Inhibitory Effects of CaCl2 and Pectin Methylesterase on Fruit Softening of Raspberry during Cold Storage

Quality of raspberry fruit experiences a rapid decline after harvest due to its vulnerable texture and high moisture content. Application of calcium chloride (CaCl2) combined with pectin methylesterase (PME) is efficient in delaying fruit softening. In this study, the effects of exogenous CaCl2 alone or in combination with PME on the structure of the cell wall, the molecular properties of pectin, and the amount of free water of raspberry during postharvest storage were investigated. The results showed that CaCl2 combined with PME treatment could maintain fruit firmness and inhibit weight loss. The treatment of CaCl2+PME maintained the cell wall structure via sustaining middle lamella integrity and reducing the activities of cell wall-degrading enzymes, such as polygalacturonase, pectin methylesterase, β-galactosidase, α-L-arabinofuranosidase, and β-xylosidase. In addition, CaCl2+PME treatment could effectively increase the content of chelate-soluble pectin (CSP) and develop a cross-linked structure between Ca2+ and CSP. Moreover, CaCl2+PME treatment was of benefit in maintaining free water content. CaCl2 in combination with PME treatment could be a promising method for inhibiting softening and maintaining the quality of postharvest raspberry during cold storage.

Anatomy and cell wall ultrastructure of sunflower stalk rind

The anatomy and ultrastructure of sunflower stalk rind are closely related to its conversion and utilization. We studied systematically the anatomy and ultrastructure of the stalk rind using light, scanning electron, transmission electron and fluorescence microscopy. The results showed that the stalk rind consisted of phloem fibers (PF), xylem fibers (XF), vessel elements (V), ground parenchyma cells (GPC), axial parenchyma cells (APC), xylem ray parenchyma cells (XRPC), and pith ray parenchyma cells (PRPC). These cell walls were divided into the middle lamella, primary wall, and secondary wall (S). It was found that the S of PF, XF and V was further divided into three layers (S1–S3), while the S of APC, GPC, XRPC and PRPC showed a non-layered cell wall organization or differentiated two (S1, S2) to seven layers (S1–S7). Our research revealed the plasmodesmata characteristics in the pit membranes (PMs) between parenchyma cells (inter-GPCs, inter-XRPCs, and inter-PRPCs). The morphology of the plasmodesmata varied with the types of parenchyma cells. The thickness and diameter of PMs between the cells (inter-Vs, V–XF, V–APC, and V–XRPC) were greater than that of PMs between parenchyma cells. The cell corners among parenchyma cells were intercellular space. The lignification degree of vessels was higher than that of parenchyma cells and fibers. The results will provide useful insights into the biological structure, conversion and utilization of sunflower stalk rind.

Mapping Pectic-Polysaccharide Epitopes in Cell Walls of Forage Chicory (Cichorium intybus) Leaves

The cell walls of forage chicory (Cichorium intybus) leaves are known to contain high proportions of pectic polysaccharides. However, little is known about the distribution of pectic polysaacharides among walls of different cell types/tissues and within walls. In this study, immunolabelling with four monoclonal antibodies was used to map the distribution of pectic polysaccharides in the cell walls of the laminae and midribs of these leaves. The antibodies JIM5 and JIM7 are specific for partially methyl-esterified homogalacturonans; LM5 and LM6 are specific for (1→4)-β-galactan and (1→5)-α-arabinan side chains, respectively, of rhamnogalacturonan I. All four antibodies labelled the walls of the epidermal cells with different intensities. JIM5 and JIM7, but not LM5 or LM6, labelled the middle lamella, tricellular junctions, and the corners of intercellular spaces of ground, xylem and phloem parenchyma. LM5, but not LM6, strongly labelled the walls of the few sclerenchyma fibres in the phloem of the midrib and lamina vascular bundles. The LM5 epitope was absent from some phloem parenchyma cells. LM6, but not LM5, strongly labelled the walls of the stomatal guard cells. The differential distribution of pectic epitopes among walls of different cell types and within walls may reflect the deposition and modification of these polysaccharides which are involved in cell wall properties and cell development.

Metabolomics Profiling of Brassinolide and Abscisic Acid In Response to High Temperature Stress

To determine the effect of abscisic acid (ABA), brassinolide (BR) and ABA+BR on grape quality under high temperature stress (HTS), various metabolites were analyzed. Compared with the control (CK), DL-tryptophan, D-raffinose, geniposidic acid, dodecanedioic acid and polyphenols were found to be higher after ABA treatment. After BR treatment, amino acids and poricoic acid B were higher than in CK. And carbohydrates and amino acids were upregulated after ABA+BR treatment. BR and ABA+BR treatment also induced higher endogenous ABA and epibrassinolide (EBR) contents. In addition, treated grape had higher soluble solid concentrations (SSC) and soluble sugar content, and delayed the degradation of middle lamella and microfibrils. Antioxidant and heat shock related genes were examined, which significantly increased in treated grape. The finding of this study suggested that ABA, BR and ABA+BR are very useful for alleviating high temperature damage by increasing the accumulation of osmotic adjustment substances, and endogenous hormones content.

Microbial Retting of Banana Pseudostem

Fibrous wastes correspond to approximately 54.3 wt.-% of a banana plant. The waste weight is mainly of bunches, stems, and leaves. These ligno-cellulosic fibre waste can be designed and tailored to yield value added products which can be exploited by the farmers, exporters and weavers. Pectin (plant cement) and hemicelluloses (22-25%) found in plant cell walls in the plate (middle lamella). These macromolecules have needs to be hydrolyzed faster without affecting the quality of fibre. The chemical and mechanical processes adopted for fibres actually affect color uptake, strength of fibre thus fetching lower price. We report a biological ecofriendly method of retting where pectinases produced by the endophytic microorganisms of banana pseudostem were used to separate the fibre bundles of cellulose.. Endophytes are microorganisms which live in close relationship with living plant tissues in a cooperative relationship delivering a few auxiliary metabolites and proteins with the possibility to hydrolyse a few plant-determined macromolecules of the host. They spend the entire or part of their lifecycle colonizing within sound tissues of the host plants.The endophytic pectinase helps to break down the cell walls and can find commercial application in extraction of fibres.Seven bacterial isolates were screened and isolated in pectinase screening agar medium [PSAM]. Pectinase producing endophytic bacteria is isolated from corm of Amorphophallus paeoniifolius reported by us are Staphylococcus sciuri, Exiguobacterium acetylicum, Exiguobacterium indicum which are good pectinase producers having specific activity of 8.26, 12.61, 6.81 respectively. Treatment of banana pseudostem with these microbes showed 91%, 94%, 96.7% loss in the total weight of stem leaving behind pure cellulose fibres.

Influence of Lignin On Plastic Flow Deformation of Wood

In this study, we clarified the influence of lignin in wood on its plastic flow deformation due to shear sliding of wood cells. Wood samples were subjected to delignification, where the lignin structure gradually changed, and characterized for their chemical and physicochemical properties, and deformability by free compression testing. The delignified wood deformed by efficient stretching and maintained its cell structures at a lower pressure compared to the untreated wood. The deformability was evaluated from two viewpoints: the initial resistance to plastic flow and final stretchability. The deformability of the delignified and untreated wood increased with increasing compressive temperature, even though the changes in molecular motility associated with the glass transition of lignin contributed minimally to the improvement in deformability. In the early stages of delignification, the molecular mass of lignin in the compound middle lamella decreased, which reduced the initial resistance to plastic flow. However, during the early stages of delignification, the stretchability of delignified wood was scarcely affected by changes in lignin. As the amount of lignin was further reduced and delignification proceeded in the vicinity of the polysaccharides, the stretchability significantly improved. The correlation between chemical and physicochemical properties and plastic flow deformability presented in this paper will be helpful for low-energy and highly productive forming of solid-state wood.

Shattering tolerance in Brassica napus L.

Brassica is the second-largest oilseed crop after Soybean. The total production of Brassica in the overall world is 71 million tons. In Pakistan, its total production per unit area is very low. Biotic and abiotic stresses mainly affect the brassica crop. In agriculture, shattering is the dispersal of crops seeds before their ripening. The pod wall shatters and breaks apart when it loses its hydration and cells split in a dehiscence zone organized at a suture between the edge of the lignified pod and the vascular tissue replum. The degeneration of middle lamella and loss of cellular cohesion in the dehiscence zone are the main reasons for pod shattering and seed losses. Grain yield losses in Brassica vary from 10 to 25 percent due to shattering. More than 400 kg has-1 or 12% seed losses can be occurred due to pod shattering under unfavorable conditions. Insect pest and disease damage also accelerate ripening and pod shattering. The main breeding techniques for developing rapeseed grain yield potential are a good knowledge and application of the morphological, physiological, and genetic basis of grain yield. Modern technologies, such as embryo rescue, marker-assisted breeding, and novel variation (mutation), may make it much simpler to introduce new rapeseed types having shattering tolerance than traditional methods. Thus, an overview of anatomical and physiological aspects and genetics of shattering is presented in the context of recent advances in molecular genetics and several agronomic managements to avoid shattering in Brassica.

Mapping lignification dynamics with a combination of chemistry, data segmentation and ratiometric analysis

This article describes a new methodology for detailed mapping of the lignification capacity of plant cell walls that we have called "REPRISAL" for REPorter Ratiometrics Integrating Segmentation for Analyzing Lignification. REPRISAL consists of the combination of three separate approaches. In the first approach, H*, G* and S* monolignol chemical reporters, corresponding to p-coumaryl alcohol, coniferyl alcohol and sinapyl alcohol, are used to label the growing lignin polymer in a fluorescent triple labelling strategy based on the sequential use of 3 main bioorthogonal chemical reactions. In the second step, an automatic parametric and/or artificial intelligence (AI) segmentation algorithm is developed that assigns fluorescent image pixels to 3 distinct cell wall zones corresponding to cell corners (CC), compound middle lamella (CML) and secondary cell walls (SCW). The last step corresponds to the exploitation of a ratiometric approach enabling statistical analyses of differences in monolignol reporter distribution (ratiometric method 1) and proportions (ratiometric method 2) within the different cell wall zones. In order to demonstrate the potential of REPRISAL for investigating lignin formation we firstly describe its use to map developmentally-related changes in the lignification capacity of WT Arabidopsis interfascicular fiber cells. We then show how it can be used to reveal subtle phenotypical differences in lignification by analyzing the Arabidopsis prx64 peroxidase mutant and provide further evidence for the implication of the AtPRX64 protein in floral stem lignification. Finally, we demonstrate the general applicability of REPRISAL by using it to map lignification capacity in poplar, flax and maize.

Histochemical Structure and Tensile Properties of Birch Cork Cell Walls

Tensile tests of birch cork were performed in the tangential direction. Birch cork in the wet state showed significantly higher extensibility and toughness than those in the oven-dried state. The histochemical structure of birch cork was investigated by microscopic observation and spectroscopic analysis. Birch cork cell walls showed a three-layered structure. In transmission electron micrographs, osmium tetroxide stained the outer and inner layers, whereas potassium permanganate stained the middle and inner layers. After chemical treatment to remove suberin and lignin, the outer and inner layers disappeared and Fourier-transformed infrared spectra showed the cellulose I pattern. Polarizing light micrographs indicated that molecular chains in the outer and inner layers were oriented perpendicular to suberin lamination, whereas those in the inner layer showed longitudinal orientation. These results suggested that the outer and inner layers mainly consist of suberin, whereas the middle layer and compound middle lamella consist of lignin, cellulose, and other polysaccharides. We hypothesized a hierarchical model of the birch cork cell wall. The lignified cell wall with helical arrangement of cellulose microfibrils is sandwiched between two suberized walls. Cellulose microfibrils in the middle layer act like a spring and bear tensile loads. In the wet state, water and cellulose in the compound middle lamella transfer tensile stress between cells. In the dried state, this stress-transferal system functions poorly and fewer cells bear stress. Suberin in the outer and inner layers prevents absolute drying to maintain mechanical properties of the bark and to bear tensile stress caused by trunk diameter growth.

Characterisation of compound middle lamella isolated by a combination of wet-beating, sedimentation, and methanol dialysis

A simple method was developed to isolate the compound middle lamella (CML) in its natural state using wet-beating, sedimentation, and methanol dialysis. The isolated CML fraction was characterised. The CML was isolated in 2.0% yield from the mature wood of Ginkgo biloba. Scanning electron microscopy (SEM) images of the isolated CML fraction showed the presence of flake-like structures. However, some small amounts of secondary wall substance was observed. The lignin content of the CML fraction was approximately 45%. Thioacidolysis reactions with the isolated CML fraction revealed that the lignin β-O-4 structure was scarce in the isolated fraction and that numerous structures with two or more consecutive condensed-type bonds were present. Results from the solid-state carbon-13 nuclear magnetic resonance (13C NMR) experiments indicated that the CML fraction had low crystallinity, indicating low cellulose content. Liquid-state NMR analysis of the lignin from the CML fraction revealed the absence of p-hydroxyphenyl (H) units.