柱花草花粉不育和可育分离群体基因组DNA分离方法优化  

An Optimized Method of Extracting Genomic DNA from Sterile and Fertile Segregated Populations in Stylosanthes guianensis

作　　者：张子雄 黄春琼[1] 刘国道[1] 

机构地区：[1]中国热带农业科学院热带作物品种资源研究所农业部华南作物基因资源与种质创制重点实验室,儋州571737 [2]海南大学农学院,海口570228

出　　处：《分子植物育种》2017年第9期3623-3628,共6页Molecular Plant Breeding

基　　金：国家自然科学基金(31402124)资助

摘　　要：本研究以柱花草(Stylosanthes guianensis)不育和可育两种分离群体为试验材料,采用优化改进后的CTAB法提取柱花草不育和可育两种分离群体的叶片的基因组DNA。试验结果表明:提取出的DNA检测浓度值373.7~604.3 ng/μL,A260/A280处于1.83~1.91,A260/A230处于1.90~2.05。所提的DNA琼脂糖凝胶的电泳条带显示清晰明亮,没有明显的拖带现象。ISSR-PCR琼脂糖凝胶电泳在不育和可育植株的条带之间具有明显的多态性条带。说明用此优化改进的CTAB法提取的目标基因组DNA的纯度高,质量好。能很好地用于分子遗传特性分析和以ISSR-PCR和SRAP-PCR为基础的分子生物学方面。该改进的试验方法简单、高效、快捷。The sterile and fertile segregated populations in Stylosanthes guianensis were used as experimental materials. The genomic DNA of the leaves from sterile and fertile segregated populations in S. guianensis was extracted by the optimized CTAB method. The results showed that the concentration of the extraction of genomic DNA ranged from 373.70 to 604.30 ng/μL. The values of A260/A280 were between 1.83-1.91 in each sample and the values of A260/A230 were ranged from 1.90 to 2.05. The extracted products had a bright band without tailing which could be observed through agarose gel electrophoresis. There were some clear and distinguishable polymorphic bands between the sterile and fertile plants by the ISSR-PCR agarose gel electrophoresis. The study indicated that genomic DNA which was obtained by the optimized CTAB method had good integrity and high purity, and it can be used in molecular genetic analysis and molecular biology research in ISSR-PCR and SRAP-PCR. The optimized CTAB method was simple, effective, and rapid.

关 键 词：柱花草(Stylosanthes guianensis) 不育和可育分离群体 DNA提取 优化CTAB 

分 类 号：S541.9[农业科学—作物学]