Ascorbic Acid (AsA, vitamin C) has multiple biological roles in plants. Although several
pathways for the novo synthesis of AsA have been reported, it is the L-Gal, also known
as the Smirnoff-Wheeler (SW), the main pathway operating in photosynthetic tissues in
plants. Although there is abundant information of how these genes are regulated at the
transcriptional level, little is known about the regulation and the compartmentation of this
pathway in higher plants. In order to investigate the localization of dynamics of proteins
involved in AsA biosynthesis in vivo at the cellular level, we have generated GFP fusions
using the genomic region of the last five genes of the pathway, expected to be localized
in the cytoplasm. These constructs have been transformed in Arabidopsis thaliana (Col-
0) plants and generated stable transgenic lines. Detailed localization and dynamics of the
proteins are currently being investigated. In parallel, we have analyzed whether these
proteins form a complex in planta using co-immunoprecipitation assays using Nicotiana
benthamiana. Our data indicate that most protein of the SW pathway show a nucleocytoplasmic
localization and that they associate in vivo. Details of the results obtained
will be presented.
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