基于分子标记技术的苦瓜育种材料遗传多样性快速分析

Rapid Analysis of Genetic Diversity of Momordica charantia Breeding Materials by Molecular Marker Technology

为了进一步缩短苦瓜分子标记技术的操作流程,最大限度促进其在分子标记辅助选择育种中的应用,利用一管式植物DNAout试剂盒法,从苦瓜种子中快速提取基因组DNA,采用11对SSR引物和13对SRAP引物分别对10份苦瓜材料基因组进行PCR扩增,并对苦瓜材料的遗传多样性进行分析。结果发现,建立的SSR和SRAP快速扩增反应体系可以获得清晰、数量稳定以及多态性丰富的条带,11对SSR引物共扩增出195条条带,其中多态性带164条,多态性条带比率85.7%。UPMGA聚类分析表明,10份材料的遗传相似系数介于0.533~0.845,平均值为0.690,在阈值0.69处,可将供试苦瓜分为2大类群。13对SRAP引物共扩增出231条条带,其中多样性条带201条,多态性条带比率87.5%。UPMGA聚类分析表明,遗传相似系数介于0.554~0.775,平均值0.662,在阈值0.66处可将供试苦瓜分为2大类群。综合2种分子标记技术的分类结果更具有说服力。

The object of this study is to further shorten the operation process of molecular marker techno-logy in bitter gourd(Momordica charantia)and maximize its application in molecular marker assisted selection breeding.In this paper,a tubular plant DNAout kit was used to fast extract genomic DNA from bitter gourd seeds,then the genomic DNA of 10 bitter gourd materials were amplified by PCR with 11 pairs of SSR primers and 13 pairs of SRAP primers,and the genetic diversity of materials was also analyzed.Results indicated that the established SSR and SRAP rapid amplification reaction system could obtain clear,stable and rich polymorphic bands.A total of 195 bands were amplified by 11 pairs of SSR primers,of which 164 bands were polymorphic,with a polymorphic band ratio of 85.7%.UPMGA cluster analysis showed that the genetic similarity coefficients of 10 materials ranged from 0.533 to 0.845,with an average of 0.690.At the threshold of 0.69,the tested materials could be divided into two groups.A total of 231 bands were amplified by 13 pairs of SRAP primers,of which 201 bands were polymorphic,and the polymorphic band rate was 87.5%.UPMGA cluster analysis showed that the genetic similarity coefficients ranged from 0.554 to 0.775,with an average of 0.662,and all materials could be divided into two groups at the threshold of 0.66.Since both markers can obtain abundant polymorphic bands,the molecular marker methodology can be applied to the genetic diversity analysis of bitter gourd.Moreover,the classification results by combining the two molecular marker techniques are more convincing.