不同玉米自交系中导入SAMS基因的转化与检测

Transformation and Detection of SAMS Genes Transferred by Different Inbred Lines of Maize

以优良玉米自交系丹598、郑58、昌7-2、掖478的茎尖为受体,采用农杆菌介导法将抗旱基因SAMS转入玉米,对转化植株进行PCR检测,共获得阳性植株35株,表明SAMS基因已经整合到玉米基因组中。同时对影响茎尖转化效率的主要因素进行研究。结果表明,侵染时间、菌液浓度、共培养时间和乙酰丁香酮对转化率影响显著。菌液OD600值为0.5～0.7,侵染15min,共培养3d,乙酰丁香酮浓度为100μmol/L时,有利于提高玉米茎尖转化率,其中昌7-2转化率最高。

The SAMS gene was transferred by the shoot apical point of different inbred lines,which included Dan598,Zheng58,Chang7-2 and Ye478 through Agrobacterium-mediated transformation.The transgenic maize were identified by PCR,and 35 positive transgenic plants were obtained,which showed that the SAMS gene has been integrated into the four different inbred lines.Besides we studied the important factors influenced the efficiency of the transformation.The results shows that the frequency of transformation were different with the various concentration of bacteria、 acetosyringone（AS） and the different time of infection.And we found that the bacteria concentration（OD6000.5-0.7）,the infection time（25 min）,the co-culture time（3 d）,and the concentration of acetosyringone（100 μmol/L） were optimal.And Chang7-2 has the highest percent conversion with the optimal condition.