摘要
目的:以高粱丝黑穗病2381(抗病)、矮四(感病)为材料,以优化SSR反应体系为目的。方法:采用CTAB法提取高粱基因组总DNA,用SSR分子标记技术对其进行多态性扩增,通过对体系中不同的Taq酶浓度、模板浓度、dNTP浓度和引物浓度的梯度分析。结果:建立并优化了的SSR-PCR反应体系为20μl反应体系:dNTP浓度为200μmol/L、Taq酶浓度为1.5U、DNA浓度为100ng和引物浓度为0.4μmol/L。达到了较理想的扩增效果。用该体系对94对SSR引物进行了筛选,其中47对引物扩增出了多态性谱带,其中Xtxp3和Xtxp13在抗感的品种间扩增出了差异谱带。结论:应用优化的SSR反应体系可以对高粱丝黑穗病基因进行分析。
Objective： It is optimized SSR reaction system of the Head Smut resistance gene on Sorghum. Result Sorghttm of 2381 （resistant parent against head smut） AM（susceptible parent against head smut） were studied. Method： DNA extraction and purification were elected and optimized. It is CTAB method. Result： SSR reaction systems were optimized. Reaction system of 20μl： dNTP concentration was 200μmol/L, Taq concentration was 1.5U, the concentration of DNA was 100ng and the concentration of primers were 0.4μmol/L. 94 SSR primers were used to screen the samples for markers linked to the head smut resistance gene of Sorghum. 47 primers were high efficient. Polymorplfism bands were amplified with 47 primers. Polymorplfism bands between resistant parent and susceptible parent were amplified with Xtxp13 and Xtxp3. Conclusion： SSR reaction system of the Head Smut Resistance gene on Sorghum was optimized.
出处
《生物技术》
CAS
CSCD
北大核心
2009年第3期37-39,共3页
Biotechnology
基金
辽宁省教育厅重点实验室项目(20060806)
公益性行业(农业)科研专项(nyhyzx07-011-03)资助
