Abstract

BackgroundSugars modulate expression of hundreds of genes in plants. Previous studies on sugar signaling, using intact plants or plant tissues, were hampered by tissue heterogeneity, uneven sugar transport and/or inter-conversions of the applied sugars. This, in turn, could obscure the identity of a specific sugar that acts as a signal affecting expression of given gene in a given tissue or cell-type.Methodology/Principal FindingsTo bypass those biases, we have developed a novel biological system, based on stem-cell-like Arabidopsis suspension culture. The cells were grown in a hormone-free medium and were sustained on xylose as the only carbon source. Using functional genomics we have identified 290 sugar responsive genes, responding rapidly (within 1 h) and specifically to low concentration (1 mM) of glucose, fructose and/or sucrose. For selected genes, the true nature of the signaling sugar molecules and sites of sugar perception were further clarified using non-metabolizable sugar analogues. Using both transgenic and wild-type A. thaliana seedlings, it was shown that the expression of selected sugar-responsive genes was not restricted to a specific tissue or cell type and responded to photoperiod-related changes in sugar availability. This suggested that sugar-responsiveness of genes identified in the cell culture system was not biased toward heterotrophic background and resembled that in whole plants.ConclusionsAltogether, our research strategy, using a combination of cell culture and whole plants, has provided an unequivocal evidence for the identity of sugar-responsive genes and the identity of the sugar signaling molecules, independently from their inter-conversions or use for energy metabolism.

Highlights

  • Changes in the environment of green plants result in constant fluctuation in the contents of endogenous sugars, which – besides being the initial carbon backbone for biological molecules represent the main energy source in both the autotrophic and heterotrophic tissues

  • In order to bypass the previously-mentioned biases inherent to studies on sugar signaling, we have developed a novel system, based on the use of a stem-cell-like suspension culture of Arabidopsis thaliana which could be grown without addition of any hormones [17]

  • TUB5 was used as endogenous reference gene in samples derived from A. thaliana cell culture after analysis of the microarray data showed no changes in the expression in response to sugar treatment

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Summary

Conclusions

Our research strategy, using a combination of cell culture and whole plants, has provided an unequivocal evidence for the identity of sugar-responsive genes and the identity of the sugar signaling molecules, independently from their inter-conversions or use for energy metabolism

Introduction
Materials and Methods
Results and Discussion

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