花椰菜温敏型雄性不育系的RAPD标记

Cauliflower Thermo-sensitive Male Sterile Gene RAPD Marker

选取温敏花椰菜不育系GS-19与GS-31杂交组合的F2高可育和高不育单株构建基因池,利用100对随机引物对其进行RAPD标记。同时,采用正交设计对其反应体系及扩增条件进行优化。试验结果表明,在25μL反应体系中含dNTPs0.625mmol/L,引物0.5μmol/L,DNA模板60ng,Taq酶1.5U,超纯水14.9μL;反应条件为94℃预变性4min,然后进行94℃变性30s,36℃退火45s,72℃延伸90s,35个循环后,再72℃延伸7min,花椰菜RAPD扩增效果较好。P21-1800为花椰菜温敏雄性不育基因的连锁标记。

F2 population derived from F1 hybrid developed from the cross GS-19/GS-31,was used in this study to build two gene pools.It has been the use of orthogonal design system and optimization of PAPD-PCR amplification conditions.And adopted by 100 pairs of random primers to marker two gene pools.The results showed that this system was 25 μL total volume contained 60 ng template DNA,0.625 mmol/L dNTPs,0.5 μmol/L random primer,1.5 U Taq ploymerase.After pre-denaturing at 94℃ for 4 min form in,under the condition of denaturing at 94℃ for 30 s,annealing at 36℃for 45 s,extension at 72℃ for 90 s,amplify for 35 cycles,and extension at 72℃ for 7 min at last,the result of amplication was good for RAPD research in heat.P21-1800 can be the linked marker of cauliflower thermo-sensitive male sterile gene.