29. エチレン応答性転写因子 (ERFs) の RNAi 法を用いた選択的発現抑制

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A plant hormone, ethylene functions in various aspects of the plant life cycle. Ethylene regulates many defensive gene expressions via GCC box, cis-acting element. ERFs (Ethylene-Responsive Factors) are transcriptional factors that bind to GCC box with a highly conserved DNA-binding domain, ERF domain. Whereas all four ERFs isolated from tobacco are induced by ethylene and exogenously applied cycloheximide (CHX), these ERFs have different activities ; e.g. ERF1,ERF2,and ERF4 are activators and ERF3 is a repressor. It is still unclear how these ERFs interact with each other in the complicated hormone signal pathway and how many genes are under the regulation of ERFs. In this study, we first tried to establish the transformants which have suppressed expression of each ERF. For this purpose, we used selective suppression by RNAi. Specific double stranded RNA expression vectors for each ERF, i.e., ERF2ir, ERF3ir, ERF4ir were constructed and introduced into tobacco (Nicotiana tabacum, cv. Samson NN). After selection on kanamycin medium, 8 ERF4ir transformants, 7 ERF3ir transformants, and 2 ERF2ir transformants were obtained. Whereas these transformants have been under investigation, we report here the result of ERF4ir transformants. ERF4ir tobacco showed normal appearance as non-transformant, whereas the mRNA of ERF4 was hardly detected even after CHX treatment. This result suggests the RNAi method is effective to suppress the expression of ERF4. When the expression of other ERFs, ERF1/2/3,and basic PR5 (osmotin) were analyzed after CHX treatment, ERF4ir transformants showed similar induction as control plant, indicating that ERF4 was not the main regulator for the expression of ERF1/2/3 and osmotin. Characterizations of other ERFir transformants are on going.
植物化学調節学会の論文
2003-10-10

29. エチレン応答性転写因子 (ERFs) の RNAi 法を用いた選択的発現抑制

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