SSR技术鉴定玉米品种真实性反应体系的优化研究

摘要

[目的]优化SSR技术鉴定玉米品种真实性反应体系.[方法]对SSR技术参数(PCR反应体系、退火温度和电泳时间)进行了优化,并对辽宁省主推的10个玉米品种进行了鉴定.[结果]最优PCR反应体系为:灭菌超纯水14.60μl,10 × Buffer(Mg2+)2.00μl,dNTPs 1.20μl,Taq酶0.20μl,正、反向引物各0.50μl及DNA原液1.00μl.退火温度和电泳时间对PCR扩增结果的影响较大,所选的引物不同,在同一条件下呈现理想电泳条带所需的最佳退火温度和电泳时间不同.[结论]该体系应用于杂交玉米品种的真实性快速鉴定是可行的.%[ Objective] The aim was to optimize SSR reaction system applied in the identification of authenticity of maize variety. [ Method] The technical parameters of SSR including PCR reaction system, annealing temperature and electrophoresis time were optimized to identify 10 major maize varieties in Liaoning Province. [ Result ] The optimum PCR reaction system was: 14.60 μl sterile ultrapure water, 2.00 μl 10 x Buffer ( Mg2 + ) , 1.20 μl dNTPs, 0. 20 μ1 Taq enzyme, 0.50 μl each of the forward and reverse primers and 1.00 μl DNA stock solution.Annealing temperature and eleetrophoresis time could greatly influenced the results of PCR amplification. The optimal annealing temperature and electrophoresis time required for the ideal electrophoresis bands under the same conditions were different when different primers were used.[ Conclusion ] The system is feasible to apply in rapid identification of authenticity of hybrid maize varieties.