In this paper, Phoenix dactylifera L. date palm seeds (RDS) were first characterized from a physicochemical point of view, in particular the proximate and ultimate composition, XRD, and SEM-EDX analyses. Based on findings RDS contains 77.4 % of total carbohydrates, 12.8 % of lignin, and a higher heating value (HHV) of 15.88 MJ kg(-1). Immuno-labeling intensity (OD) of non-cellulosic RDS fraction was also determined thanks to the ELISA technique after the dissolution of polysaccharides in ionic liquid showing the presence of mannans and xyloglucans and the absence of arabinans in date seeds, a significant amount of homogalacturonans were also detected. The thermal decomposition and kinetic of date palm seeds (RDS) using TGA-DTG, and DSC measurements were then investigated. The RDS samples were heated in the temperature ranges 25-600 degrees C at 5, 10, 15, and 20 degrees C/min. The decomposition of hemicelluloses, cellulose, and lignin, active pyrolysis of RDS, was held in a range of temperature 160-450 degrees C. The kinetic parameters such as activation energy (E-a) and pre-exponential factor (A) were determined for two degradation steps by using iso-conversional model-free methods. The E-a and pre-exponential factor obtained by the Kissinger method are 212 kJ mol(-1) and 7.8x10(19) min(-1) for the first decomposition interval (I1) and 172 kJ mol(-1) and 5.96x10(13) min(-1) for the second decomposition interval I2. The same average parameters calculated by model-free methods are 126-156 kJ mol(-1) and 180-213 kJ mol(-1) and 3.36x10(14) and 7.07x10(17) min(-1) respectively for the first and second intervals of thermal decomposition. Indicating that activation energy decreases in the final stages of the process and that the energy required for hemicelluloses degradation is lower than that of cellulose. The most probable reaction functions have been determined for these two stages, by Coats-Redfern (CR) and Criado methods, leading to considerably improved calculation performance over the entire conversion range. The pyrolysis reaction models of RDS are described by reaction, second order F2 for cellulose and hemicelluloses. With the Arrhenius parameters obtained from the fitting model of CR, we attempt to reconstruct the temperature-dependent mass conversion curves and have resulted in generally acceptable results. Based on the Arrhenius parameter values, obtained by the Kissinger equation, the changes in entropy, enthalpy, Gibbs free energy, and lifetime predictions have been estimated for the thermal degradation processes of RDS.

Molecular cues linked to heartwood formation open new (complementary) perspectives to genetic breeding programs of Douglas-fir, a tree species largely cultivated in Europe for the natural durability and civil engineering properties of its wood. RNAs from a single genotype of Douglas-fir, extracted from three distinct wood zones (outer sapwood, inner sapwood and transition zone) at four vegetative seasons to generate an extensive RNA-seq dataset used to apprehend the in-wood dynamic and seasonality of heartwood formation in this hardwood model species. Previously published data collected on somatic embryos of the same genotype could be merged with the present dataset to upgrade grade the Douglas-fir reference transcriptome.

The wood polysaccharide composition, a new analytical method, based on ionic liquid dissolution of low amount of biomass coupled with an ELISA essay of polysaccharides. In the present work, we synthesized and tested several imidazolium and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) based ILs for their ability to solubilize Douglas-fir wood while preserving the wall polymer integrity. The couple times-temperatures have been essayed for wood dissolution. Then their efficiency for wood biomass dissolution was compared to the impact of IL on storing and/or destroy polysaccharides. Thanks to the ELISA technique with a set of mAbs against epitopes of the main hemicellulose, pectin, and protein families of cell wall components. Wood destructuration at 80˚C with the 1-ethyl-3-methylimidazolium bromide represents a good compromise of wood dissolution efficiency and low polysaccharide destruction.

This work aims to extrapolate the energy conversion of Raw Loquat Kernels (RLK) by pyrolysis under the inert atmosphere. For this purpose, the main physicochemical composition of the RLK was first investigated. The proximate analysis shows that RLK has a relatively low moisture content, an absence of heavy metals, and the presence of an important amount of organic matter. In order to evaluate polysaccharides composition, RLK was dissolved in 1-ethyl-3-methylimidazolium bromide [Emim]Br ionic liquid; the resulting solutions are then immuno-labeled with monoclonal Antibodies (mAbs) against plant cell wall polysaccharides epitopes. Polysaccharides have been then analyzed using the ELISA technique with a set of 14 mAbs against hemicellulose and pectin. The LM19 immuno-labeling was more intense after 3 days of treatment at 80 °C, followed by 2 hours at 100 °C, indicating the important amount of un-esterified Homogalacturonans among RLK polysaccharides. Xray diffraction pattern indicates the presence of a small amount of crystalline cellulose. Surface functional groups and morphology of RLK were also investigated. RLK was then subjected to thermal analyses TG/DTG, DTA, and DSC under an inert atmosphere. The kinetic parameters were determined by the Coats-Redfern method. The highest activation energies were found in the first thermal degradation region where the main pyrolysis reaction took place, and the largest weight loss occurred, with energy values running from 24.34 to 117.06 KJ/mol.

Somatic embryogenesis techniques have been developed for most coniferous species, but only using very juvenile material. To extend the techniques’ scope, better integrated understanding of the key biological, physiological and molecular characteristics of embryogenic state is required. Therefore, embryonal masses (EMs) and non-embryogenic calli (NECs) have been compared during proliferation at multiple levels. EMs and NECs originating from a single somatic embryo (isogenic lines) of each of three unrelated genotypes were used in the analyses, which included comparison of the lines’ anatomy by transmission light microscopy, transcriptomes by RNAseq Illumina sequencing, proteomes by free-gel analysis, contents of endogenous phytohormones (indole-3-acetic acid, cytokinins and ABA) by LC-MS analysis, and soluble sugar contents by HPLC. EMs were characterized by upregulation (relative to levels in NECs) of transcripts, proteins, transcription factors and active cytokinins associated with cell differentiation accompanied by histological, carbohydrate content and genetic markers of cell division. In contrast, NECs were characterized by upregulation (relative to levels in EMs) of transcripts, proteins and products associated with responses to stimuli (ABA, degradation forms of cytokinins, phenols), oxidative stress (reactive oxygen species) and carbohydrate storage (starch). Sub-Network Enrichment Analyses that highlighted functions and interactions of transcripts and proteins that significantly differed between EMs and NECs corroborated these findings. The study shows the utility of a novel approach involving integrated multi-scale transcriptomic, proteomic, biochemical, histological and anatomical analyses to obtain insights into molecular events associated with embryogenesis and more specifically to the embryogenic state of cell in Douglas-fir.

To explore poorly understood differences between primary and subsequent somatic embryogenic lines of plants, we induced secondary (2ry) and tertiary (3ry) lines from cotyledonary somatic embryos (SEs) of two Douglas-fir genotypes: SD4 and TD17. The 2ry lines exhibited significantly higher embryogenic potential (SE yields) than the 1ry lines initiated from zygotic embryos (SD4, 2155 vs 477; TD17, 240 vs 29 g− 1 f.w.). Moreover, we observed similar differences in yield between 2ry and 3ry lines of SD4 (2400 vs 3921 g− 1 f.w.). To elucidate reasons for differences in embryogenic potential induced by repetitive somatic embryogenesis we then compared 2ry vs 1ry and 2ry vs 3ry lines at histo-cytological (using LC-MS/MS) and proteomic levels.

The composition of sapwood, transition zone and heartwood extracts from Pseudotsuga menziesii was studied using LC/ESI–MS/MS. Based on retention times, and MS fragmentation patterns, 40 phenolic compounds, 11 terpenes and 7 fatty and organic acids were identified or accordingly characterized. The flavonoids taxifolin-O-hexoside and the corresponding aglycon taxifolin, dihydrokaempferol, pinocembrin, pinobanksin, quercetin and flavan-3-ols, catechin and epicatechin were the main compounds detected in wood collected in the fall. Taxifolin derivatives were identified as monomers, dimers and flavonolignans. Comparative metabolite profiling of the three zones showed differences in metabolite distribution. Flavonoids were distributed in the 3 zones. Lignans, flavonolignans, phenolic acid derivatives and terpenes accumulated mainly in the transition zone while tannins were detected only in sapwood. Fatty and organic acids were present in sapwood and the transition zone. Multiple Reaction Monitoring (MRM) was used for the quantification of main metabolites of extracts in order to observe seasonal variations in the wood. The relative quantity of 35 selected compounds in each zone was discussed. The lowest metabolite content was usually observed in winter whereas flavonoids were found in high quantities in heartwood either in spring or autumn.

The objective of this study is to estimate the influence of mixed mode on the global fracture energy. The experimental tests were carried out using three Douglas genotypes. In order to evaluate the fracture parameters the Wedge Splitting Tests were performed. The fracture parameters were estimated from the optical measurements coupled with numerical and analytical approaches. Firstly, the fracture energy was estimated from the work of fracture calculated from the splitting force - Crack Opening Displacement curve. In this case, the Crack Opening Displacement was measured by means of Mark Tracking method. Secondly, the fracture parameters and, consequently the fracture energy was evaluated using an integrated mixed mode approach. In this second case, the experimental measurements were performed by Digital Image Correlation. The fracture energy corresponding to opening and shear modes was calculated by using an adjustment procedure coupled with Stress Intensity Factor calculation and the phase angle. This second approach allowing the calculation of fracture energy without the influence of experimental noises. The results show that the proposed integrated mixed mode approach allows evaluate the part of each mode in the fracture process. (C) 2017 Elsevier Ltd. All rights reserved.

Dans le cadre de cette etude, nous presentons une approche originale de la durabilite en proposant une transversalite entre patrimoine genetique de differentes familles de Douglas et durabilite en structure en employant, comme ‘marqueur’, le comportement hygroscopique du materiau. Cette transversalite entre biologie et mecanique du bois a pour but d’accroitre ce degre de durabilite afin de limiter le recours systematique a ces traitements de preservation souvent agressifs. Ce travail se presente sous forme d'une etude statistique avec ambition de mettre en exergue des genotypes en lien avec un faible coefficient de diffusion et des isothermes de sorption a humidite d’equilibre basse.

Les substances humiques sont connues pour être des produits naturels pouvant être utilisés comme bio-fertilisant. Dans ce travail nous avons testé des molécules issus d’un traitement thermo-mécanique de déchets de sciure du bois de peuplier (SHB: substance humique du bois, BoisValor). Les SHB ont servi d’amendement organique à une culture de Lantana camara. Dans ce travail nous démontrons que les SHB à l’instar des SH (substances humiques fossiles) sont des biofertilisants pour les cultures horticoles. Leur utilisation améliore la croissance et le développement de l’appareil végétatif et réduit le temps de culture avant floraison.

L’arsenic (As) est un polluant métalloïde normalement présent dans l’environnement. Co-produit de l’extraction du minerai d’or, l’arsenic constitue le principal polluant des terrils miniers comme le terril de la mine de Chéni en Haute Vienne (87). Au cours de ce travail, nous avons démontré qu’il est possible de re-végétaliser un terril riche en As (8 g.kg-1 de substrat) grâce à un apport de terre exempte de polluants. Ce traitement diminue à la fois la contamination des plantes, mais améliore également la rétention de l’eau dans le sol, diminuant ainsi l’expression des contraintes hydriques. La stabilisation des sols par les plantes est importante car elle permettra de réduire l'érosion du terril et, donc, la fuite de polluant vers les cours d’eau bordant cette friche industrielle.

Pseudotsuga menziesii (Mirb) Franco, called Douglas fir or Oregon pine, is a species of Pinaceae family, native to North America. It was introduced in Europe in the nineteenth century from poorly documented genotypes. Its heartwood has the ability to resist to attacks caused by pathogens due to the presence of metabolites which display antioxidant, antifungal and insecticide activities [1, 2]. Chemical markers present in the three areas of its wood, namely sapwood, transition zone and heartwood were identified. In the meantime, their metabolic profiles were compared in order to establish a correlation between the presence of metabolites and genotype of trees. Wood sample was ground to a fine powder. The extraction was carried out by solvents of increasing polarity and the mixture acetone/H2O (70/30) provides the best yield. Optimal conditions for liquid solid extraction with the same solvent were found using accelerated solvent extractor at 100 ° C. The composition of each extract was determined by LC-UV. Qualitative analysis of the metabolite profile of extracts obtained from the three areas of wood was performed using tandem mass spectrometry (LC/MS and LC/MS/MS). Mass spectra data were recorded using electrospray ionization in the negative mode. Compounds were identified using the m/z ratio and their fragmentation patterns. Phenolic compounds were predominant with flavonoids followed by terpenes. Taxifolin and quercetin were found to be the most abundant metabolites present in the three areas along with pinobanskin, myricetin and dihydrokaempferol. More than sixty-four compounds were identified in heartwood, forty in transition zone and sixty in sapwood. The developed method in LC/MS/MS allowed the separation of isomers for taxifolin and its derivatives which are aglycons or O-glycosylated. They showed the same fragmentations and m/z at different retention times.

The plant cell wall is a complex 3D network composed of polysaccharides, lignin and proteins. The knowledge of the structure and content of each cell wall polymer is a prerequisite to understand their functions during plant development and adaptation but also to optimise their industrial applications. The analysis of cell wall compounds is complicated by their multiple molecular interactions. In this review, we present numerous methods to purify, characterise and quantify proteins, polysaccharides and lignin from the wall. Two kinds of approaches are detailed: the first presents in vitro methods which involve the breakdown of the molecular linkages between polymers thanking to chemical, physical and/or enzymatic treatments. The second approach describes in situ methods that allow the cell wall polymer characterisation thanking to many analytical techniques coupled with microscopy. If microscopy is the common point of all of them, their development is associated with improvement of analytical techniques, increasing their power of resolution.

The protein fraction of extracellular polymeric substance (EPS) from two anaerobic granular sludge samples was characterized with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and a far western blotting method. SDS-PAGE was used with various staining applications to obtain a protein (silver), glycoprotein [periodic acid–Shiff's (PAS)] or proteoglycan-like (Alcian blue at pH 2.5 (carboxylic group) or 1 (sulphated group)) fingerprint. The fingerprints of the EPS denatured protein from the two sludge samples differed. Some proteins are specific to Soluble (S) or Bound (B)-EPS (20–100 kDa). Denatured proteins with a polysaccharide moieties characterization are more present in B-EPS. Glycoproteins with α-d-mannosyl and/or α-d-glucosyl (90, 50, 40 kDa) were detected. Proteoglycan-like and sulphated proteoglycan-like substances are also detected, mainly in B-EPS. A 68 kDa sulphated proteoglycan-like substance contains two glucidic residue types: α-d-mannosyl and/or α-d-glucosyl and N-acetyl-β-d-glucosamine. Such heteroproteins are present around the membrane as well as the surface-layer from Archaea and from some bacteria. The glycoprotein and sulphated proteoglycan-like substance are assumed to contribute to anaerobic granule strength, thanks to their ability to perform interactions of various nature (ionic, hydrophobic, Ca2+ as divalent cation bridging, etc.).

The arabinogalactan proteins (AGPs) are O-glycosylated proteins consisting of less than 10 % proteins involved in many aspects of plant growth and development. Our work focuses on the development of a sensitive and easy method for monitoring partially purified AGP. Fluorescence excitation-emission matrix (EEM) spectrophotometry was used to select excitation-emission wavelengths of arabic gum and plant βglucosyl Yariv AGP for the SEC fluorescence system. Commercial as plant AGP were then separated by high performance size exclusion chromatography (SEC) on two columns in series (Agilent bio SEC 100 Å and 300 Å) coupled with 215 nm UV and 221/350 nm Ex/Em fluorescence detection. Separate molecules were then collected and analyzed by biochemical and immunological techniques, as well as by mass spectrometry. Here we demonstrated that AGP could be well separated and monitored by SEC UV/fluorescence proceeding. Biochemical, immunological and mass spectrometry confirmed that the analyzed peaks correspond to AGP molecules. If this method was not able to separate all the AGP expressed in plant, it was suitable to monitor β-glucosyl Yariv AGP evolution during plant development, such as in cocoa embryogenesis.

Exopolymeric substances (EPS) were extracted by EDTA from activated and anaerobic granular sludge. Due to the presence of EDTA in EPS extract, interferences were pointed out for the characterization of EPS by means of the colorimetric methods and fluorescence spectroscopy. Other methods have been investigated to characterize the EPS protein fraction. Size exclusion chromatography (SEC), performed at a fluorescence excitation–emission matrix of 221/360 nm (tryptophan protein-like substances) for detection, was suitable and allowed obtaining a fingerprint of the protein-like substance fractions and determining apparent molecular weight (MW). Polyacrylamide gel electrophoresis (PAGE) was performed under either native or denaturing conditions. Various staining applications after EPS migration are effective in obtaining a protein (silver staining) or glycoprotein (PAS staining) fingerprint or MW distribution. SEC and PAGE are both appropriate techniques for the qualitative characterization of protein fractions from EPS extracted by EDTA and moreover differentiate EPS according to sludge origin and type.

We studied the distribution of wall ingrowth (WI) polymers by probing thin sections of companion cells specialized as transfer cells in minor veins of Medicago sativa cv Gabès blade with affinity probes and antibodies specific to polysaccharides and glycoproteins. The wall polymers in the controls were similar in WIs and in the primary wall but differently distributed. The extent of labeling in these papillate WIs differed for JIM5 and JIM7 homogalacturonans but was in the same range for LM5 and LM6 rhamnogalacturonans and xyloglucans. These data show that WI enhancement probably requires arabinogalactan proteins (JIM8) mainly localized on the outer part of the primary wall and WIs. By comparison, NaCl-treated plants exhibited cell wall polysaccharide modifications indicating (1) an increase in unesterified homogalacturonans (JIM5), probably implicated in Na(+) binding and/or polysaccharide network interaction for limiting turgor variations in mesophyll cells; (2) enhancement of the xyloglucan network with an accumulation of fucosylated xyloglucans (CCRC-M1) known to increase the capacity of cellulose binding; and (3) specific recognition of JIM8 arabinogalactan proteins that could participate in both wall enlargement and cohesion by increasing the number of molecular interactions with the other polymers. In conclusion, the cell wall polysaccharide distribution in enlarged WIs might (1) participate in wall resistance to sequestration of Na(+), allowing a better control of hydric homeostasis in mesophyll cells to maintain metabolic activity in source leaves, and (2) maintain tolerance of M. sativa to NaCl.