Upon heat stress processing of ribosomal RNA precursors into mature rRNAs is compromised after cleavage at primary P site in Arabidopsis thaliana.

Transcription and processing of 45S rRNAs in the nucleolus are keystones of ribosome biogenesis. While these processes are severely impacted by stress conditions in multiple species, primarily upon heat exposure, we lack information about the molecular mechanisms allowing sessile organisms without a temperature-control system, like plants, to cope with such circumstances. We show that heat stress disturbs nucleolar structure, inhibits pre-rRNA processing and provokes imbalanced ribosome profiles in Arabidopsis thaliana plants. Notably, the accuracy of transcription initiation and cleavage at the primary P site in the 5'ETS (5' External Transcribed Spacer) are not affected but the levels of primary 45S and 35S transcripts are, respectively, increased and reduced. In contrast, precursors of 18S, 5.8S and 25S RNAs are rapidly undetectable upon heat stress. Remarkably, nucleolar structure, pre-rRNAs from major ITS1 processing pathway and ribosome profiles are restored after returning to optimal conditions, shedding light on the extreme plasticity of nucleolar functions in plant cells. Further genetic and molecular analysis to identify molecular clues implicated in these nucleolar responses indicate that cleavage rate at P site and nucleolin protein expression can act as a checkpoint control towards a productive pre-rRNA processing pathway.

Biochemical differences in the skin of two blueberries (Vaccinium corymbosum) varieties with contrasting firmness: Implication of ions, metabolites and cell wall related proteins in two developmental stages.

The pursuit of firmer and better-quality blueberries is a continuous task that aims at a more profitable production. To this end it is essential to understand the biological processes linked to fruit firmness, which may diverge among tissues. By contrasting varieties with opposing firmness, we were able to elucidate events that, taking place at immature stage, lay the foundation to produce a firmer ripe fruit. A deep analysis of blueberry skin was carried out, involving diverse comparative approaches including proteomics and metabolomics coupled to immunolocalization assays. In'O'Neal' (low firmness) enhanced levels of aquaporins, expansins and pectin esterases at the green stage were found to be critical in distinguishing it from 'Emerald' (high firmness). The latter featured higher levels of ABA, low methyl esterified pectins in tricellular junctions and high levels of catechin at this stage. Meanwhile, in 'Emerald' 's ripe fruit epicarp, several mechanisms of cell wall reinforcement such as calcium and probably boron bridges, appear to be more prominent than in 'O'Neal'. This study highlights the importance of cell wall reorganization and structure, abundance of specific metabolites, water status, and hormonal signalling in connection to fruit firmness. These findings result particularly valuable in order to improve the fertilization procedures or in the search of molecular markers related with firmness.