光照培养的籼稻成熟胚高效再生及转化的研究

Transformation,Inducing and Regeneration of Callus Initiated from Mature Embryos of Indica

以常规籼系明恢86、七山占、中优早8的成熟胚为外植体,通过根癌农杆菌介导法,建立了一种光照培养条件下的新型籼稻成熟胚高效再生和转化体系。结果表明,籼稻在MS+4%麦芽糖+2 mg/L 2,4-D+0.5 mg/L KT+1 mg/L IAA+0.5 mg/L 6-BA+2.878 g/L L-脯氨酸+1 g/L水解酪蛋白,于32℃持续光照10 d左右,可获得愈伤组织的平均诱导率约为84.9%;愈伤组织在MS+2 mg/L KT+1 mg/L IAA+2 mg/L 6-BA+1 g/L水解酪蛋白的培养基上再生率达到80.4%。建立在上述再生体系基础上的GUS基因的瞬时表达率为55.8%。

In this study we established an efficient indica regeneration system initiated from mature embryos of Minghui 86,Qishanzhan and Zhongyouzao8.The results showed that MS medium was the most suitable for callus induction and differentiation.4% maltose could produce the highest rate of callus induction and differentiation.Adding 2 mg/L 2,4-D,0.5 mg/L KT,1 mg/L IAA,0.5 mg/L 6-BA,2.878 g/L L-proline,1 g/L casein hydrolysate in the callus induction medium,it were continuous lighted for 7 days at 32℃,2 mg/L KT,1 mg/L IAA,2 mg/L 6-BA,1 g/L casein hydrolysate supplemented in differentiation medium could significantly improve the callus induction rate and regeneration rate.It was also under continuous light at 32℃.The average induction rate was 84.9% and regeneration rate was 80.4%.By the Agrobacterium-mediated transformation method,hyg-resistant calli were obtained,using indica Minghui 86 as a genetic transformation receptor.GUS staining with resistant calli showed that the reporter gene had been integrated into the rice chromosome and had expressed.This transformation method based on the above-mentioned regeneration system increased the transient expression efficiency to 55.8%.