Wall Ultrastructure Research Articles

Honey Bee Pollen in Meagre (Argyrosomus regius) Juvenile Diets: Effects on Growth, Diet Digestibility, Intestinal Traits, and Biochemical Markers Related to Health and Stress.

Simple SummaryRecently, several studies have focused on the use of nutraceuticals and honey bee products to improve the welfare and sustainability of animal husbandry. Honey bee pollen is rich in bioactive substances, presenting a strong antioxidant activity with possible positive effects on growth performance and non-specific immune responses in reared fish. Despite its favorable characteristics, the addition of honey bee pollen to a meagre (Argyrosomus regius) diet in our trial resulted in a reduction of growth performances and diet digestibility, histological alterations of intestinal morphology, and high levels of biomolecular stress markers, probably due to its complex ultrastructure, which is indigestible for monogastric animals. These negative effects could be overcome by using bioactive component extraction methods and thus eliminating the indigestible fractions. Our results confirmed the general assumption that it should always be considered necessary to test nutraceutical additives of natural origin in a given species in order to verify the effective positive action and exclude any negative repercussions on animal health.This research aimed to evaluate the effects of the inclusion of honey bee pollen (HBP) in meagre (Argyrosoumus regius) juveniles’ diets on growth performance, diet digestibility, intestinal morphology, and immunohistochemistry. Furthermore, stress-related molecular markers and biochemical blood profile of fish were evaluated, together with mineral trace and toxic element concentration in pollen and diets. Specimens of meagre (360) of 3.34 ± 0.14 g initial body weight, were randomly allocated to twelve 500 L circular tanks (30 fish per tank). Four diets were formulated: a control diet and three experimental diets with 1%, 2.5%, and 4% of HBP inclusion. All the growth parameters and crude protein and ether extract digestibility coefficients were negatively linearly affected by increased HBP inclusion (p < 0.0001). Histology of medium intestine showed slight signs of alterations in group HPB1 and HPB2.5 compared to control. Fish from HBP4 group showed severe alterations at the intestinal mucosa level. Immunohistochemical detection of TNF-α in the medium intestine showed the presence of TNF-α+ cells in the lamina propria, which resulted in accordance with the increased level of the TNF-α protein detected by immunoblotting in the liver. This stress situation was confirmed by the increased hepatic level of HSP70 (p < 0.05) in fish fed the HBP4 diet and by the linear decrease of total serum protein levels in HBP-containing diets (p < 0.0001). These negative effects can be related to the ultrastructure of the bee pollen grain walls, which make the bioactive substances unavailable and can irritate the intestine of a carnivorous fish such as meagre.

Wall ultrastructure of the Permian pollen grain Lueckisporites virkkiae Potonié et Klaus 1954 emend. Clarke : Evidence for botanical affinity

Permian spore–pollen assemblages are dominated by striate bisaccate pollen grains. The botanical affinity of many of these is unknown, which limits their value in ecological reconstruction and biostratigraphy. Lueckisporites virkkiae Potonié et Klaus 1954 emend. Clarke 1965 is a bitaeniate bisaccate pollen grain of uncertain botanical affinity that dominates late Permian Euramerican assemblages, but also bridges phytogeographical provinces. It is classified into three “variants,” which capture a spectrum of morphological variability, but further complicates matters by questioning whether it comprises a natural taxon. Here we report on a TEM analysis of the gross structure and wall ultrastructure of L. virkkiae variants from the upper Permian (Lopingian) of Kimberley, Nottinghamshire, UK. The variants have the same gross structure and wall ultrastructure. The exine consists of a three-layered ektexine composed of a thin tectum, an alveolate infratectum and a thin foot layer, subtended by a solid endexine. Alveolae are either small and rounded, columellate-like, or irregularly shaped, depending on whether the exine is viewed in transverse or longitudinal section. Sacci are protosaccate, with narrow and irregular, or large and voluminous, endoreticulations. Similarities in gross structure and wall ultrastructure suggest that the current taxonomic grouping is correct with the variants representing the range of variation within a single species. We suggest that the differences in morphology are subtle and attributable to harmomegarthy. Comparisons with fossil and modern conifer pollen grains suggest an affinity with conifers. Co-occurrence with the Zechstein conifer macrofossil Pseudovoltzia liebeana (Geinitz) Florin 1880 (Majonicaceae) indicates this as a possible source.

Palynological analysis of the genus Dryopteris Adans. (Dryopteridaceae) in Argentina.

The spore morphology and wall ultrastructure of Dryopteris filix-mas, D. patula and D. wallichiana from Argentina were studied using light microscope, scanning and transmission electron microscope. The study was carried out with herbarium material from Argentine institutions. Equatorial diameters, polar diameters and laesura length were measured. The spores are monoletes with a rugate ornamentation. The folds are short to long, inflate, irregular in shape and size, and varying from subglobose to elongate. The perispore surface is rugulate. The exospore of all the species analyzed is two-layered in section. Simple and branched channels are also present. The perispore is composed of two layers, the inner one forms the ornamentation and the outer covers all the outer and inner surfaces. Some abnormalities, such as globose, triangular or twisted spores were observed. The morphology and ultrastructure of the species are very similar. The differences observed are related to the length and thickness of the perispore folds. The characteristics of these spores would not provide relevant information to differentiate species or sections within the genus, but can provide information for phylogenetic studies as well as for alterations in the biological cycles.

Analyzing Plant Cell Wall Ultrastructure by Scanning Near-Field Optical Microscopy (SNOM).

The importance of lignocellulosic materials in various research areas including for example biorefinery processes and new biomaterials is constantly rising. Therefore, a detailed knowledge on the macromolecular assembly of plant cell walls is needed. However, despite the tremendous progress in the structural and chemical analysis of plant cell walls there are still uncertainties concerning the ultrastructure. This is either due to Rayleigh criterion based limitations of the analytical methods or the relatively low chemical information gained with high spatial resolution techniques. In this chapter scanning near field optical microscopy (SNOM) is presented as a powerful tool for the subdiffraction limited chemical and structural characterization of plant cell walls.

Anatomy of the Windmill Palm (Trachycarpus fortunei) and Its Application Potential

The windmill palm (Trachycarpus fortunei (Hook.) H. Wendl.) is widely distributed and is an important potential source of lignocellulosic materials. The lack of knowledge on the anatomy of the windmill palm has led to its inefficient use. In this paper, the diversity in vascular bundle types, shape, surface, and tissue proportions in the leaf sheaths and stems were studied with digital microscopy and scanning electron microscope (SEM). Simultaneously, fiber dimensions, fiber surfaces, cell wall ultrastructure, and micromechanics were studied with atomic force microscopy (AFM) and a nanoindenter. There is diversity among vascular bundles in stems and leaf sheaths. All vascular bundles in the stems are type B (circular vascular tissue (VT) at the edge of the fibrous sheath (FS)) while the leaf sheath vascular bundles mostly belong to type C (aliform (VT) at the center of the (FS), with the wings of the (VT) extending to the edge of the vascular bundles). In addition, variation among the vascular bundle area and tissue proportion in the radial direction of the stems and different layers of the leaf sheaths is also significant. Microscopically, the fibers in the stem are much wider and longer than that in the leaf sheath. The secondary walls of stem fibers are triple layered while those in the leaf sheath are double layered. The indentation modulus and hardness of the cell wall of leaf sheath fibers are higher than that of the stem. An independent sample t-test also showed a significant difference between stems and leaf sheaths. All this indicates that windmill palm stems and leaf sheaths are two different materials and have different application prospects.

Correlation of mechanical properties of peach slices with cell wall polysaccharides and cell morphology during hot air predrying

The mechanical properties, cell wall polysaccharides, cell morphology, and cell wall ultrastructure of peach slices during hot air predrying were investigated. A continuous decrease was observed with respect to the elastic modulus and maximum stress, and total pectin, hemicellulose, and cellulose contents of the cell wall in the peach samples with an increase in the drying time. However, an increasing trend was seen in the viscosity index and fractal dimension of the cell cross-sectional area. The elastic modulus and maximum stress were positively correlated and grouped with the cellulose content and total pectin content. The viscosity index was close to the fractal dimension of the cell cross-sectional area and was negatively correlated with the content of cell wall polysaccharides. In this study, the correlations of mechanical properties with cell wall polysaccharides and cell morphology during hot air predrying were constructed for guiding the textural quality improvement of hot air dried products. Practical applications Hot air drying is often used in the dehydrated products of fruits and vegetables. In this study, peach fruit was selected to study the mechanical properties of peach slices during the hot air predrying process by observing the cell wall polysaccharides, the morphology, and microstructure of cells. Their changing trend and correlations were revealed. The results of this study can be useful for the process optimization and textural quality improvement of hot air dried fruits and vegetables.

Relationship of wood cell wall ultrastructure to bacterial degradation of wood

ABSTRACT This review presents information on the relationship of ultrastructure and composition of wood cell walls, in order to understand how wood degrading bacteria utilise cell wall components for their nutrition. A brief outline of the structure and composition of plant cell walls and the degradation patterns associated with bacterial degradation of wood cell walls precedes the description of the relationship of cell wall micro- and ultrastructure to bacterial degradation of the cell wall. The main topics covered are cell wall structure and composition, patterns of cell wall degradation by erosion and tunnelling bacteria, and the relationship of cell wall ultrastructure and composition to wood degradation by erosion and tunnelling bacteria. Finally, pertinent information from select recent studies employing molecular approaches to identify bacteria which can degrade lignin and other wood cell wall components is presented, and prospects for future investigations on wood degrading bacteria are explored.

Deterioration of the cell wall in waterlogged wooden archeological artifacts, 2400 years old

ABSTRACT The relationship between the cell wall ultrastructure of waterlogged wooden archeological artifacts and the state of water bound to cell walls and free in voids is fundamental to develop consolidating and drying technologies. Herein, a lacquer-wooden ware and a boat-coffin dating 4th century BC were selected as representative artifacts to study. Wood anatomy results indicated that they belonged to Idesia sp. and Machilus sp., respectively. They exhibited a typical spongy texture, as revealed by SEM observations, and their water contents had increased significantly. Solid state NMR, Py-GC/MS, imaging FTIR microscopy and 2D-XRD results demonstrated that the deterioration resulted from the partial cleavages of both polysaccharide backbones and cellulose hydrogen-bonding networks, almost complete elimination of acetyl side chains of hemicellulose, the partial depletion of β-O-4 interlinks, as well as oxidation and demethylation/demethoxylation of lignin. These further caused the disoriented arrangement of crystalline cellulose, and the decrease in cellulose crystallite dimensions and crystallinity. In consequence, mesopores and macropores formed, and the number of moisture-adsorbed sites and their accessibility increased. Moreover, results on free water deduced by the changes of pore structure and the maximum monolayer water capacity achieved by the GAB model indicated that water in waterlogged archeological wooden artifacts was mainly free water in mesopores.

Wood cell wall ultrastructure The key to understanding wood properties and behaviour

ABSTRACT In the last 100 years, major advances have been made in understanding wood cell wall ultrastructure in tracheids, fibres, vessels and parenchyma and its relationship with xylem function and wood properties. This review will focus on how the development of imaging techniques and their application to wood cell walls has led to an understanding of cell wall organisation and the relationship between micro and macro scale properties in wood and wood-based materials. Topics such as wood formation, wood chemistry and reaction wood have recently been reviewed elsewhere and are considered only briefly in this review. Two features of wood cell walls have dominated the literature; orientation and layering of cellulose which determines the longitudinal stiffness of wood, and the distribution (topochemistry) of lignin which determines compression strength and pulping properties.

Unravelling the consequences of ultra-fine milling on physical and chemical characteristics of flax fibres

In recent years, lignocellulosic biomass has been increasingly used in various applications. While for many of them the plant materials require coarse milling, some new applications for green chemistry, bio-energy and bio-packaging necessitate comminution to obtain very finely calibrated particles (below 200μm in size). This milling step is not inconsequential for lignocellulosic materials and can influence the physical and chemical characteristics of the powder. However, these different effects are still poorly understood. In this work, we study the impact of ultra-fine milling on the physico-chemical properties of plant fibres. Flax was chosen for this study because of its well-described structure and biochemical composition, making it a model material. Our main results evidence a strong impact of ball milling on flax fibre aspect ratio but also on cell wall ultrastructure and composition. Cellulose content and crystallinity significantly decrease with milling time, leading to higher water sorption and lower thermal stability.

New insights on the paleobiology, biostratigraphy and paleogeography of the pre-Sturtian microfossil index taxon Cerebrosphaera

Important biological and geological events occurred during the early to middle Neoproterozoic. Among diversifying eukaryotic assemblages, populations of Cerebrosphaera, a distinctive and robust organic-walled vesicular microfossil (acritarch), show restricted stratigraphic distribution in several late Tonian to early Cryogenian worldwide successions. Here, we report the first occurrence of this taxon in Africa, in the Bouenza Subgroup (Republic of the Congo), enlarging its paleogeographic distribution and biostratigraphic significance. We also attempt to determine its biological affinity, using a combined analytical approach on specimens from the Kanpa and Hussar formations, Australia, and from the Svanbergfjellet Formation, Spitsbergen. Morphological and quantitative analyses were performed using light microscopy and scanning electron microscopy. The analyses show fine-scale morphological details and a morphological continuum between the former species Cerebrosphaera ananguae and Cerebrosphaera buickii, confirming their synonymy as proposed by a recently revised taxonomy. These observations also highlighted the presence of a thin external envelope, previously reported but formerly described and illustrated here for the first time. The characteristics of this envelope, the large diameter range of the vesicles, and the absence of excystment structure, suggest that Cerebrosphaera was a metabolically active growing cell. Ultrastructural analyses performed with TEM revealed a complex multilayered wall ultrastructure. The molecular composition and thermal maturity of the organic walls were estimated using Infrared and Raman microspectroscopies. The wall of Cerebrosphaera has a highly aromatic composition with short/highly branched aliphatic chains. The complex morphology and wall ultrastructure, combined with the large size (not a criterion by itself) of Cerebrosphaera, confirm its eukaryotic nature. Comparison with strikingly similar modern analogues permits to suggest a possible affinity to stem metazoan eggs, based on morphology and ultrastructure, but the chemical composition is unlike known biopolymers. This hypothesis is also consistent with estimates from molecular clocks. If confirmed, our results would provide an older direct evidence for stem metazoans than the Cryogenian biomarker and Ediacaran body fossil records. Our study reveals that Cerebrosphaera populations are important for Neoproterozoic biostratigraphy, but also participated to the diversification of eukaryotes in worldwide connected oceans.

Effects of nitric oxide fumigation treatment on retarding cell wall degradation and delaying softening of winter jujube (Ziziphus jujuba Mill. cv. Dongzao) fruit during storage

Winter jujube (Ziziphus jujuba Mill. cv. Dongzao) fruit were fumigated with 20 μL L−1 of nitric oxide (NO) for 3 h and then stored at 0 ± 1 ºC with 90–95 % relative humidity for 75 d. The effects of NO fumigation on changes in fruit firmness, activities of cell wall degrading enzymes, the compositions of the cell walls, as well as the cell wall ultrastructure in the winter jujube fruit were investigated during storage. The results showed that the decline in fruit firmness, contents of CDTA-soluble pectin (CSP), Na2CO3-soluble pectin (SSP), hemicellulose, and cellulose, as well as the increase in water-soluble pectin (WSP) of the winter jujube fruit could be delayed by NO treatment relative to the control. NO treatment also suppressed the activities of pectin methylesterase (PME; EC 3.1.1.11), polygalacturonase (PG; EC 3.2.1.15), β-galactosidase (β-Gal; EC 3.2.1.23) and cellulase (EC 3.2.1.4) during storage. Furthermore, ultrastructural observations revealed that NO treatment delayed the disruption of cell wall structure of the winter jujube fruit. These results indicated that retarding the disassembly of cell walls polysaccharides via inhibition of an array of cell wall degrading enzymes activities, which in turn would help maintain the intactness of the cell walls, might be one possible mechanism by which NO treatment can help prevent softening of the winter jujube fruit.

Cell wall polysaccharides degradation and ultrastructure modification of apricot during storage at a near freezing temperature

The effects of near freezing temperature (NFT) storage at −1.9 °C on cell wall degradation of ‘Shushanggan’ apricot was studied comparing to 0 °C and 5 °C storage. Our results indicated that NFT storage strongly inhibited the solubilization of Na2CO3-soluble pectin and cellulose, by the suppression of cell wall modifying enzymes (polygalacturonase, β-Galactosidase, pectin methyl esterase and cellulase) and related genes expressions. The loss of side chains was the main modification in CDTA (Cyclohexane-diamine-tetraacetic Acid)-soluble pectin during storage and made the main contribution to the softening of apricot, while the loss of side chain was suppressed by NFT storage. Microscopic observation showed that NFT storage delayed the degradation of pectin fraction and protected cell wall structure from loosing. This study proves that NFT storage is an effective technology to suppress the cell wall polysaccharides degradation and ultrastructure modification of apricot.

Postantibiotic and Sub-MIC Effects of Exebacase (Lysin CF-301) Enhance Antimicrobial Activity against Staphylococcus aureus.

CF-301 (exebacase) is a recombinantly produced bacteriophage-derived lysin (cell wall hydrolase) and is the first agent of this class to enter clinical development in the United States for treating bacteremia including endocarditis due to Staphylococcus aureus Whereas rapid bactericidal activity is the hallmark in vitro and in vivo response to CF-301 at exposures higher than the MIC, prolonged antimicrobial activity, mediated by cell wall damage, is predicted at concentrations less than the MIC. In the current study, a series of in vitro pharmacodynamic parameters, including the postantibiotic effect (PAE), postantibiotic sub-MIC effect (PA-SME), and sub-MIC effect (SME), were studied to determine how short-duration and sub-MIC CF-301 exposures affect the growth of surviving staphylococci and extend its antimicrobial activity. Mean PAE, PA-SME, and SME values up to 4.8, 9.3, and 9.8 h, respectively, were observed against 14 staphylococcal strains tested in human serum; growth delays were extended by 6 h in the presence of daptomycin. Exposures to CF-301 at sub-MIC levels as low as 0.001× to 0.01× MIC (∼1 to 10 ng/ml) resulted in aberrant cell wall ultrastructure, increased membrane permeability, dissipation of membrane potential, and inhibition of virulence phenotypes, including agglutination and biofilm formation. A mouse thigh infection model designed to study the PAE was used to confirm our findings and demonstrate in vivo growth delays of ≥19.3 h. Our findings suggest that at CF-301 concentrations less than the MIC during therapeutic use, sustained reductions in bacterial fitness and virulence may substantially enhance efficacy.

Dynamic Changes in Arabinogalactan-Protein, Pectin, Xyloglucan and Xylan Composition of the Cell Wall During Microspore Embryogenesis in Brassica napus.

Microspore embryogenesis is a manifestation of plant cell totipotency whereby new cell walls are formed as a consequence of the embryogenic switch. In particular, the callose-rich subintinal layer created immediately upon induction of embryogenesis was recently related to protection against stress. However, little is currently known about the functional significance of other compositional changes undergone by the walls of embryogenic microspores. We characterized these changes in Brassica napus at different stages during induction of embryogenic microspores and development of microspore-derived embryos (MDEs) by using a series of monoclonal antibodies specific for cell wall components, including arabinogalactan-proteins (AGPs), pectins, xyloglucan and xylan. We used JIM13, JIM8, JIM14 and JIM16 for AGPs, CCRC-M13, LM5, LM6, JIM7, JIM5 and LM7 for pectins, CCRC-M1 and LM15 for xyloglucan, and LM11 for xylan. By transmission electron microscopy and quantification of immunogold labeling on high-pressure frozen, freeze-substituted samples, we profiled the changes in cell wall ultrastructure and composition at the different stages of microspore embryogenesis. As a reference to compare with, we also studied in vivo microspores and maturing pollen grains. We showed that the cell wall of embryogenic microspores is a highly dynamic structure whose architecture, arrangement and composition changes dramatically as microspores undergo embryogenesis and then transform into MDEs. Upon induction, the composition of the preexisting microspore intine walls is remodeled, and unusual walls with a unique structure and composition are formed. Changes in AGP composition were related to developmental fate. In particular, AGPs containing the JIM13 epitope were massively excreted into the cell apoplast, and appeared associated to cell totipotency. According to the ultrastructure and the pectin and xyloglucan composition of these walls, we deduced that commitment to embryogenesis induces the formation of fragile, plastic and deformable cell walls, which allow for cell expansion and microspore growth. We also showed that these special walls are transient, since cell wall composition in microspore-derived embryos was completely different. Thus, once adopted the embryogenic developmental pathway and far from the effects of heat shock exposure, cell wall biosynthesis would approach the structure, composition and properties of conventional cell walls.

New insights in the male anatomy, spermatophore formation, and sperm structure in Atyidae: The red cherry shrimp Neocaridina davidi

AbstractThis study aims to analyze the functional anatomy of the male reproductive system in Neocaridina davidi, a very popular ornamental species of caridean shrimp. Mature males were cold‐anaesthetized and their reproductive systems were dissected for histological and histochemical analysis, while the spermatozoa and spermatophore wall ultrastructure were analyzed under transmission electron microscopy. The male reproductive system consisted of two coiled testes, which were continuous with the vasa deferentia. Testes were positioned on the dorsal side of the cephalothorax above the hepatopancreas, and comprised seminiferous tubules where spermatogenesis occurred. Each vas deferens (VD) was a long tube dorsolaterally positioned with respect to the hepatopancreas, and increased in diameter at the distal end. Three regions could be recognized in the VD: proximal, middle, and distal. The proximal region had a cylindrical epithelium with secretory cells. The middle region (or typhlosole) had a dorsal fold (or typhlosole) with a thick columnar epithelium and high secretory activity. The spermatophore was a continuous cord with three acellular layers, which were mainly characterized by the presence of neutral glycoconjugates and proteins. The sperm morphology was distinct from the inverted cup‐shaped spermatozoa observed in the majority of caridean shrimps. The spermatozoa in specimens of N. davidi were spherical in shape, with a cross‐striated, single, short spike, and arranged in clusters of three or four sperm cells. The composition of the spermatophore, and the arrangement and form of the spermatozoa, seem to be unique in comparison to other species of Caridea.

Current achievements in the mechanically pretreated conversion of plant biomass.

At present, "mechanochemistry" is synonymous with "grinding," according to the views of a significant number of scientists and technologists. Often, one comes across the opinion that "the less the particle size, the better." The cases of considering chemical reactions occurring during pretreatment, as well as considering changes in the ultrastructure of cell walls are extremely rare. Also, the wrong choice of the type of mechanical impact and the equipment used in most cases leads to excessive consumption of electrical energy and reduce economic efficiency. The review presents the currently available published data on mechanically activated processes for the pretreatment of plant materials and shows that when using mechanical treatment, it is necessary to look more closely at the phenomena occurring, rather than reducing everything to the production of fine and ultrafine powders. As a result of mechanical action, active surface radicals can form, hydrothermal chemical processes can occur, and mechanocomposites can form. The role of interphase processes, changes in surface chemistry, related dimensional effects, and the disordering of the crystal structure and amorphization should be taken into account. In addition, the physicochemical insights in mechanical pretreatment makeit possible to more efficiently use the energy delivered to the material, and, consequently, increase the economic efficiency of the activation process.

Quadricoccopsis gen. nov., a new genus of Quadricoccus-like algae in Oocystaceae from China (Trebouxiophyceae, Chlorophyta)

Members of Quadricoccus-like algae are characterized by oval to ellipsoid cells adherent to the bowl-shaped or stretched, empty mother cell walls and are common in phytoplankton of inland waters. To date, the morphologically similar genera Quadricoccus and Lobocystis are accepted for this group and the former had been phylogenetic positioned in Oocystaceae. In this study, seven strains of Quadricoccus-like algae were identified and successfully cultured in the laboratory. Light and electron microscope observations and phylogenetic analysis revealed that the strains represent three different species within a new genus, described here as Quadricoccopsis gen. nov. It differed from genus Quadricoccus by characteristically cell adherent mode that two pairs of daughter cells connected to the mother cell remnant respectively by the pole and median portion and distinguished from Lobocystis by variable 2-4-8 autospores, characteristically cell adherent mode and only found in limnetic water. The three new species, described here as Q. simplex, Q. parva and Q. glomerata, differed in cell size, colony morphology and autospore number. Phylogenetic analysis revealed the genus Quadricoccopsis in Oocystaceae and a close relationship with Oocystidium, which is far away from the phylogenetic position of Quadricoccus in one of the granulated clades. The Quadricoccus-like algae were, therefore, proved to be a paraphyly. Furthermore, apart from Planctonema-like algae, the Oocystaceae characteristic cell wall ultrastructure, which is multi-layered with cellulose fibrils in each layer perpendicular to those of the adjoining layer, is not shown again in genus Quadricoccopsis. Different cell wall substructures may be related to the colony formation mechanism. Whether the ultrastructure criterion is applied to all the Oocystaceae needs to be re-evaluated and, further, the definition of this family should be discussed.

Evolution of flax cell wall ultrastructure and mechanical properties during the retting step

Flax retting is a major bioprocess in the cultivation and extraction cycle of flax fibres. The aim of the present study is to improve the understanding of the evolution of fibre properties and ultrastructure caused by this process at the plant cell wall scale. Initially, investigations of the mechanical performances of the flax cell walls by Atomic Force Microscopy (AFM) in Peak Force mode revealed a significant increase (+33%) in the cell wall indentation modulus with retting time. Two complementary structural studies are presented here, namely using X-Ray Diffraction (XRD) and solid state Nuclear Magnetic Resonance (NMR). An estimation of the cellulose crystallinity index by XRD measurements, confirmed by NMR, shows an increase of 8% in crystallinity with retting mainly due to the disappearance of amorphous polymer. In addition, NMR investigations show a compaction of inaccessible cell wall polymers, combined with an increase in the relaxation times of the C4 carbon. This densification provides a structural explanation for the observed improvement in mechanical performance of the secondary wall of flax fibres during the field retting process.

Calicioid lichens and fungi in amber – Tracing extant lineages back to the Paleogene

Calicioid lichens and fungi are a polyphyletic grouping of tiny ascomycetes that accumulate a persistent spore mass (mazaedium) on top of their usually well-stalked ascomata (‘mazaediate fungi’). In addition to extant forms, six fossils of the group were previously known from European Paleogene amber. Here we report nine new fossils and analyze the preserved features of all fossils to assess their applicability for dating molecular phylogenies. Many fossils are extremely well preserved, allowing detailed comparisons with modern taxa. SEM investigation reveals that even fine details of ascospore wall ultrastructure correspond to those seen in extant specimens. All fossils can confidently be assigned to modern genera: three to Calicium (Caliciaceae, Lecanoromycetes), five to Chaenotheca (Coniocybaceae, Coniocybomycetes), six to Chaenothecopsis (Mycocaliciaceae, Eurotiales), and one to Phaeocalicium (Mycocaliciaceae, Eurotiales). Several Calicium and Chaenotheca fossils are assignable to specific lineages within their genera, while the Chaenothecopsis fossils demonstrate the extent of intraspecific variation within one such lineage. Some features in the morphology of Chaenotheca succina nov. sp. seem to be ancestral as they have not been reported from modern species of the genus.