【摘要】 为探讨收集保存的毛叶枣种质的亲缘关系,运用正交设计建立优化毛叶枣ISSR反应体系。结果表明:即25μL PCR体系中含2.5μL的10×PCR Buffer、3.75μL的2.5 mmol/L dNTPs、1.5μL的0.5 U/μL Taq酶、1.5μL的5μm/L引物,0.75μL的100 ng/μL DNA。然后利用ISSR标记对21份毛叶枣材料进行分类研究,并从100条引物中筛选13条扩增条带数量丰富、清晰、稳定的引物用于亲缘关系分析。结果显示:共扩增条带98条,其中多态性条带78条,多态性百分率85.7%;根据ISSR扩增结果,21份材料遗传相似系数范围为0.51~0.90。通过UPGMA法进行聚类分析,在遗传相似系数0.67水平,21份毛叶枣种质可分为3类:第I类为蜜丝枣1个种质;第Ⅱ类均来自台湾品种群,有17个种质;第Ⅲ类滇刺枣野生种、缅甸长果种和缅甸圆果种。大多数品种间相似性系数在0.67以上,说明保存种质的遗传多样性较低。更多还原

【Abstract】 A genetic analysis of 21 diverse Zizyphus mauritiana accessions was performed using ISSR marker.The orthogonal design was used to optimize the ISSR-PCR amplification system on Z.mauritiana.The optimal components of the PCR system(25 μL)were: 2.5 μL of 10×PCR buffer,3.75 μL of 2.5 mmol/L dNTPs,1.5 μL of 0.5 U/μL TaqDNA ploymerase,1.5 μL of 5 μm/L primer,0.75 μL of 100 ng/μL DNA and 15 μL ddH2O2.Twenty-one Z.mauritiana species were classified by ISSR makers.Thirteen primers were selected from 100 primers and used for ISSR amplification.Ninety-eight bands were generated,78 of which were polymorphic bands(the percentage of polymorphic band,PPB=85.7%).The genetic similarity coefficient was 0.51 to 0.90 according to ISSR amplification.The clustering dendrogram was constructed by UPGMA method to explore the taxonomic relationship in 21 Z.mauritiana materials.At 0.67 level of similarity coefficient,the 21 Z.mauritiana materials were classified as three groups: groupⅠ: var.Misi;groupⅡ: 17 varieties from Taiwan;group Ⅲ: var.Diancizao,var.Miandianyuanguo and var.Miandianchangguo.The similarity coefficient was above 0.67 level in most varieties,suggesting that the genetic diversity was low in the 21 Z.mauritiana species.were above 0.67 level in most varieties,suggested that genetic diversity was low in 21 Zizyphus mauritiana species.更多还原