农杆菌介导的茄子愈伤组织遗传转化研究

Genetic transformation of eggplant callus mediated by agrobacterium tumefaciens

以茄子愈伤组织为材料,在优化愈伤组织成苗培养基的基础上,采用正交试验,进行农杆菌介导的茄子愈伤组织遗传转化研究.结果显示,在MS+6-BA(1 mg.L-1)+IAA(0.2 mg.L-1)+KT(1.5 mg.L-1)的培养基上茄子的愈伤组织诱导成苗率最高.结果表明,影响茄子愈伤组织转化的主要因素是预培养时间,其次是侵染时间,再次是共培养时间,并且预培养9 d,侵染20 min,共培养1 d时转化率最高.分子检测表明,11株阳性植株中有7株成功扩出bar基因序列,说明这7株为转基因植株,转基因植株占阳性植株的比率为63.6%.

Eggplant callus was used as materials. Medium for regenerate was optimized and transformation was designed by orthogonal test. The results showed that regeneration rate was the highest under the condition of the medium of MS＋6-BA（1mg·L^-1） ＋ IAA（0.2 mg·L^-1） ＋ KT（1.5mg·L^-1） and the transformation ratio was the highest when pre-cuhure for nine days in advance, invading for 20 min and co-culture for one day. The factors affecting transformation rate was as follows： the preculture time was the most important, next was invading time, and co-culture time was the last. The molecular determination indicated that seven plantlets among eleven positive plantlets were amplified bar gene, which indicated that these seven plantlets has been successfully transformed. The rate of transgenic plantlet to positive plantlets was 63.6%.