YS型小麦温敏不育基因的RAPD标记

Tagging Wheat Thermo-sensitive Male Sterile Gene of YS Type A3314 by RAPD Analysis

选取温敏小麦不育系A3314与Silverstar杂交组合的F2高可育和高不育单株构建基因池,利用500对随机引物对其进行多态性分析,同时对其反应体系和扩增条件进行优化。试验结果表明,在25μL反应体系中使用50 ng的模板DNA,2 mmol/L Mg2+,0.2 mmol/L dNTPS,0.2μmol/L引物,1 UTaq酶;反应条件为94℃预变性5 min,然后进行94℃变性45 s,37℃结合45 s,72℃延伸90 s,40个循环后,再72℃延伸10 min,小麦RAPD扩增效果较好;S310750为小麦温敏不育基因的连锁标记。

F2 population derived from F1 hybrid developed from the cross A3314/Silverstar, was used in this study. Fertile bulk was constructed by polling equal amount of 15 highly fertile lines. Sterile bulk was obtained by polling equal amount of 15 highly sterile lines. A3314 and Silverstar, were analyzed with 500 pairs of random primers. And optimization of the RAPD reaction system of wheat. It showed that this system was 25 μL total volume contained 50 ng template DNA, 2 mmol/L Mg2＋ ,0.2 mmol/L dNTPS,0.2 μmol/L random primer, 1 U Taq ploymerase. After pre-denaturing at 94℃ for 5 min, under the condition of denaturing at 94℃ for 45 s, annealing at 37 ℃ for 45 s, extension at 72℃ for 90 s, amplify- ing for 40 cycles, and extension at 72℃ for 10 min at last, the result of amplication was good for RAPD research in wheat. And S310750 can be the linked marker of wheat thermo-sensitive male sterile gene.