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

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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.