Editorial: Plastid Proteostasis: Relevance of Translation, and Post-Translational Modifications

Due to their bacterial endosymbiotic origin plastids are organelles with both nuclear-encoded and plastid-encoded proteins. Therefore, a highly integrated modulation of gene expression between the nucleus and the plastome is needed in plant cell development. Plastids have retained for the most part a prokaryotic gene expression machinery but, differently from prokaryotes and eukaryotes, they have largely abandoned transcriptional control and switched to predominantly translational control of their gene expression. Some transcriptional regulation is known to occur, but the coordinate expression between the nucleus and the plastome takes place mainly through translational regulation. However, the regulatory mechanisms of plastid gene expression (PGE) are mediated by intricate plastid-nuclear interactions and are still far from being fully understood. Although, for example, translational autoregulation mechanisms in algae have been described for subunits of heteromeric protein complexes and termed control by epistasy of synthesis (CES), only few autoregulatory proteins have been identified in plant plastids. It should be noted of course that PGE in C. reinhardtii is different from that in plants in many aspects. Another example of investigation in this research area is to understand the interactions that occur during RNA binding between nucleus-encoded RNA-binding proteins and the respective RNA sequences, and how this influences the translation initiation process. In addition to this, the plastid retains a whole series of mechanisms for the preservation of its protein balance (proteostasis), including specific proteases, as well as molecular chaperones and enzymes useful in protein folding. After synthesis, plastid proteins must rapidly fold into stable three dimensional structures and often undergo co- and posttranslational modifications to perform their biological mission, avoiding aberrant folding, aggregation and targeting with the help of molecular chaperones and proteases. We believe that this topic is highly interesting for many research areas because the regulation of PGE is not only of wide interest for plant biologists but has also biotechnological implications. Indeed, plastid transformation turns out to be a very promising tool for the production of recombinant proteins in plants, yet some limitations must still be overcome and we believe that this is mainly due to our limited knowledge of the mechanisms in plastids influencing the maintenance of proteostasis

VviAGL11 self-regulates and targets hormone- and secondary metabolism-related genes during seed development

: VviAGL11, the Arabidopsis SEEDSTICK homolog, has been proposed to have a causative role in grapevine stenospermocarpy. An association between a mutation in the coding sequence (CDS) and the seedless phenotype was reported, however, no working mechanisms have been demonstrated yet. We performed a deep investigation of the full VviAGL11 gene sequence in a collection of grapevine varieties belonging to several seedlessness classes that revealed three different promoter-CDS combinations. By investigating the expression of the three VviAGL11 alleles, and by evaluating their ability to activate the promoter region, we observed that VviAGL11 self-activates in a specific promoter-CDS combination manner. Furthermore, by transcriptomic analyses on ovule and developing seeds in seeded and seedless varieties and co-expression approaches, candidate VviAGL11 targets were identified and further validated through luciferase assay and in situ hybridization. We demonstrated that VviAGL11 Wild Type CDS activates Methyl jasmonate esterase and Indole-3-acetate beta-glucosyltransferase, both involved in hormone signaling and Isoflavone reductase, involved in secondary metabolism. The dominant-negative effect of the mutated CDS was also functionally ectopically validated in target induction. VviAGL11 was shown to co-localize with its targets in the outer seed coat integument, supporting its direct involvement in seed development, possibly by orchestrating the crosstalk among MeJA, auxin, and isoflavonoids synthesis. In conclusion, the VviAGL11 expression level depends on the promoter-CDS allelic combination, and this will likely affect its ability to activate important triggers of the seed coat development. The dominant-negative effect of the mutated VviAGL11 CDS on the target genes activation was molecularly validated. A new regulatory mechanism correlating VviAGL11 haplotype assortment and seedlessness class in grapevine is proposed

Comparative 454 pyrosequencing of transcripts from two olive genotypes during fruit development

<p>Abstract</p> <p>Background</p> <p>Despite its primary economic importance, genomic information on olive tree is still lacking. 454 pyrosequencing was used to enrich the very few sequence data currently available for the <it>Olea europaea </it>species and to identify genes involved in expression of fruit quality traits.</p> <p>Results</p> <p>Fruits of <it>Coratina</it>, a widely cultivated variety characterized by a very high phenolic content, and <it>Tendellone</it>, an oleuropein-lacking natural variant, were used as starting material for monitoring the transcriptome. Four different cDNA libraries were sequenced, respectively at the beginning and at the end of drupe development. A total of 261,485 reads were obtained, for an output of about 58 Mb. Raw sequence data were processed using a four step pipeline procedure and data were stored in a relational database with a web interface.</p> <p>Conclusion</p> <p>Massively parallel sequencing of different fruit cDNA collections has provided large scale information about the structure and putative function of gene transcripts accumulated during fruit development. Comparative transcript profiling allowed the identification of differentially expressed genes with potential relevance in regulating the fruit metabolism and phenolic content during ripening.</p

A New Perspective for Vineyard Terroir Identity: Looking for Microbial Indicator Species by Long Read Nanopore Sequencing

Grapevine is one of the most important fruit crops worldwide, being Portugal one of the top wine producers. It is well established that wine sensory characteristics from a particular region are defined by the physiological responses of the grapevine to its environment and thus, the concept of terroir in viticulture was established. Among all the factors that contribute to terroir definition, soil microorganisms play a major role from nutrient recycling to a drastic influence on plant fitness (growth and protection) and of course wine production. Soil microbiome from four different terroirs in Quinta dos Murças vineyard was analysed through long-read Oxford Nanopore sequencing. We have developed an analytical pipeline that allows the identification of function, ecologies, and indicator species based on long read sequencing data. The Douro vineyard was used as a case study, and we were able to establish microbiome signatures of each terroir.info:eu-repo/semantics/publishedVersio

Gene expression signature induced by grape intake in healthy subjects reveals wide-spread beneficial effects on peripheral blood mononuclear cells

Abstract Using a transcriptomic approach, we performed a pilot study in healthy subjects to evaluate the changes in gene expression induced by grape consumption. Blood from twenty subjects was collected at baseline (T0), after 21 days of grape-rich diet (T1) and after one-month washout (T2). Gene expression profiling of peripheral blood mononuclear cells from six subjects identified 930 differentially expressed transcripts. Gene functional analysis revealed changes (at T1 and/or T2) suggestive of antithrombotic and anti-inflammatory effects, confirming and extending previous finding on the same subjects. Moreover, we observed several other favourable changes in the transcription of genes involved in crucial processes such as immune response, DNA and protein repair, autophagy and mitochondrial biogenesis. Finally, we detected significant changes in many long non-coding RNAs genes, whose regulatory functions are being increasingly appreciated. Altogether, our data suggest that a grape diet may exert its beneficial effects by targeting different strategic pathways

High Levels of Variation Within Gene Sequences of Olea europaea L.

Gene sequence variation in cultivated olive (Olea europaea L. subsp. europaea var. europaea), the most important oil tree crop of the Mediterranean basin, has been poorly evaluated up to now. A deep sequence analysis of fragments of four genes, OeACP1, OeACP2, OeLUS and OeSUT1, in 90 cultivars, revealed a wide range of polymorphisms along all recognized allele forms and unexpected allele frequencies and genotype combinations. High linkage values among most polymorphisms were recorded within each gene fragment. The great sequence variability corresponded to a low number of alleles and, surprisingly, to a small fraction of genotype combinations. The distribution, frequency, and combination of the different alleles at each locus is possibly due to natural and human pressures, such as selection, ancestrality, or fitness. Phylogenetic analyses of allele sequences showed distant and complex patterns of relationships among cultivated olives, intermixed with other related forms, highlighting an evolutionary connection between olive cultivars and the O. europaea subspecies cuspidata and cerasiformis. This study demonstrates how a detailed and complete sequence analysis of a few gene portions and a thorough genotyping on a representative set of cultivars can clarify important issues related to sequence polymorphisms, reconstructing the phylogeny of alleles, as well as the genotype combinations. The identification of regions representing blocks of recombination could reveal polymorphisms that represent putatively functional markers. Indeed, specific mutations found on the analyzed OeACP1 and OeACP2 fragments seem to be correlated to the fruit weight

Proteome Regulation during Olea europaea Fruit Development

Widespread in the Mediterranean basin, Olea europaea trees are gaining worldwide popularity for the nutritional and cancer-protective properties of the oil, mechanically extracted from ripe fruits. Fruit development is a physiological process with remarkable impact on the modulation of the biosynthesis of compounds affecting the quality of the drupes as well as the final composition of the olive oil. Proteomics offers the possibility to dig deeper into the major changes during fruit development, including the important phase of ripening, and to classify temporal patterns of protein accumulation occurring during these complex physiological processes.In this work, we started monitoring the proteome variations associated with olive fruit development by using comparative proteomics coupled to mass spectrometry. Proteins extracted from drupes at three different developmental stages were separated on 2-DE and subjected to image analysis. 247 protein spots were revealed as differentially accumulated. Proteins were identified from a total of 121 spots and discussed in relation to olive drupe metabolic changes occurring during fruit development. In order to evaluate if changes observed at the protein level were consistent with changes of mRNAs, proteomic data produced in the present work were compared with transcriptomic data elaborated during previous studies.This study identifies a number of proteins responsible for quality traits of cv. Coratina, with particular regard to proteins associated to the metabolism of fatty acids, phenolic and aroma compounds. Proteins involved in fruit photosynthesis have been also identified and their pivotal contribution in oleogenesis has been discussed. To date, this study represents the first characterization of the olive fruit proteome during development, providing new insights into fruit metabolism and oil accumulation process

Research Topic: Plastid Proteostasis: Relevance of Transcription, Translation, and Post-translational Modifications

and plastid-encoded proteins. Therefore, a highly integrated modulation of gene expression between the nucleus and the plastome is needed in plant cell development. Plastids have retained for the most part a prokaryotic gene expression machinery but, differently from prokaryotes and eukaryotes, they have largely abandoned transcriptional control and switched to predominantly An image of the adaxial side of a sugar beet leaf taken with a conventional fluorescence microscope. The green color is due to autofluorescence of chlorophyll in the chloroplasts. In the box, the optical section of a tobacco protoplast taken with a confocal microscope (at left) and a representation of the chloroplasts structure based on a transmission electron microscopy (TEM) image (at right) is shown. The drawings inside the TEM image represent translation of chloroplast mRNA by 70S ribosomes. Image: Michele Bellucci. Frontiers in Plant Science 3 December 2017 | Plastid Proteostasis translational control of their gene expression. Some transcriptional regulation is known to occur, but the coordinate expression between the nucleus and the plastome takes place mainly through translational regulation. However, the regulatory mechanisms of plastid gene expression (PGE) are mediated by intricate plastid-nuclear interactions and are still far from being fully understood. Although, for example, translational autoregulation mechanisms in algae have been described for subunits of heteromeric protein complexes and termed control by epistasy of synthesis (CES), only few autoregulatory proteins have been identified in plant plastids. It should be noted of course that PGE in C. reinhardtii is different from that in plants in many aspects. Another example of investigation in this research area is to understand the interactions that occur during RNA binding between nucleus-encoded RNA-binding proteins and the respective RNA sequences, and how this influences the translation initiation process. In addition to this, the plastid retains a whole series of mechanisms for the preservation of its protein balance (proteostasis), including specific proteases, as well as molecular chaperones and enzymes useful in protein folding. After synthesis, plastid proteins must rapidly fold into stable three dimensional structures and often undergo co- and posttranslational modifications to perform their biological mission, avoiding aberrant folding, aggregation and targeting with the help of molecular chaperones and proteases. We believe that this topic is highly interesting for many research areas because the regulation of PGE is not only of wide interest for plant biologists but has also biotechnological implications. Indeed, plastid transformation turns out to be a very promising tool for the production of recombinant proteins in plants, yet some limitations must still be overcome and we believe that this is mainly due to our limited knowledge of the mechanisms in plastids influencing the maintenance of proteostasis

Nicotiana attenuata SIPK, WIPK, NPR1, and Fatty Acid-Amino Acid Conjugates Participate in the Induction of Jasmonic Acid Biosynthesis by Affecting Early Enzymatic Steps in the Pathway1[W][OA]

Wounding and herbivore attack elicit the rapid (within minutes) accumulation of jasmonic acid (JA) that results from the activation of previously synthesized biosynthetic enzymes. Recently, several regulatory factors that affect JA production have been identified; however, how these regulators affect JA biosynthesis remains at present unknown. Here we demonstrate that Nicotiana attenuata salicylate-induced protein kinase (SIPK), wound-induced protein kinase (WIPK), nonexpressor of PR-1 (NPR1), and the insect elicitor N-linolenoyl-glucose (18:3-Glu) participate in mechanisms affecting early enzymatic steps of the JA biosynthesis pathway. Plants silenced in the expression of SIPK and NPR1 were affected in the initial accumulation of 13-hydroperoxy-linolenic acid (13-OOH-18:3) after wounding and 18:3-Glu elicitation by mechanisms independent of changes in 13-lipoxygenase activity. Moreover, 18:3-Glu elicited an enhanced and rapid accumulation of 13-OOH-18:3 that depended partially on SIPK and NPR1 but was independent of increased 13-lipoxygenase activity. Together, the results suggested that substrate supply for JA production was altered by 18:3-Glu elicitation and SIPK- and NPR1-mediated mechanisms. Consistent with a regulation at the level of substrate supply, we demonstrated by virus-induced gene silencing that a wound-repressed plastidial glycerolipase (NaGLA1) plays an essential role in the induction of de novo JA biosynthesis. In contrast to SIPK and NPR1, mechanisms mediated by WIPK did not affect the production of 13-OOH-18:3 but were critical to control the conversion of this precursor into 12-oxo-phytodienoic acid. These differences could be partially accounted for by reduced allene oxide synthase activity in WIPK-silenced plants

Identification and gene expression analysis of AUX1 influencing adventitious root induction in olive cuttings (Olea europaea L.)

Olive is one of the most important fruit crops throughout the Mediterranean Basin, mainly propagated by cuttings. The adventitious root development is a key stage in vegetative propagation however the low rooting capacity of some cultivars severely affects the efficiency of olive clonal propagation. Auxin Influx Carrier gene (AUX1), plays a key role in lateral root formation in many plant species promoting the export of IAA from newly developing leaves to lateral root primordia. Putative olive homologues were amplified by using degenerate primers designed on the conserved regions of AUX1 transcripts identified in other plants. Transcript and amino acid sequences in root (OeAUX1R) and base of cutting (OeAUX1B) were different causes of polymorphisms relating to possible distinct roles in these tissues. In order to investigate the gene expression patterns, Real-time PCR was performed on cuttings during the rooting stage collected from genotypes characterized by high and low rooting ability. Moreover, the gene expression was investigated on different olive tissues. Preliminary results showed that the expression of OeAUX1B and OeAUX1R in base of cuttings and roots of the high-rooting genotype were higher which suggests the hypothesis of the involvement of OeAUX1 in olive rooting. Bioinformatics analysis revealed that AUX1 gene had 8 exons in olive and the sequence of this gene in plant was conserved during evolution