Aloe vera mucilage loaded gelatin electrospun fibers contained in polylactic acid coaxial system and polylactic acid and poly(e-caprolactone) tri-layer membranes for tissue engineering.

BACKGROUND: Polymeric electrospun mats have been used as scaffolds in tissue engineering for the development of novel materials due to its characteristics. The usage of synthetic materials has gone in decline due to environmental problems associated with their synthesis and waste disposal. Biomaterials such as biopolymers have been used recently due to good compatibility on biological applications and sustainability. OBJECTIVE: The purpose of this work is to obtain novel materials based on synthetic and natural polymers for applications on tissue engineering. METHODS: Aloe vera mucilage was obtained, chemically characterized, and used as an active compound contained in electrospun mats. Polymeric scaffolds were obtained in single, coaxial and tri-layer structures, characterized and evaluated in cell culture. RESULTS: Mucilage loaded electrospun fibers showed good compatibility due to formation of hydrogen bonds between polymers and biomolecules from its structure, evidenced by FTIR spectra and thermal properties. Cell viability test showed that most of the obtained mats result on viability higher than 75%, resulting in nontoxic materials, ready to be used on scaffolding applications. CONCLUSION: Mucilage containing fibers resulted on materials with potential use on scaffolding applications due to their mechanical performance and cell viability results.

Structure, mechanical and adhesive properties of the cellulosic mucilage in Ocimum basilicum seeds.

Plant seeds and fruits, like those of Ocimum basilicum, develop a mucilaginous envelope rich in pectins and cellulosic fibers upon hydration. This envelope promotes adhesion for attachment to soils and other substrates for dispersal and protection of the seed for a safe germination. Initially at hydration, the mucilage envelope demonstrates low adhesion and friction, but shows increasing adhesive and frictional properties during dehydration. However, the mechanisms underlying the cellulose fiber arrangement and the mechanical properties, especially the elasticity modulus of the mucilage envelope at different hydration conditions are not fully known. In this study, which is based on scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and light microscopy, the structure of the seed coat and arrangement of the cellulose fibers of basil seeds were characterized. Moreover, we performed pull-off force measurements to estimate adhesive properties and JKR-tests to estimate E-modulus of the mucilage at different hydration levels. Microscopy results demonstrate that cellulose fibers are split at their free ends into smaller fibrils, which might enhance the adhesive properties of the mucilage. Adhesive forces in contact increased during dehydration and reached maximum of 33 mN shortly before complete dehydration. The E-modulus of the mucilage changed from 1.4 KPa in water to up to 2.1 MPa in the mucilage at the maximum of its adhesion performance. Obtained results showed hydrogel-like mechanical properties during dehydration and cellulose fiber structures similar to the nanofibrous systems in other organisms with strong adhesive properties. STATEMENT OF SIGNIFICANCE: This paper reveals the hierarchical cellulose fiber structure in Ocimum basilicum's mucilaginous seed coat, suggesting increased fiber splitting towards the end, potentially enhancing adhesion contact areas. Mechanical tests explore elasticity modulus and adhesion force during various hydration stages, crucial as these properties evolve with mucilage desiccation. A rare focus on mucilaginous seed coat mechanical properties, particularly cellulose-reinforced fibers, provides insight into the hydrogel-like mucilage of plant seeds. Adhesion forces peak just before complete desiccation and then decline rapidly. As mucilage water content decreases, the E-modulus rises, displaying hydrogel-like properties during early dehydration stages with higher water content. This study might bring the focus to plant seeds as inspiration for biodegradable glues and applications for hydrogel research.

Effects of dispersing media on the rheological and tribological properties of basil seed mucilage-based thickened liquids.

The development of thickening powders for the management of dysphagia is imperative due to the rapid growth of aging population and prevalence of the dysphagia. One promising thickening agent that can be used to formulate dysphagia diets is basil seed mucilage (BSM). This work investigates the effects of dispersing media, including water, milk, skim milk, and apple juice, on the rheological and tribological properties of the BSM-thickened liquids. Shear rheology results revealed that the thickening ability of BSM in these media in ascending order is milk < skim milk ≈ apple juice < water. On the other hand, extensional rheology demonstrated that the longest filament breakup time was observed when BSM was dissolved in milk, followed by skim milk, water, and apple juice. Furthermore, tribological measurements showed varying lubrication behavior, depending on the BSM concentration and dispersing media. Dissolution of BSM in apple juice resulted in the most superior lubrication property compared with that in other dispersing media. Overall, this study provides insights on BSM's application as a novel gum-based thickening powder in a range of beverages and emphasizes how important it is for consumers to have clear guidance for the use of BSM in dysphagia management.

Encapsulation of sunflower and flaxseed oils using Opuntia (Cactaceae) mucilage as a core-shell material through coacervation methods: A study on formulation, characterization, and in vitro digestion.

Sunflower oil (SFO) and Flaxseed oil (FSO) were microencapsulated using simple and complex coacervation techniques with Opuntia (Cactaceae) mucilage (Mu) and with a combination of Mu with chitosan (Chit). The encapsulation efficiency (EE) of SFO and FSO in emulsions using Mu/Chit shells was 96.7% and 97.4%, respectively. Morphological studies indicated successful entrapment of oils in core shells with particle sizes ranging from 1396 ± 42.4 to 399.8 ± 42.3 nm. The thermogravimetric analyses demonstrated enhanced core protection with thermal stability noted for microcapsules regardless of encapsulation method. The stability of the microcapsules, during in vitro digestion was studied. The obtained results revealed that the microcapsules are intact in oral conditions and have a slow release of oil over stomach digestion and rapid release in the small intestine. The results showed that Mu and Mu/Chit coacervates can be used as effective carrier systems to encapsulate sensitive ingredients and functional oils.

Investigation on Fermented Milk Quality after the Addition of Flaxseed Mucilage and the Use of Lactobacillus delbrueckii subsp. bulgaricus and Lactiplantibacillus plantarum AG9.

BACKGROUND: Flaxseed mucilage (FSM) is one of the healthy components of flaxseed. FSM is an example of a material that can be used in the food, cosmetic, and pharmaceutical industries due to its rheological properties. FSM consists mainly of two polysaccharides, arabinoxylan, and rhamnogalacturonan I, and it also contains protein components and minerals. The prospect of using FSM in food is due to its gelling, water binding, emulsifying, and foaming properties. In addition, valuable natural sources of phenolic compounds such as lignans, phenolic acids, flavonoids, phenylpropanoids, and tannins are partially extracted from flaxseed in FSM. These antioxidant components have pharmacological properties, including anti-diabetic, anti-hypertensive, immunomodulatory, anti-inflammatory and neuroprotective properties. A combination of FSM and lactobacilli in dairy foods can improve their functional properties. This study aimed to develop dairy products by adding of FSM and using two lactic acid bacteria (LAB). FSM (0.2%) was used as an ingredient to improve both the texture and antioxidant properties of the product. METHODS: Skim milk was fermented with 0.2% flaxseed mucilage using Lactobacillus delbrueckii subs. bulgaricus and the probiotic Lactiplantibacillus plantarum AG9. The finished fermented milk products were stored at 4 °C for 14 days. Quantitative chemical, textural, and antioxidant analyses were carried out. RESULTS: Adding 0.2% FSM to the dairy product stimulated the synthesis of lactic acid. FSM increased the viscosity and water-holding capacity of L. bulgaricus or L. bulgaricus/L. plantarum AG9 fermented milk products. Combining these starter strains with FSM promoted the formation of a hard, elastic, resilient casein matrix in the product. When only L. plantarum AG9 was used for the fermentation, the dairy product had a high syneresis and a low viscosity and firmness; such a product is inferior in textural characteristics to the variant with commercial L. bulgaricus. The addition of FSM improved the textural properties of this variant. The use of L. plantarum AG9 and FSM makes it possible to obtain a fermented milk product with the highest content of polyphenolic compounds, which have the highest antioxidant properties and stimulate lipase and α-glucosidase inhibitor synthesis. Combining of L. bulgaricus and L. plantarum AG9 in the starter (20% of the total mass of the starter) and adding of 0.2% FSM is the optimal combination for obtaining a dairy product with high textural and antioxidant properties. CONCLUSIONS: The physicochemical properties (viscosity, syneresis, water holding capacity, texture) and antioxidant properties of fermented milk were improved. In the future, as part of the work to investigate the functional properties of dairy products with FSM, studies will be conducted using in in vivo models.

Characterization and performance evaluation of colorimetric pH-sensitive indicator based on Ò -carrageenan/quince seed mucilage hydrogel as freshness/spoilage monitoring of rainbow trout fillet.

The aim of research was to fabricate a novel indicator by using κ-carrageenan and quince seed mucilage (QSM) hydrogels and red cabbage anthocyanin. The porosity of the hydrogel was controlled using different ratios of κ-carrageenan(C):QSM(Q) (C90:Q10, C70:Q30, and C50:Q50). The hardness of hydrogels decreased from 28.6 ± 0.3 N for C90Q10 to 11.0 ± 1.0 N for C50Q50 sample. However, according to field emission scanning electron microscopy (FE-SEM) analysis, the C50R50 sample had the best morphology with smooth surface and uniform interconnected porous network. Hydrogen bonding interactions among anthocyanins, QSM, and κ-carrageenan were confirmed by Fourier transforms infrared (FT-IR) spectroscopy. The indicator showed a color variation from red to yellow over the pH range of 2-12. Also, the indicator exhibited high sensitivity to ammonia vapors (SRGB = 115%) and good color stability. The C50QRA indicator was used for monitoring rainbow trout fillet spoilage and revealed a visually-detectable color change from red to green upon detecting total volatile basic nitrogen (TVB-N) content produced throughout storage at 4 °C. Generally, the halochromic hydrogel developed in this research can be suggested as a more sensitive and accurate freshness indicator than conventional indicator solid supports.

Evaluation of microwave irradiated Polyacrylamide grafted Opuntia leaf mucilage graft copolymer (OPM-g-PAM) as effective controlled release polymer for release of Rosuvastastin as model drug.

Controlled drug delivery systems offer numerous advantages. This research evaluates Opuntia leaf mucilage grafted with polyacrylamide (OPM-g-PAM) as a promising controlled-release polymer. PAM chains were grafted onto the backbone of OPM using a microwave-assisted method. Optimization of the best grade was based on % grafting efficiency and intrinsic viscosity, followed by extensive physical and analytical characterizations. Analytical characterizations revealed semicrystalline nature of the biomaterial. SEM and AFM observations revealed rough and porous surfaces, indicating effective grafting. Swelling behavior showed maximum sensitivity at pH 7, with reduced swelling at higher sodium chloride concentrations. A comparative study of % drug release of Rosuvastatin over 24 h showed that the optimized grade controlled drug release effectively, achieving 78.5 % release compared to 98.8 % for GF-3. The release data fitted the Korsmeyer-Peppas model, with an "n" value of 0.8334, indicating non-Fickian (anomalous) diffusion. Bacterial biodegradability studies confirmed the high biodegradability of the graft copolymer. In vitro acute toxicity tests showed no toxicity, as confirmed by histopathological studies of heart, liver, and kidney. Overall, the results indicate that OPM-g-PAM is a highly promising material for use in drug delivery systems, demonstrating potential as a novel controlled-release polymer.

Mimosa/quince seed mucilage-co-poly (methacrylate) hydrogels for controlled delivery of capecitabine: Simulation studies, characterization and toxicological evaluation.

This research focused on developing pH-regulated intelligent networks using quince and mimosa seed mucilage through aqueous polymerization to sustain Capecitabine release while overcoming issues like short half-life, high dosing frequency, and low bioavailability. The resulting MSM/QSM-co-poly(MAA) hydrogel was evaluated for several parameters, including complex structure formation, stability, pH sensitivity, morphology, and elemental composition. FTIR, DSC, and TGA analyses confirmed the formation of a stable, complex cross-linked network, demonstrating excellent stability at elevated temperatures. SEM analysis revealed the hydrogels' smooth, fine texture with porous surfaces. PXRD and EDX results indicated the amorphous dispersion of Capecitabine within the network. The QMM9 formulation achieved an optimal Capecitabine loading of 87.17 %. The gel content of the developed formulations ranged from 65.21 % to 90.23 %. All formulations exhibited excellent swelling behavior, with ratios between 65.91 % and 91.93 % at alkaline pH. In vitro dissolution studies indicated that up to 98 % of Capecitabine was released after 24 h at pH 7.4, demonstrating the potential for sustained release. Furthermore, toxicological evaluation in healthy rabbits confirmed the system's safety, non-toxicity, and biocompatibility.

Lactoferrin-chia seed mucilage complex coacervates for intestinal delivery of quercetin and fortification of set yogurt.

This study aimed to develop complex coacervates utilizing lactoferrin (LF) and chia seed mucilage (CSM) for promoting intestinal delivery of quercetin (Q) and fortification of set yogurt. Three cross-linkers, including calcium chloride (CC), transglutaminase (TG), and polyphenolic complex (HP), were used to further reinforce the coacervate network. Cross-linked coacervates had higher values of coacervate yield, encapsulation efficiency, and loading capacity. They efficiently preserved Q under gastric condition (⁓87%-99%), with CSM-TG-Q-LF being most effective for intestinal delivery of Q. Moreover, digested pellets of the cross-linked coacervates displayed better antioxidant activity than the uncross-linked coacervates with CSM-TG-Q-LF pellets showing maximum bioactivity. The Q-loaded coacervates demonstrated superior assembly in the yogurt matrix compared to the unencapsulated Q. Moreover, the coacervate systems, especially CSM-TG-Q-LF significantly improved the textural properties of yogurt and the stability of Q in it. Therefore, CSM-TG-LF is a promising carrier to promote intestinal delivery and food application of hydrophobic molecules.

Enhancing mechanical and water barrier properties of starch film using chia mucilage.

Biodegradable packaging materials are increasingly being investigated due to rising concerns about food safety and environmental conservation. This study examines the incorporation of chia mucilage (CM) into starch-based films using the casting method, aiming to understand its effects on the structure and functionality of the films. CM, an anionic heteropolysaccharide, is hypothesized to enhance the mechanical and barrier properties of the films through polymer interactions and hydrogen bonding. Our findings confirm that CM incorporation results in films with uniformly smooth surfaces, indicating high compatibility and homogeneity within the starch matrix. Notably, CM improves film transparency and crystallinity. Mechanical assessments show a remarkable elevation in tensile strength, soaring from 5.21 MPa to 12.38 MPa, while elongation at break decreases from 61.73 % to 31.42 %, indicating a trade-off between strength and flexibility. Additionally, water solubility decreases from 57.97 % to 41.40 %, and water vapor permeability is reduced by 30 % with CM loading. These results highlight the role of CM in facilitating the formation of a dense, interconnected polymeric network within the starch matrix. Given the soluble dietary fiber nature of CM, the CS/CM (corn starch/chia mucilage) blended films are expected to be safe for food packaging and applicable as edible films with health benefits.

Potential use of extracted flax seed mucilage in the construction of macroporous cryo-scaffolds.

Mucilage is a natural source of polysaccharides that has recently attracted attention for use in biomaterial production. It attracts attention with its easy and fast extraction, biocompatibility, high water retention capacity, and biodegradability. Although there are studies on the characterization of mucilage obtained from different plant sources, the interaction of this polymer with other polymers and its potential to form new biomaterials have not yet been sufficiently investigated. Based on this, in this study, the potential of mucilage extracted from flaxseed for the production of cryogels for tissue engineering applications was demonstrated. Firstly, yield, basic physicochemical properties, morphology, and surface charge-dependent isoelectric point determination studies were carried out for the characterization of the extracted mucilage. The successful preparation of mucilage was evaluated for the construction of cryo-scaffolds and 3D, spongy, and porous structures were obtained in the presence of chitosan and polyvinyl alcohol polymers. A heterogeneous morphology with interconnected macro and micro porosity in the range of approximately 85-115 m pore diameter was exhibited. Due to the high hydrophilic structure of the mucilage, which is attached to the structure with weak hydrogen bonds, the contact angle values of the scaffolds were obtained below 80° and they showed the ability to absorb 1000 times their dry weight in approximately 30 min. As a preliminary optimization study for the evaluation of mucilage in cryogel formation, this work introduced a new construct to be developed as wound dressing scaffold for deep and chronic wounds.

Application of edible nanocomposites from chitosan/fenugreek seed mucilage/selenium nanoparticles for protecting lemon from green mold.

Green (Penicillium digitatum) mold can severely endanger the citrus fruits production and quality. Targeting the protection of lemon fruits from green mold infestations with nanobiotechnology approach, the fenugreek seed mucilage (FM) was extracted and exploited for biosynthesis of selenium (SeNPs) nanoparticles; their nanocomposites (NCs) with chitosan (CT) was constructed and employed as antifungal materials and edible coating (ECs) to protect lemon fruits against green mold. The nanoparticles formation and conjugations were verified by infrared (FTIR) analysis and electron microscopy. The FM-synthesized SeNPs had particles average of 8.35 nm, were the NCs of them with CT had size mean of 49.33 nm and charged with +22.8 mV. The CT/FM/SeNPs composite exhibited superior antifungal actions toward P. digitatum isolates, up to 32.2 mm inhibition diameter and 12.5 mg/mL inhibitory concentration, which exceeded the actions of imazilil. The microscopic screening of exposed P. digitatum to NCs clarified their mycelial destructive action within 30 h. The coating of infected lemons with fabricated NCs led to complete elimination of green mold development after 10 days of coating, without any infestation remarks. The innovative fabrication of NCs from CT/FM/SeNPs is strongly suggested to protect citrus crops from green mold and preserve fruits quality.

Stimuli-responsive glucuronoxylan polysaccharide from quince seeds for biomedical, food packaging, and environmental applications.

Mucilage is a gelatinous mixture of polysaccharides secreted from the seed coat and/or pericarp of many plant seeds when soaked in water. Mucilage affected seed germination while maintaining hydration levels during scarcity. Cydonia oblonga (quince) seeds are natural hydrocolloids extruding biocompatible mucilage mainly composed of polysaccharides. Quince seed mucilage (QSM) has fascinated researchers due to its applications in the food and pharmaceutical industries. On a commercial scale, QSM preserved the sensory and physiochemical properties of various products such as yogurt, desserts, cakes, and burgers. QSM is responsive to salts, pH, and solvents and is mainly investigated as edible coatings in the food industry. In tablet formulations, modified and unmodified QSM as a binder sustained the release of various drugs such as cefixime, capecitabine, diclofenac sodium, theophylline, levosulpiride, diphenhydramine, metoprolol tartrate, and acyclovir sodium. QSM acted as a reducing and capping agent to prepare nanoparticles for good antimicrobial resistance, photocatalytic characteristics, and wound-healing potential. The present review discussed the extraction optimization, chemical composition, stimuli-responsiveness, and viscoelastic properties of mucilage. The potential of mucilage in edible films, tissue engineering, and water purification will also be discussed.

Seed coating with phages for sustainable plant biocontrol of plant pathogens and influence of the seed coat mucilage.

Pathogens resistant to classical control strategies pose a significant threat to crop yield, with seeds being a major transmission route. Bacteriophages, viruses targeting bacteria, offer an environmentally sustainable biocontrol solution. In this study, we isolated and characterized two novel phages, Athelas and Alfirin, which infect Pseudomonas syringae and Agrobacterium fabrum, respectively, and included the recently published Pfeifenkraut phage infecting Xanthomonas translucens. Using a simple immersion method, phages coated onto seeds successfully lysed bacteria post air-drying. The seed coat mucilage (SCM), a polysaccharide-polymer matrix exuded by seeds, plays a critical role in phage binding. Seeds with removed mucilage formed five to 10 times less lysis zones compared to those with mucilage. The podovirus Athelas showed the highest mucilage dependency. Phages from the Autographiviridae family also depended on mucilage for seed adhesion. Comparative analysis of Arabidopsis SCM mutants suggested the diffusible cellulose as a key component for phage binding. Long-term activity tests demonstrated high phage stability on seed surfaces and significantly increasing seedling survival rates in the presence of pathogens. Using non-virulent host strains enhanced phage presence on seeds but also has potential limitations. These findings highlight phage-based interventions as promising, sustainable strategies for combating pathogen resistance and improving crop yield.

Application of seed mucilage as functional biopolymer in meat product processing and preservation.

Meat products consumption is rising globally, but concerns about sustainability, fat content, and shelf life. Synthetic additives and preservatives used for extending the shelf life of meat often carry health and environmental drawbacks. Seed mucilage, natural polysaccharides, possesses unique functional properties like water holding, emulsifying, and film forming, offering potential alternatives in meat processing and preservation. This study explores the application of seed mucilage from diverse sources (e.g., flaxseed, psyllium, basil) in various meat and meat products processing and preservation. Mucilage's water-holding and emulsifying properties can potentially bind fat and decrease the overall lipid content in meat and meat-based products. Moreover, antimicrobial and film-forming properties of mucilage can potentially inhibit microbial growth and reduce oxidation, extending the shelf life. This review emphasizes the advantages of incorporating mucilage into processing and coating strategies for meat and seafood products.

Microwave-assisted extraction of mucilage from juá: Characterization and antioxidant activity.

Microwave-assisted extraction of mucilage from juá was investigated using response surface methodology. The optimal conditions for extraction were a power of 300 W, an extraction time of 240 s, a pH of 8.0, and a water/sample ratio of 1/6, which achieved a 26.43% yield. The monosaccharide composition and antioxidant activity of the mucilage from juá fruits from different regions of Caatinga were investigated. The fruits from Agreste Paraibano showed the highest mucilage extraction yield (18.64%) compared to that of fruits from Mata Paraibana (MP) (12.37%), Borborema (BB) (11.47%), and Sertão Paraibano (8.31%) (p < 0.05). Glucose (32.8%-50.8%) and arabinose (19.3%-32.9%) were the main monosaccharides found in juá mucilage. The mucilage from fruits in the MP presented the highest antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl and oxygen radical absorbance capacity assays. Our results demonstrated the potential for the future exploration and application of juá mucilage in the food industry. PRACTICAL APPLICATION: Juá (Ziziphus joazeiro Mart.) mucilage contains phenolic compounds and antioxidant activity, and its extraction by MAE is efficient, as it contributed to a higher yield.

Structure, chemical modification, and functional applications of mucilage from Mimosa pudica seeds - A review.

Mimosa pudica (MP) is an ornamental plant due to seismonastic movements that close leaves and fall petioles in response to touch, wind, light, heat, cold, and vibration. The seeds of MP secrete smart, biocompatible, and non-toxic mucilage that has captivated researchers due to its widespread use in various fields such as pharmaceuticals and biotechnology. The mucilage is responsive to pH, salt solutions, and solvents and acts as a binder in tablet formulations for targeted drug delivery. The mucilage is chemically modifiable via acetylation, succinylation, and graft polymerization. Chemically modified MP mucilage appeared supersorbent for heavy metal ion uptake. Nanoparticles synthesized using mucilage as a reducing and capping agent displayed significant antimicrobial and wound-healing potential. Crosslinking of mucilage using citric acid as a crosslinking agent offers a sustained release of drugs. The present review is aimed to discuss extraction optimization, structure, modification, and the stimuli-responsive nature of mucilage. The review article will cover the potential of mucilage as emulsifying, suspending, bio-adhesive, gelling, and thickening agent. The role of mucilage as a capping and reducing agent for nanoparticles will also be discussed.

Fenugreek seed mucilage-based active edible films for extending fresh fruit shelf life: Antimicrobial and physicochemical properties.

Trigonella foenum-graecum, known as fenugreek, belongs to the leguminous family of wild growth in Western Asia, Europe, the Mediterranean, and Asia; its ripe seeds contain a pool of bioactive substances with great potential in the food industry and medicine. In this study, fenugreek seed mucilage (FSM) was extracted and characterized in its structural properties by X-ray diffraction, nuclear magnetic resonance, and high-performance liquid chromatography. Then, the applicability of FSM as an antimicrobial agent was demonstrated via the development of novel, active, edible FSM-based biofilms containing carboxymethyl cellulose and rosemary essential oil (REO). Incorporating REO in the biofilms brought about specific changes in Fourier-transform infrared spectra, affecting thermal degradation behavior. Scanning electron microscopy and atomic force microscopy morphography showed an even distribution of REO and smoother surfaces in the loaded films. Besides, the solubility tests evidenced a reduction in water solubility with increasing REO concentration from 1 to 3 wt%. The biological assay evidenced the antimicrobial activity of REO-loaded biofilms against Staphylococcus aureus and Escherichia coli. Finally, whole apples were dip-coated with FSM-based solutions to showcase future edible systems. The REO-loaded biofilms extended the shelf life of apples to 30 days, demonstrating their potential for sustainable and active coatings.

Brassica napus BnaA09.MYB52 enhances seed coat mucilage accumulation and tolerance to osmotic stress during seed germination in Arabidopsis thaliana.

Seed coat mucilage plays an important role in promoting seed germination under adversity. Previous studies have shown that Arabidopsis thaliana MYB52 (AtMYB52) can positively regulate seed coat mucilage accumulation. However, the role of Brassica napus MYB52 (BnaMYB52) in accumulation of seed coat mucilage and tolerance to osmotic stress during seed germination remains largely unknown. We cloned the BnaA09.MYB52 coding domain sequence from B. napus cv ZS11, identified its conserved protein domains and elucidated its relationship with homologues from a range of plant species. Transgenic plants overexpressing BnaA09.MYB52 in the A. thaliana myb52-1 mutant were generated through Agrobacterium-mediated transformation and used to assess the possible roles of BnaA09.MYB52 in accumulation of seed coat mucilage and tolerance to osmotic stress during seed germination. Subcellular localization and transcriptional activity assays demonstrated that BnaA09.MYB52 functions as a transcription factor. RT-qPCR results indicate that BnaA09.MYB52 is predominantly expressed in roots and developing seeds of B. napus cv ZS11. Introduction of BnaA09.MYB52 into myb52-1 restored thinner seed coat mucilage in this mutant to levels in the wild type. Consistently, expression levels of three key genes participating in mucilage formation in developing seeds of myb52-1 were also restored to wild type levels by overexpressing BnaA09.MYB52. Furthermore, BnaA09.MYB52 was induced by osmotic stress during seed germination in B. napus, and ectopic expression of BnaA09.MYB52 successfully corrected sensitivity of the myb52-1 mutant to osmotic stress during seed germination. These findings enhance our understanding of the functions of BnaA09.MYB52 and provide a novel strategy for future B. napus breeding.

Expression of an endo-rhamnogalacturonase from Aspergillus aculeatus enhances release of Arabidopsis transparent mucilage.

Mucilage is a gelatinous and sticky hydrophilic polysaccharide released from epidermal cells of seed coat after the hydration of mature seeds and is composed primarily of unbranched rhamnogalacturonan I (RG-I). In this study, we produced a recombinant endo-RG-I hydrolase from Aspergillus aculeatus (AaRhgA) in the fission yeast Schizosaccharomyces pombe and examined its substrate preference for pyridylaminated (PA) RG-I with the various degrees of polymerization (DP). Recombinant AaRhgA requires PA-RG-I with a DP of 10 or higher for its hydrolase activity. We heterologously expressed the AarhgA gene under the strong constitutive promoter, cauliflower mosaic virus 35S promoter, in Arabidopsis thaliana. In a series of biochemical analyses of each mucilage fraction released from the water-imbibed seeds of the transgenic plants, we found the enhanced deposition of the transparent mucilage layer that existed in the peripheral regions of the adherent mucilage and was not stained with ruthenium red. This study demonstrated the feasibility of manipulating the mucilage organization by heterologous expression of the endo-RG-I hydrolase.