•The wheat grain central endosperm cavity is filled with a gel-like structure.•The gel-like structure autofluoresces under UV excitation.•It contains fluorescent, water-absorbing and gel-forming ...feruloylated arabinoxylans.•High and stable cavity water content suggests a role in regulating grain hydration.
Cereal grains provide a substantial part of the calories for humans and animals. The main quality determinants of grains are polysaccharides (mainly starch but also dietary fibers such as arabinoxylans, mixed-linkage glucans) and proteins synthesized and accumulated during grain development in a specialized storage tissue: the endosperm.
In this study, the composition of a structure localized at the interface of the vascular tissues of the maternal plant and the seed endosperm was investigated. This structure is contained in the endosperm cavity where water and nutrients are transferred to support grain filling. While studying the wheat grain development, the cavity content was found to autofluoresce under UV light excitation. Combining multispectral analysis, Fourier-Transform infrared spectroscopy, immunolabeling and laser-dissection coupled with wet chemistry, we identified in the cavity arabinoxylans and hydroxycinnamic acids. The cavity content forms a “gel” in the developing grain, which persists in dry mature grain and during subsequent imbibition. Microscopic magnetic resonance imaging revealed that the gel is highly hydrated.
Our results suggest that arabinoxylans are synthesized by the nucellar epidermis, released in the cavity where they form a highly hydrated gel which might contribute to regulate grain hydration.
•Dry chemo-mechanical pretreatments impact biomass crystallinity and macroporosity.•Dry chemo-mechanical pretreatment increased the carboxylate production rate by RWS.•Dry chemo-mechanical pretreated ...biomass enhances the microbial xylanase activity.•Initial biomass macroporosity correlated with the biological acidogenic potential.
Innovative dry chemo- and chemo-mechanical pretreatments form an interesting approach for modifying the native physico-chemical composition of lignocellulose facilitating its microbial conversion to carboxylates. Here, the impact of four dry-pretreatment conditions on the microbial transformation of wheat straw was assessed: milling to 2mm and 100µm, and NaOH chemical impregnation at high substrate concentrations combined with milling at 2mm and 100µm. Pretreatment effect was assessed in the light of substrate structure and composition, its impact on the acidogenic potential and the major enzyme activities of a rumen-derived microbial consortium RWS. Chemo-mechanical pretreatment strongly modified the substrate macroporosity. The highest carboxylate production rate was reached after dry chemo-mechanical treatment with NaOH at 100µm. A positive impact of the dry chemo-mechanical treatment on xylanase activity was observed also. These results underline that increasing substrate macroporosity by dry chemo-mechanical pretreatment had a positive impact on the microbial acidogenic potential.
Cereal crop by-products are a promising source of renewable raw material for the production of biofuel from lignocellulose. However, their enzymatic conversion to fermentable sugars is detrimentally ...affected by lignins. Here the characterization of the Brachypodium Bd5139 mutant provided with a single nucleotide mutation in the caffeic acid O-methyltransferase BdCOMT6 gene is reported. This BdCOMT6-deficient mutant displayed a moderately altered lignification in mature stems. The lignin-related BdCOMT6 gene was also found to be expressed in grains, and the alterations of Bd5139 grain lignins were found to mirror nicely those evidenced in stem lignins. The Bd5139 grains displayed similar size and composition to the control. Complementation experiments carried out by introducing the mutated gene into the AtCOMT1-deficient Arabidopsis mutant demonstrated that the mutated BdCOMT6 protein was still functional. Such a moderate down-regulation of lignin-related COMT enzyme reduced the straw recalcitrance to saccharification, without compromising the vegetative or reproductive development of the plant.
Starch and fibre can be extracted, using wet or dry processes, from a variety of grain legumes and used as ingredients for food. a-Galactosides can be isolated during wet processes from the soluble ...extract. Starch isolates or concentrates are mostly produced from peas, whereas dietary fibre fractions from peas and soyabean are commercially available. The physico-chemical characteristics of fibre fractions very much depend on their origin, outer fibres being very cellulosic whereas inner fibres contain a majority of pectic substances. Inner fibres are often used as texturing agents whereas outer fibres find their main uses in bakery and extruded products, where they can be introduced to increase the fibre content of the food. Most investigations on impacts on health have been performed on soyabean fibres. When positive observations were made on lipaemia, glucose tolerance or faecal excretion, they were unfortunately often obtained after non-realistic daily doses of fibres. Legume starches contain a higher amount of amylose than most cereal or tuber starches. This confers these starches a lower bioavailability than that of most starches, when raw or retrograded. Their low glycaemic index can be considered as beneficial for health and especially for the prevention of diseases related to insulin resistance. When partly retrograded, these starches can provide significant amount of butyrate to the colonic epithelium and may help in colon cancer prevention. a-Galactosides are usually considered as responsible for flatus but their apparent prebiotic effects may be an opportunity to valorize these oligosaccharides.
Monoclonal antibodies were raised against rhamnogalacturonan I backbone, a pectin domain, using Arabidopsis thaliana seed mucilage-derived rhamnogalacturonan I oligosaccharides—BSA conjugates. Two ...monoclonal antibodies, designated INRA-RU1 and INRA-RU2, selected for further characterization, were specific for the backbone of rhamnogalacturonan I, displaying no binding activity against the other pectin domains i.e. homogalacturonans, galactans or arabinans. A range of oligosaccharides was prepared by enzymatic digestion of rhamnogalacturonan I isolated from Arabidopsis thaliana seed mucilage and from sugar beet pectin, purified by low-pressure chromatography and characterized by high-performance anion-exchange chromatography and mass spectrometry. These rhamnogalacturonan I oligomers were used to characterize the binding site of the two monoclonal antibodies by competitive inhibition. Both INRA-RU1 and INRA-RU2 showed maximal binding to the →2)-α-l-rhamnosep-(1→4)-α-d-galacturonic acid p-(1→₇ structural motif but differed in their minimum binding requirement. INRA-RU2 required at least two disaccharide (rhamnose-galacturonic acid) repeats for the antibody to bind, while INRA-RU1 required a minimum of six disaccharide repeats. Furthermore, the binding capacity of INRA-RU1 decreased steeply as the number of disaccharide repeats go beyond seven. Each of these antibodies reacted with hairy regions isolated from sugar beet pectin. Immunofluorescence microscopy indicated that both antibodies can be readily used to detect rhamnogalacturonan I epitopes in various cell wall samples.
Apple texture is one of the critical quality features for the consumer. Texture depends on several factors that are difficult to control and which change with freezing. To better understand the ...mechanisms involved in the texture degradation of apple tissues following freezing/thawing, our approach was to combine mechanical properties, cellular structure and cell wall composition measurements on fresh and thawed apples (Granny Smith) after three different freezing protocols (at −20
°C, −80
°C and −196
°C). This work highlighted the interest of applying macrovision and image texture analysis to quantify the freezing effects on cellular structure and ice crystal size. Freezing at −20
°C and after immersion into liquid nitrogen were the protocols affecting the most fruit texture leading to cell membrane breakage resulting in cell wall collapse and tissue breakage, respectively, which accounted for the mechanical behaviour of the samples. All freezing protocols induced vacuole burst showing that the turgor pressure preservation remains critical during the freezing process.
•Lignins were detected earlier than previously reported in developing wheat grain.•Alkali-releasable ferulate increased during development of grain outer layers.•Ferulate and lignin monomers ...releasable from pericarp decreased at grain maturity.•Resistant bonds between cell wall polymers probably form at maturity.
Important biological, nutritional and technological roles are attributed to cell wall polymers from cereal grains. The composition of cell walls in dry wheat grain has been well studied, however less is known about cell wall deposition and modification in the grain outer layers during grain development.
In this study, the composition of cell walls in the outer layers of the wheat grain (Triticum aestivum Recital cultivar) was investigated during grain development, with a focus on cell wall phenolics. We discovered that lignification of outer layers begins earlier than previously reported and long before the grain reaches its final size. Cell wall feruloylation increased in development. However, in the late stages, the amount of ferulate releasable by mild alkaline hydrolysis was reduced as well as the yield of lignin-derived thioacidolysis monomers. These reductions indicate that new ferulate-mediated cross-linkages of cell wall polymers appeared as well as new resistant interunit bonds in lignins. The formation of these additional linkages more specifically occurred in the outer pericarp.
Our results raised the possibility that stiffening of cell walls occur at late development stages in the outer pericarp and might contribute to the restriction of the grain radial growth.
•Wheat tissues had specific reproducible autofluorescence profiles using macro imaging.•Wheat pericarp and aleurone layer fluoresce differently in air or water.•Autofluorescence of wheat tissues is ...similar before and after grinding.•Multispectral fluorescence imaging can help identifying the histological origins.
Many plant tissues can be observed thanks to the autofluorescence of their cell wall components. Multispectral autofluorescence imaging at the macroscopic scale is a rapid efficient way of observing samples with a large field of view (>1cm2) and a nice resolution (<3μm per pixel). The objective of this work was to evaluate autofluorescence multispectral imaging at the macroscopic scale to identify the tissular origin of particles through their autofluorescence profiles. The pericarp and aleurone layer of wheat grain were used to compare contrasted autofluorescence profiles measured in cross sections and particles. Air and water mounting media were tested. Principal component analysis and variance analysis showed that the autofluorescence properties were retained from cross sections to particles. The mounting media modified the autofluorescence profiles and water was found to be better to observe both particles and sections. These results show that multispectral autofluorescence imaging at the macroscopic scale can be used to identify the histological origin of particles.
Solid-state fermentation is a potential technology for developing lignocellulosic biomass-based biorefineries. This work dealt with solid-state fermentation for carboxylates production from corn ...stover, as building blocks for a lignocellulosic feedstock-based biorefinery. The effect of extrusion pretreatment, together with the action of a microbial consortia and hydrolytic enzymes as biotic triggers, was investigated on corn stover conversion, microbial metabolic pathways, and populations. The extrusion caused changes in the physical and morphological characteristics, without altering the biochemical composition of the corn stover. Extrusion also led to remarkable differences in the composition of the indigenous microbial population of the substrate. Consequently, it affected the structure of community developed after fermentation and the substrate conversion yield, which increased by 118% (from 23 ± 4 gCOD/kgVSi obtained with raw substrate to 51 ± 1 gCOD/kgVSi with extruded corn stover) with regard to self-fermentation experiments. The use of activated sludge as inoculum further increased the total substrate conversion into carboxylates, up to 60 ± 2 gCOD/kgVSi, and shaped the microbial communities (mainly composed of bacteria from the
Clostridia
and
Bacteroidia
classes) with subsequent homogenization of the fermentation pathways. The addition of hydrolytic enzymes into the reactors further increased the corn stover conversion, leading to a maximum yield of 142 ± 1 gCOD/kgVSi. Thus, extrusion pretreatment combined with the use of an inoculum and enzyme addition increased by 506% corn stover conversion into carboxylates. Beside biomass pretreatment, the results of this study indicated that biotic factor greatly impacted solid-state fermentation by shaping the microbial communities and related metabolic pathways.
Mannan is a class of cell wall polysaccharides widespread in the plant kingdom. Mannan structure and properties vary according to species and organ. The cell walls of cereal grains have been ...extensively studied due to their role in cereal processing and to their beneficial effect on human health as dietary fiber. Recently, we showed that mannan in wheat (Triticum aestivum) grain endosperm has a linear structure of β-1,4-linked mannose residues. The aim of this work was to study the biosynthesis and function of wheat grain mannan. We showed that mannan is deposited in the endosperm early during grain development, and we identified candidate mannan biosynthetic genes expressed in the endosperm. The functional study in wheat was unsuccessful therefore our best candidate genes were expressed in heterologous systems. The endosperm-specificTaCslA12 gene expressed in Pichia pastoris and in an Arabidopsis thaliana mutant depleted in glucomannan led to the production of wheat-like linear mannan lacking glucose residues and with moderate acetylation. Therefore, this gene encodes a mannan synthase and is likely responsible for the synthesis of wheat endosperm mannan.
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