNA,可以满足SSR及SRAP扩增的需要。