玉米RAPD程序优化研究及其初步探讨  

Studies on the Protocol Optimum and Preliminary Applification RAPD in Maize

作　　者：傅骏骅[1] 李连城[1] 刘新芝[1] 彭泽斌[1] 黄长玲 

机构地区：[1]中国农业科学院作物育种栽培研究所,北京100081

出　　处：《作物学报》1997年第1期56-61,共6页Acta Agronomica Sinica

基　　金：攀登计划资助课题

摘　　要：本文以黄早四、汶黄、Mo17等我国主要玉米自交系为材料,使用国产PCR仪,对玉米RAPD分析中PCR过程的DNA模板浓度,扩增反应的循环数进行了研究,并针对国产PCR扩增仪(PCR—90AD)的特点,确定了DNA变性,引物和模板DNA结合,引物沿模板延伸3个步骤的时间。实验结果表明玉米PCR过程使用10～15ng的模板DNA,在94℃模板变性时间1分30秒,36℃引物退火2分钟,72℃引物延伸2分30秒,扩增循环35～40周期条件下扩增效果较好,使用这一RAPD程序研究不同自交系间遗传差异,结果与系谱法基本一致,而同一自交系的10个单株之间,RAPD带型是一致的。本实验建立了适合玉米RAPD分析的PCR程序及条件,为RAPD分析法应用于玉米的遗传研究打下了良好的基础。Using maize inbreds Huang zau 4 et al. as experimental material, we studied the template concentration and the number of amplified cycle, and determined the time for DNA denaturing step, primer annealing step, and primer extension step in Reserch Program Thermal Controller (PCR-90 AD) made in China. The results indicated that under the condition of template concentration ranging between 10-50 ng DNA, denaturing at 94°C for 1 min and 30 sec, primer annealing at 36°C for 2 min, primer extension at 72°C for 2 min and 30 sec, and the number of cycles from 35 to 40, the amplification was good for PCR program in maize. The genetic distance among different inbreds and the genetic variation in each maize inbred were tested by the RAPD markers. The results showed that therse was no genetic variation with in the inbred , but the genetic difference among indreds was found, which is consistent with the pedigree analysis. RAPD pattern founded in this experiment lays a good foundation for application of RAPD in maize genetic research.

关 键 词：RAPD 玉米 自交系 

分 类 号：S513.035.1[农业科学—作物学]