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In response to an open problem raised by S. Rabinowitz, we prove that $$ \begin{align*} \begin{array} [c]{l} \left( \left( x^{2}+y^{2}\right) {}^{2}+8y\left( y^{2}-3x^{2}\right) \right) {}^{2}+432y\left( y^{2}-3x^{2}\right) \left( 351-10\left( x^{2}+y^{2}\right) \right) \\ =567^{3}+28\left( x^{2}+y^{2}\right) {}^{3}+486\left( x^{2}+y^{2}\right) \left( 67\left( x^{2}+y^{2}\right) -567\times18\right) \end{array} \end{align*} $$ is the equation of a plane convex curve of constant width.

To determine whether a beat-by-beat cardiovascular index (CARDEAN: cardiovascular depth of analgesia, Alpha-2 Ltd, Lyon, France) reduces the incidence of tachycardia in ASA I–III patients undergoing orthopaedic surgery. A total of 76 patients were prospectively randomized into (1) a control group or (2) the CARDEAN group, in which the nurse anaesthetist was blinded to CARDEAN application. In addition to conventional signs, an external observer instructed the nurse anaesthetist to administer sufentanil 0.1 µg kg−1 when the CARDEAN crossed a threshold (≥ 60). The primary outcome was the incidence of tachycardia (> 120% of reference heart rate, HR). Non-invasive blood pressure (BP), electrocardiogram (ECG), O2 saturation-photoplethysmography and the bispectral index (40

The non-animal peroxidases belong to a superfamily of oxidoreductases that reduce hydrogen peroxide and oxidize numerous substrates. Since their initial characterization in 1992, a number of studies have provided an understanding of the origin and evolution of this protein family. Here, we report a comprehensive evolutionary analysis of non-animal peroxidases using integrated in silico and biochemical approaches. Thanks to the availability of numerous genomic sequences from more than 2500 species belonging to 14 kingdoms together with expert and comprehensive annotation of peroxidase sequences that have been centralized in a dedicated database, we have been able to use phylogenetic reconstructions to increase our understanding of the evolutionary processes underlying the diversification of non-animal peroxidases. We analysed the distribution of all non-animal peroxidases in more than 200 eukaryotic organisms in silico. First, we show that the presence or absence of non-animal peroxidases correlates with the presence or absence of certain organelles or with specific biological processes. Examination of almost 2000 organisms determined that ascorbate peroxidases (APxs) and cytochrome c peroxidases (CcPs) are present in those containing chloroplasts and mitochondria, respectively. Plants, which contain both organelles, are an exception and contain only APxs without CcP. Class II peroxidases (CII Prxs) are only found in fungi with wood-decay and plant-degradation abilities. Class III peroxidases (CIII Prxs) are only found in streptophyte algae and land plants, and have been subjected to large family expansion. Biochemical activities of APx, CcP, and CIII Prx assessed using protein extracts from 30 different eukaryotic organisms support the distribution of the sequences resulting from our in silico analysis. The biochemical results confirmed both the presence and classification of the non-animal peroxidase encoding sequences.

Streptomycetes are Gram-positive actinobacteria largely represented in the plant root microbiota. The genetic determinants involved in the presence of Streptomyces in the rhizosphere are mostly unknown but can rely on the ability to release phytohormones, degrade plant cell-wall polysaccharides and produce specialised metabolites. Here we sequenced the genome of the rhizospheric and plant defence-stimulating strain Streptomyces sp. AgN23. We found out that it belongs to the soil and plant root dwelling S. violaceusniger clade. The genome annotation of AgN23 revealed the ability of the bacterium to synthesise auxin, a major regulator of plant development, to degrade plant cell wall with a large repertoire of carbohydrate degrading enzymes and to produce antimicrobials (rustmicin, mediomycin, niphimycin, nigericin) and plant bioactive compounds (nigericin, echosides, elaiophylin) through a set of biosynthetic gene clusters. We also found that these genomic features are well-conserved among members of the S. violaceusniger clade. In addition, AgN23 display original events of biosynthetic gene clusters acquisitions and losses which may account for its beneficial effect on plants. Taken together, our work supports the hypothesis that hydrolytic enzymes and specialised metabolites repertoires underpin the interaction of bacteria belonging to the S. violaceusniger clade with plant roots within the rhizosphere.Impact statementStreptomycetes are filamentous Gram-positive bacteria universally found around and within host plant tissues. These actinobacteria have been extensively investigated for their tremendous ability to produce diverse specialised metabolites (e.g., antibiotics). By contrast their impact on host plant physiology is widely neglected. Whether specific lineage of Streptomyces colonise host plant and what are the underlying molecular mechanisms is poorly documented. Here we report a chromosome-scale assembly of AgN23 genome, a Streptomyces sp. strain previously characterised for its ability to activate the plant immune system. This reference sequence enabled us to position AgN23 in the S. violaceusniger clade from which several representatives have been isolated worldwide from the rhizosphere of unrelated plants. Comparative genomic studies suggest that S. violaceusniger spp. produce a prominent CAZyome with expansion of plant cell wall degrading enzymes families and a conserved specialised metabolism acting on host plant physiology and its rhizospheric microbiota. These genomic features may underly S. violaceusniger spp. adaptation to the rhizopsheric niche.Data summaryThe raw reads sequences of AgN23 genome are available at NCBI on the Sequence Read Archive portal for PacBio and MiSeq data (SRR13990229 and SRR14028548 respectively). The Genome assembly is available on the NCBI nucleotide portal under the accession NZ_CP007153.1. This genome sequence was uploaded on the MicroScope platform for genome annotation and analysis (https://mage.genoscope.cns.fr/microscope/home/index.php) [1]. The RNA-seq raw reads are archived in the NCBI Bioproject PRJNA745930. The following eight supplementary tables are included in the online version of this article.Supplementary Information 1: Genomes used in this study. The accession number used from the NCBI portal, name, size, number of contigs as well as the level of completeness of the assembly are indicated.Supplementary Information 2: List of the single copy core genes used by autoMLST to build the phylogenetic tree in Figure 1.Supplementary information 3: Annotation of AgN23 full chromosome. For each gene the frame of translation, sequence length and position on the chromosome are indicated. All genes were annotated according to the Microscope platform, see materials and methods. In addition, the expression for each gene is reported in transcripts per million (TPM) based on the the RNA-seq data from three biological replicates.Supplementary Information 4: Genomes having a Mash-based estimated ANI (Average Nucleotide Identity) superior or egal to 80% according to autoMLST.Supplementary Information 5: Prediction of the CAZyme encoding genes using HMMER dbCAN2. The genes are sorted according their CAZy families. For each gene, the begin position on the chromosome, the CAZy category, the annotation, the expression level in transcripts per million (TPM) and the predicted targets of the putative enzymes are described.Supplementary Information 6: Gene identified by antiSMASH in the region containing a biosynthetic gene cluster. Expression levels in transcripts per million (TPM) are indicated for each gene. Annotated central bioynthetic genes are indicated as Y. Those are the ones used for the calculation of mean BGC expression in Table 2.Supplementary Information 7: Annotation of AgN23 genes putatively involved in biosynthetic pathways for Auxins related phytohomones. Expression levels in transcripts per million (TPM) are indicated for each gene. The genes were detected by blasting reference KEGG sequences for each KEGG ONTOLOGY against AgN23 genes. A cut off of 70% identity and 40% coverage was applied to detect positive hits. These biosynthetic pathways and the KEGG ONTOLOGY are indicated in column F and G.Supplementary Information 8: Inspection of BiG-FAM hits with AgN23 BGCs to identify homologous BGCs found outside the S. violaceusniger clade. BiG-FAM distance higher than 900 were excluded from the analysis.The authors confirm all supporting data, code and protocols have been provided within the article or through supplementary data files.

Classical (real) hedgehogs can be regarded as the geometrical realiza-tions of formal di¤erences of convex bodies in R n+1. Like convex bodies, hedgehogs can be identi…ed with their support functions. Adopting a pro-jective viewpoint, we prove that any holomorphic function h : C n ! C can be regarded as the 'support function' of a complex hedgehog H h in C n+1. In the same vein, we introduce the notion of evolute of such a hedgehog H h in C 2 , and a natural (but apparently hitherto unknown) notion of complex curvature, which allows us to interpret this evolute as the locus of the centers of complex curvature. It is of course permissible to think that the development of a 'Brunn-Minkowski theory for complex hedgehogs' (replacing Euclidean volumes by symplectic ones) might be a promising way of research. We give …rst two results in this direction. We next return to real hedgehogs in R 2n endowed with a linear complex structure. We introduce and study the notion of evolute of a hedgehog. We particularly focus our attention on R 4 endowed with a linear Kahler structure determined by the datum of a pure unit quaternion. In parallel, we study the symplectic area of the images of the oriented Hopf circles under hedgehog parametrizations and introduce a quaternionic curvature function for such an image. Finally, we consider brie ‡y the convolution of hedgehogs, and the particular case of hedgehogs in R 4n regarded as a hyperkahler vector space.

Eucalypts are the most planted trees worldwide, but most of them are frost sensitive. Overexpressing transcription factors for CRT-repeat binding factors (CBFs) in transgenic Eucalyptus confer cold resistance both in leaves and stems. While wood plays crucial roles in trees and is affected by environmental cues, its potential role in adaptation to cold stress has been neglected. Here, we addressed this question by investigating the changes occurring in wood in response to the overexpression of two CBFs, taking advantage of available transgenic Eucalyptus lines. We performed histological, biochemical, and transcriptomic analyses on xylem samples. CBF ectopic expression led to a reduction of both primary and secondary growth, and triggered changes in xylem architecture with smaller and more frequent vessels and fibers exhibiting reduced lumens. In addition, lignin content and syringyl/guaiacyl (S/G) ratio increased. Consistently, many genes of the phenylpropanoid and lignin branch pathway were upregulated. Most of the features of xylem remodeling induced by CBF overexpression are reminiscent of those observed after long exposure of Eucalyptus trees to chilling temperatures. Altogether, these results suggest that CBF plays a central role in the cross-talk between response to cold and wood formation and that the remodeling of wood is part of the adaptive strategies to face cold stress.

Eucalypts are the most planted trees worldwide, but most of them are frost sensitive. Overexpressing transcription factors for CRT-repeat binding factors (CBFs) in transgenic Eucalyptus confer cold resistance both in leaves and stems. While wood plays crucial roles in trees and is affected by environmental cues, its potential role in adaptation to cold stress has been neglected. Here, we addressed this question by investigating the changes occurring in wood in response to the overexpression of two CBFs, taking advantage of available transgenic Eucalyptus lines. We performed histological, biochemical, and transcriptomic analyses on xylem samples. CBF ectopic expression led to a reduction of both primary and secondary growth, and triggered changes in xylem architecture with smaller and more frequent vessels and fibers exhibiting reduced lumens. In addition, lignin content and syringyl/guaiacyl (S/G) ratio increased. Consistently, many genes of the phenylpropanoid and lignin branch pathway were upregulated. Most of the features of xylem remodeling induced by CBF overexpression are reminiscent of those observed after long exposure of Eucalyptus trees to chilling temperatures. Altogether, these results suggest that CBF plays a central role in the cross-talk between response to cold and wood formation and that the remodeling of wood is part of the adaptive strategies to face cold stress.

Pythium oligandrum is a soil born free living oomycete able to parasitize fungi and oomycetes prey, including important plant and animals pathogens. Pythium oligandrum can colonize endophytically the root tissues of diverse plants where it induces plant defenses. Here we report the first long-read genome sequencing of a P. oligandrum strain sequenced by PacBio technology. Sequencing of genomic DNA loaded onto six SMRT cells permitted the acquisition of 913,728 total reads resulting in 112X genome coverage. The assembly and polishing of the genome sequence yielded180 contigs (N50 = 1.3 Mb; L50 = 12). The size of the genome assembly is 41.9 Mb with a longest contig of 2.7 Mb and 15,007 predicted protein-coding genes among which 95.25% were supported by RNAseq data, thus constituting a new Pythium genome reference. This data will facilitate genomic comparisons of Pythium species that are commensal, beneficial or pathogenic on plant, or parasitic on fungi and oomycete to identify key genetic determinants underpinning their diverse lifestyles. In addition comparison with plant pathogenic or zoopathogenic species will illuminate genomic adaptations for pathogenesis toward widely diverse hosts.

Background and aims The plant Hirtella physophora, the ant Allomerus decemarticulatus and a fungus, Trimmatostroma sp., form a tripartite association. The ants manipulate both the plant trichomes and the fungus to build galleries under the stems of their host plant used to capture prey. In addition to its structural role, the fungus also improves nutrient uptake by the host plant. But it still remains unclear whether the fungus plays an indirect or a direct role in transferring nutrients to the plant. This study aimed to trace the transfer of N from the fungus to the plant's stem tissue. Methods Optical microscopy and transmission electron microscopy (TEM) were used to investigate the presence of fungal hyphae in the stem tissues. Then, a 15N-labelling experiment was combined with a nanoscale secondary-ion mass spectrometry (NanoSIMS 50) isotopic imaging approach to trace the movement of added 15N from the fungus to plant tissues. Key results The TEM images clearly showed hyphae inside the stem tissue in the cellular compartment. Also, fungal hyphae were seen perforating the wall of the parenchyma cell. The 15N provisioning of the fungus in the galleries resulted in significant enrichment of the 15N signature of the plant's leaves 1 d after the 15N-labelling solution was deposited on the fungus-bearing trap. Finally, NanoSIMS imaging proved that nitrogen was transferred biotrophically from the fungus to the stem tissue. Conclusions This study provides evidence that the fungi are connected endophytically to an ant-plant system and actively transfer nitrogen from 15N-labelling solution to the plant's stem tissues. Overall, this study underlines how complex the trophic structure of ant-plant interactions is due to the presence of the fungus and provides insight into the possibly important nutritional aspects and tradeoffs involved in myrmecophyte-ant mutualisms.

Adipose tissue, as the main energy storage organ and through its endocrine activity, is interconnected with all physiological functions. It plays a fundamental role in energy homeostasis and in the development of metabolic disorders. Up to now, this tissue has been analysed as a pool of different cell types with very little attention paid to the organization and putative partitioning of cells. Considering the absence of a complete picture of the intimate architecture of this large soft tissue, we developed a method that combines tissue clearing, acquisition of autofluorescence or lectin signals by confocal microscopy, segmentation procedures based on contrast enhancement, and a new semi-automatic image analysis process, allowing accurate and quantitative characterization of the whole 3D fat pad organization. This approach revealed the unexpected anatomic complexity of the murine subcutaneous fat pad. Although the classical picture of adipose tissue corresponds to a superposition of simple and small ellipsoidal lobules of adipose cells separated by mesenchymal spans, our results show that segmented lobules display complex 3D poly-lobular shapes. Despite differences in shape and size, the number of these poly-lobular subunits is similar from one fat pad to another. Finally, investigation of the relationships of these subunits between each other revealed a never-described organization in two clusters with distinct molecular signatures and specific vascular and sympathetic nerve densities correlating with different browning abilities. This innovative procedure reveals that subcutaneous adipose tissue exhibits a subtle functional heterogeneity with partitioned areas, and opens new perspectives towards understanding its functioning and plasticity.

The non-animal peroxidases belong to a superfamily of oxidoreductases that reduce the hydrogen peroxide and oxidize numerous substrates. Since their initial characterization in 1992, several advances have provided an understanding into the origin and evolutionary history of this family of proteins. Here, we report for the first time an exhaustive evolutionary analysis of non-animal peroxidases using integratedin silicoand biochemical strategies. Thanks to the availability of numerous genomic sequences from many species belonging to different kingdoms together with expert and exhaustive annotation of peroxidase sequences centralized in a dedicated database, we have deepened our understanding of the evolutionary process underlying non-animal peroxidases through phylogenetic reconstructions. We analysed the distribution of all non-animal peroxidases in more than 200 eukaryotic organismsin silico. First, we show that the presence or absence of non-animal peroxidases can be correlated with the presence or absence of certain organelles or with specific biological processes. Examining a wide range of organisms, we confirmed that ascorbate peroxidases (APx) and cytochromes c peroxidases (CcP) were detected respectively in chloroplast and mitochondria containing organisms. Plants, which contain both organelles, are an exception and contain only APxs without CcP. Class III peroxidases (CIII Prx) were only detected in plants and Class II peroxidases (CII Prx) in fungi related to wood decay and plant degradation.Moreover, we demonstrate that biochemical activities (APx, CcP and CIII Prx) assayed in protein extracts obtained from 30 different eukaryotic organisms strongly support the distribution of the sequences resulting from ourin silicoanalysis. The biochemical results confirmed both the presence and classification of non-animal peroxidase encoding sequences.

Streptomycetes are soil-dwelling, filamentous actinobacteria and represent a prominent bacterial clade inside the plant root microbiota. The ability of streptomycetes to produce a broad spectrum of antifungal metabolites suggests that these bacteria could be used to manage plant diseases. Here, we describe the identification of a soil

International audience; Plant cell walls are made of polysaccharidic-proteinaceous complex matrices. Molecular interactions governing their organization remain understudied. We take advantage of the highly dynamic cell walls of Arabidopsis seed mucilage secretory cells to propose a hierarchical multi-molecular interaction model within a cell wall domain. We show that the PECTINMETHYLESTERASE INHIBITOR6 activity creates a partially demethylesterified pectin pattern acting as a platform allowing positioning of PEROXIDASE36 in a remote primary cell wall domain during early development. This allows triggering the loosening of this domain during later development, in turn leading to proper physiological function upon mature seed imbibition and germination. We anticipate that this pioneer example of molecular scaffold within a cell wall domain is more widespread through other combinations of the individual molecular players all belonging to large multigenic families. These results highlight the role of cell wall polysaccharide-protein interactions in the organization of cell wall domains.

International audience; Streptomycetes are soil-dwelling, filamentous actinobacteria and represent a prominent bacterial clade inside the plant root microbiota. The ability of streptomycetes to produce a broad spectrum of antifungal metabolites suggests that these bacteria could be used to manage plant diseases. Here, we describe the identification of a soil Streptomyces strain named AgN23 which strongly activates a large array of defense responses when applied on Arabidopsis thaliana leaves. AgN23 increased the biosynthesis of salicylic acid, leading to the development of salicylic acid induction deficient 2 (SID2)-dependent necrotic lesions. Size exclusion fractionation of plant elicitors secreted by AgN23 showed that these signals are tethered into high molecular weight complexes. AgN23 mycelium was able to colonize the leaf surface, leading to plant resistance against Alternaria brassicicola infection in wild-type Arabidopsis plants. AgN23-induced resistance was found partially compromised in salicylate, jasmonate, and ethylene mutants. Our data show that Streptomyces soil bacteria can develop at the surface of plant leaves to induce defense responses and protection against foliar fungal pathogens, extending their potential use to manage plant diseases.

Defense pathways and stress responses induced under Cd stress were illustrated in roots of hydroponically grown Medicago truncatula seedlings. Actually, the ascorbate-glutathione and antioxidative system, secondary metabolism events including peroxidases, phenolic compounds, and lignification launching, and developmental modifications were described. Cd (100 μM) initially increased reactive oxygen species, enhanced antioxidative (total SOD, CAT, and PRX) and ascorbate-glutathione-related metabolism enzymes (APX and MDAR), except in A17 and TN1.11. In agreement with peroxidase enhancement, physiological measurement and in situ observation illustrated soluble phenolic compound accumulation under Cd treatment. However, lignification was restricted to recently created protoxylem elements established in the root tip area, usually constituting the elongation zone. Cell death was increased. In the absence of necrotic reactions, developmental changes including lignin deposition, increase in cellulose and pectin contents, intercellular meatus, and condensed and deformed hairs were noticed in Cd-treated roots.

We give, under appropriate regularity assumptions, a strengthening of the Aleksandrov–Fenchel inequality in the form of a stability estimate.

International audience; Cryogenic transmission electron microscopy of high-pressure freezing (HPF) samples is a well-established technique for the analysis of liquid containing specimens. This technique enables observation without removing water or other volatile components. The HPF technique is less used in scanning electron microscopy (SEM) due to the lack of a suitable HPF specimen carrier adapter. The traditional SEM cryotransfer system (PP3000T Quorum Laughton, East Sussex, UK; Alto Gatan, Pleasanton, CA, USA) usually uses nitrogen slush. Unfortunately, and unlike HPF, nitrogen slush produces water crystal artefacts. So, we propose a new HPF specimen carrier adapter for sample transfer from HPF system to cryogenic-scanning electronic microscope (Cryo-SEM). The new transfer system is validated using technical two applications, a stearic acid in hydroxypropyl methylcellulose solution and mice myocardium. Preservation of samples is suitable in both cases. Cryo-SEM examination of HPF samples enables a good correlation between acid stearic liquid concentration and acid stearic occupation surface (only for homogeneous solution). For biological samples as myocardium, cytoplasmic structures of cardiomyocyte are easily recognized with adequate preservation of organelle contacts and inner cell organization. We expect this new HPF specimen carrier adapter would enable more SEM-studies using HPF.

Hedgehogs are (possibly singular and self-intersecting) hypersurfaces that describe Minkowski differences of convex bodies in $\mathbb{R}^{n+1}$. They are the natural geometrical objects when one seeks to extend parts of the Brunn-Minkowski theory to a vector space which contains convex bodies. In this paper, we prove that in every generic path of hedgehogs performing the eversion of the sphere in $\mathbb{R}^{3}$, there exists a hedgehog that has positive swallowtails. This study was motivated by an open problem raised in 1985 by Langevin, Levitt, and Rosenberg.

Plant non-specific lipid transfer proteins (nsLTPs) belong to a complex multigenic family implicated in diverse physiological processes. However, their function and mode of action remain unclear probably because of functional redundancy. Among the different roles proposed for nsLTPs, it has long been suggested that they could transport cuticular precursor across the cell wall during the formation of the cuticle, which constitutes the first physical barrier for plant interactions with their aerial environment. Here, we took advantage of the Arabidopsis thaliana etiolated hypocotyl model in which AtLTP2 was previously identified as the unique and abundant nsLTP member in the cell wall proteome, to investigate its function. AtLTP2 expression was restricted to epidermal cells of aerial organs, in agreement with the place of cuticle deposition. Furthermore, transient AtLTP2-TagRFP over-expression in Nicotiana benthamiana leaf epidermal cells resulted in its localization to the cell wall, as expected, but surprisingly also to the plastids, indicating an original dual trafficking for a nsLTP. Remarkably, in etiolated hypocotyls, the atltp2-1 mutant displayed modifications in cuticle permeability together with a disorganized ultra-structure at the cuticle-cell wall interface completely recovered in complemented lines, whereas only slight differences in cuticular composition were observed. Thus, AtLTP2 may not play the historical purported nsLTP shuttling role across the cell wall, but we rather hypothesize that AtLTP2 could play a major structural role by maintaining the integrity of the adhesion between the mainly hydrophobic cuticle and the hydrophilic underlying cell wall. Altogether, these results gave new insights into nsLTP functions.

International audience; Hydathodes are water pores found on leaves of a wide range of vascular plants and are the sites of guttation. We report here on the detailed anatomy of cauliflower (Brassica oleracea) and Arabidopsis (Arabidopsis thaliana) hydathodes. Hydathode surface presents pores resembling stomata giving access to large cavities. Beneath, the epithem is composed of a lacunar and highly vascularized parenchyma offering a direct connection between leaf surface and xylem vessels. Arabidopsis hydathode pores were responsive to ABA and light similar to stomata. The flg22 flagellin peptide, a well-characterized elicitor of plant basal immunity, did not induce closure of hydathode pores in contrast to stomata. Because hydathodes are natural infection routes for several pathogens, we investigated hydathode infection by the adapted vascular phytopathogenic bacterium Xanthomonas campestris pv campestris (Xcc), the causal agent of black rot disease of Brassicaceae. Microscopic observations of hydathodes six days postinoculation indicated a digestion of the epithem cells and a high bacterial multiplication. Postinvasive immunity was shown to limit pathogen growth in the epithem and is actively suppressed by the type III secretion system and its effector proteins. Altogether, these results give a detailed anatomic description of Brassicaceae hydathodes and highlight the efficient use of this tissue as an initial niche for subsequent vascular systemic dissemination of Xcc in distant plant tissues.

Power consumption and high compute density are the key factors to be considered when building a compute node for the upcoming Exascale revolution. Current architectural design and manufacturing technologies are not able to provide the requested level of density and power efficiency to realise an operational Exascale machine. A disruptive change in the hardware design and integration process is needed in order to cope with the requirements of this forthcoming computing target. This paper presents the ExaNoDe H2020 research project aiming to design a highly energy efficient and highly integrated heterogeneous compute node targeting Exascale level computing, mixing low-power processors, heterogeneous co-processors and using advanced hardware integration technologies with the novel UNIMEM Global Address Space memory system. This work was supported by the ExaNoDe project that has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 671578. The work presented in this paper reflects only authors’ view and the European Commission is not responsible for any use that may be made of the information it contains.

SUMMARY Downy mildew of sunflower is caused by Plasmopara halstedii (Farlow) Berlese & de Toni. Plasmopara halstedii is an obligate biotrophic oomycete pathogen that attacks annual Helianthus species and cultivated sunflower, Helianthus annuus. Depending on the sunflower developmental stage at which infection occurs, the characteristic symptoms range from young seedling death, plant dwarfing, leaf bleaching and sporulation to the production of infertile flowers. Downy mildew attacks can have a great economic impact on sunflower crops, and several Pl resistance genes are present in cultivars to protect them against the disease.Nevertheless, some of these resistances have been overcome by the occurrence of novel isolates of the pathogen showing increased virulence. A better characterization of P. halstedii infection and dissemination mechanisms,and the identification of the molecular basis of the interaction with sunflower, is a prerequisite to efficiently fight this pathogen. This review summarizes what is currently known about P. halstedii, provides new insights into its infection cycle on resistant and susceptible sunflower lines using scanning electron and light microscopy imaging, and sheds light on the pathogenicity factors of P. halstedii obtained from recent molecular data. Taxonomy: Kingdom Stramenopila; Phylum Oomycota; Class Oomycetes; Order Peronosporales; Family Peronosporaceae; Genus Plasmopara; Species Plasmopara halstedii. Disease symptoms: Sunflower seedling damping off, dwarfing of the plant, bleaching of leaves, starting from veins, and visible white sporulation, initially on the lower side of cotyledons and leaves. Plasmopara halstedii infection may severely impact sunflower seed yield. Infection process: In spring, germination of overwintered sexual oospores leads to sunflower root infection. Intercellular hyphae are responsible for systemic plant colonization and the induction of disease symptoms.Under humid and fresh conditions, dissemination structures are produced by the pathogen on all plant organs to release asexual zoosporangia.These zoosporangia play an important role in pathogen dissemination, as they release motile zoospores that are responsible for leaf infections on neighbouring plants. Disease control: Disease control is obtained by both chemical seed treatment (mefenoxam) and the deployment of dominant major resistance genes, denoted Pl. However, the pathogen has developed fungicide resistance and has overcome some plant resistance genes. Research for more sustainable strategies based on the identification of the molecular basis of the interaction are in progress. Useful websites: http://www.heliagene.org/HP, http://lipm -helianthus.toulouse.inra.fr/dokuwiki/doku.php?id=start, https:// www.heliagene.org/PlasmoparaSpecies (soon available).

Objectives: Needle decompression of tension pneumothorax in soldiers of the French infantry has a risk for failure when the standard procedure that involves the insertion of a 14-gauge, 5-cm catheter into the 2nd intercostal space (ICS) is used. This study measured the chest wall thickness (CWT) to assess whether this approach is appropriate. Methods: CWT was measured by ultrasound in 122 French soldiers at the 2nd and 4th ICSs on both the right and left sides. Results: CWT was measured at 4.19 cm (±0.96 cm) at the 2nd ICS and 3.00 cm (±0.91 cm) at the 4th ICS (p < 0.001). CWT was greater than 5 cm in 24.2% of cases at the 2nd ICS and 4.9% of cases at the 4th ICS (p < 0.001). Conclusions: This study suggests a high risk of failure when using the technique currently taught in the French army. A lateral approach into the 4th ICS could decrease this risk. The results of this study must be validated in patients presenting tension pneumothorax.

Oxidative disorders were triggered in the presence of Cd toxicity in early seedling growth of six Medicago truncatula genotypes. Interactions between root growth inhibition, cadmium uptake, as well as the occurrence of oxidative injury suggest differential responses of the genotypes, with susceptible or tolerant accessions. ROS enhancement was observed in situ and H₂O₂ content was measured, that did not seem related to tolerance or susceptibility. Oxidative burst impact on cell membrane integrity was analyzed in agreement with MDA content and glucose exudation, which suggest an active role of this burst in susceptible lines. Transcriptional changes in response to cadmium treatment were analyzed on target genes involved in (1) ROS-scavenging enzymes (superoxide dismutase (SOD; EC1.15.1.1) and peroxidase (PRX; EC 1.11.1.7)), (2) reduced glutathione (γ-Glu-Cys-Gly, GSH) metabolism (glutathione-S-transferase (GST; EC: 2.5.1.18) and glutathione reductase (GR; EC 1.8.1.7)), and (3) metal-chelating metabolism (PCS). The susceptible line shows no response or non-timely gene expression patterns. This research work gave an overview of the deleterious effects and oxidative injury of cadmium stress in Medicago truncatula. Oxidative defense efficiency and gene upregulation should explain relative tolerance in tested genotypes.

Hydathodes are water pores found on leaves of a wide range of vascular plants and are the sites of guttation. We report here on the detailed anatomy of cauliflower (

Junctional hemorrhage (i.e., between the trunk and limbs) are too proximal for a tourniquet and difficult to compress. These hemorrhages are responsible for 20% of preventable deaths by bleeding on the battlefield. The majority of these involve the groin area. Devices allowing a proximal compression for arterial axes have been recently developed.The purpose of this study was to compare the use of two junctional- tourniquet models, the Combat Ready Clamp (CRoC®) and the SAM® Junctional Tourniquet (SJT), in simulated out-of-hospital trauma care when tourniquets were ineffective to stop the arterial flow.During our clinical study, 84 healthy volunteers wearing battle dress performed a physical exercise to come approximate the operational context. The volunteers were randomly divided into two groups according to the device (the CRoC or SJT) used as supplement to a tourniquet self-applied to the root of the thigh. The primary study end point was the complete interruption of popliteal arterial flow, measured with Doppler auscultation. Time to effectiveness and subjective questionnaire data to evaluate the devices' application were also collected.Junctional device effectiveness was almost 90% for both the CRoC and the SJT, and did not differ between them, either used with a tourniquet (ρ = .36) or alone (ρ = .71). The time to effectiveness of the SJT was significantly shorter than that of the CRoC (ρ = .029).The SJT and the CRoC were equally effective. The SJT was faster to apply and preferred by the users. Our study provides objective evidence to the French Tactical Casualty Care Committee for improving junctional hemorrhage treatment.

article in press; International audience; Eucalyptus are of tremendous economic importance being the most planted hardwoods worldwide for pulp and paper, timber and bioenergy. The recent release of the Eucalyptus grandis genome sequence pointed out many new candidate genes potentially involved in secondary growth, wood formation or lineage-specific biosynthetic pathways. Their functional characterization is, however, hindered by the tedious, time-consuming and inefficient transformation systems available hitherto for eucalypts. To overcome this limitation, we developed a fast, reliable and efficient protocol to obtain and easily detect co-transformed E. grandis hairy roots using fluorescent markers, with an average efficiency of 62%. We set up conditions both to cultivate excised roots in vitro and to harden composite plants and verified that hairy root morphology and vascular system anatomy were similar to wild-type ones. We further demonstrated that co-transformed hairy roots are suitable for medium-throughput functional studies enabling, for instance, protein subcellular localization, gene expression patterns through RT-qPCR and promoter expression, as well as the modulation of endogenous gene expression. Down-regulation of the Eucalyptus cinnamoyl-CoA reductase1 (EgCCR1) gene, encoding a key enzyme in lignin biosynthesis, led to transgenic roots with reduced lignin levels and thinner cell walls. This gene was used as a proof of concept to demonstrate that the function of genes involved in secondary cell wall biosynthesis and wood formation can be elucidated in transgenic hairy roots using histochemical, transcriptomic and biochemical approaches. The method described here is timely because it will accelerate gene mining of the genome for both basic research and industry purposes.

We consider Gauss rigidity and Gauss infinitesimal rigidity for hedgehogs of \({\mathbb{R}^{3}}\) (regarded as Minkowski differences of closed convex surfaces of \({\mathbb{R}^{3}}\) with positive Gaussian curvature). Besides, we prove under an appropriate differentiability condition that whenever we perform a deformation of a hedgehog so that its curvature function remains constant, its algebraic volume also remains constant.

Summary Two C-repeat binding factor genes (EguCBF1a ⁄ b), isolated from E. gunnii and differentially cold-regulated, were constitutively overexpressed in a cold-sensitive Eucalyptus hybrid. In addition to the expected improvement on freezing tolerance, some resulting transgenic lines (EguCBF1a-OE and EguCBF1b-OE) exhibited a decrease in stomata density and an over-accumulation of anthocyanins also observed to a lesser extent in a cold-acclimated control plant. Given that the induction of five putative CBF target genes was observed in CBF-overexpressing lines as well as in the coldacclimated control line, these phenotypes might be related to cold acclimation. In comparison with the control plant, the most altered transgenic line (EguCBF1a-OE A1 line), exhibited reduced growth and better water retention capacity. This modified phenotype includes reduced leaf area and thickness associated with a decrease in cell size, as well as a higher oil gland density and a wax deposition on the cuticle. Surprisingly, the EguCBF1b-OE B9 line, with a level of transgene expression equivalent to the A1 line, showed a less marked phenotype, suggesting a difference in transactivation efficiency between EguCBF1A and B factors. The features of these transgenic lines provide the first signs of adaptive mechanisms controlled by CBF transcription factors in an evergreen broad-leaved tree. These data also open new prospects towards genetic improvement on Eucalyptus for freezing tolerance.

Resume En introduisant une notion de derivation des surfaces convexes de R 3 dans l'espace de Lorentz–Minkowski R 3 , 1 , nous donnons pour les surfaces convexes de R 3 un equivalent naturel d'une majoration du deficit isoperimetrique des courbes convexes de R 2 en termes d'aire algebrique de leur developpee. Nous etablissons en outre une serie d'inegalites geometriques pour les focales des surfaces convexes.

By combining Zinnia elegans in vitro tracheary element genomics with reverse genetics in Arabidopsis, we have identified a new upstream component of secondary wall formation in xylary and interfascicular fibers. Walls are thin 1 (WAT1), an Arabidopsis thaliana homolog of Medicago truncatula NODULIN 21 (MtN21), encodes a plant-specific, predicted integral membrane protein, and is a member of the plant drug/metabolite exporter (P-DME) family (transporter classification number: TC 2.A.7.3). Although WAT1 is ubiquitously expressed throughout the plant, its expression is preferentially associated with vascular tissues, including developing xylem vessels and fibers. WAT1:GFP fusion protein analysis demonstrated that WAT1 is localized to the tonoplast. Analysis of wat1 mutants revealed two cell wall-related phenotypes in stems: a defect in cell elongation, resulting in a dwarfed habit and little to no secondary cell walls in fibers. Secondary walls of vessel elements were unaffected by the mutation. The secondary wall phenotype was supported by comparative transcriptomic and metabolomic analyses of wat1 and wild-type stems, as many transcripts and metabolites involved in secondary wall formation were reduced in abundance. Unexpectedly, these experiments also revealed a modification in tryptophan (Trp) and auxin metabolism that might contribute to the wat1 phenotype. Together, our data demonstrate an essential role for the WAT1 tonoplast protein in the control of secondary cell wall formation in fibers.

Fusarium wilt is a soil-borne disease caused by formae specialis of Fusarium oxysporum on a large number of cultivated and wild plants. The susceptibility of the model legume plant Medicago truncatula to Fusarium oxysporum was studied by root-inoculating young plants in a miniaturised hydroponic culture. Among eight tested M. truncatula lines, all were susceptible to F. oxysporum f.sp. medicaginis, the causal agent of Fusarium wilt in alfalfa. However, a tolerant line, F83005.5, and a susceptible line, A17, could be distinguished by scoring the disease index. The fungus was transformed with the GFP marker gene and colonisation of the plant roots was analysed by epifluorescence and confocal microscopy. A slightly atypical pattern of root colonisation was observed, with massive fungal growth in the cortex. Although colonisation was not significantly different between susceptible and tolerant plants, the expression of some defence-related genes showed discrimination between both lines. A study with 10 strains from various host-plants indicated that M. truncatula was a permissive host to F. oxysporum.

Although cell wall remodeling is an essential feature of plant growth and development, the underlying molecular mechanisms are poorly understood. This work describes the characterization of Arabidopsis (Arabidopsis thaliana) plants with altered expression of ARAF1, a bifunctional α-l-arabinofuranosidase/β-d-xylosidase (At3g10740) belonging to family 51 glycosyl-hydrolases. ARAF1 was localized in several cell types in the vascular system of roots and stems, including xylem vessels and parenchyma cells surrounding the vessels, the cambium, and the phloem. araf1 T-DNA insertional mutants showed no visible phenotype, whereas transgenic plants that overexpressed ARAF1 exhibited a delay in inflorescence emergence and altered stem architecture. Although global monosaccharide analysis indicated only slight differences in cell wall composition in both mutant and overexpressing lines, immunolocalization experiments using anti-arabinan (LM6) and anti-xylan (LM10) antibodies indicated cell type-specific alterations in cell wall structure. In araf1 mutants, an increase in LM6 signal intensity was observed in the phloem, cambium, and xylem parenchyma in stems and roots, largely coinciding with ARAF1 expression sites. The ectopic overexpression of ARAF1 resulted in an increase in LM10 labeling in the secondary walls of interfascicular fibers and xylem vessels. The combined ARAF1 gene expression and immunolocalization studies suggest that arabinan-containing pectins are potential in vivo substrates of ARAF1 in Arabidopsis.

To successfully colonize their host, pathogens produce effectors that can interfere with host cellular processes. Here we investigated the function of CRN13 candidate effectors produced by plant pathogenic oomycetes and detected in the genome of the amphibian pathogenic chytrid fungus Batrachochytrium dendrobatidis (BdCRN13). When expressed in Nicotiana, AeCRN13, from the legume root pathogen Aphanomyces euteiches, increases the susceptibility of the leaves to the oomycete Phytophthora capsici. When transiently expressed in amphibians or plant cells, AeCRN13 and BdCRN13 localize to the cell nuclei, triggering aberrant cell development and eventually causing cell death. Using Forster resonance energy transfer experiments in plant cells, we showed that both CRN13s interact with nuclear DNA and trigger plant DNA damage response (DDR). Mutating key amino acid residues in a predicted HNH-like endonuclease motif abolished the interaction of AeCRN13 with DNA, the induction of DDR and the enhancement of Nicotiana susceptibility to P. capsici. Finally, H2AX phosphorylation, a marker of DNA damage, and enhanced expression of genes involved in the DDR were observed in A. euteiches-infected Medicago truncatula roots. These results show that CRN13 from plant and animal eukaryotic pathogens promotes host susceptibility by targeting nuclear DNA and inducing DDR.

In the Euclidean plane ℝ2, we define the Minkowski difference 𝒦–𝓛 of two arbitrary convex bodies 𝒦, 𝓛 as a rectifiable closed curve ℋh⊂ ℝ2that is determined by the differenceh=h𝒦–h𝓛of their support functions. This curve ℋhis called the hedgehog with support functionh. More generally, the object of hedgehog theory is to study the Brunn–Minkowski theory in the vector space of Minkowski differences of arbitrary convex bodies of Euclidean space ℝn+1, defined as (possibly singular and self-intersecting) hypersurfaces of ℝn+1. Hedgehog theory is useful for: (i) studying convex bodies by splitting them into a sum in order to reveal their structure; (ii) converting analytical problems into geometrical ones by considering certain real functions as support functions. The purpose of this paper is to give a detailed study of plane hedgehogs, which constitute the basis of the theory. In particular: (i) we study their lengthmeasures and solve the extension of the Christoffel–Minkowski problemto plane hedgehogs; (ii) we characterize support functions of plane convex bodies among support functions of plane hedgehogs and support functions of plane hedgehogs among continuous functions; (iii) we study the mixed area of hedgehogs in ℝ2and give an extension of the classical Minkowski inequality (and thus of the isoperimetric inequality) to hedgehogs.

In n-dimensional Euclidean space $ \mathbb{R}^n+1 $ , an N-hedgehog is the envelope of a family of cooriented hyperplanes including exactly N hyperplanes with given unit normal vector. Hedgehogs (or 1-hedgehogs) with C2 support function can be interpreted as differences of convex bodies of class $ C_{+}^{2} $ . We studied in [5] the extension to hedgehogs of classical geometrical inequalities for convex bodies. On some spaces of hedgehogs, the Aleksandrov-Fenchel inequality is reversed and an inner product is defined in terms of mixed volume [5]. This paper gives a similar study for N-hedgehogs in $ \mathbb{R}^2(N\in\mathbb{N}^\ast) $ . We give a geometrical interpretation of the Sturm-Hurwitz theorem and a new proof of this theorem for C 2-functions. We also consider the Minkowski problem for N-hedgehogs.

LetC be a convex curve of constant width and of classC 4 + . It is known thatC has at least 6 vertices and its interior contains either a point through which infinitely many normals pass or an open set of points through each of which pass at least 6 normals. If all its vertices are nondegenerate, then: (i)C has exactly 6 vertices if, and only if, its evolute is the boundary of a topological disc through each interior point of which pass at least 6 normals; (ii) ifC has more than 6 vertices, then there exists an open set of points through each of which pass at least 10 normals. The proof: (i) expresses the number of normals passing through a point as a function of the index with respect to the evolute; (ii) relates this index to the number of singularities of the evolute (i.e. of vertices). Furthermore, we give formulas for counting singularities of generic hedgehogs in ℝ2 and ℝ3.

Ce travail avait pour objectif de rationaliser les prescriptions d’examens immuno-hematologiques (IH) dans 3 chirurgies: cholecystectomie (CHOL) (cœlioscopie, laparotomie), appendicectomie (APP) (Mac Burney, cœlioscopie), cure de hernie parietale (HERN) (toutes techniques). Le DIM a fourni la liste des patients operes entre 2011 et 2015 (code CCAM) qui a ete confrontee a celle des receveurs extraite du logiciel Cursus (Guyot Walser) et des donnees IH extraites du logiciel Synergy (Technidata). Les dossiers des patients transfuses entre 3 j avant et 15 jours apres la chirurgie ont ete revus. Sur 772 CHOL, 10 ont ete transfuses: 5 CHOL contingentes d’une chirurgie a procedures multiples (CPM), 4 CHOL isolees non programmees, 1 cas de CHOL isolee programmee (plaie de l’artere cystique). Sur 434 APP, 2 ont ete transfusees dans un cadre de CPM. Sur 857 HERN, 6 ont ete transfuses: 2 HERN inscrites dans une CPM, 4 HERN isolees programmees (2 transfusions preoperatoires pour anemie, 2 postoperatoires pour complication d’un traitement anticoagulant). Au final, en dehors des CPM, le risque transfusionnel a ete estime a 0,2 % pour CHOL, 0 % pour APP et 0,5 % pour HERN. Un bilan IH (groupage et RAI) avait ete prescrit dans pres de 100 % des APP, 70 % des CHOL et moins de 10 % des HERN. En decembre 2016, la suppression du bilan IH systematique dans ces 3 chirurgies a ete validee et les prescripteurs concernes–anesthesistes et urgentistes–ont ete informes. Une comparaison entre les chirurgies realisees en 2017 et les 30 derniers actes realises en 2016 montre une reduction des prescriptions de bilan IH en cas d’APP (71 % vs. 17 %– p