Comparative analysis of reporter gene expression regulated by constitutive promoter for woody plant transformation

利用植物原生质体检测了由已知组成型启动子调控的GFP基因的瞬时表达，以比较启动子在几种植物组织来源的原生质体中的活性。目的是明确木本植物转化中的有效启动子。在电穿孔研究中，除已明确的7种植物外，还对白骨壤属、海桑属等4种红树林原生质体的分离条件进行了新的研究。在35 S启动子（E12 Ω）和玉米ubiquitin启动子的修饰下，落叶松（Larix）原生质体表达GFP，而35 S启动子不能表达GFP。E12 Ω启动子也可用于木榄（红树林）。为了在mRNA水平上比较启动子的活性，建立了基于qRT-PCR的评价体系，并构建了含EL 2 Ω启动子的木本植物生物成像质粒载体。

The transient expression of GFP gene, which regulated by known constitutive promoter in plant, was examined with protoplasts in order to compare the promoter activity in each of protoplast derived from several plant tissues. It was aimed to clarify the efficient promoter in woody plant transformation. For the electroporation study, isolation condition of protoplast was newly clarified in four kinds of mangrove including Avicennia and Sonneratia genus in addition to the previously clarified seven plants. The protoplast of Larix (conifer) showed strong GFP expression by modified 35S (El2Ω) and maize ubiquitin promoter, but not 35S promoter. El2Ω promoter were also available for Bruguiera (mangrove). To compare the promoter activity at the mRNA level, qRT-PCR-based evaluation system was developed, and the plasmid vector for bio-imaging of woody plant was constructed with EL2Ω promoter.