柱花草RAPD品种鉴定方法

针对柱花草不易从种子表型进行品种区分、品种真实性缺乏有效的早期鉴定技术,尝试建立柱花草种子RAPD品种鉴别方法.选取11个柱花草品种,采用改良的CTAB法从种子中提取总DNA.在优化的RAPD反应条件下,从100条随机引物中筛选出4条构建指纹图谱.4条引物总扩增带45条,其中多态性带42条,多态带比率达93.3%.通过聚类分析,对指纹图谱进行可靠性评估.11个品种的聚类情况与其亲缘关系相符合,表明依据图谱上的DNA带进行品种鉴定是可信的.本研究首次为柱花草种子的品种鉴定提供了完整的分子标记检测方法.

Stylo RAPD Variety Identification Technique

Varieties of Stylo were difficult to be distinguished by observing morphological characteristics of seeds, so the variety authenticity can not be guaranteed at the earliest growth stage. RAPD method was proposed in variety identification of Stylo seeds. 11 Stylo varieties were used in the study, DNA was extracted from seeds by modified CTAB method. With optimized RAPD procedure, 100 random primers were screened and 4 of them were used in fingerprints constructing. A total of 45 DNA bands were amplified with the 4 primers, 42 of them were polymorphic, the percentage of polymorphic bands reached 93.3%. The validity of the fingerprints were assessed by clustering analysis. The Results showed the 11 varieties grouped were accord with their relationships and the DNA bands can be used in Stylo variety identificating. For the first time, a stylo seeds variety authenticity testing method was established based on molecular markers.