正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

基于SRAP标记的莲种质资源遗传多样性分析

利用SRAP标记对来自国内外莲主要产区在20世纪初期育出、并在莲育种中具有重要利用价值的40份莲属材料进行了分析。11对引物在40份材料中共扩增了184条条带,多态性条带127条,多态性频率为69.0%,表明莲属具有丰富的遗传多样性。聚类分析结果表明,莲属种质资源可分为3大类群,这与传统意义上的形态学分类相一致。且40份莲品种(材料)表现出很强的地域相似性,这说明来自同一个地域内的品种(材料)分化程度低,遗传多样性不丰富,极易造成品种单一化。     

Analysis of genetic variation in eggplant and related Solanum species using sequence-related amplified polymorphism markers

Collection and characterization of all sorts of germplasm resources are required for the development of new cultivars. Molecular characterization is more reliable than morphological characterization. Here, we employed sequence-related amplified polymorphism (SRAP) markers to evaluate genetic variation in a diverse collection of 56 Solanum accessions. Fifty-five SRAP primer combinations were used and a total of 635 polymorphic bands were observed. Cluster analysis by the unweighted pair-group method with arithmetic averages based on similarity matrices indicated that there were three clusters: (i) S. melongena; (ii) S. aethiopicum; (iii) S. surattense. The coefficients of genetic similarity among all the accessions ranged from 0.04 to 0.96 with an average of 0.73, and averaged 0.78 among S. melongena accessions originated from China, indicating extensive genetic variation. These results demonstrated that SRAP can be efficiently used to estimate genetic diversity and analyze phylogenetic relationship.     

基于表型和SRAP 标记的切花菊品种遗传多样性分析

 利用45 个表型性状和SRAP 标记分析56 个切花菊品种的遗传多样性。表型变异分析结果 表明：21 个性状表现出品种内一致性高及品种间特异性强;主成分分析发现,主成分贡献值较大的性状 有花序直径、花序类型和瓣型等花部性状,其次是叶部和茎杆性状,说明所选用的切花菊品种在分类时 应以花部性状为主,叶部和茎杆性状为辅;表型性状基于遗传距离UPGMA 聚类,将56 个切花菊品种分 为平瓣类、匙瓣类、桂瓣类、匙瓣-平瓣类和管瓣类,聚类结果大致按照花径-瓣型-花型分类。14 对 SRAP 引物组合扩增56 个切花菊品种的DNA,共扩增出454 条带,其中多态性带423 条,占扩增带数的 93.17%,多态性含量PIC 值在0.72 ~ 0.89 之间,平均为0.82,说明切花菊品种在分子水平上具有丰富的 遗传多样性。基于SRAP 标记的UPGMA 聚类分析显示：品种间遗传相似系数在0.64 ~ 0.97 之间,将56 个切花菊品种分为平瓣类-匙瓣、桂瓣类和管瓣类,聚类结果大致按照花径-瓣型-花型分类。Mantel 检验相关性系数为0.682,两种聚类结果有相似之处,均能很好体现试验切花菊品种间的遗传关系。     

菜薹抽薹性状的ISSR和SRAP分析

以早抽薹品种CT07和晚抽薹品种T0601为亲本构建F2分离群体,采用分离集团混合分析法 (Bulked segregant analysis,BSA)筛选与菜薹抽薹基因连锁的分子标记.用45条ISSR(Inter-simple sequence repeat,简单重复序列间扩增)引物和20对SRAP(Sequence related amplified polymorphism,序列相关扩增多态性)引物组合进行PCR扩增筛选,其中UBC834、UBC876、E13M4和E5M7对亲本和DNA池的扩增图谱表明,4个引物的差异条带均出现在早抽薹品种CT07和早现蕾(抽薹)池中.经F2代群体单株验证,4个标记均与早抽薹基因相连锁,且标记E13M4最近,距离为16.8 cM,这为菜薹抽薹基因的定位和分子标记辅助育种奠定了基础.     

SRAP分子标记在棉花遗传育种中的应用

SRAP(Sequence-related Amplified Polymorphism)是1项基于PCR技术的新型分子标记技术,具有操作简便、稳定、高效、高共显性、便于测序目标片断等特点。介绍了SRAP分子标记技术的原理和特点,并综述其在棉花遗传连锁图谱的构建、遗传多样性分析、基因定位等方面的研究进展和应用前景。     

八仙花SRAP反应体系的建立与优化

为建立适合八仙花SRAP-PCR分子标记技术体系,以八仙花栽培品种H.macrophylla‘Lavbla’为材料,通过单因子实验分别研究了模板DNA、dNTPs、Mg2 、Taq酶浓度和引物用量对八仙花SRAP扩增反应的影响,确立了八仙花SRAP分析的最佳反应体系为25μl:模板DNA60ng、Mg2 2.0mmol/L、dNTPs0.7mmol/L、Taq酶0.5U、引物0.7μmol/L×2、10×PCRBuffer2.5μl,该反应条件下八仙花SRAP扩增条带清晰,多态性丰富。     

Method of Constructing Core Collection for Malus sieversii in Xinjiang, China Using Molecular Markers

The method for constructing core collection ofMalus sieversii based on molecular marker data was proposed. According to 128 SSR allele of 109 M. sieversii, an allele preferred sampling strategy was used to construct M. sieversii core collection, using the UPGMA （unweighted pair-group average method） cluster method according to Nei ＆ Li, SM, and Jaccard genetic distances, by stepwise clustering, and compared with the random sampling strategy. The number of lost allele and t-test of Nei＇s gene diversity and Shannon＇s information index were used to evaluate the representative core collections. The results showed that compared with the random sampling strategy, allele preferred sampling strategy could construct more representative core collections. SM, difference for construction of M. sieversii core collection. Jaccard, and Nei ＆ Li genetic distances had no significant SRAP （sequence-related amplified polymorphism） data and morphological data showed that allele preferred sampling strategy was a good sampling strategy for constructing core collection of M. sieversii. Allele preferred sampling strategy combined with SM, Jaccard, and Nei ＆ Li genetic distances using stepwise clustering was the suitable method for constructing M. sieversii core collection.     

基于SRAP的辣木种质资源遗传多样性和亲缘关系分析

为探讨辣木(Moringa spp.)种质资源间的遗传关系,采用相关序列扩增多态性(sequence-related amplified polymorphism,SRAP)分子标记方法对18份辣木种质资源的遗传多样性和亲缘关系进行分析.结果表明,从170对引物中筛选出13对多态性较高的引物组合,共扩增出104条清晰稳...     

无核葡萄种质亲缘关系的SRAP研究

为充分了解及更合理地利用无核葡萄的种质资源,辅助无核葡萄育种,本研究采用SRAP分子标记技术,对23 个国内外主流无核葡萄品种的种质亲缘关系进行分析。结果表明：从45 对SRAP引物中筛选出38 对多态性引物,共扩增出236 条谱带,其中171 条为多态性谱带,多态性谱带百分率为72.4%。遗传相似分析显示,供试材料间遗传相似系数的变化范围为0.58~0.90,遗传距离在1.0313~4.9643 范围内变化。在遗传相似系数为0.61 处,可将供试材料分为2 个大类群,在相似系数为0.66 处,Ⅰ类群和Ⅱ类群均可分为2个亚群。说明23个品种总体来讲亲缘关系较远,特别欧美杂种具有较高的遗传多样性,部分欧亚种品种间亲缘关系较近,杂交育种时应注意亲本选择。