Abscisic acid- and ethylene-induced abscission of yellow lupine flowers is mediated by jasmonates.

The appropriate timing of organ abscission determines plant growth, development, reproductive success, and yield in relation to crop species. Among these, yellow lupine is an example of a crop species that loses many fully developed flowers, which limits the formation of pods with high-protein seeds and affects its economic value. Lupine flower abscission, similarly to the separation of other organs, depends on a complex regulatory network functioning in the cells of the abscission zone (AZ). In the present study, genetic, biochemical, and cellular methods were used to highlight the complexity of the interactions among strong hormonal stimulators of abscission, including abscisic acid (ABA), ethylene, and jasmonates (JAs) precisely in the AZ cells, with all results supporting that the JA-related pathway has an important role in the phytohormonal cross-talk leading to flower abscission in yellow lupine. Based on obtained results, we conclude that ABA and ET have positive influence on JAs biosynthesis and signaling pathway in time-dependent manner. Both phytohormones changes lipoxygenase (LOX) gene expression, affects LOX protein abundance, and JA accumulation in AZ cells. We have also shown that the signaling pathway of JA is highly sensitive to ABA and ET, given the accumulation of COI1 receptor and MYC2 transcription factor in response to these phytohormones. The results presented provide novel information about the JAs-dependent separation of organs and provide insight and details about the phytohormone-related mechanisms of lupine flower abscission.

Differential expression of genes in olive leaves and buds of ON- versus OFF-crop trees.

Alternate bearing (AB) refers to the tendency of trees to have an irregular crop load from 1 year (ON) to the next year (OFF). Despite its economic importance, it is not fully understood how gene networks and their related metabolic pathways may influence the irregular bearing in olive trees. To unravel molecular mechanisms of this phenomenon in olive (cv. Conservalia), the whole transcriptome of leaves and buds from ON and OFF-trees was sequenced using Illumina next generation sequencing approach. The results indicated that expressed transcripts were involved in metabolism of carbohydrates, polyamins, phytohormones and polyphenol oxidase (POD) related to antioxidant system. Expression of POD was increased in leaf samples of ON- versus OFF-trees. The expression pattern of the greater number of genes was changed more in buds than in leaves. Up-regulation of gene homologues to the majority of enzymes that were involved in photorespiration metabolism pathway in buds of ON-trees was remarkable that may support the hypotheses of an increase in photorespiratory metabolism in these samples. The results indicated changes in expression pattern of homologous to those taking part of abscisic acid and cytokinin synthesis which are connected to photorespiration. Our data did not confirm expression of homologue (s) to those of chlorogenic acid metabolism, which has been addressed earlier that have a probable role in biennial bearing in olive. Current findings provide new candidate genes for further functional analysis, gene cloning and exploring of molecular basses of AB in olive.

Insight into the cellular effects of nitrated phospholipids: Evidence for pleiotropic mechanisms of action.

Nitrated phospholipids have been recently identified in biological systems and showed to display anti-oxidant and anti-inflammatory potential in models of inflammation in vitro. Here, we have explored the effects of nitrated 1-palmitoyl-2-oleyl-phosphatidyl choline (NO2-POPC) in cellular models. We have observed that NO2-POPC, but not POPC, induces cellular changes consisting in cytoskeletal rearrangement and cell shrinking, and ultimately, loss of cell adhesion or impaired cell attachment. NO2-POPC releases NO in vitro and induces accumulation of NO in cells. Nevertheless, the effects of NO2-POPC are not superimposable with those of NO donors, which points to distinctive mechanisms of action. Notably, they show a stronger parallelism, although not complete overlap, with the effects of nitrated fatty acids. Interestingly, redistribution of vimentin by NO2-POPC is attenuated in a C328S mutant, thus indicating that this residue may be a target for direct or indirect modification in NO2-POPC-treated cells. Additionally, NO2-POPC interacts with several typical lipoxidation targets in vitro, including vimentin and PPARγ constructs, likely through cysteine residues. Therefore, nitrated phospholipids emerge as potential novel electrophilic lipid mediators with selective actions.

S-nitroso- and nitro- proteomes in the olive (Olea europaea L.) pollen. Predictive versus experimental data by nano-LC-MS.

The data presented here are related to the research article entitled "Generation of nitric oxide by olive (Olea europaea L.) pollen during in vitro germination and assessment of the S-nitroso- and nitro-proteomes by computational predictive methods" doi:10.1016/j.niox.2017.06.005 (Jimenez-Quesada et al., 2017) [1]. Predicted cysteine S-nitrosylation and Tyr-nitration sites in proteins derived from a de novo assembled and annotated pollen transcriptome from olive tree (Olea europaea L.) were obtained after using well-established predictive tools in silico. Predictions were performed using both default and highly restrictive thresholds. Numerous gene products identified with these characteristics are listed here. An experimental validation of the data, consisting in nano-LC-MS (Liquid Chromatography-Mass Spectrometry) determination of olive pollen proteins after immunoprecipitation with antibodies to anti-S-nitrosoCys and anti-3-NT (NitroTyrosine) allowed identification of numerous proteins subjected to these two post-translational modifications, which are listed here together with information regarding their cross-presence among the predictions.

The influence of abscisic acid on the ethylene biosynthesis pathway in the functioning of the flower abscission zone in Lupinus luteus.

Flower abscission is a highly regulated developmental process activated in response to exogenous (e.g. changing environmental conditions) and endogenous stimuli (e.g. phytohormones). Ethylene (ET) and abscisic acid (ABA) are very effective stimulators of flower abortion in Lupinus luteus, which is a widely cultivated species in Poland, Australia and Mediterranean countries. In this paper, we show that artificial activation of abscission by flower removal caused an accumulation of ABA in the abscission zone (AZ). Moreover, the blocking of that phytohormone's biosynthesis by NDGA (nordihydroguaiaretic acid) decreased the number of abscised flowers. However, the application of NBD - an inhibitor of ET action - reversed the stimulatory effect of ABA on flower abscission, indicating that ABA itself is not sufficient to turn on the organ separation. Our analysis revealed that exogenous ABA significantly accelerated the transcriptional activity of the ET biosynthesis genes ACC synthase (LlACS) and oxidase (LlACO), and moreover, strongly increased the level of 1-aminocyclopropane-1-carboxylic acid (ACC) - ET precursor, which was specifically localized within AZ cells. We cannot exclude the possibility that ABA mediates flower abscission processes by enhancing the ET biosynthesis rate. The findings of our study will contribute to the overall basic knowledge on the phytohormone-regulated generative organs abscission in L. luteus.

Temperature and pyoverdine-mediated iron acquisition control surface motility of Pseudomonas putida.

Pseudomonas putida KT2440 is unable to swarm at its common temperature of growth in the laboratory (30 degrees C) but exhibits surface motility similar to swarming patterns in other Pseudomonas between 18 degrees C and 28 degrees C. These motile cells show differentiation, consisting on elongation and the presence of surface appendages. Analysis of a collection of mutants to define the molecular determinants of this type of surface movement in KT2440 shows that while type IV pili and lipopolysaccharide O-antigen are requisites flagella are not. Although surface motility of flagellar mutants was macroscopically undistinguishable from that of the wild type, microscopy analysis revealed that these mutants move using a distinct mechanism to that of the wild-type strain. Mutants either in the siderophore pyoverdine (ppsD) or in the FpvA siderophore receptor were also unable to spread on surfaces. Motility in the ppsD strain was totally restored with pyoverdine and partially with the wild-type ppsD allele. Phenotype of the fpvA strain was not complemented by this siderophore. We discuss that iron influences surface motility and that it can be an environmental cue for swarming-like movement in P. putida. This study constitutes the first report assigning an important role to pyoverdine iron acquisition in en masse bacterial surface movement.

Biochemical characterization and cellular localization of 11S type storage proteins in olive (Olea europaea L.) seeds.

The composition of seed storage proteins (SSPs) in olive endosperm and cotyledon has been analyzed. Precursor forms of these proteins are made up of individual proteins, which have been purified to homogeneity and further named p1-p5 (20.5, 21.5, 25.5, 27.5, and 30 kDa, respectively). N-terminal sequences of p1 and p2 proteins displayed relevant homology to the basic subunit of the 11S family of plant SSPs (legumins). Two-dimensional polyacrylamide gel electrophoresis experiments allowed us to verify the basic character of p1 and p2 and the acidic character of p3, p4, and p5 proteins. In addition, the putative presence of highly similar isoforms or posttranslational modifications of these polypeptides was detected. As a result, a model describing the putative association of p1-p5 proteins into subunits of alpha(acidic)/beta(basic) type has been proposed. Solubility experiments have shown that the majority of these olive seed proteins from the 11S storage protein family are extracted with aqueous alcohol and only partially with water and diluted saline solutions, therefore suggesting their similarity to prolamines. Moreover, no visible differences were found in either subunit composition or 11S proteins mass among six olive cultivars examined. This result suggests that the synthesis of storage proteins is highly conserved in this plant species. By using a rabbit antiserum raised to p1 protein, the proteins have also been immunolocalized in olive seed tissues, showing that they accumulate in conspicuous protein bodies present in both the endosperm and the cotyledon.