Abstract

The accumulation and localization of dehydrin-like proteins were analyzed in the embryonic axes of beech (Fagus sylvatica L.) seeds during maturation between the 12th and 19th week after flowering and in the dry state after 2 years of storage. Three dehydrin-like proteins were reported using an antibody specific to the K-segment, and their subcellular localization was investigated using an immunocytochemistry approach. All dehydrin-like proteins were phosphorylated, and two, with molecular masses of 26 and 44 kDa, accumulated in mature seeds. During seed maturation, dehydrin-like proteins were frequently detected inside the round or segmented nuclei, along the cell plasma membrane and associated with small vesicles. Moreover, they were found to be attached to the mitochondrial and amyloplast envelopes. These proteins were also located in the cytosol and were associated with membrane structures throughout the cytoplasm. Conversely, in the embryonic axes of stored seeds, fewer specific locations were found; dehydrin-like proteins were associated mostly with amyloplasts, and were detected to a lesser extent in the nuclei, vacuoles, and other cytoplasmic structures. Bioinformatic tools were used to predict the putative beech dehydrin sites that interact with DNA, proteins, and lipid membranes to highlight the reported subcellular locations. Here, we present and discuss the possible roles of dehydrin-like proteins in seeds with respect to their subcellular localizations.

Highlights

  • The analyzed cells came from the roothypocotyl axis, because we demonstrated that the accumulating ROS that spread from the root cap to the root apical meristem in embryonic axes of F. sylvatica seeds are key factors that affect the success of long-term storage as well as Brassica napus root is the most susceptible tissue to water deficit stress soon after germination (BagniewskaZadworna 2008)

  • Three dehydrin-like proteins were detected in protein extracts of common beech (F. sylvatica L.) embryonic axes during seed maturation beginning from the 12th up to the 19th week after flowering (WAF) (Fig. 2a)

  • There were no specific dehydrin localizations associated with specific beech seed developmental stages, and there were no differences at the subcellular level in all analyzed tissues from root-hypocotyl axis; for this work we focused on the cortical region

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Summary

Introduction

We present and discuss the possible roles of dehydrin-like proteins in seeds with respect to their subcellular localizations. Late embryogenesis abundant (LEA) proteins, including dehydrins, are hydrophilic molecules accumulated during development and water stress (reviewed in Rorat 2006; Battaglia and others 2008; Hara 2010; Battaglia and Covarrubias 2013). The structural and functional characteristics of LEA proteins are being investigated to understand their role during the adaptive response to water deficit in plants (Rorat 2006; Battaglia and others 2008; Hara 2010; Battaglia and Covarrubias 2013). We decided to investigate their potential role in cell protection during natural dehydration during seed development and seed storage

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