Dai Z, An G
Institute of Biological Chemistry, Washington State University, Pullman 99164-6340, USA.
Plant Physiol. 1995 Dec;109(4):1191-7. doi: 10.1104/pp.109.4.1191.
Transgenic tobacco (Nicotiana tabacum L.) plants carrying a fusion between the nopaline synthase (nos) promoter and chloramphenicol acetyltransferase (CAT) reporter gene (caf) were tested for their response to treatment with H2O2. The nos promoter-driven CAT activity increased significantly by addition of H2O2, reaching the maximum level at 15 mM. Kinetic analysis for CAT activity showed that induction by H2O2 was similar to that of methyl jasmonate (MJ), but was much slower than induction by salicylic acid (SA). Time-course experiments for mRNA level also revealed that the response to H2O2 treatment was similar to that of MJ. The nos promoter displayed a rapid and transient induction of mRNA with SA treatment, with the maximum levels occurring at 3 h, whereas the levels induced by H2O2 or MJ treatment increased continuously during the 11-h experimental period. The antioxidants N-acetyl-L-cysteine and catechol did not alter the SA effect. The responses of the nos promoter to H2O2, MJ, and wounding were significantly reduced by deletions of the CAAT box region and the sequence between -112 and -101. However, these deletions did not significantly alter the SA response. This suggests that H2O2 may have a different mechanism from that of SA for inducing nos promotor activity.
对携带胭脂碱合成酶(nos)启动子与氯霉素乙酰转移酶(CAT)报告基因(caf)融合体的转基因烟草(Nicotiana tabacum L.)植株进行了过氧化氢(H2O2)处理反应测试。添加H2O2后,nos启动子驱动的CAT活性显著增加,在15 mM时达到最高水平。CAT活性的动力学分析表明,H2O2诱导与茉莉酸甲酯(MJ)相似,但比水杨酸(SA)诱导慢得多。mRNA水平的时间进程实验也表明,对H2O2处理的反应与MJ相似。SA处理时,nos启动子显示出mRNA的快速瞬时诱导,在3小时达到最高水平,而H2O2或MJ处理诱导的水平在11小时实验期内持续增加。抗氧化剂N-乙酰-L-半胱氨酸和儿茶酚不会改变SA的作用。CAAT框区域以及-112至-101之间的序列缺失后,nos启动子对H2O2、MJ和创伤的反应显著降低。然而,这些缺失并没有显著改变SA反应。这表明H2O2诱导nos启动子活性的机制可能与SA不同。