ALBA DNA/RNA-binding proteins form an ancient family, which in eukaryotes diversified into two Rpp25-like and Rpp20-like subfamilies. In most studied model organisms, their function remains unclear, but they are usually associated with RNA metabolism, mRNA translatability and stress response. In plants, the enriched number of ALBA family members remains poorly understood. Here, we studied ALBA dynamics during reproductive development in Arabidopsis at the levels of gene expression and protein localization, both under standard conditions and following heat stress. In generative tissues, ALBA proteins showed the strongest signal in mature pollen where they localized predominantly in cytoplasmic foci, particularly in regions surrounding the vegetative nucleus and sperm cells. Finally, we demonstrated the involvement of two Rpp25-like subfamily members ALBA4 and ALBA6 in RNA metabolism in mature pollen supported by their co-localization with poly(A)-binding protein 3 (PABP3). Collectively, we demonstrated the engagement of ALBA proteins in male reproductive development and the heat stress response, highlighting the involvement of ALBA4 and ALBA6 in RNA metabolism, storage and/or translational control in pollen upon heat stress. Such dynamic re-localization of ALBA proteins in a controlled, developmentally and environmentally regulated manner, likely reflects not only their redundancy but also their possible functional diversification in plants.

CNRS LGDP UMR5096 58 Av Paul Alduy 66860 Perpignan France

Imaging Facility Institute of Experimental Botany of the Czech Academy of Sciences Rozvojová 263 165 02 Prague 6 Czech Republic

Institute of Biophysics of the Czech Academy of Sciences Královopolská 612 00 Brno Czech Republic

Laboratory of Pollen Biology Institute of Experimental Botany of the Czech Academy of Sciences Rozvojová 263 165 02 Prague 6 Czech Republic

LGDP UMR5096 Université de Perpignan via Domitia 58 Av Paul Alduy 66860 Perpignan France

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Regulatory dynamics of gene expression in the developing male gametophyte of Arabidopsis

Telomerase Interaction Partners-Insight from Plants

Heat stress response mechanisms in pollen development