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he oomycete pathogen Aphanomyces astaci, also known as "crayfish plague", is an obligate fungal-like parasite of freshwater crustaceans and is considered responsible for the ongoing decline of native European crayfish populations. A. astaci is thought to secrete a wide array of effectors and enzymes that facilitate infection, however their molecular mechanisms have been poorly characterized. Here, we report the identification of AA15 lytic polysaccharide monooxygenases (LPMOs) as a new group of secreted virulence factors in A. astaci. We show that this enzyme family has greatly expanded in A. astaci compared to all other oomycetes, and that it may facilitate infection through oxidative degradation of crystalline chitin, the most abundant polysaccharide found in the crustacean exoskeleton. These findings reveal new roles for LPMOs in animal-pathogen interactions, and could help inform future strategies for the protection of farmed and endangered species.

Main conclusion: Methyl-jasmonate induces large increases in p-coumarate linked to arabinoxylan in Brachypodium and in abundance of GT61 and BAHD family transcripts consistent with a role in synthesis of this linkage. Jasmonic acid (JA) signalling is required for many stress responses in plants, inducing large changes in the transcriptome, including up-regulation of transcripts associated with lignification. However, less is known about the response to JA of grass cell walls and the monocot-specific features of arabinoxylan (AX) synthesis and acylation by ferulic acid (FA) and para-coumaric acid (pCA). Here, we show that methyl-jasmonate (MeJA) induces moderate increases in FA monomer, > 50% increases in FA dimers, and five–sixfold increases in pCA ester-linked to cell walls in Brachypodium callus. Direct measurement of arabinose acylated by pCA (Araf-pCA) indicated that most or all the increase in cell-wall pCA was due to pCA ester-linked to AX. Analysis of the RNA-seq transcriptome of the callus response showed that these cell-wall changes were accompanied by up-regulation of members of the GT61 and BAHD gene families implicated in AX decoration and acylation; two BAHD paralogues were among the most up-regulated cell-wall genes (seven and fivefold) after 24 h exposure to MeJA. Similar responses to JA of orthologous BAHD and GT61 transcripts are present in the RiceXPro public expression data set for rice seedlings, showing that they are not specific to Brachypodium or to callus. The large response of AX-pCA to MeJA may, therefore, indicate an important role for this linkage in response of primary cell walls of grasses to JA signalling.

Lignocellulose forms the structural framework of woody plant biomass and represents the most abundant carbon source on the Planet. Turnover of woody biomass is a critical component of the planetary carbon cycle, and the enzymes involved are of increasing industrial importance as industry moves away from fossil to renewable carbon resources. Shipworms are marine bivalve molluscs that digest wood and play a key role in global carbon recycling by reprocessing plant biomass in the oceans. Previous studies suggest that wood digestion in shipworms is dominated by enzymes produced by endosymbiotic bacteria found in the animal’s gills, while little is known about the identity and function of endogenous enzymes produced by shipworms. Using a combination of meta-transcriptomic, proteomic, imaging and biochemical analyses, we reveal a complex digestive system dominated by uncharacterized enzymes that are secreted by a specialized digestive gland and that accumulate in the cecum, where wood digestion occurs. Using a combination of transcriptomics, proteomics and microscopy, we show that the digestive proteome of the shipworm Lyrodus pedicellatusis mostly comprised of enzymes produced by the animal itself, with a small but significant contribution from symbiotic bacteria. The digestive proteome is dominated by a novel 300 kDa multi-domain glycoside hydrolase that accounts for more than half of the total protein content in the cecum and functions in the hydrolysis of β-1,4-glucans, the most abundant polymers in wood. These studies allow an unprecedented level of insight into an unusual and ecologically important process for wood recycling in the marine environment, and open up new biotechnological opportunities in the mobilization of sugars from lignocellulosic biomass.

Up to 33% of hemp by mass can be lost in the form of dust during processing for fibre production. Heptane Soxhlet extractions and supercritical carbon dioxide extractions (scCO2) of hemp dust samples yielded significant quantities of high value lipophilic molecules including fatty acids, policosanols (fatty alcohols), fatty aldehydes, hydrocarbons, sterols, triterpenoids and cannabinoids (cannabidiol (CBD)). Dust collected from different stages of the mechanical processing of hemp fibres gave rise to lipophilic extractives with varying compositions, thus making the isolation and purification of these compounds easier. Of particular interest is CBD (5832.5 ± 118.9 μg/g of dust), which has attracted much attention for clinical-level studies due to its therapeutic efficacy in the treatment of a variety of central nervous system (CNS) disorders. Factorial experimental design was carried out to optimise the scCO2 extraction, with 350 bar and 50 °C yielding the selective extraction of higher value components.

Background: Using globally abundant crop residues as a carbon source for energy generation and renewable chemicals production stands out as a promising solution to reduce current dependency on fossil fuels. In nature, such as in compost habitats, microbial communities efficiently degrade the available plant biomass using a diverse set of synergistic enzymes. However, deconstruction of lignocellulose remains a challenge for industry due to recalcitrant nature of the substrate and the inefficiency of the enzyme systems available, making the economic production of lignocellulosic biofuels difficult. Metatranscriptomic studies of microbial communities can unveil the metabolic functions employed by lignocellulolytic consortia and identify new biocatalysts that could improve industrial lignocellulose conversion. Results: In this study, a microbial community from compost was grown in minimal medium with sugarcane bagasse sugarcane bagasse as the sole carbon source. Solid-state nuclear magnetic resonance was used to monitor lignocellulose degradation; analysis of metatranscriptomic data led to the selection and functional characterization of several target genes, revealing the first glycoside hydrolase from Carbohydrate Active Enzyme family 11 with exo-1,4-β-xylanase activity. The xylanase crystal structure was resolved at 1.76 Å revealing the structural basis of exo-xylanase activity. Supplementation of a commercial cellulolytic enzyme cocktail with the xylanase showed improvement in Avicel hydrolysis in the presence of inhibitory xylooligomers. Conclusions: This study demonstrated that composting microbiomes continue to be an excellent source of biotechnologically important enzymes by unveiling the diversity of enzymes involved in in situ lignocellulose degradation.

Microbial communities are essential to a wide range of ecologically and industrially important processes. To control or predict how these communities function, we require a better understanding of the factors which influence microbial community productivity. Here, we combine functional resource use assays with a biodiversity-ecosystem functioning (BEF) experiment to determine whether the functional traits of constituent species can be used to predict community productivity. We quantified the abilities of 12 bacterial species to metabolise components of lignocellulose and then assembled these species into communities of varying diversity and composition to measure their productivity growing on lignocellulose, a complex natural substrate. A positive relationship between diversity and community productivity was caused by a selection effect whereby more diverse communities were more likely to contain two species that significantly improved community productivity. Analysis of functional traits revealed that the observed selection effect was primarily driven by the abilities of these species to degrade β-glucan. Our results indicate that by identifying the key functional traits underlying microbial community productivity we could improve industrial bioprocessing of complex natural substrates.

Silicon (Si) is taken up from the soil as monosilicic acid by plant roots, transported to leaves and deposited as phytoliths, amorphous silica (SiO2) bodies, which are a key component of anti-herbivore defense in grasses. Silicon transporters have been identified in many plant species, but the mechanisms underpinning Si transport remain poorly understood. Specifically, the extent to which Si uptake is a passive process, driven primarily by transpiration, or has both passive and active components remains disputed. Increases in foliar Si concentration following herbivory suggest plants may exercise some control over Si uptake and distribution. In order to investigate passive and active controls on Si accumulation, we examined both genetic and environmental influences on Si accumulation in the forage grass Festuca arundinacea. We studied three F. arundinacea varieties that differ in the levels of Si they accumulate. Varieties not only differed in Si concentration, but also in increases in Si accumulation in response to leaf damage. The varietal differences in Si concentration generally reflected differences in stomatal density and stomatal conductance, suggesting passive, transpiration-mediated mechanisms underpin these differences. Bagging plants after damage was employed to minimize differences in stomatal conductance between varieties and in response to damage. This treatment eliminated constitutive differences in leaf Si levels, but did not impair the damage-induced increases in Si uptake: damaged, bagged plants still had more leaf Si than undamaged, bagged plants in all three varieties. Preliminary differential gene expression analysis revealed that the active Si transporter Lsi2 was highly expressed in damaged unbagged plants compared with undamaged unbagged plants, suggesting damage-induced Si defenses are regulated at gene level. Our findings suggest that although differences in transpiration may be partially responsible for varietal differences in Si uptake, they cannot explain damage-induced increases in Si uptake and deposition, suggesting that wounding causes changes in Si uptake, distribution and deposition that likely involve active processes and changes in gene expression. Introduction