The Oomycete Microbe-Associated Molecular Pattern, Arachidonic Acid, and an Ascophyllum nodosum-Derived Plant Biostimulant Induce Defense Metabolome Remodeling in Tomato.

Arachidonic acid (AA) is an oomycete-derived microbe-associated molecular pattern (MAMP) capable of eliciting robust defense responses and inducing resistance in plants. Similarly, Ascophylum nodosum extract (ANE) from the brown seaweed A. nodosum, a plant biostimulant that contains AA, can also prime plants for defense against pathogen challenges. A previous parallel study comparing the transcriptomes of AA- and ANE-root-treated tomatoes demonstrated significant overlap in transcriptional profiles, a shared induced resistance phenotype, and changes in the accumulation of various defense-related phytohormones. In this work, untargeted metabolomic analysis via liquid chromatography-mass spectrometry was conducted to investigate the local and systemic metabolome-wide remodeling events elicited by AA and ANE root treatment in tomatoes. Our study demonstrated AA and ANE's capacity to locally and systemically alter the metabolome of tomatoes with enrichment of chemical classes and accumulation of metabolites associated with defense-related secondary metabolism. AA- and ANE-root-treated plants showed enrichment of fatty acyl-glycosides and strong modulation of hydroxycinnamic acids and derivatives. Identification of specific metabolites whose accumulation was affected by AA and ANE treatment revealed shared metabolic changes related to ligno-suberin biosynthesis and the synthesis of phenolic compounds. This study highlights the extensive local and systemic metabolic changes in tomatoes induced by treatment with a fatty acid MAMP and a seaweed-derived plant biostimulant with implications for induced resistance and crop improvement.

Mechanistic insights into the liquefaction stage of enzyme-mediated biomass deconstruction.

Effective enzyme-mediated viscosity reduction, disaggregation, or "liquefaction," is required to overcome the rheological challenges resulting from the fibrous, hygroscopic nature of lignocellulosic biomass, particularly at the high solids loadings that will be required for an economically viable process. However, the actual mechanisms involved in enzyme-mediated liquefaction, as determined by viscosity or yield stress reduction, have yet to be fully resolved. Particle fragmentation, interparticle interaction, material dilution, and water-retention capacity were compared for their ability to quantify enzyme-mediated liquefaction of model and more realistic pretreated biomass substrates. It was apparent that material dilution and particle fragmentation occurred simultaneously and that both mechanisms contributed to viscosity/yield stress reduction. However, their relative importance was dependent on the nature of the biomass substrate. Interparticle interaction and enzyme-mediated changes to these interactions was shown to have a significant effect on slurry rheology. Liquefaction was shown to result from the combined action of material dilution, particle fragmentation, and alteration of interactions at particle surfaces. However, the observed changes in water retention capacity did not correlate with yield stress reduction. The relative importance of each mechanism was significantly influenced by the nature of the biomass substrate and its physicochemical properties. An ongoing challenge is that mechanisms, such as refining, which enhance enzyme accessibility to the cellulosic component of the substrate, are detrimental to slurry rheology and will likely impede enzyme-mediated liquefaction when high substrate concentrations are used.

Protection of Marine Mammals.

Within the European Defense Agency (EDA), the Protection of Marine Mammals (PoMM) project, a comprehensive common marine mammal database essential for risk mitigation tools, was established. The database, built on an extensive dataset collection with the focus on areas of operational interest for European navies, consists of annual and seasonal distribution and density maps, random and systematic sightings, an encyclopedia providing knowledge on the characteristics of 126 marine mammal species, data on marine mammal protection areas, and audio information including numerous examples of various vocalizations. Special investigations on marine mammal acoustics were carried out to improve the detection and classification capabilities.

Alkaline extract of the seaweed Ascophyllum nodosum stimulates arbuscular mycorrhizal fungi and their endomycorrhization of plant roots.

Ascophyllum nodosum extracts (ANE) are well-established plant biostimulants that improve stress tolerance and crop vigour, while also having been shown to stimulate soil microbes. The intersection of these two stimulatory activities, and how they combine to enhance plant health, however, remains poorly understood. In the present study, we aimed to evaluate: (1) the direct effect of ANE on the arbuscular mycorrhizal fungus Rhizophagus irregularis, and (2) whether ANE influences endomycorrhization in plants. ANE enhanced development of R. irregularis in vitro, showing greater spore germination, germ tube length, and hyphal branching. Greenhouse-grown Medicago truncatula drench-treated with ANE formed mycorrhizal associations faster (3.1-fold higher mycorrhization at week 4) and grew larger (29% greater leaf area by week 8) than control plants. Foliar applications of ANE also increased root colonization and arbuscular maturity, but did not appear to enhance plant growth. Nonetheless, following either foliar or drench application, M. truncatula genes associated with establishment of mycorrhizae were expressed at significantly higher levels compared to controls. These results suggest that ANE enhances mycorrhization through both direct stimulation of arbuscular mycorrhizal fungus growth and through stimulation of the plant's accommodation of the symbiont, together promoting the establishment of this agriculturally vital plant-microbe symbiosis.

Enzyme-Mediated Lignocellulose Liquefaction Is Highly Substrate-Specific and Influenced by the Substrate Concentration or Rheological Regime.

The high viscosities/yield stresses of lignocellulose slurries makes their industrial processing a significant challenge. However, little is known regarding the degree to which liquefaction and its enzymatic requirements are specific to a substrate's physicochemical and rheological properties. In the work reported here, the substrate- and rheological regime-specificities of liquefaction of various substrates were assessed using real-time in-rheometer viscometry and offline oscillatory rheometry when hydrolyzed by combinations of cellobiohydrolase (Trichoderma reesei Cel7A), endoglucanase (Humicola insolens Cel45A), glycoside hydrolase (GH) family 10 xylanase, and GH family 11 xylanase. In contrast to previous work that has suggested that endoglucanase activity dominates enzymatic liquefaction, all of the enzymes were shown to have at least some liquefaction capacity depending on the substrate and reaction conditions. The contribution of individual enzymes was found to be influenced by the rheological regime; in the concentrated regime, the cellobiohydrolase outperformed the endoglucanase, achieving 2.4-fold higher yield stress reduction over the same timeframe, whereas the endoglucanase performed best in the semi-dilute regime. It was apparent that the significant differences in rheology and liquefaction mechanisms made it difficult to predict the liquefaction capacity of an enzyme or enzyme cocktail at different substrate concentrations.

Laccase-mediated hydrophilization of lignin decreases unproductive enzyme binding but limits subsequent enzymatic hydrolysis at high substrate concentrations.

One of the predominant mechanisms by which lignin restricts effective enzymatic deconstruction of lignocellulosic materials is the unproductive adsorption of enzymes. Although this inhibition can be partially mitigated through hydrophilization of lignin during thermochemical pretreatment, these types of treatments could potentially worsen slurry rheology, consequently making it more difficult to process the material at high substrate concentrations. In the work reported here, laccases were used to specifically modify lignin hydrophilicity within steam-pretreated substrate via in situ phenolic compound grafting. While lignin hydrophilization reduced unproductive enzyme adsorption, high-solids hydrolysis efficiency decreased significantly due to mass transfer limitations. It was apparent that low-solids hydrolysis experiments were a poor predictor of substrate digestibility at high-solids conditions and that substrate-water interactions impacted both substrate digestibility and slurry rheology.