高RA甜叶菊辅助选择育种EST-SSR分子标记开发
毕业论文关键词：甜叶菊；EST-SSR分子标记；遗传多样性；RA；育种26229
Development of EST - SSR Molecular Marker for Selection of High RA Stevia
Abstract： Based on the EST-SSR molecular marker, the relationship between EST-SSR and the content of RA in stevia was studied. 85 EST-SSR primers were designed and the PCR products the polymorphism was 31, and the polymorphism ratio was 71.7%. The polymorphism of 31 stevia leaves was analyzed by PAGE with agarose gel and polyacrylamide gel. UPGMA cluster analysis based on genetic similarity coefficient showed that the three major groups of Stevia were clustered into two groups, and 23 materials were clustered into 5 major branches. In the selected 31 primers, the amplified fragments of four high RA stevia leaves were absent compared with other stevia leaves at the position of 180bp. Therefore, primers 31 were expected to be selected for screening high RA stevia material EST-SSR primers.
Key words: Stevia rebaudiana；EST-SSR molecular markers；genetic persity；RA；breeding
目录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 1
1 材料与方法 2
1. 1 实验材料、试剂及仪器 2
1．1．1 实验材料 2
1．1．2 实验试剂 2
1．1．3 实验仪器 2
1．2 实验方法 2
1．2．1 甜叶菊采集 2
1．2．2 DNA提取 3
1．2．3 PCR扩增 3
1．2．3．1 PCR扩增前实验准备 3
1．2．3．2 PCR扩增反应体系及反应条件 3
1．2．3．3 PCR产物的电泳分离 4
2 结果与分析 4
2．1 DNA提取结果 4
2．2 PCR反应结果 5
2．2．1 以全池为模板PCR结果 5
2．2．2 以半池为模板PCR结果 6
2．2．3 以单个材料为模板PCR结果 10
2．3 遗传多样性分析 13
2.3.1 三类糖苷含量不同的甜叶菊类群间及个体间的遗传距离和聚类分析 13
2.3.2 甜叶菊遗传多样性分析 15
2.3.3 甜叶菊糖苷含量鉴别用引物 15
3 讨论 16
致谢 16
参考文献 16
图1 甜叶菊基因组DNA 4
图2 以全池为模板PCR结果 5
图3 以半池为模板PCR结果（1） 6
图3 以半池为模板PCR结果（2） 6
图3 以半池为模板PCR结果（3） 7
图3以半池为模板PCR结果（4） 7
图4 以半池为模板PAGE结果 9
图5 以单个材料为模板PCR结果（1） 10
图5 以单个材料为模板PCR结果（2） 10
图6 甜叶菊基因组DNA重提结果 11
图7 重提DNA的PCR结果 11
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