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以猕猴桃属(Actinidia Lindl.)不同种幼嫩叶片为材料,建立了基因组DNA提取的改良SDS法,在此基础上采用正交试验设计和单因素分析相结合的方法,建立了适合猕猴桃SRAP分析的优化体系,即在20μL总的反应体系中包括:DNA(40 ng·μL~(-1))1μL、Taq DNA酶(5 U·μL~(-1))0.2μL、d NTPs(2.5 mmol·L~(-1))1.4μL、引物(10μmol·L~(-1))各1.5μL、Mg~(2+)(25 mmol·L~(-1))2.0μL、10×缓冲液2.5μL、dd H2O 9.9μL。利用该体系对32份猕猴桃品种资源进行遗传多样性和亲缘关系分析,结果表明14条引物共扩增出275个多态性位点,多态性百分率为100%,SRAP可以作为猕猴桃资源亲缘关系研究的有效标记;在遗传相似系数0.73水平处,供试材料可区分为4组,分别是中华猕猴桃、美味猕猴桃、黑蕊猕猴桃和毛花猕猴桃组。聚类结果表明中华猕猴桃与美味猕猴桃有着非常近的亲缘关系,毛花猕猴桃与中华猕猴桃之间的亲缘关系较远,黑蕊猕猴桃与美味猕猴桃之间亲缘关系可能较近。
Based on Actinidia Lindl. Different kinds of young leaves, the improved SDS method for genomic DNA extraction was established. On the basis of this, orthogonal experimental design and single factor analysis were combined to establish the SRAP analysis method suitable for kiwifruit The optimized reaction system consisted of 1 μL of DNA (40 ng · μL -1), 0.2 μL of Taq DNase (2.5 U · μL -1), 2.5 μL of dNTPs · L -1, 1.5 μL each of primers (10 μmol·L -1), 2.0 μL Mg 2+ (25 mmol·L -1), 10 × buffer 2.5 μL, dd H2O 9.9 μL. The genetic diversity and genetic relationship of 32 kiwifruit cultivars were analyzed by using this system. The results showed that 275 polymorphic loci were amplified by 14 primers, and the percentage of polymorphism was 100%. SRAP could be used as kiwifruit resource kinship The effective markers of the study; genetic similarity coefficient 0.73 level, the test material can be divided into four groups, namely Chinese kiwi fruit, kiwi fruit, kiwifruit and kiwifruit group. The clustering results showed that kiwifruit and kiwifruit in China had a very close genetic relationship. The kiwifruit between kiwifruit and Chinese gooseberry were far away from each other, and the relationship between kiwifruit and kiwi fruit might be closer.