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39 páginas.- 14 figuras.- 134 referencias ; Lignin is largely produced by the oxidative radical coupling of the three monolignols, p-coumaryl, coniferyl, and sinapyl alcohols, which form the p-hydroxyphenyl, guaiacyl, and syringyl lignin units in the lignin polymer. However, it is increasingly appreciated that other "unconventional" phenolic compounds can also behave as lignin monomers participating in oxidative radical reactions during the lignification process and becoming integrated into the lignin polymer in some plants. Various phenolic compounds have been found incorporated into the lignin, including phenolic compounds derived from the monolignol biosynthetic pathway (e.g., monolignol ester conjugates, caffeyl and 5-hydroxyconiferyl alcohols, hydroxycinnamaldehydes, among others). More fascinating still was the discovery in some plants of a series of phenolic compounds arising from other polyphenolic biosynthetic pathways, including flavonoids (tricin, dihydrotricin, naringenin, naringenin chalcone), hydroxystilbenes (resveratrol, piceatannol, and isorhapontigenin, and their O-glucosylated counterparts piceid, astringin, and isorhapontin), and ferulic acid amides (feruloyltyramine, and diferuloylputrescine), that are integrated into the lignin polymer. All of these discoveries indicate that the lignin structure is far more complex than previously thought, that the definition of lignin should not be restricted to a polymer derived solely from the three traditional monolignols, and that other unconventional lignin monomers must be considered, thus expanding the established definition of lignin. The overriding factor remains, however, that lignification, the process of polymerization to the lignin polymer, allows this compositional flexibility because it is a simple chemical process. © 2022 Elsevier Ltd ; JCdR, JRe and AG were funded by the Spanish Project AGL2017-83036-R and PID2020-118968RB-100 (financed by Ministerio de Ciencia e Innovación, MCIN, Agencia Estatal de Investigación, AEI, and European Regional Development Fund, ERDF, "A way of making Europe"), the Regional Andalusian Government (Consejerı ́a de Transformación Económica, Industria, Conocimiento y Universidades, Junta de Andalucıía, project P20-00017), and CSIC (project PIE 202040E185). JRa and HK were funded by the DOE Great Lakes Bioenergy Research Center (DOE Office of Science DE-SC0018409 ; Peer reviewed

12 páginas.- 6 figuras.- 2 tablas.- 56 referencias.-The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation ; Rice (Oryza sativa L.) straw is a highly abundant, widely available, and low cost agricultural waste that can be used as a source to extract valuable phytochemicals of industrial interest. Hence, in the present work, the chemical composition of the lipophilic compounds present in rice straw was thoroughly characterized by gas chromatography and mass spectrometry using medium-length high-temperature capillary columns, which allowed the identification of a wide range of lipophilic compounds, from low molecular weight fatty acids to high molecular weight sterols esters, sterol glucosides, or triglycerides in the same chromatogram. The most abundant lipophilic compounds in rice straw were fatty acids, which accounted for up to 6,400 mg/kg (41.0% of all identified compounds), followed by free sterols (1,600 mg/kg; 10.2%), sterol glucosides (1,380 mg/kg; 8.8%), fatty alcohols (1,150 mg/kg; 7.4%), and triglycerides (1,140 mg/kg; 7.3%), along with lower amounts of high molecular weight wax esters (900 mg/kg; 5.8%), steroid ketones (900 mg/kg; 5.8%), monoglycerides (600 mg/kg; 3.8%), alkanes (400 mg/kg; 2.6%), diglycerides (380 mg/kg; 2.4%), sterol esters (380 mg/kg; 2.4%), tocopherols (340 mg/kg; 2.2%), and steroid hydrocarbons (60 mg/kg; 0.4%). This information is of great use for the valorization of rice straw to obtain valuable lipophilic compounds of interest for the nutraceutical, pharmaceutical, cosmetic, and chemical industries. Moreover, this knowledge is also useful for other industrial uses of rice straw, as in pulp and papermaking, since some lipophilic compounds are at the origin of the so-called pitch deposits during pulping. Copyright © 2022 Rosado, Marques, Rencoret, Gutiérrez and del Río. ; This work was supported by the projects AGL2017-83036-R and PID2020-118968RB-I00 (funded by MCIN/AEI/10.13039/501100011033 and, as appropriate, by "ERDF A way of making Europe") and the Regional Andalusian Government, Consejería de Transformación Económica, Industria, Conocimiento y Universidades/FEDER (project P20-00017). MJR acknowledges the Spanish Ministry of Science, Innovation and Universities for a FPI fellowship (PRE2018-083267). ; Peer reviewed

Lipophilic extractives from wood and nonwood fibers exert a negative impact in pulp and paper manufacture causing pitch deposits. We have shown the effectiveness of the laccase-mediator system in removing pulp lipids regardless the pulping process and the raw material used. The enzymatic treatments were performed using a fungal laccase from Pycnoporus cinnabarinus and 1-hydroxybenzotriazole (HBT) as mediator. Gas chromatography-mass spectrometry of extracts from the enzymatically-treated pulps revealed that most of the lipids (including free and conjugated sitosterol, fatty and resin acids, and triglycerides) were removed. Improved pulp brightness and decreased kappa number were also observed. Then, the chemistry of the reactions of lipophilic extractives with laccase-HBT was studied using model alkanes, fatty alcohols, fatty acids, resin acids, free sterols, sterol esters and triglycerides. The laccase alone modified some unsaturated lipids, however, the most rapid and extensive removal was obtained in the presence of mediator. Different degradation patterns were observed, and several oxidation products were identified. Lignin related phenols were tested as alternative to synthetic mediators for lipid removal from eucalypt pulp. Over 90% removal of free and conjugated sitosterol, similar to that attained with HBT, was obtained with some of them. A positive effect on pulp brightness and kappa number was also obtained, especially after a peroxide stage. ; This work was supported by the Spanish projects AGL2008-00709, BIO2007-28719-E and BIO2008-01533, and the BIORENEW project of the European Union (contract NMP2-CT-2006-026456). INRA (Marseille, France) is acknowledged for the P. cinnabarinus strain, and Beldem (Andenne, Belgium) for laccase production. ENCE (Pontevedra, Spain), UPM Kymmene (Valkeakoski, Finland), and CELESA (Tortosa, Spain), and UPC (Terrassa, Spain) are acknowledged for the eucalypt pulp, spruce TMP pulp and flax pulp samples, respectively. ; Peer reviewed

14 páginas.-- 10 figuras.-- 1 tabla.-- 46 referencias.-- Author's Choice—Final version free via Creative Commons CC-BY license. ; Versatile peroxidase (VP) is a high redox-potential peroxidase of biotechnological interest that is able to oxidize phenolic and non-phenolic aromatics, Mn2 , and different dyes. The ability of VP from Pleurotus eryngii to oxidize water-soluble lignins (softwood and hardwood lignosulfonates) is demonstrated here by a combination of directed mutagenesis and spectroscopic techniques, among others. In addition, direct electron transfer between the peroxidase and the lignin macromolecule was kinetically characterized using stopped-flow spectrophotometry. VP variants were used to show that this reaction strongly depends on the presence of a solvent-exposed tryptophan residue (Trp-164). Moreover, the tryptophanyl radical detected by EPR spectroscopy of H2O2-activated VP (being absent from the W164S variant) was identified as catalytically active because it was reduced during lignosulfonate oxidation, resulting in the appearance of a lignin radical. The decrease of lignin fluorescence (excitation at 355 nm/emission at 400 nm) during VP treatment under steady-state conditions was accompanied by a decrease of the lignin (aromatic nuclei and side chains) signals in one-dimensional and two-dimensional NMR spectra, confirming the ligninolytic capabilities of the enzyme. Simultaneously, size-exclusion chromatography showed an increase of the molecular mass of the modified residual lignin, especially for the (low molecular mass) hardwood lignosulfonate, revealing that the oxidation products tend to recondense during the VP treatment. Finally, mutagenesis of selected residues neighboring Trp-164 resulted in improved apparent second-order rate constants for lignosulfonate reactions, revealing that changes in its protein environment (modifying the net negative charge and/or substrate accessibility/binding) can modulate the reactivity of the catalytic tryptophan ; This work was supported by the INDOX (KBBE-2013-613549) European Union project, the HIPOP (BIO2011-26694), NOESIS (BIO2014-56388-R), and BIORENZYMERY (AGL2014-53730-R) projects of the Spanish Ministry of Economy and Competitiveness (MINECO) co-financed by FEDER funds, and the PRIN 2009-STNWX3 project of the Italian Ministry of Education, Universities and Research (MIUR). ; Supported by an FPI Fellowship of the Spanish MINECO.-- Supported by a contract of the CSIC project 201440E097.-- Supported by a Ramón y Cajal contract of the Spanish MINECO ; Peer reviewed

19 páginas.- 9 figuras.- referencias ; Tomato varieties resistant to the bacterial wilt pathogen Ralstonia solanacearum have the ability to restrict bacterial movement in the plant. Inducible vascular cell wall reinforcements seem to play a key role in confining R. solanacearum into the xylem vasculature of resistant tomato. However, the type of compounds involved in such vascular physico-chemical barriers remain understudied, while being a key component of resistance. Here we use a combination of histological and live-imaging techniques, together with spectroscopy and gene expression analysis to understand the nature of R. solanacearum-induced formation of vascular coatings in resistant tomato. We describe that resistant tomato specifically responds to infection by assembling a vascular structural barrier formed by a ligno-suberin coating and tyramine-derived hydroxycinnamic acid amides. Further, we show that overexpressing genes of the ligno-suberin pathway in a commercial susceptible variety of tomato restricts R. solanacearum movement inside the plant and slows disease progression, enhancing resistance to the pathogen. We propose that the induced barrier in resistant plants does not only restrict the movement of the pathogen, but may also prevent cell wall degradation by the pathogen and confer anti-microbial properties, effectively contributing to resistance. ; Research is funded by MCIN/AEI/10.13039/501100011033 (NSC, MV), MCIN/AEI/PID2019-110330GB-C21 (MF, OS), MCIN/AEI/PID2020-118968RBI00 (JR), through the 'Severo Ochoa Programme for Centres of Excellence in R&D' (SEV-2015-0533, CEX2019-000917 and CEX2019-000902-S funded by MCIN/AEI/ 10.13039/501100011033), and by the Spanish National Research Council (CISC) pie-201620E081 (JR, AG) and the Generalitat de Catalunya (2017SGR765 grant). AK is the recipient of a Netaji Subhas – Indian Council of Agricultural Research International Fellowship. SS acknowledges financial support from DOC-FAM, European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement no. 754397. This work was also supported by the CERCA Program/Generalitat de Catalunya. ; Peer reviewed

Coumarate 3-hydroxylase (C3H) catalyzes a key step of the synthesis of the two main lignin subunits, guaiacyl (G) and syringyl (S) in dicotyledonous species. As no functional data are available in regards to this enzyme in monocotyledonous species, we generated C3H1 knock-down maize plants. The results obtained indicate that C3H1 participates in lignin biosynthesis as its down-regulation redirects the phenylpropanoid flux: as a result, increased amounts of p-hydroxyphenyl (H) units, lignin-associated ferulates and the flavone tricin were detected in transgenic stems cell walls. Altogether, these changes make stem cell walls more degradable in the most C3H1-repressed plants, despite their unaltered polysaccharide content. The increase in H monomers is moderate compared to C3H deficient Arabidopsis and alfalfa plants. This could be due to the existence of a second maize C3H protein (C3H2) that can compensate the reduced levels of C3H1 in these C3H1-RNAi maize plants. The reduced expression of C3H1 alters the macroscopic phenotype of the plants, whose growth is inhibited proportionally to the extent of C3H1 repression. Finally, the down-regulation of C3H1 also increases the synthesis of flavonoids, leading to the accumulation of anthocyanins in transgenic leaves. ; Funding: This work was supported by the Spanish "Ministerio de Economía y Competitividad" [AGL2011-30545-C02-01 to D.C.-R., AGL2011-30545-C02-02 to A.E., and AGL2011-25379 (co-financed by FEDER funds) to J.C.R.] and received financial support from the CONSOLIDER-INGENIO program [CSD2007-00036] from the Spanish Ministerio de Ciencia e Innovación. This work was carried out within the framework of the "Xarxa de Referència de Biotecnologia" (XarBa) from the Autonomous Government of Catalonia. R.S. was financed by the postdoctoral contracts "Isidro Parga Pondal" supported by the Autonomous Government of Galicia and the European Social Fund and "Ramón y Cajal", supported by the Spanish Ministry of Economy and Competitiveness and the University of Vigo. J.R. thanks CSIC for a JAE-DOC contract of the program "Junta para la Ampliación de Estudios" co-financed by the European Social Fund. ; Peer Reviewed

31 páginas.- 14 figuras.- 92 referencias.- Supplementary Materials: The following are available online at https://www.mdpi.com/article/10.3390/jof7060426/s1 .- Acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). ; Pleurotus eryngii is a grassland-inhabiting fungus of biotechnological interest due to its ability to colonize non-woody lignocellulosic material. Genomic, transcriptomic, exoproteomic, and metabolomic analyses were combined to explain the enzymatic aspects underlaying wheat–straw transformation. Up-regulated and constitutive glycoside–hydrolases, polysaccharide–lyases, and carbohydrate–esterases active on polysaccharides, laccases active on lignin, and a surprisingly high amount of constitutive/inducible aryl–alcohol oxidases (AAOs) constituted the suite of extracellular enzymes at early fungal growth. Higher enzyme diversity and abundance characterized the longer-term growth, with an array of oxidoreductases involved in depolymerization of both cellulose and lignin, which were often up-regulated since initial growth. These oxidative enzymes included lytic polysaccharide monooxygenases (LPMOs) acting on crystalline polysaccharides, cellobiose dehydrogenase involved in LPMO activation, and ligninolytic peroxidases (mainly manganese-oxidizing peroxidases), together with highly abundant H2O2-producing AAOs. Interestingly, some of the most relevant enzymes acting on polysaccharides were appended to a cellulose-binding module. This is potentially related to the non-woody habitat of P. eryngii (in contrast to the wood habitat of many basidiomycetes). Additionally, insights into the intracellular catabolism of aromatic compounds, which is a neglected area of study in lignin degradation by basidiomycetes, were also provided. The multiomic approach reveals that although non-woody decay does not result in dramatic modifications, as revealed by detailed 2D-NMR and other analyses, it implies activation of the complete set of hydrolytic and oxidative enzymes characterizing lignocellulose-decaying basidiomycetes. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. ; This research was funded by the GENOBIOREF (BIO2017-86559-R) project of the Spanish Ministry of Science and Innovation (co-financed by FEDER funds) to F.J.R.-D. and S.C.; and by the Consejo Superior de Investigaciones CientÍficas (PIE-202120E019 to A.T.M. and PIE-201620E081 to A.G.). The work conducted by the JGI, a DOE Office of Science User Facility, is supported by the Office of Science of the U.S. DOE under contract DE-AC02-05CH11231. Part of this material is also based upon work supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research (BER) under the Early Career Award Program to D.S. This work was authored in part by the National Renewable Energy Laboratory, operated by Alliance for Sustainable Energy, LLC, for the U.S. Department of Energy (DOE) under Contract No. DE-AC36-08GO28308. The views expressed in the article do not necessarily represent the views of the DOE or the U.S. Government. The U.S. Government retains and the publisher, by accepting the article for publication, ; Peer reviewed