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利用农杆菌介导的原位转化方法将植物抗旱相关转录因子BcWRKY2基因导入优良玉米自交系郑58、K10、黄早四和Mo17。采用正交设计对玉米品种、菌液浓度、乙酰丁香酮浓度(AS)、真空处理时间和表面活性剂Silwet-77浓度进行了优化。研究结果表明,各因素对转化率影响顺序依次是菌液浓度>品种>AS浓度>真空处理时间>Silwet-77浓度。确定了最优转化条件为品种郑58、真空处理时间20min、菌液浓度为OD600=0.9、AS浓度100μmol/L、Silwet-77浓度0.05%。PCR及RT-PCR分子检测试验说明,BcWRKY2基因已经整合到玉米基因组中,并且得到表达。
Agrobacterium tumefaciens - mediated in situ transformation of plant drought - related transcription factor BcWRKY2 gene into good maize inbred lines Zheng 58, K10, Huang Zaisi Mo17. Orthogonal design was used to optimize corn variety, bacterial concentration, acetosyringone concentration (AS), vacuum treatment time and surfactant Silwet-77 concentration. The results showed that the order of influence of various factors on the conversion rate was bacterial concentration> variety> AS concentration> vacuum treatment time> Silwet-77 concentration. The optimal transformation conditions were identified as Zheng 58, vacuum treatment time 20min, OD600 = 0.9, AS concentration 100μmol / L and Silwet-77 concentration 0.05%. PCR and RT-PCR molecular test showed that the BcWRKY2 gene has been integrated into the maize genome and expressed.