垦鉴稻10号遗传转化体系的建立

Establishment of Genetic Transformation System in Kenjiandao No.10

农杆菌介导的遗传转化技术,可优化水稻遗传转化体系。为研究农杆菌介导的遗传转化,以垦鉴稻10号去壳种子作为试验材料,研究了75%乙醇、0.1%升汞和5%次氯酸钠不同配比及处理时间消毒后的效果;利用试验筛选的最佳消毒条件,分别以MS培养基、B5培养基、N6培养基以及NMB培养基附加不同浓度2,4-D,筛选诱导愈伤的最佳培养基,并进行愈伤组织的转化。结果表明:诱导愈伤的最佳消毒处理组合为75%酒精1 min+0.1%升汞10 min+5%次氯酸钠4 min;经过方差分析NMB培养基附加2.5 mg·L^-12,4-D对该品种愈伤组织的诱导效果明显优于其他培养基,诱导的愈伤组织大而洁白,硬度适中。将含p3300-ubi-BhGolS1-Tnos重组子的根癌农杆菌与愈伤组织共同培养,经检测,得到转化成功的愈伤组织。

Agrobacterium-mediated genetic transformation technology can optimize rice genetic transformation system.In order to study agrobacterium-mediated genetic transformation,taking Kenjiandao No.10 as the experimental materials,the effect of different matching and treatment time on the mature embryo callus were studied using 75%alcohol,0.1%mercuric chloride and 5%sodium hypochlorite as disinfectants.Use MS,B5,N6 and NMB as selective medium and add different concentrations of plant hormones to find out the best medium to inductive callus.The results showed that,the best callus induction disinfection treatment combination was:75%alcohol 1 min+0.1%mercuric chloride 10 min+5%sodium hypochlorite 4 min;after analysis of variance NMB medium with 2.5 mg·L^-12,4-D was better than the other medium in callus induction,and the induced callus were larger with white color and moderate hardness.After cultivating Agrobacterium tumefaciens plasmid with vector p3300-ubi-BhGolS1-Tnos and callus together,the successful transformation of callus were obtained.