Accumulation pattern of dehydrins during sugarcane (var. SP80.3280) somatic embryogenesis

Abstract

The objective of the present study was to determine dehydrin protein levels in sugarcane var. SP80- 3280 during somatic embryogenesis. Dehydrins from embryogenic and non-embryogenic cell cultures were analyzed using western blot and in situ immunolocalization microscopy. Both techniques employ antibodies raised against a highly conserved lysine-rich 15-amino acid sequence termed the K-domain, which is extensively used to recognize proteins immunologically related to the dehydrin family. In embryogenic cultures, western blot analysis of the heat-stable protein fraction revealed eleven major bands ranging from 52 to 17 kDa. They were already visible on the first days, gradually increasing until reaching peak values around day 14, when organogenesis begins, to later decrease in concurrence with the appearance of green plantlets (around day 28). These fluctuations indicate that this pattern of accumulation is under developmental control. Dehydrins were mainly immunolocalized in the nuclei. A phosphatase treatment of protein extracts caused a mobility shift of the 52, 49, and 43 kDa dehydrin bands suggesting a putative modulation mechanism based on protein phosphorylation. In sugarcane embryogenic cultures, presence of dehydrins is a novel finding. Dehydrins were absent in non-embryogenic cultures. The novel findings regarding accumulation, nuclear localization, and phosphorylation of dehydrins provide a starting point for further research on the role of these proteins in the induction and/or maintenance of embryogenesis. Key message The novel findings regarding accumulation, nuclear localization, and phosphorylation of dehydrins provide a starting point for further research on the role of these proteins in the induction and/or maintenance of embryogenesis.