a b s t r a c tTolerance to drought remains poorly described for Jatropha curcas accessions from different geographical and climatic origins. To address this issue we studied the response of two J. curcas accessions, one from Indonesia (wet tropical climate) and the other from Cape Verde islands (semi-arid climate). Potted seedlings (with 71 days) of both accessions were subjected to continuous well watered conditions (control) or to a drought stress period followed by re-watering. To mimic natural conditions in which drought stress develops gradually, stress was imposed progressively by reducing irrigation (10% reduction every 2 days, on a weight base), for a period of 28 days, until a field capacity of 15% (maximum stress) was achieved, followed by one week under well-watered conditions. We measured soil and plant water status, growth and biomass partitioning, leaf morphology, leaf gas exchange and chlorophyll a fluorescence. Both accessions maintained high leaf relative water content (70-80%) even at maximum stress. Net photosynthesis (A n ) was not affected by mild to moderate stress but it abruptly dropped at severe stress. This was due to reduced stomatal conductance, which showed earlier decline than A n . Plant growth (stem elongation, leaf emergence and total leaf area) was reduced, minimizing water loss, but no significant differences were found between accessions. Drought stress did not reduce chlorophyll contents but led to reduced chlorophyll a/b. Both accessions showed fast recovery of both stomatal and photochemical parameters suggesting a good tolerance to water stress. Both J. curcas accessions showed a-dehydrationavoidant behaviour, presenting a typical water saving strategy due to strict stomatal regulation, regardless of their provenance.

Calcium-dependent protein kinases (CDPKs) are involved in plant tolerance mechanisms to abiotic stresses. Although CDPKs are recognized as key messengers in signal transduction, the specific role of most members of this family remains unknown. Here we test the hypothesis that OsCPK17 plays a role in rice cold stress response by analyzing OsCPK17 knockout, silencing, and overexpressing rice lines under low temperature. Altered OsCPK17 gene expression compromises cold tolerance performance, without affecting the expression of key cold stress-inducible genes. A comparative phosphoproteomic approach led to the identification of six potential in vivo OsCPK17 targets, which are associated with sugar and nitrogen metabolism, and with osmotic regulation. To test direct interaction, in vitro kinase assays were performed, showing that the sucrose phosphate synthase OsSPS4, and the aquaporin OsPIP2;1/OsPIP2;6 are phosphorylated by OsCPK17 in a calcium-dependent manner. Altogether, our data indicates that OsCPK17 is required for a proper cold stress response in rice, likely affecting the activity of membrane channels and sugar metabolism.Rice production is severely affected by different abiotic stresses, including cold. Cold perception is mediated by calcium signals that activate kinases to elicit the adequate cellular response. In this work, we show the involvement of the rice calciumdependent protein kinase 17 (OsCPK17) in such a process. We show that altered OsCPK17 gene expression in transgenic lines affects cold tolerance performance, This article is protected by copyright. All rights reserved.and that OsCPK17 targets proteins are associated with osmotic regulation, and sugar and nitrogen metabolism.

Plants can cope with adverse environmental conditions through the activation of stress response signalling pathways, in which the proteasome seems to play an important role. However, the mechanisms underlying the proteasome-mediated stress response in rice are still not fully understood. To address this issue, we have identified a rice E3-ubiquitin ligase, OsHOS1, and characterized its role in the modulation of the cold stress response. Using a RNA interference (RNAi) transgenic approach we found that, under cold conditions, the RNAi::OsHOS1 plants showed a higher expression level of OsDREB1A. This was correlated with an increased amount of OsICE1, a master transcription factor of the cold stress signalling. However, the up-regulation of OsDREB1A was transient and the transgenic plants did not show increased cold tolerance. Nevertheless, we could confirm the interaction of OsHOS1 with OsICE1 by Yeast-Two hybrid and bi-molecular fluorescence complementation in Arabidopsis protoplasts. Moreover, we could also determine through an in vitro degradation assay that the higher amount of OsICE1 in the transgenic plants was correlated with a lower amount of OsHOS1. Hence, we could confirm the involvement of the proteasome in this response mechanism. Taken together our results confirm the importance of OsHOS1, and thus of the proteasome, in the modulation of the cold stress signalling in rice.

Jatropha curcas, a multipurpose plant attracting a great deal of attention due to its high oil content and quality for biofuel, is recognized as a drought-tolerant species. However, this drought tolerance is still poorly characterized. This study aims to contribute to uncover the molecular background of this tolerance, using a combined approach of transcriptional profiling and morphophysiological characterization during a period of water-withholding (49 d) followed by rewatering (7 d). Morphophysiological measurements showed that J. curcas plants present different adaptation strategies to withstand moderate and severe drought. Therefore, RNA sequencing was performed for samples collected under moderate and severe stress followed by rewatering, for both roots and leaves. Jatropha curcas transcriptomic analysis revealed shoot- and root-specific adaptations across all investigated conditions, except under severe stress, when the dramatic transcriptomic reorganization at the root and shoot level surpassed organ specificity. These changes in gene expression were clearly shown by the down-regulation of genes involved in growth and water uptake, and up-regulation of genes related to osmotic adjustments and cellular homeostasis. However, organ-specific gene variations were also detected, such as strong up-regulation of abscisic acid synthesis in roots under moderate stress and of chlorophyll metabolism in leaves under severe stress. Functional validation further corroborated the differential expression of genes coding for enzymes involved in chlorophyll metabolism, which correlates with the metabolite content of this pathway.