Chickpea is a grain legume that enhances soil fertility and represents an important source of green proteins for human health. It is typically cultivated in marginal areas with limited water availability. The aim of this study was to shed light into the molecular mechanisms of drought tolerance of this legume. First, the physiological response of nine stable genotype lines in control and drought-stress conditions was assessed. Two of these genotypes (Desi PI5980808 and Kabuli Flip07 318 C) showed opposite physiological responses to drought stress. Desi PI5980808 displayed a reduced chlorophyll content and an unaltered concentration of osmolytes (proline and soluble sugars) under drought stress. Kabuli Flip07 318 C did not show any reduction in photosynthesis and chlorophyll content, but a significant increase of proline and soluble sugars was observed under the drought stress. To identify genes and molecular mechanisms involved in drought tolerance, RNA-seq was performed in control conditions and after one week of drought stress in these two contrasting genotypes. The genotype with higher drought sensitivity showed more intense changes in gene expression than the genotype with less sensitivity, up-regulating genes involved in photophosphorylation process (transferases, oxygen lyases and oxidoreductases), hormones (brassinosteroids, abscisic acid and gibberellin response), solute transporters, nutrient uptake, and cell wall properties (cellulose synthases, hemicellulose synthases, poligalacturonases, pectate lyases). Small number of up-regulated genes in the genotype with lower drought sensitivity included those involved in chromatin modifications. These results will be helpful for further studies aiming at identifying genes and molecular markers to be used in breeding strategies to develop chickpea cultivars more resilient to water stress.

Arundo donax L. is a C3 fast-growing grass that yields high biomass under stress. To elucidate its ability to produce biomass under high salinity, we investigated short/long-term NaCl responses of three ecotypes through transcriptional, metabolic and DNA methylation profiling of leaves and roots. Prolonged salt treatment discriminated the sensitive ecotype 'Cercola' from the tolerant 'Domitiana' and 'Canneto' in terms of biomass. Transcriptional and metabolic responses to NaCl differed between the ecotypes. In roots, constitutive expression of ion transporter and stress-related transcription factors' genes was higher in 'Canneto' and 'Domitiana' than 'Cercola' and 21-day NaCl drove strong up-regulation in all ecotypes. In leaves, unstressed 'Domitiana' confirmed higher expression of the above genes, whose transcription was repressed in 'Domitiana' but induced in 'Cercola' following NaCl treatment. In all ecotypes, salinity increased proline, ABA and leaf antioxidants, paralleled by up-regulation of antioxidant genes in 'Canneto' and 'Cercola' but not in 'Domitiana', which tolerated a higher level of oxidative damage. Changes in DNA methylation patterns highlighted a marked capacity of the tolerant 'Domitiana' ecotype to adjust DNA methylation to salt stress. The reduced salt sensitivity of 'Domitiana' and, to a lesser extent, 'Canneto' appears to rely on a complex set of constitutively activated defences, possibly due to the environmental conditions of the site of origin, and on higher plasticity of the methylome. Our findings provide insights into the mechanisms of adaptability of A. donax ecotypes to salinity, offering new perspectives for the improvement of this species for cultivation in limiting environments.

The effects of UV-B irradiation and Biomin (a natural substance extracted from coal with active ingredients of humic acids) on the content of endogenous polyamines spermine, spermidine and putrescine, and free amino acids in shoots and roots of young triticale seedlings were investigated. Biomin was added to the nutrient medium 3 days prior to UVB irradiation. The seedlings were treated with 7.7 kJ m(-2) day(-1) UV-B light for 4 days. The exposure to UV-B increased total free amino acids, while Biomin application alone did not affect considerably their content. The treatment with UV-B or Biomin alone provoked augmentation of conjugated and bound polyamine (PA) fractions. Data suggest that Biomin alleviates the negative consequences of UV-B stress, manifested by the normalized amino acid and polyamine amounts in the UV-B + Biomin-treated plants. This study demonstrates the protective effect of Biomin on triticale plants against UV-B irradiation, which could be related to alterations in PAs and amino acids pools.

Stimulated production of secondary phenolic metabolites and proline was studied by using cell cultures of artichoke [Cynara cardunculus L. subsp. scolymus (L.) Hayek] submitted to nutritional stress. Artichoke cell cultures accumulated phenolic secondary metabolites in a pattern similar to that seen in artichoke leaves and heads (capitula). This paper shows that both callus and cell suspension cultures under nutritional stress accumulated phenolic compounds and proline, at the same time their biomass production was negatively affected by nutrient deficiency. The results obtained strongly suggest that plant tissues respond to nutrient deprivation by a defensive costly mechanism, which determines the establishment of a mechanism of trade-off between growth and adaptive response. Furthermore, the results of this research suggest that perception of abiotic stress and increased phenolic metabolites are linked by a sequence of biochemical processes that also involves the intracellular free proline and the oxidative pentose phosphate pathway. The main conclusion of this paper is that, once calli and cell suspension cultures respond to nutrient deficiency, in acclimated cells the establishment of a negative correlation between primary metabolism (growth) and secondary metabolism (defence compounds) is observed.

Background Drought is a major constraint for plant growth and crop productivity that is receiving an increased attention due to global climate changes. Chloroplasts act as environmental sensors, however, only partial information is available on stress-induced mechanisms within plastids. Here, we investigated the chloroplast response to a severe drought treatment and a subsequent recovery cycle in tomato through physiological, metabolite and proteomic analyses. Results Under stress conditions, tomato plants showed stunted growth, and elevated levels of proline, abscisic acid (ABA) and late embryogenesis abundant gene transcript. Proteomics revealed that water deficit deeply affects chloroplast protein repertoire (31 differentially represented components), mainly involving energy-related functional species. Following the rewatering cycle, physiological parameters and metabolite levels indicated a recovery of tomato plant functions, while proteomics revealed a still ongoing adjustment of the chloroplast protein repertoire, which was even wider than during the drought phase (54 components differentially represented). Changes in gene expression of candidate genes and accumulation of ABA suggested the activation under stress of a specific chloroplast-to-nucleus (retrograde) signaling pathway and interconnection with the ABA-dependent network. Conclusions Our results give an original overview on the role of chloroplast as enviromental sensor by both coordinating the expression of nuclear-encoded plastid-localised proteins and mediating plant stress response. Although our data suggest the activation of a specific retrograde signaling pathway and interconnection with ABA signaling network in tomato, the involvement and fine regulation of such pathway need to be further investigated through the development and characterization of ad hoc designed plant mutants.

The addition of acetone to rac-2-methylsulfinyl-benzaldehyde, catalyzed by secondary amines (morpholine, piperidine, L-proline, L-prolinamide and (S)-(-)-alpha,alpha-diphenyl-2-pyrrolidinemethanol trimethylsilyl ether), was realized. Interesting aspects, such as 1,4-asymmetric induction by the remote sulfinyl group of the aldehyde but also the stereochemical control and kinetic resolution of the racemic substrate by the chiral organocatalyst were examined. Finally, an unexpected stereoselective retro-aldol reaction catalyzed by L-proline was pointed out. (C) 2016 Elsevier Ltd. All rights reserved.

Tomato is a major crop in the Mediterranean basin, where the cultivation in the open field is often vulnerable to drought. In order to adapt and survive to naturally occurring cycles of drought stress and recovery, plants employ a coordinated array of physiological, biochemical, and molecular responses. Transcriptomic studies on tomato responses to drought and subsequent recovery are few in number. As the search for novel traits to improve the genetic tolerance to drought increases, a better understanding of these responses is required. To address this need we designed a study in which we induced two cycles of prolonged drought stress and a single recovery by rewatering in tomato. In order to dissect the complexity of plant responses to drought, we analyzed the physiological responses (stomatal conductance, CO2 assimilation, and chlorophyll fluorescence), abscisic acid (ABA), and proline contents. In addition to the physiological and metabolite assays, we generated transcriptomes for multiple points during the stress and recovery cycles. Cluster analysis of differentially expressed genes (DEGs) between the conditions has revealed potential novel components in stress response. The observed reduction in leaf gas exchanges and efficiency of the photosystem PSII was concomitant with a general down-regulation of genes belonging to the photosynthesis, light harvesting, and photosystem I and II category induced by drought stress. Gene ontology (GO) categories such as cell proliferation and cell cycle were also significantly enriched in the down-regulated fraction of genes upon drought stress, which may contribute to explain the observed growth reduction. Several histone variants were also repressed during drought stress, indicating that chromatin associated processes are also affected by drought. As expected, ABA accumulated after prolonged water deficit, driving the observed enrichment of stress related GOs in the up-regulated gene fractions, which included transcripts putatively involved in stomatal movements. This transcriptomic study has yielded promising candidate genes that merit further functional studies to confirm their involvement in drought tolerance and recovery. Together, our results contribute to a better understanding of the coordinated responses taking place under drought stress and recovery in adult plants of tomato.

A selection of amino acids, namely arginine, proline and tyrosine previously irradiated to 3.2 mega-Gray in the solid state and analyzed by differential scanning calorimetry (DSC) and optical rotatory dispersion (ORD) were analyzed in the present work by mass spectrometry with the purpose to identify the radiolysis products and validate the results obtained previously with DSC and ORD. The radiolysis of amino acids is a top-down approach of a research program designed to assess the radiolysis resistance of these molecules for 4.6 x 10(9) years once buried in primitive bodies of the Solar System.

Efficient utilization of saline land for food cultivation can increase agricultural productivity and rural income. To obtain information on the salt tolerance/susceptibility of wild chicory (Cichorium intybus L.), the influence of salinity (0-260 mM NaCl) on chicory seed germination and that of two salinity levels of irrigation water (100 and 200 mM NaCl) on plant growth, antioxidative enzyme activity, and accumulation of proline and malondialdehyde (MDA) were investigated. The trials were performed outdoors, in pots placed under a protective glass covering, for two consecutive years. Seeds showed a high capacity to germinate in saline conditions. The use of 100 mM NaCl solution resulted in 81 % germination, whereas seed germinability decreased below 40 % using salt concentrations above 200 mM NaCl. Wild chicory showed tolerance to medium salinity (100 mM NaCl), whereas a drastic reduction in biomass was observed when 200 mM NaCl solution was used for irrigation. MDA, present in higher amounts in leaves than in roots, decreased in both tissues under increasing salinity. Proline content increased remarkably with the level of salt stress, more so in roots than in leaves. In salt stress conditions, the activity of antioxidant enzymes (APX, CAT, POD, SOD) was enhanced. The electrophoretic patterns of the studied enzymes showed that the salinity of irrigation water affected only the intensity of bands, but did not activate new isoforms. Our results suggest that wild chicory is able to grow in soil with moderate salinity by activating antioxidative responses both in roots and leaves.

Micropropagation of Origanum vulgare L. by shoot buds, as a potential model system for studying carbon skeleton diversion from growth to secondary metabolism as adaptive response to nutrient deficiency, has been performed. In addition, the antioxidant phenolic compounds, produced by shoots under nutritional stress or in response to exogenously added proline, have been studied. Caffeic acid, rosmarinic acid, and lithospermic acid B have been isolated in oregano shoot cultures by reversed-phase high-performance liquid chromatography, and their structures have been elucidated by tandem mass spectrometry. Both nutritional stress, which in turn causes a moderate increase of constitutive free proline, and exogenous proline affect growth and antioxidant phenolic content of oregano shoots, compared to control. The role of proline, and the associated redox cycle, as a form of metabolic signaling based on a transfer of redox potential amongst interacting cell pathways, which in turn elicit phenolic metabolism via stimulated carbon flux through oxidative pentose phosphate pathway, is discussed. Furthermore, the potential use of oregano tissue and callus cultures as a new strategy to enable the production of useful secondary metabolites on a commercial scale is also discussed.

Energy dependence of the proline/glutamate antiporter in rat kidney mitochondria has been investigated by means of both spectroscopic measurements and isotopic techniques, using either normal or [C-14]glutamate-loaded mitochondria. The sensitivity of the proline/glutamate antiport to the ionophores valinomycin and nigericin, under conditions in which Delta Psi and Delta pH are selectively affected, shows that the exchange is energy dependent. Measurements of both membrane potential and proton movement across the mitochondrial membrane suggest that proline/glutamate antiport is driven by the electrochemical proton gradient via the Delta Psi dependent proline/glutamate translocator and Delta pH-dependent glutamate/ OH- carrier. Such a carrier provides for re-uptake of glutamate that has already passed out of the mitochondria in exchange with incoming proline, made possible by the existence of a separate pool of glutamate in the intermembrane space, directly shown by means of HPLC measurements.