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.     

青钱柳种质资源多样性SRAP初步分析

为了利用分子标记方法评价青钱柳种质资源的遗传多样性,本文对青钱柳DNA的提取、SRAP扩增体系重要参数进行了优化,运用优化体系筛选多态性引物,并用1对引物对青钱柳9个种源进行了遗传多样性初步分析。研究结果建立了适于青钱柳SRAP的扩增体系;从110对SRAP引物中筛选出了13对多态性引物,运用1对引物组合Me7＋Em2扩增获得21个多态性位点,多态率达100%。遗传多样性分析表明有效等位基因数（Ne）为1.3429,平均Shannon＇s信息指数（I）为0.3687,Nei＇s基因多样性指数（H）为0.2267,种源的遗传分化指数Gst为0.1983;聚类分析结果表明9个种源在遗传距离0.100处聚为3类,聚类结果和地理距离之间呈现较高的相关性。本文的研究结果表明所建立的青钱柳种质资源库具有广泛的遗传多样性,为进一步的开发利用提供了条件。     

Pattern of genetic diversity of cultivated and non-cultivated mashua, <Emphasis Type="Italic">Tropaeolum tuberosum</Emphasis>, in the Cusco region of Perú

This paper analyzes the genetic diversity of mashua cultivars grown in six communities in the Cusco region of Perú, of non-cultivated forms collected in the same region, and a sample of cultivars from the germplasm collection at the International Potato Center (CIP). From the DNA fingerprinting generated by SRAP markers, it is clear that mashua is a genetically variable crop with a range of similarity ranging from 65 to 99%. The widest range of variation was found for the most isolated community, Sayllafaya. Another important finding was that most of the non-cultivated accessions are likely feral races resulting from escape to cultivation rather than wild relatives. In general the range of variation of the cultivars from the communities and their feral relatives are not represented in the cultivars sampled of the collection maintained at the International Potato Center, Perú.     

猴头菌DNA提取条件及SRAP- PCR体系的

以猴头菌(Hericiumerinaceus)菌丝体为试验材料,建立最优的DNA提取条件和SRAP- PCR扩增体系.结果表明:改进的SDS- CTAB法能较好地满足猴头菌属DNA的提取,优化后SRAP- PCR反应体系经过验证,该体系可用于猴头菌的遗传多样性分析和品种鉴定.     

番茄品种分子指纹的SRAP分析

[目的]利用SRAP技术对3个番茄样品进行品种鉴定。[方法]以3个番茄样品(瓯秀806、瓯秀201和一个待确认品种)的新鲜叶片为试验材料,通过改良的CTAB法提取DNA,对其进行SRAP分析并构建供试样品间的聚类树状图。[结果]利用设计的54对引物扩增出多态性位点103个,分析表明3号番茄品种确认为瓯秀806。[结论]该试验结果表明应用SRAP分析技术可以对番茄品种进行有效的鉴定。     

应用SRAP与ISSR分析烟草种质资源遗传多样性及遗传演化关系

应用ISSR与SRAP两种分子标记,研究国内外96份烟草种质的遗传多样性及不同栽培类型种质的遗传演化关系。表明烟属种间具有丰富的遗传多样性,种间的遗传相似性(GS)在0.28~0.58之间,遗传分化系数(Gst)为0.83。普通栽培种品种间遗传多样性水平较低,品种间的遗传相似性在0.61~0.99之间,栽培种内的遗传多样性为烤烟>晒晾烟>白肋烟>香料烟。当相似系数在0.67作切割线时,基于2种标记的96份烟草种质资源的聚类结果为,(1)普通烟草栽培品种材料91份聚在同一大类,而黄花烟、黏烟草、浅波烟草、哥西氏烟草、香甜烟草5个种也分别为单独的个类,同普通烟草栽培种类群完全区别开来;(2)从进化上看,烤烟和晒晾烟间的遗传进化关系最近,香料烟和黄花烟之间的亲缘关系较远;普通烟草栽培种中国内外来源的烟草品种亲缘关系极其相近,遗传分化现象甚微;(3)2种分子标记虽然原理不同,但分析结果趋势相近(r=0.68,P=1.000)。     

华山松SRAP-PCR反应体系的优化

为建立华山松SRAP-PCR反应体系,研究先采用L16(45)正交设计对影响华山松SRAP反应的5个因子(DNA模板浓度、Mg2+浓度、dNTPs浓度、引物浓度、Taq酶)在4个水平上进行优化试验。结果表明:各因素对SRAP反应的影响依次为:Mg2+>dNTPs>Taq酶>DNA模板>引物;对SRAP反应结果影响较大的3个因子(Mg2+浓度、dNTPs浓度、Taq酶浓度)进行单因素试验;确立华山松SRAP反应最佳体系为:20μL的PCR体系中含有Mg2+2.2mmol/L、dNTPs0.25mmol/L、DNA模板60ng、Taq酶0.8mol/s、引物0.8μmol/L。将该体系用于华山松的SRAP扩增能获得稳定、清晰的多态性条带。     

甘蓝型油菜黄籽高含油量育种资源遗传多样性分析

以2份白菜型油菜为对照,用SRAP标记研究了57份甘蓝型油菜黄籽高油育种资源的遗传关系。在遗传相似系数0.682处,可将59份材料分为3类:两份白菜型油菜聚为一类;甘蓝型油菜除选系Y58单独聚为一类外,其他所有选系归为另一类,显示黄籽高油育种资源遗传基础较窄。在遗传相似性系数0.738处,又可将57份甘蓝型油菜分为6个亚类,其中53份材料归为两个大的亚类,系谱或亲缘关系较近的材料一般聚在同一亚类,且按系谱关系比地理来源划分明显,提示地理来源配制强优势黄籽高油杂交组合可靠性可能不如按系谱来源。     

基于毛细管电泳技术的牡丹SRAP-PCR反应体系优化及引物筛选

以牡丹叶片DNA为模板,对SRAP-PCR反应程序进行研究,确定适合的反应程序,即94℃预变性5 min;94℃变性1 min,33℃退火1 min,72℃延伸1 min,共5个循环;随后94℃变性1 min,52℃退火1 min,72℃延伸1 min,共35个循环;最后72℃延伸10 min。基于毛细管电泳技术,采用正交设计L18(37)对牡丹SRAP-PCR反应体系的5因素(Taq酶,Mg2+,模板DNA,dNTP,引物)3个水平进行了优化,构建了牡丹SRAP最佳反应体系:模板DNA 50 ng,dNTP 0.25 mmol/L,Mg2+浓度2.5 mmol/L,引物浓度0.4μmol/L,TaqDNA聚合酶0.5 U,总体积为25μL。各因素对扩增结果影响程度均不同:dNTPs>引物>DNA模板>Mg2+>Taq酶。运用该体系从756个SRAP引物组合中筛选出多态性好、条带清晰的26个引物组合,并证明了该体系稳定可靠。该体系的建立以及引物组合的确定为今后利用SRAP分子技术进行牡丹的相关研究奠定了科学基础。