栽培高粱叶绿体DNA的酶切片段分析

RFLP Analysis of Cultivated Sorghum Chloroplast Male Sterility Special DNA

实验选取了具优良性状的高粱 ( Sorghum bicolor) 4品种、两细胞质雄性不育系 ,采用改进的高盐低 p H值法对其叶绿体 DNA进行提取、纯化、限制性内切酶酶切分析 .结果表明 :改进的高盐低 p H值法无论在 DNA的得率及质量上都有一定的提高 ,足以满足 RFL P或其他酶切方法的需要 ;高粱栽培品种间的叶绿体 DNA变异很少 ,反映了各族间极其相近的亲缘关系 ;在可育胞质品种与胞质雄性不育系的叶绿体 DN A之间明显存在两类酶切式样的差异 ,在雄性不育系的叶绿体 DNA中存在一可能的点突变和另一可能的大断缺失 ,该高粱叶绿体 DNA的变异是否参与雄性不育性的形成尚需进一步的实验加以证实 .

Four elite varieties and two cytoplasm male sterile lines of sorghum were chosen to be analyzed. Chloroplast DNA was extracted from leaf tissues with a high salt and low pH buffer.Chloroplast DNA extracted with this method needs no further purification,and can directly be digested with six restriction enzymes. The result shows that closer affinity exists among these basic traces,which exhibits the conservatism in chloroplast DNA evolution.In particular, it obviously exhibits two types of restriction patterns between fertile and CMS lines in ApaI and PstI digestions.A probable large deletion and another probable site mutation were sorted out in the CMS lines.It needs further research to prove if these cpDNA variations were involved in CMS formation or not during sorghum evolution.