Grapevine (Vitis vinifera L.) promoter analysis by biolistic-mediated transient transformation of cell suspensions

Abstract

Expression of alcohol dehydrogenase promoters from Vitis vinifera L. and Arabidopsis thaliana was investigated in homologous and heterologous cell systems after bombardment of chimeric genes. The effect of several parameters including wounding, DNA delivery and construct type on luciferase/beta-glucuronidase activities was evaluated. In parallel, alcohol dehydrogenase activity was assayed in normal and gaseous nitrogen-treated cells. Compared to A. thaliana, results showed large differences in reporter gene activity according to the Adh promoter leader sequence and the cell suspension system. The V. vinifera cell system was found to be appropriate to investigate Adh promoter functioning. A novel protocol, based on transient genetic transformation of grape-One cells by biolistic is proposed to study grapevine promoter expression, either in air or in response to anaerobiosis.

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