农杆菌介导的小麦成熟胚遗传转化影响因素分析

为解决农杆菌介导的小麦成熟胚遗传转化效率较低的难题,以春小麦Bobwhite成熟胚为外植体,分析植物激素、愈伤组织预培养时间、侵染时间和共培养阶段干燥处理对小麦成熟胚愈伤组织遗传转化的影响,并在遗传转化处理中辅以3个组织再生相关基因(TaSERK1、TaSERK2、TaNiR)的qRT-PCR分析。结果表明,愈伤组织预培养阶段添加适量的2,4-D和Picloram均可诱导成熟胚愈伤组织的形成,但是2mg·L~(-1)Picloram培养基中愈伤组织的胚萌发率为20.78%,远高于2,4-D诱导的胚萌发率(11.38%);共培养阶段对农杆菌侵染的愈伤组织进行干燥处理能适当提高愈伤组织的转化效率;qRT-PCR分析发现再生相关基因TaNiR相对表达量的增加能适当提高愈伤组织的转化率。

Factors Influencing Agrobacterium Mediated Transformation in Mature Embryo of Wheat (Triticum astivum)

To solve the problem of Agrobacterium mediated transformation in mature embryo of wheat (Triticum astivum),spring wheat ‘Bobwhite’ was used as receptor.The effects of plant hormones,the pre-culture time,Agrobacterium mediated transformation time and co-culture methods on wheat mature embryo callus induction and differentiation were analysed.During the transformation,three regeneration related genes (TaSERK1、TaSERK2,TaNiR) were analysed by qRT-PCR.The results showed that adding 2,4-D and Picloram could induce the formation of mature embryo callus,but the germination rate of callus induced by 2 mg · L-1 picloram(20.78％) is higher than that induced by 2,4-D(11.38 ％).Dry processing in co-culture stage could obviously improve the transformation efficiency of callus.It was found that the change of the relative expression regeneration related genes (TaNiR ) could increase the transformation rate of callus by qRT-PCR.