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New Phytologist - Lipo‐chitooligosaccharide signalling blocks a rapid pathogen‐induced ROS burst without impeding immunity

Molecular signals released by microbes at the surface of plant roots and leaves largely determine host responses, notably by triggering either immunity or symbiosis. How these signalling pathways cross‐talk upon coincident perception of pathogens and symbionts is poorly described in plants forming symbiosis.

Nitrogen fixing symbiotic Rhizobia spp. and Arbuscular Mycorrhizal fungi produce Lipo‐chitooligosaccharides (LCOs) to initiate host symbiotic programs. In Medicago truncatula roots, the perception of LCOs leads to reduced efflux of Reactive Oxygen Species (ROS). In contrast, pathogen perception generally triggers a strong ROS burst and activates defence gene expression.

Here we show that incubation of M. truncatula seedlings with culture filtrate (CF) of the legume pathogen Aphanomyces euteiches alone or simultaneously with Sinorhizobium meliloti LCOs, resulted in a strong ROS release. However, this response was completely inhibited if CF was added after pre‐incubation of seedlings with LCOs. In contrast, expression of immunity‐associated genes in response to CF and disease resistance to A. euteiches remained unaffected by LCO treatment of M. truncatula roots.

Our findings suggest that symbiotic plants evolved ROS inhibition response to LCOs to facilitate early steps of symbiosis whilst maintaining parallel defence mechanisms toward pathogens.

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PLoS Biology - Nitrogen fixation in a landrace of maize is supported by a mucilage-associated diazotrophic microbiota

PLoS Biology - Nitrogen fixation in a landrace of maize is supported by a mucilage-associated diazotrophic microbiota | LRSV Publications | Scoop.it

Plants are associated with a complex microbiota that contributes to nutrient acquisition, plant growth, and plant defense. Nitrogen-fixing microbial associations are efficient and well characterized in legumes but are limited in cereals, including maize. We studied an indigenous landrace of maize grown in nitrogen-depleted soils in the Sierra Mixe region of Oaxaca, Mexico. This landrace is characterized by the extensive development of aerial roots that secrete a carbohydrate-rich mucilage. Analysis of the mucilage microbiota indicated that it was enriched in taxa for which many known species are diazotrophic, was enriched for homologs of genes encoding nitrogenase subunits, and harbored active nitrogenase activity as assessed by acetylene reduction and 15N2 incorporation assays. Field experiments in Sierra Mixe using 15N natural abundance or 15N-enrichment assessments over 5 years indicated that atmospheric nitrogen fixation contributed 29%–82% of the nitrogen nutrition of Sierra Mixe maize.

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Nature Plants - Fern genomes elucidate land plant evolution and cyanobacterial symbioses

Nature Plants - Fern genomes elucidate land plant evolution and cyanobacterial symbioses | LRSV Publications | Scoop.it

Ferns are the closest sister group to all seed plants, yet little is known about their genomes other than that they are generally colossal. Here, we report on the genomes of Azolla filiculoides and Salvinia cucullata(Salviniales) and present evidence for episodic whole-genome duplication in ferns—one at the base of ‘core leptosporangiates’ and one specific to Azolla. One fern-specific gene that we identified, recently shown to confer high insect resistance, seems to have been derived from bacteria through horizontal gene transfer. Azolla coexists in a unique symbiosis with N2-fixing cyanobacteria, and we demonstrate a clear pattern of cospeciation between the two partners. Furthermore, the Azolla genome lacks genes that are common to arbuscular mycorrhizal and root nodule symbioses, and we identify several putative transporter genes specific to Azolla–cyanobacterial symbiosis. These genomic resources will help in exploring the biotechnological potential of Azolla and address fundamental questions in the evolution of plant life.

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Nature Plants - Oak genome reveals facets of long lifespan

Nature Plants - Oak genome reveals facets of long lifespan | LRSV Publications | Scoop.it

Oaks are an important part of our natural and cultural heritage. Not only are they ubiquitous in our most common landscapes1 but they have also supplied human societies with invaluable services, including food and shelter, since prehistoric times2. With 450 species spread throughout Asia, Europe and America3, oaks constitute a critical global renewable resource. The longevity of oaks (several hundred years) probably underlies their emblematic cultural and historical importance. Such long-lived sessile organisms must persist in the face of a wide range of abiotic and biotic threats over their lifespans. We investigated the genomic features associated with such a long lifespan by sequencing, assembling and annotating the oak genome. We then used the growing number of whole-genome sequences for plants (including tree and herbaceous species) to investigate the parallel evolution of genomic characteristics potentially underpinning tree longevity. A further consequence of the long lifespan of trees is their accumulation of somatic mutations during mitotic divisions of stem cells present in the shoot apical meristems. Empirical4 and modelling5 approaches have shown that intra-organismal genetic heterogeneity can be selected for6and provides direct fitness benefits in the arms race with short-lived pests and pathogens through a patchwork of intra-organismal phenotypes7. However, there is no clear proof that large-statured trees consist of a genetic mosaic of clonally distinct cell lineages within and between branches. Through this case study of oak, we demonstrate the accumulation and transmission of somatic mutations and the expansion of disease-resistance gene families in trees.

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BMC Biology - Genomics analysis of Aphanomyces spp. identifies a new class of oomycete effector associated with host adaptation 

Oomycetes are a group of filamentous eukaryotic microorganisms that have colonized all terrestrial and oceanic ecosystems, and they include prominent plant pathogens. The Aphanomyces genus is unique in its ability to infect both plant and animal species, and as such exemplifies oomycete versatility in adapting to different hosts and environments. Dissecting the underpinnings of oomycete diversity provides insights into their specificity and pathogenic mechanisms. By carrying out genomic analyses of the plant pathogen A. euteiches and the crustacean pathogen A. astaci, we show that host specialization is correlated with specialized secretomes that are adapted to the deconstruction of the plant cell wall in A. euteiches and protein degradation in A. astaci. The A. euteiches genome is characterized by a large repertoire of small secreted protein (SSP)-encoding genes that are highly induced during plant infection, and are not detected in other oomycetes. Functional analysis revealed an SSP from A. euteiches containing a predicted nuclear-localization signal which shuttles to the plant nucleus and increases plant susceptibility to infection. Collectively, our results show that Aphanomyces host adaptation is associated with evolution of specialized secretomes and identify SSPs as a new class of putative oomycete effectors.
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Chemosphere - Influence of arbuscular mycorrhizal fungi on antimony phyto-uptake and compartmentation in vegetables cultivated in urban gardens

Chemosphere - Influence of arbuscular mycorrhizal fungi on antimony phyto-uptake and compartmentation in vegetables cultivated in urban gardens | LRSV Publications | Scoop.it
1. Context Urban areas are often contaminated with various forms of persistent metal (loid) and emerging contaminants such as antimony (Sb). Thus, in the context of urban agriculture where sustainable practices such as biofertilizers application (arbuscular mycorrhizal fungi, AMF) could improve nutrient transfer from the soil to the vegetables, the effect of AMF on metal (loid) mobility and human bioaccessibility is still poorly known. 

 2. Methods The role of AMF in Sb uptake by lettuce and carrot grown in artificial substrate spiked with different Sb chemical species was investigated. Plants were grown under hydroponic conditions and half of the treatments received a concentrated spore solution to obtain mycorrhized and non-mycorrhized plants. Three weeks before harvest, plants were exposed to 10 mg.L−1 of either Sb2O3 or KSbO-tartrate (KSb). 

 3. Results The presence of AMF significantly increased its accumulation in carrots (all organs) with higher accumulation in roots. In lettuce, accumulation appeared to be dependent on the Sb chemical species. Moreover, it was observed for the first time that AMF changed the human bioaccessible fraction of Sb in edible organs. 

 4. Implications The present results highlight a possible risk of Sb transfer from soil to edible plants cultivated in soil naturally containing AMF propagules, or when AMF are added as biofertilizers. After validating the influence of soil environment and AMF on Sb behavior in the field, these results should be considered in health risk assessments.
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GigaScience - 10KP: A Phylodiverse Genome Sequencing Plan 

GigaScience - 10KP: A Phylodiverse Genome Sequencing Plan  | LRSV Publications | Scoop.it

Understanding plant evolution and diversity in a phylogenomic context is an enormous challenge, due in part to limited availability of genome-scale data across phylodiverse species. The 10KP (10,000 Plants) Genome Sequencing Project will sequence and characterize representative genomes from every major clade of embryophytes, green algae, and protists (excluding fungi) within the next five years. By implementing (and continuously improving) leading-edge sequencing technologies and bioinformatics tools, 10KP will catalogue the genome content of plant and protist diversity, and make these data freely available as an enduring foundation for future scientific discovery and application. 10KP is structured as an international consortium, open to the global community, including botanical gardens, plant research institutes, universities, and private industry. Our immediate goal is to establish a policy framework for this endeavour, the principles of which are outlined here.

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New Phytol - High intraspecific genome diversity in the model arbuscular mycorrhizal symbiont Rhizophagus irregularis

New Phytol - High intraspecific genome diversity in the model arbuscular mycorrhizal symbiont Rhizophagus irregularis | LRSV Publications | Scoop.it

Arbuscular mycorrhizal fungi (AMF) are known to improve plant fitness through the establishment of mycorrhizal symbioses. Genetic and phenotypic variations among closely related AMF isolates can significantly affect plant growth, but the genomic changes underlying this variability are unclear. To address this issue, we improved the genome assembly and gene annotation of the model strain Rhizophagus irregularis DAOM197198, and compared its gene content with five isolates of R. irregularis sampled in the same field. All isolates harbor striking genome variations, with large numbers of isolate-specific genes, gene family expansions, and evidence of interisolate genetic exchange. The observed variability affects all gene ontology terms and PFAM protein domains, as well as putative mycorrhiza-induced small secreted effector-like proteins and other symbiosis differentially expressed genes. High variability is also found in active transposable elements. Overall, these findings indicate a substantial divergence in the functioning capacity of isolates harvested from the same field, and thus their genetic potential for adaptation to biotic and abiotic changes. Our data also provide a first glimpse into the genome diversity that resides within natural populations of these symbionts, and open avenues for future analyses of plant–AMF interactions that link AMF genome variation with plant phenotype and fitness.

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Current Protein & Peptide Science: Plant Cell Wall Proteomics as a Strategy to Reveal Candidate Proteins Involved in Extracellular Lipid Metabolism

Current Protein & Peptide Science: Plant Cell Wall Proteomics as a Strategy to Reveal Candidate Proteins Involved in Extracellular Lipid Metabolism | LRSV Publications | Scoop.it

Plant cell walls are composite structures surrounding cells and involved in both mechanical support and perception of their environment. They are mainly composed of polysaccharides (90-95% of their mass) and proteins (5-10%). The cell wall proteins (CWPs) contribute to the arrangements and modifications of polymer networks and to cell-to-cell communication. The structure and composition of cell walls are not uniform in the whole plants, but rather specialized in different cell types to fulfil different functions. As examples, two kinds of cells are covered with extracellular structures composed of lipids: epidermal cells of aerial organs synthesize a cuticle on their outside surface whereas endodermal root cells form a suberin surrounding strip. In both cases, these particular hydrophobic layers contribute to the protection of the cells against attacks by pathogens or abiotic stresses and regulate physiological processes. If the intracellular biosynthesis of the molecules forming these layers starts to be welldescribed, the mechanisms of their assembly beyond the plasma membrane remain largely unknown. In this review, this issue is addressed on the basis on cell wall proteomics data which has allowed the identification of many CWPs possibly related to lipid metabolism during the last years, in particular in Arabidopsis thaliana and tomato. These data are combined with transcriptomics and genetics studies. The main known roles of extracellular proteins related to lipid metabolism are discussed.

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J Exp Bot - Medicago truncatula GRAS protein RAD1 supports arbuscular mycorrhiza symbiosis and Phytophthora palmivora susceptibility 

J Exp Bot - Medicago truncatula  GRAS protein RAD1 supports arbuscular mycorrhiza symbiosis and  Phytophthora palmivora  susceptibility  | LRSV Publications | Scoop.it

The roots of most land plants are colonized by symbiotic arbuscular mycorrhiza (AM) fungi. To facilitate this symbiosis, plant genomes encode a set of genes required for microbial perception and accommodation. However, the extent to which infection by filamentous root pathogens also relies on some of these genes remains an open question. Here, we used genome-wide association mapping to identify genes contributing to colonization of Medicago truncatula roots by the pathogenic oomycete Phytophthora palmivora. Single-nucleotide polymorphism (SNP) markers most significantly associated with plant colonization response were identified upstream of RAD1, which encodes a GRAS transcription regulator first negatively implicated in root nodule symbiosis and recently identified as a positive regulator of AM symbiosis. RAD1 transcript levels are up-regulated both in response to AM fungus and, to a lower extent, in infected tissues by P. palmivora where its expression is restricted to root cortex cells proximal to pathogen hyphae. Reverse genetics showed that reduction of RAD1 transcript levels as well as a rad1 mutant are impaired in their full colonization by AM fungi as well as by P. palmivora. Thus, the importance of RAD1 extends beyond symbiotic interactions, suggesting a general involvement in M. truncatula microbe-induced root development and interactions with unrelated beneficial and detrimental filamentous microbes.

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Bioinform Proteomics  - Plant cell wall proteomics : An assessment twenty years after launching

Bioinform Proteomics  - Plant cell wall proteomics : An assessment twenty years after launching | LRSV Publications | Scoop.it
Cell walls are complex structures surrounding plant cells. They provide not only mechanical support and protection against environmental changes, but also a mean for cell-to-cell communication. They are mainly constituted of polysaccharides (about 90% of their mass) and proteins. Cell wall proteins (CWPs) play critical roles because they contribute to the plasticity of the cell wall architecture during development and in response to biotic and abiotic environmental changes. Their systematic identification has started in the 2000’s with the sequencing of the genome of the Arabidopsis thaliana model plant and the development of adapted mass spectrometry (MS) technologies. Since then, many other plants have been studied among which plants of agronomical interest. The description of cell wall proteomes has fully benefited not only from the improvement of MS technologies, but also from better sample preparation and peptide separation prior to MS analysis. Bioinformatics has also played critical roles by designing software allowing protein identification, annotation and quantification, as well creating MS data repositories.
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PNAS - Molecular link between auxin and ROS-mediated polar growth

Root hair polar growth is endogenously controlled by auxin and sustained by oscillating levels of reactive oxygen species (ROS). These cells extend several hundred-fold their original size toward signals important for plant survival. Although their final cell size is of fundamental importance, the molecular mechanisms that control it remain largely unknown. Here we show that ROS production is controlled by the transcription factor RSL4, which in turn is transcriptionally regulated by auxin through several auxin response factors (ARFs). In this manner, auxin controls ROS-mediated polar growth by activating RSL4, which then up-regulates the expression of genes encoding NADPH oxidases (also known as RESPIRATORY BURST OXIDASE HOMOLOG proteins) and class III peroxidases, which catalyze ROS production. Chemical or genetic interference with ROS balance or peroxidase activity affects root hair final cell size. Overall, our findings establish a molecular link between auxin and ROS-mediated polar root hair growth.

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Nature Protocol - Detection of nucleic acid–protein interactions in plant leaves using fluorescence lifetime imaging microscopy

DNA-binding proteins (DNA-BPs) and RNA-binding proteins (RNA-BPs) have critical roles in living cells in all kingdoms of life. Various experimental approaches exist for the study of nucleic acid–protein interactions in vitro and in vivo, but the detection of such interactions at the subcellular level remains challenging. Here we describe how to detect nucleic acid–protein interactions in plant leaves by using a fluorescence resonance energy transfer (FRET) approach coupled to fluorescence lifetime imaging microscopy (FLIM). Proteins of interest (POI) are tagged with a GFP and transiently expressed in plant cells to serve as donor fluorophore. After sample fixation and cell wall permeabilization, leaves are treated with Sytox Orange, a nucleic acid dye that can function as a FRET acceptor. Upon close association of the GFP-tagged POI with Sytox-Orange-stained nucleic acids, a substantial decrease of the GFP lifetime due to FRET between the donor and the acceptor can be monitored. Treatment with RNase before FRET–FLIM measurements allows determination of whether the POI associates with DNA and/or RNA. A step-by-step protocol is provided for sample preparation, data acquisition and analysis. We describe how to calibrate the equipment and include a tutorial explaining the use of the FLIM software. To illustrate our approach, we provide experimental procedures to detect the interaction between plant DNA and two proteins (the AeCRN13 effector from the oomycete Aphanomyces euteiches and the AtWRKY22 defensive transcription factor from Arabidopsis). This protocol allows the detection of protein–nucleic acid interactions in plant cells and can be completed in <2 d.
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ESPR - Vermicompost addition influences symbiotic fungi communities associated with leek cultivated in metal-rich soils

ESPR - Vermicompost addition influences symbiotic fungi communities associated with leek cultivated in metal-rich soils | LRSV Publications | Scoop.it

In the context of urban agriculture, where soils are frequently contaminated with metal(loid)s (TM), we studied the influence of vermicompost amendments on symbiotic fungal communities associated with plants grown in two metal-rich soils. Leek (Allium porrum L.) plants were grown with or without vermicompost in two metal-rich soils characterized by either geogenic or anthropogenic TM sources, to assess the influence of pollutant origin on soil-plant transfer. Fungal communities associated with the leek roots were identified by high throughput Illumina MiSeq and TM contents were measured using mass spectrometry. Vermicompost addition led to a dramatic change in the fungal community with a loss of diversity in the two tested soils. This effect could partially explain the changes in metal transfer at the soil-AMF-plant interface. Our results suggest being careful while using composts when growing edibles in contaminated soils. More generally, this study highlights the need for further research in the field of fungal communities to refine practical recommendations to gardeners.

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Cell - The Chara Genome: Secondary Complexity and Implications for Plant Terrestrialization

Cell - The Chara Genome: Secondary Complexity and Implications for Plant Terrestrialization | LRSV Publications | Scoop.it

 

Land plants evolved from charophytic algae, among which Charophyceae possess the most complex body plans. We present the genome of Chara braunii; comparison of the genome to those of land plants identified evolutionary novelties for plant terrestrialization and land plant heritage genes. C. braunii employs unique xylan synthases for cell wall biosynthesis, a phragmoplast (cell separation) mechanism similar to that of land plants, and many phytohormones. C. braunii plastids are controlled via land-plant-like retrograde signaling, and transcriptional regulation is more elaborate than in other algae. The morphological complexity of this organism may result from expanded gene families, with three cases of particular note: genes effecting tolerance to reactive oxygen species (ROS), LysM receptor-like kinases, and transcription factors (TFs). Transcriptomic analysis of sexual reproductive structures reveals intricate control by TFs, activity of the ROS gene network, and the ancestral use of plant-like storage and stress protection proteins in the zygote.

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New Phytol - Arbuscular mycorrhizal fungi: intraspecific diversity and pangenomes

New Phytol - Arbuscular mycorrhizal fungi: intraspecific diversity and pangenomes | LRSV Publications | Scoop.it

Arbuscular mycorrhizal fungi (AMF) are ubiquitous plant symbionts with an intriguing population biology. Conspecific AMF strains can vary substantially at the genetic and phenotypic levels, leading to direct and quantifiable variation in plant growth. Recent studies have shown that high intraspecific diversity is very common in AMF, and not only found in model species. Studies have also revealed how the phenotype of conspecific isolates varies depending on the plant host, highlighting the functional relevance of intraspecific phenotypic plasticity for the AMF ecology and mycorrhizal symbiosis. Recent work has also demonstrated that conspecific isolates of the model AMF Rhizophagus irregularis harbor large and highly variable pangenomes, highlighting the potential role of intraspecific genome diversity for the ecological adaptation of these symbionts.

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Science - Phylogenomics reveals multiple losses of nitrogen-fixing root nodule symbiosis

Science - Phylogenomics reveals multiple losses of nitrogen-fixing root nodule symbiosis | LRSV Publications | Scoop.it
The root nodule symbiosis of plants with nitrogen-fixing bacteria impacts global nitrogen cycles and food production but is restricted to a subset of genera within a single clade of flowering plants. To explore the genetic basis for this scattered occurrence, we sequenced the genomes of ten plant species covering the diversity of nodule morphotypes, bacterial symbionts and infection strategies. In a genome-wide comparative analysis of a total of 37 plant species, we discovered signatures of multiple independent loss-of-function events in the indispensable symbiotic regulator NODULE INCEPTION ( NIN ) in ten out of 13 genomes of non-nodulating species within this clade. The discovery that multiple independent losses shaped the present day distribution of nitrogen-fixing root nodule symbiosis in plants reveals a phylogenetically wider distribution in evolutionary history and a so far underestimated selection pressure against this symbiosis.
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Current Opinion in Plant Biology - Symbiosis genes for immunity and vice versa

Current Opinion in Plant Biology - Symbiosis genes for immunity and vice versa | LRSV Publications | Scoop.it
Highlights 
• Several LysM receptor proteins have a dual function in symbiosis and immunity. 
• Symbiotic transcription factors can also control pathogen infection. • Genes of hormonal pathways regulate both pathogen and symbiont colonisation. 
• Crosstalk between symbiosis and immunity also occurs within the flavonoid pathway. 

Basic molecular knowledge on plant–pathogen interactions has largely been gained from reverse and forward genetics in Arabidopsis thaliana. However, as this model plant is unable to establish endosymbiosis with mycorrhizal fungi or rhizobia, plant responses to mutualistic symbionts have been studied in parallel in other plant species, mainly legumes. The resulting analyses led to the identification of gene networks involved in various functions, from microbe recognition to signalling and plant responses, thereafter assigned to either mutualistic symbiosis or immunity, according to the nature of the initially inoculated microbe. The increasing development of new pathosystems and genetic resources in model legumes and the implementation of reverse genetics in plants such as rice and tomato that interact with both mycorrhizal fungi and pathogens, have highlighted the dual role of plant genes previously thought to be specific to mutualistic or pathogenic interactions. The next challenges will be to determine whether such genes have similar functions in both types of interaction and if not, whether the perception of microbial compounds or the involvement of specific plant signalling components is responsible for the appropriate plant responses to the encountered microorganisms.
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Current Opinion in Plant Biology - What have we learnt from studying the evolution of the arbuscular mycorrhizal symbiosis?

Current Opinion in Plant Biology - What have we learnt from studying the evolution of the arbuscular mycorrhizal symbiosis? | LRSV Publications | Scoop.it
Highlights 
• The molecular mechanisms governing the ability of plants to engage in the AM symbiosis originated in the first land plants. 
• A core set of symbiotic genes has been conserved in all studied AM host plants. 
• Loss of a core set of symbiotic genes correlates with the irreversible loss of the AM symbiosis. 
• AM loss could be linked to the evolution of novel nutrient acquisition strategies. 

The arbuscular mycorrhizal (AM) symbiosis is a nearly ubiquitous association formed by most land plants. Numerous insights into the molecular mechanisms governing this symbiosis have been obtained in recent years leading to the identification of a core set of plant genes essential for successful formation of the AM symbiosis by angiosperm hosts. Recent phylogenetic analyses indicate that while the origin of some of these symbiotic genes predated the first land plants, the rest appeared through processes including de novo evolution and gene duplication that occurred specifically in the land plants. Purifying selection on this core gene set has been maintained over millions of years of plant evolution to conserve the AM symbiosis. However, several independent losses of this association have been recorded in numerous embryophyte lineages. In these lineages, potential compensatory mechanisms have been identified that could have helped these plants overcome the adversities imposed by the loss of the AM symbiosis. This review will focus on the processes governing the conservation of the AM symbiosis in the land plant lineage.
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Proteomics - Cell Wall Proteome of Sugarcane Young and Mature Leaves and Stems

Proteomics - Cell Wall Proteome of Sugarcane Young and Mature Leaves and Stems | LRSV Publications | Scoop.it
By characterizing the cell wall proteomes of different sugarcane organs (leaves and stems) at two developmental stages (young vs mature/apical vs basal), it is possible to address unique characteristics in each of them. Four-month-old leaves show a higher proportion of oxido-reductases and proteins related to lipid metabolism (LM), besides a lower proportion of proteins acting on polysaccharides, in comparison to 4-month-old internodes. It is possible to note that sugarcane leaves and young stems have the highest LM rate than all species, which is assumed to be linked to cuticle formation. The data generated enrich the number of cell wall proteins (CWPs) identified in sugarcane, reaching 277. To our knowledge, sugarcane has now the second higher coverage of monocot CWP in plants.
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Biofouling - Metabolome and proteome changes between biofilm and planktonic phenotypes of the marine bacterium Pseudoalteromonas lipolytica TC8

A number of bacteria adopt various lifestyles such as planktonic free-living or sessile biofilm stages. This enables their survival and development in a wide range of contrasting environments. With the aim of highlighting specific metabolic shifts between these phenotypes and to improve the overall understanding of marine bacterial adhesion, a dual metabolomics/proteomics approach was applied to planktonic and biofilm cultures of the marine bacterium Pseudoalteromonas lipolytica TC8. The liquid chromatography mass spectrometry (LC-MS) based metabolomics study indicated that membrane lipid composition was highly affected by the culture mode: phosphatidylethanolamine (PEs) derivatives were over-produced in sessile cultures while ornithine lipids (OLs) were more specifically synthesized in planktonic samples. In parallel, differences between proteomes revealed that peptidases, oxidases, transcription factors, membrane proteins and the enzymes involved in histidine biosynthesis were over-expressed in biofilms while proteins involved in heme production, nutrient assimilation, cell division and arginine/ornithine biosynthesis were specifically up-regulated in free-living cells.
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J Exp Bot - Plant extracellular vesicles are incorporated by a fungal pathogen and inhibit its growth

J Exp Bot - Plant extracellular vesicles are incorporated by a fungal pathogen and inhibit its growth | LRSV Publications | Scoop.it

Extracellular vesicles (EV) are membrane particles released by cells into their environment and are considered to be key players in intercellular communication. EV are produced by all domains of life but limited knowledge about EV in plants is available, although their implication in plant defense has been suggested. We have characterized sunflower EV and tested whether they could interact with fungal cells. EV were isolated from extracellular fluids of seedlings and characterized by transmission electron microscopy and proteomic analysis. These nanovesicles appeared to be enriched in cell wall remodeling enzymes and defense proteins. Membrane-labeled EV were prepared and their uptake by the phytopathogenic fungus Sclerotinia sclerotiorum was verified. Functional tests further evaluated the ability of EV to affect fungal growth. Spores treated with plant EV showed growth inhibition, morphological changes, and cell death. Conclusive evidence on the existence of plant EV is presented and we demonstrate their ability to interact with and kill fungal cells. Our results introduce the concept of cell-to-cell communication through EV in plants.

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Proteomics - Cell wall proteome analysis of Arabidopsis thaliana mature stems

Proteomics - Cell wall proteome analysis of Arabidopsis thaliana mature stems | LRSV Publications | Scoop.it
Plant stems carry flowers necessary for species propagation and need to be adapted to mechanical disturbance and environmental factors. The stem cell walls are different from other organs and can modify their rigidity or viscoelastic properties for the integrity and the robustness required to withstand mechanical impacts and environmental stresses. Plant cell wall is composed of complex polysaccharide networks also containing cell wall proteins (CWPs) crucial to perceive and limit the environmental effects. The CWPs are fundamental players in cell wall remodeling processes, and today, only 86 have been identified from the mature stems of the model plant Arabidopsis thaliana. With a destructive method, this study has enlarged its coverage to 302 CWPs. This new proteome is mainly composed of 27.5% proteins acting on polysaccharides, 16% proteases, 11.6% oxido-reductases, 11% possibly related to lipid metabolism and 11% of proteins with interacting domains with proteins or polysaccharides. Compared to stem cell wall proteomes already available (Brachypodium distachyon, Sacharum officinarum, Linum usitatissimum, Medicago sativa), that of A. thaliana stems has a higher proportion of proteins acting on polysaccharides and of proteases, but a lower proportion of oxido-reductases.
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Frontiers in Plant Science - Proline Hydroxylation in Cell Wall Proteins: Is It Yet Possible to Define Rules?

Frontiers in Plant Science - Proline Hydroxylation in Cell Wall Proteins: Is It Yet Possible to Define Rules? | LRSV Publications | Scoop.it

Cell wall proteins (CWPs) play critical and dynamic roles in plant cell walls by contributing to developmental processes and response to environmental cues. Since the CWPs go through the secretion pathway, most of them undergo post-translational modifications (PTMs) which can modify their biological activity. Glycosylation is one of the major PTMs of CWPs and refers to N-glycosylation, O-glycosylation and glypiation. Each of these PTMs occurs in different amino acid contexts which are not all well defined. This article deals with the hydroxylation of Pro residues which is a prerequisite for O-glycosylation of CWPs on hydroxyproline (Hyp) residues. The location of Hyp residues is well described in several structural CWPs, but yet rarely described in other CWPs. In this article, it is studied in detail in five Arabidopsis thaliana proteins using mass spectrometry data: one of them (At4g38770, AtPRP4) is a structural CWP containing 32.5% of Pro residues arranged in typical motifs, the others are either rich (27–28%, At1g31580 and At2g10940) or poor (6–8%, At1g09750 and At3g08030) in Pro residues. The known rules of Pro hydroxylation allowed a good prediction of Hyp location in AtPRP4. However, they could not be applied to the other proteins whatever their Pro content. In addition, variability of the Pro hydroxylation patterns was observed within some amino acid motifs in all the proteins and new patterns of Pro hydroxylation are described. Altogether, this work shows that Hyp residues are present in more protein families than initially described, and that Pro hydroxylation patterns could be different in each of them. This work paves the way for completing the existing Pro hydroxylation code.

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Scientific Reports - PRR2, a pseudo-response regulator, promotes salicylic acid and camalexin accumulation during plant immunity

Scientific Reports - PRR2, a pseudo-response regulator, promotes salicylic acid and camalexin accumulation during plant immunity | LRSV Publications | Scoop.it

Calcium signalling mediated by Calmodulin (CaM) and calmodulin-like (CML) proteins is critical to plant immunity. CaM and CML regulate a wide range of target proteins and cellular responses. While many CaM-binding proteins have been identified, few have been characterized for their specific role in plant immunity. Here, we report new data on the biological function of a CML-interacting partner, PRR2 (PSEUDO-RESPONSE REGULATOR 2), a plant specific transcription factor. Until now, the physiological relevance of PRR2 remained largely unknown. Using a reverse genetic strategy in A. thaliana, we identified PRR2 as a positive regulator of plant immunity. We propose that PRR2 contributes to salicylic acid (SA)-dependent responses when challenged with the phytopathogenic bacterium Pseudomonas syringae. PRR2 is transcriptionally upregulated by SA and P. syringae, enhances SA biosynthesis and SA signalling responses; e.g. in response to P. syringae, PRR2 induces the production of SA and the accumulation of the defence-related protein PR1. Moreover, PRR2 overexpressing lines exhibit an enhanced production of camalexin, a phytoalexin that confers enhanced resistance against pathogens. Together, these data reveal the importance of PRR2 in plant immune responses against P. syringae and suggest a novel function for this particular plant specific transcription factor in plant physiology.

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