Journal of Taiwan Agricultural Research

Author:Yi-Wen Wang, Hui-Chen Lai, Hsiao-Ying Hsieh, Chang-Sheng Wang*, and Da-Gin Lin*

Abstract:

      The aim of this study is to construct novel expression system driving reporter genes, AcGFP and ZsGFP, for functional evaluation of rice genes. Two constructs, pCAMBIA35SAcGFP and pCAMBIA35SZsGFP, were completed and then introduced into rice genomes by Agrobacterium-mediated transformation. The results displayed that the transformed calli of two transgenic lines present green fluorescence after culture for 14 days. The radicles and roots of 35S::ZsGFP lines presented green fluorescence in the seed lings at 14 days after germination. At the same time, the radicles and endosperm aleurone layer of 35S::ZsGFP lines also displayed green fluorescence. However, the coleoptiles and spires of two transgenic lines did not be observed green fluorescence since the interference of chlorophyll fluorescence. The 35S::ZsGFP lines showed stronger fluorescence than those of 35S::ZsGFP since the poor expression efficiency of 35S::ZsGFP system. Finally, AcGFP was applied to analyze the activities of maize Ubi-1, rice GluB5 and GluC promoters. The results displayed that transformed calli and shoots of Ubi-1::AcGFP lines could express green fluorescence. The transformed calli and shoots of GluB5::AcGFP and GluC::AcGFP lines also expressed green fluorescence. These results suggest that maize Ubi-1, rice GluB5 and GluC promoters could drive  AcGFP and produce functional fluorescent proteins. Therefore, the expression systems established in this study can provide as selectable markers in the initial stages of plant transformation and are applicable in the development of transgenic platforms and functional analysis of plant genes.

Key words:Green fluorescent protein gene, Reporter, Promoter, Expression system, Agrobacterium-mediated transformation, Rice

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