Physiochemical interaction between osmotic stress and a bacterial exometabolite promotes plant disease.

Various microbes isolated from healthy plants are detrimental under laboratory conditions, indicating the existence of molecular mechanisms preventing disease in nature. Here, we demonstrated that application of sodium chloride (NaCl) in natural and gnotobiotic soil systems is sufficient to induce plant disease caused by an otherwise non-pathogenic root-derived Pseudomonas brassicacearum isolate (R401). Disease caused by combinatorial treatment of NaCl and R401 triggered extensive, root-specific transcriptional reprogramming that did not involve down-regulation of host innate immune genes, nor dampening of ROS-mediated immunity. Instead, we identified and structurally characterized the R401 lipopeptide brassicapeptin A as necessary and sufficient to promote disease on salt-treated plants. Brassicapeptin A production is salt-inducible, promotes root colonization and transitions R401 from being beneficial to being detrimental on salt-treated plants by disturbing host ion homeostasis, thereby bolstering susceptibility to osmolytes. We conclude that the interaction between a global change stressor and a single exometabolite from a member of the root microbiome promotes plant disease in complex soil systems.

Fava bean (Vicia faba L.) protein concentrate added to beef burgers improves the bioaccessibility of some free essential amino acids after in vitro oral and gastrointestinal digestion.

The influence of partial replacement of animal protein by plant-based ingredients on the protein digestibility of beef burgers was investigated. Beef burgers were supplemented with fava bean protein concentrate (FB) or a mixture of FB and flaxseed flour (FBFS), both processed by extrusion, at different levels: 0 (control), 10, 15, and 20 % (w/w). A pilot sensory analysis was conducted to select the percentage of flour inclusion for further assays: control, 10 % FB, and 10 % FBFS. Protein digestibility, amino acid profile, and protein secondary structure of these burgers after in vitro oral and gastrointestinal digestion were studied. In vitro boluses were prepared with the AM2 masticator, simulating normal mastication, and static in vitro digestion of boluses was performed according to the INFOGEST method. Inclusion of 10 % FB in beef burgers did not alter their flavour or tenderness compared to the control, whereas tenderness and juiciness scored slightly higher for the 10 % FBFS burgers compared to 15 % and 20 % FBFS ones. Poor lipid oxidative stability during storage was observed with 10 % FBFS burgers. Total protein content was significantly higher (p < 0.05) in 10 % FB burgers than in control burgers after in vitro oral digestion. Additionally, 10 % FB burgers presented higher amounts of free essential amino acids like isoleucine, leucine, phenylalanine, and valine at the end of digestion, as well as methionine, tyrosine, and histidine. Partial substitution of meat protein by 10 % FB improves the nutritional profile of beef burgers, without altering their sensory qualities.

Genome-resolved metatranscriptomics reveals conserved root colonization determinants in a synthetic microbiota.

The identification of processes activated by specific microbes during microbiota colonization of plant roots has been hampered by technical constraints in metatranscriptomics. These include lack of reference genomes, high representation of host or microbial rRNA sequences in datasets, or difficulty to experimentally validate gene functions. Here, we recolonized germ-free Arabidopsis thaliana with a synthetic, yet representative root microbiota comprising 106 genome-sequenced bacterial and fungal isolates. We used multi-kingdom rRNA depletion, deep RNA-sequencing and read mapping against reference microbial genomes to analyse the in planta metatranscriptome of abundant colonizers. We identified over 3,000 microbial genes that were differentially regulated at the soil-root interface. Translation and energy production processes were consistently activated in planta, and their induction correlated with bacterial strains' abundance in roots. Finally, we used targeted mutagenesis to show that several genes consistently induced by multiple bacteria are required for root colonization in one of the abundant bacterial strains (a genetically tractable Rhodanobacter). Our results indicate that microbiota members activate strain-specific processes but also common gene sets to colonize plant roots.

Single Seed Microbiota: Assembly and Transmission from Parent Plant to Seedling.

The seed acts as the primary inoculum source for the plant microbiota. Understanding the processes involved in its assembly and dynamics during germination and seedling emergence has the potential to allow for the improvement of crop establishment. Changes in the bacterial community structure were tracked in 1,000 individual seeds that were collected throughout seed developments of beans and radishes. Seeds were associated with a dominant bacterial taxon that represented more than 75% of all reads. The identity of this taxon was highly variable between the plants and within the seeds of the same plant. We identified selection as the main ecological process governing the succession of dominant taxa during seed filling and maturation. In a second step, we evaluated the seedling transmission of seed-borne taxa in 160 individual plants. While the initial bacterial abundance on seeds was not a good predictor of seedling transmission, the identities of the seed-borne taxa modified the phenotypes of seedlings. Overall, this work revealed that individual seeds are colonized by a few bacterial taxa of highly variable identity, which appears to be important for the early stages of plant development. IMPORTANCE Seeds are key components of plant fitness and are central to the sustainability of the agri-food system. Both the seed quality for food consumption and the seed vigor in agricultural settings can be influenced by the seed microbiota. Understanding the ecological processes involved in seed microbiota assembly will inform future practices for promoting the presence of important seed microorganisms for plant health and productivity. Our results highlighted that seeds were associated with one dominant bacterial taxon of variable taxonomic identity. This variety of dominant taxa was due to (i) spatial heterogeneity between and within plants and (ii) primary succession during seed development. According to neutral models, selection was the main driver of microbial community assembly for both plant species.

Beneficial Effects of Linseed Supplementation on Gut Mucosa-Associated Microbiota in a Physically Active Mouse Model of Crohn's Disease.

The Western diet, rich in lipids and in n-6 polyunsaturated fatty acids (PUFAs), favors gut dysbiosis observed in Crohn's disease (CD). The aim of this study was to assess the effects of rebalancing the n-6/n-3 PUFA ratio in CEABAC10 transgenic mice that mimic CD. Mice in individual cages with running wheels were randomized in three diet groups for 12 weeks: high-fat diet (HFD), HFD + linseed oil (HFD-LS-O) and HFD + extruded linseed (HFD-LS-E). Then, they were orally challenged once with the Adherent-Invasive Escherichia coli (AIEC) LF82 pathobiont. After 12 weeks of diet, total energy intake, body composition, and intestinal permeability were not different between groups. After the AIEC-induced intestinal inflammation, fecal lipocalin-2 concentration was lower at day 6 in n-3 PUFAs supplementation groups (HFD-LS-O and HFD-LS-E) compared to HFD. Analysis of the mucosa-associated microbiota showed that the abundance of Prevotella, Paraprevotella, Ruminococcus, and Clostridiales was higher in the HFD-LS-E group. Butyrate levels were higher in the HFD-LS-E group and correlated with the Firmicutes/Proteobacteria ratio. This study demonstrates that extruded linseed supplementation had a beneficial health effect in a physically active mouse model of CD susceptibility. Additional studies are required to better decipher the matrix influence in the linseed supplementation effect.

Seed microbiota revealed by a large-scale meta-analysis including 50 plant species.

Seed microbiota constitutes a primary inoculum for plants that is gaining attention owing to its role for plant health and productivity. Here, we performed a meta-analysis on 63 seed microbiota studies covering 50 plant species to synthesize knowledge on the diversity of this habitat. Seed microbiota are diverse and extremely variable, with taxa richness varying from one to thousands of taxa. Hence, seed microbiota presents a variable (i.e. flexible) microbial fraction but we also identified a stable (i.e. core) fraction across samples. Around 30 bacterial and fungal taxa are present in most plant species and in samples from all over the world. Core taxa, such as Pantoea agglomerans, Pseudomonas viridiflava, P. fluorescens, Cladosporium perangustum and Alternaria sp., are dominant seed taxa. The characterization of the core and flexible seed microbiota provided here will help uncover seed microbiota roles for plant health and design effective microbiome engineering.

Beneficial Effects of High Intensity Interval Training and/or Linseed Oil Supplementation to Limit Obesity-Induced Oxidative Stress in High Fat Diet-Fed Rats.

High-intensity interval training (HIIT) and linseed oil (LO) supplementation are effective strategies to reduce obesity-induced oxidative stress. Our aim was to determine whether the HIIT + LO combination prevents obesity-induced oxidative stress in high fat diet (HFD)-fed rats. HFD-fed 8-week-old, male, Wistar rats were subdivided in four groups: HFD, LO (2% of sunflower oil replaced with 2% of LO in the HFD), HIIT (4 days/week for 12 weeks), and HIIT + LO. Wistar rats fed a low-fat diet (LFD) were used as controls. Epididymal and subcutaneous adipose tissue, gastrocnemius muscle, liver, and plasma samples were collected to measure oxidative stress markers (AOPP, oxLDL), antioxidant (SOD, CAT, and GPx activities) and pro-oxidant (NOx and XO) enzyme activities. Compared with the LFD, the HFD altered the pro/antioxidant status in different tissues (increase of AOPP, oxLDL, SOD and catalase activities in plasma, and SOD activity increase in liver and decrease in adipose tissues) but not in gastrocnemius. LO upregulated CAT activity and decreased NOx in liver. HIIT alleviated HFD negative effects in liver by reducing SOD and NOx activities. Moreover, the HIIT + LO combination potentiated SOD activity upregulation in subcutaneous tissue. HIIT and LO supplementation have independent beneficial effects on the pro/antioxidant balance. Their association promotes SOD activity in subcutaneous adipose tissue.

High-Intensity Interval Training and α-Linolenic Acid Supplementation Improve DHA Conversion and Increase the Abundance of Gut Mucosa-Associated Oscillospira Bacteria.

Obesity, a major public health problem, is the consequence of an excess of body fat and biological alterations in the adipose tissue. Our aim was to determine whether high-intensity interval training (HIIT) and/or α-linolenic acid supplementation (to equilibrate the n-6/n-3 polyunsaturated fatty acids (PUFA) ratio) might prevent obesity disorders, particularly by modulating the mucosa-associated microbiota. Wistar rats received a low fat diet (LFD; control) or high fat diet (HFD) for 16 weeks to induce obesity. Then, animals in the HFD group were divided in four groups: HFD (control), HFD + linseed oil (LO), HFD + HIIT, HFD + HIIT + LO. In the HIIT groups, rats ran on a treadmill, 4 days.week-1. Erythrocyte n-3 PUFA content, body composition, inflammation, and intestinal mucosa-associated microbiota composition were assessed after 12 weeks. LO supplementation enhanced α-linolenic acid (ALA) to docosahexaenoic acid (DHA) conversion in erythrocytes, and HIIT potentiated this conversion. Compared with HFD, HIIT limited weight gain, fat mass accumulation, and adipocyte size, whereas LO reduced systemic inflammation. HIIT had the main effect on gut microbiota ß-diversity, but the HIIT + LO association significantly increased Oscillospira relative abundance. In our conditions, HIIT had a major effect on body fat mass, whereas HIIT + LO improved ALA conversion to DHA and increased the abundance of Oscillospira bacteria in the microbiota.

Temporal dynamics of bacterial communities during seed development and maturation.

Seed microbiota acts as a starting point for the assembly of the plant microbiota and contributes to successful plant establishment. To date, the order and timing of microbial taxa immigration during seed development and maturation remained unknown. We investigated the temporal dynamics of seed bacterial communities in bean and radish. A high phylogenetic turnover was observed for both plant species with few taxa associated with all seed developmental stages. Greater heterogeneity in communities structure within each stage was observed for radish. While, about one-third of radish seed bacterial taxa were detected in buds, flowers and fruits, very few taxa seem to be transmitted by the floral route in bean. In the latter species, bacterial populations belonging to the P. fluorescens species complex were found either in buds, flowers and fruits or in seeds. The relative phylogenetic proximity of these bacterial populations combined with their habitat specificity led us to explore the genetic determinants involved in successful seed transmission in bean. Comparative genomic analyses of representatives bacterial strains revealed dozens of coding sequences specifically associated with seed-transmitted strains. This study provided a first glimpse on processes involved in seed microbiota assembly, which could be used for designing plant-beneficial microbial consortia.

Contribution of connective tissue components, muscle fibres and marbling to beef tenderness variability in longissimus thoracis, rectus abdominis, semimembranosus and semitendinosus muscles.

BACKGROUND: The present study aimed to identify relationships between components of intramuscular connective tissue, proportions of the different fiber types, intramuscular fat and sensory tenderness of beef cooked at 55 °C. Accordingly, four muscles differing in their metabolic and contractile properties, as well as in their collagen content and butcher value, were obtained from dairy and beef cattle of several ages and sexes and were then used to create variability. RESULTS: Correlation analyses and/or stepwise regressions were applied on Z-scores to identify the existing and robust associations. Tenderness scores were further categorized into tender, medium and tough classes using unsupervised learning methods. The findings revealed a muscle-dependant role with respect to tenderness of total and insoluble collagen, cross-links, and type IIB + X and IIA muscle fibers. The longissimus thoracis and semitendinosus muscles that, in the present study, were found to be extreme in their tenderness potential were also very different from each other and from the rectus abdominis (RA) and semimembranosus (SM). RA and SM muscles were very similar regarding their relationship for muscle components and tenderness. A relationship between marbling and tenderness was only present when the results were analysed irrespective of all factors of variation of the experimental model relating to muscle and animal type. CONCLUSION: The statistical approaches applied in the present study using Z-scores allowed identification of the robust associations between muscle components and sensory beef tenderness and also identified discriminatory variables of beef tenderness classes. © 2020 Society of Chemical Industry.

Effect of linseed fed as rolled seeds, extruded seeds or oil on fatty acid rumen metabolism and intestinal digestibility in cows.

Linseed, a source of linolenic acid, is used in ruminant diets to increase polyunsaturated fatty acids (FA) in animal products. Seed processing is known to have an impact on FA rumen metabolism, but few data are available for linseed. We studied the effect of linseed lipid on ruminal metabolism and intestinal digestibility in cows. Three modes of linseed processing: rolled linseed (RL), extruded linseed (EL) and linseed oil plus linseed meal (LO), supplemented at 7.5% of DM intake, were compared to a control diet (C). Duodenal flows, intestinal digestibility and plasma composition were determined. The duodenal flow of linolenic acid was similar among diets. The sum of t10 and t11-18:1, which were coeluted, was increased with lipid-supplemented diets and represented more than 60% of trans 18:1 for EL and LO diets. The main 18:2 isomers were c9, c12 and t11, c15 among the non-conjugated isomers, and t11, t13 among CLA. Linseed supplementation increased the duodenal flow of unsaturated intermediates of biohydrogenation, and this effect was more pronounced for extruded seeds and oil than for rolled seeds. For most 18-carbon FA, intestinal digestibility was slightly higher for C and LO diets than for RL and EL. Plasma concentrations of non-conjugated 18:2 and linolenic acid were similar among the lipid-supplemented diets. Within diet, profiles of 18:1 isomers (except c9) remained very similar between duodenal and plasma FA.

Effects of introducing linseed in livestock diet on blood fatty acid composition of consumers of animal products.

Reducing the ratio between essential fatty acids: C18:2 n-6/C18:3 n-3 down to 5 is recommended by Nutritional Guidelines. We studied the fatty acid (FA) changes in consumers' plasma following changes in livestock diet. First, a zootechnical study introduced 5% of extruded linseed into the diet of livestock to replace other oleaginous ingredients, and on an iso-nutritional values basis. The products from linseed-fed animals contained more n-3 fatty acids (precursor alpha-linolenic and derivatives obtained by elongations and desaturations) than control animal products (issued from animals fed without linseed), and more conjugated linoleic acids (CLA). The n-6/n-3 ratio was reduced by 54% in butter, 60% in meat and 86% in eggs. Following this, a double-blind, randomised, cross-over clinical study involving 75 healthy volunteers compared plasma and erythrocyte FA profiles in consumers of animal products (from livestock fed the linseed diet or from livestock fed standard diet). It showed modifications in the FA composition of the experimental human regimen with more C18:3 n-3 (1.65 vs. 0.75 g/day), and more n-3 derivatives. The C18:2 n-6/C18:3 n-3 ratio decreased (7 vs. 15). In volunteers' plasma, C18:3 n-3 increased in the essay group (0.93 vs. 0.44% of the FA), so did n-3 derivatives and CLA. The n-6/n-3 ratio decreased from 14.3 to 10.2. In erythrocytes, C20:5 n-3 increased in the essay group (0.59 vs. 0.45%) and so did C22:6 n-3. The n-6/n-3 ratio decreased in parallel from 4.2 to 3.8. Without any changes in consumers' eating habits, foodstuffs from animals fed linseed diets induced significant modifications of human plasma and erythrocyte fatty acid composition (comparable to that noted under the 'Cretan' diet) and a sharp increase in CLA.