Cell Wall Degradation Research Articles

PpERF-CRF3 selected by transcriptomic analysis plays key roles in the regulation of ABA alleviating chilling injury in peach fruit

Abscisic acid (ABA) is widely utilized to mitigate chilling injury (CI) of fruit. However, the molecular mechanism of ABA alleviates CI in peach fruit remain unclear. Herein, 10−4 M ABA treatment significantly mitigated the CI of peach fruit by reducing relative conductivity and malondialdehyde content, while increasing proline and endogenous ABA content. Transcriptomic analysis indicated that an abundant number of differentially expressed genes were altered by ABA treatment, which primarily enriched pathways such as plant hormone signal transduction, glycerophospholipid metabolism and phenylpropanoid biosynthesis. RNA-Seq results indicate that ABA modulates the transcription of genes involved in auxin, ABA and ethylene signal transduction, as well as in cell wall degradation, antioxidant, fatty acid desaturation and proline metabolism. RT-qPCR confirmed the RNA-Seq results, ABA treatment induced the transcription of proline metabolism related genes (PpP5CR2, PpP5CS, PpP5CS1) and PpERF-CRF3. Particularly noteworthy, as a nuclear protein, PpERF-CRF3 activated the expression of PpP5CR2 and PpP5CS by directly binding to their promoters and over-expression PpERF-CRF3 increased proline content and enhanced PpP5CR2 and PpP5CS expression. Overall, these findings suggest that ABA mitigates CI in peach fruit may be by mediating these pathways, and PpERF-CRF3 potentially involves this process by stimulating the expression of genes related to proline synthesis.

The DYRKP1 kinase regulates cell wall degradation in Chlamydomonas by inducing matrix metalloproteinase expression.

The cell wall of plants and algae is an important cell structure that protects cells from changes in the external physical and chemical environment. This extracellular matrix, composed of polysaccharides and glycoproteins, must be constantly remodeled throughout the life cycle. However, compared to matrix polysaccharides, little is known about the mechanisms regulating the formation and degradation of matrix glycoproteins. We report here that a plant kinase belonging to the DUAL-SPECIFICITY TYROSINE PHOSPHORYLATION-REGULATED KINASE (DYRK) family present in all eukaryotes regulates cell wall degradation after mitosis of Chlamydomonas reinhardtii by inducing the expression of matrix metalloproteinases (MMPs). Without the plant DYRK kinase (DYRKP1), daughter cells cannot disassemble parental cell walls and remain trapped inside for more than 10 days. On the other hand, the DYRKP1 complementation line shows normal degradation of the parental cell wall. Transcriptomic and proteomic analyses indicate a marked down-regulation of MMP gene expression and accumulation, respectively, in the dyrkp1 mutants. The mutants deficient in MMPs retain palmelloid structures for a longer time than the background strain, like dyrkp1 mutants. Our findings show that DYRKP1, by ensuring timely MMP expression, enables the successful execution of the cell cycle. Altogether, this study provides insight into the life cycle regulation in plants and algae.

Tomato Synaptotagmin F accelerates fruit ripening, shortens fruit shelf-life and increases susceptibility to Penicillium expansum

Synaptotagmins (SYTs), initially identified as calcium sensors for regulating synaptic vesicle exocytosis and endocytosis in mammalian neurons, play crucial role in biotic and abiotic stresses in plants. However, the function of SYTs in fruit ripening is unclear. In this study, a tomato Synaptotagmins gene, SlSYTF, was found to accelerate tomato fruit ripening. SlSYTF encodes an endoplasmic reticulum localized protein whose transcription is continuously enhanced during fruit ripening. Overexpression SlSYTF in tomato resulted in accelerated ripening progress, increased carotenoid content as well as decreased the firmness of tomato fruit, whereas the mutant slsytf-c exhibited the opposite phenotype. Importantly, SlSYTF could increase ethylene production by activating the expression of ethylene synthesis genes and prompt cell wall degradation by increasing pectinase and cellulase activities. Nevertheless, the accelerated cell wall degradation and thinned cuticle due to SlSYTF results in reduced shelf life and pathogen resistance. Collectively, we revealed a new SYTs gene that plays a dual role in tomato fruit ripening and pathogen response. This finding may shed new light on the relationship between maturation and immunity.

A vacuolar invertase gene SlVI modulates sugar metabolism and postharvest fruit quality and stress resistance in tomato

Abstract Sugars act as signaling molecules to modulate various growth processes and enhance plant tolerance to various abiotic and biotic stresses. Moreover, sugars contribute to the post-harvest flavor in fleshy fruit crops. To date, the regulation of sugar metabolism and its effect in plant growth, fruit ripening, postharvest quality and stress resistance remains not fully understood. In this study, we investigated the role of tomato gene encoding a vacuolar invertase, hydrolyzing sucrose to glucose and fructose. SlVI is specifically expressed during the tomato fruit ripening process. We found that overexpression of SlVI resulted in increased leaf size and early flowering, while knockout of SlVI led to increased fruit sucrose content, enhanced fruit firmness, and elevated resistance of postharvest fruit to Botrytis cinerea. Moreover, the content of naringenin and total soluble solids was significantly increased in SlVI knockout fruit at postharvest stage. Transcriptome analysis showed a negative feedback regulation triggered by sucrose accumulation in SlVI knockout fruit resulting in a downregulation of BAM3 and AMY2, which are critical for starch degradation. Moreover, genes associated with cell wall, cutin, wax, and flavonoid biosynthesis and pathogen resistance were upregulated in SlVI knockout fruit. Conversely, the expression levels of genes involved in cell wall degradation were decreased in knockout fruit. These results are consistent with the enhanced postharvest quality and resistance. Our findings not only provide new insights into the relationship between tomato fruit sucrose content and postharvest fruit quality, but also suggest new strategies to enhance fruit quality and extend postharvest shelf life.

Transcriptional reprogramming of tomato (Solanum lycopersicum L.) roots treated with humic acids and filter sterilized compost tea

BackgroundTo counteract soil degradation, it is important to convert conventional agricultural practices to environmentally sustainable management practices. To this end, the application of biostimulants could be considered a good strategy. Compost, produced by the composting of biodegradable organic compounds, is a source of natural biostimulants, such as humic acids, which are naturally occurring organic compounds that arise from the decomposition and transformation of organic residues, and compost tea, a compost-derived liquid formulated produced by compost water-phase extraction. This study aimed to determine the molecular responses of the roots of tomato plants (cv. Crovarese) grown under hydroponic conditions and subjected to biostimulation with humic substances (HSs) and filtered sterile compost tea (SCT).ResultsThe 13C CPMAS NMR of humic acids (HA) and SCT revealed strong O-alkyl-C signals, indicating a high content of polysaccharides.Thermochemolysis identified over 100 molecules, predominantly from lignin, fatty acids, and biopolymers. RNA-Seq analysis of tomato roots treated with HA or SCT revealed differentially expressed genes (DEGs) with distinct patterns of transcriptional reprogramming.Notably, HA treatment affected carbohydrate metabolism and secondary metabolism, particularly phenylpropanoids and flavonoids, while SCT had a broader impact on hormone and redox metabolism. Both biostimulants induced significant gene expression changes within 24 h, including a reduction in cell wall degradation activity and an increase in the expression of hemicellulose synthesis genes, suggesting that the treatments prompted proactive cell wall development.ConclusionsThe results demonstrate that HS and SCT can mitigate stress by activating specific molecular mechanisms and modifying root metabolic pathways, particularly those involved in cell wall synthesis. However, gene regulation in response to these treatments is complex and influenced by various factors. These findings highlight the biostimulatory effects of HS and SCT, suggesting their potential application in crop biofertilization and the development of innovative breeding strategies to maximize the benefits of humic substances for crops. Further research is needed to fully elucidate these mechanisms across various contexts and plant species.

Genome-wide survey of the bipartite structure and pathogenesis-related genes of Neostagonosporella sichuanensis, a causal agent of Fishscale bamboo rhombic-spot disease.

Bamboo resources have garnered significant global attention due to their excellent capacity for regeneration and high yield. Rhombic-spot disease, a substantial threat to fishscale bamboo (Phyllostachys heteroclada), is primarily caused by Neostagonosporella sichuanensis. This study first reported the genome assemblies and characteristics of two N. sichuanensis isolates using PacBio and Illumina sequencing platforms. The genomes of N. sichuanensis strain SICAUCC 16-0001 and strain SICAUCC 23-0140, with sizes of 48.0 Mb and 48.4 Mb, respectively, revealed 10,289 and 10,313 protein-coding genes. Additionally, they contained 34.99 and 34.46% repetitive sequences within AT-rich regions, with notable repeat-induced point mutation activity. Comparative genome analysis identified 1,049 contracted and 45 expanded gene families in the genome of N. sichuanensis, including several related to pathogenicity. Several gene families involved in mycotoxin metabolism, secondary metabolism, sterol biosynthesis and transport, and cell wall degradation were contracted. Compared to most analyzed necrotrophic, hemibiotrophic, and phaeosphaeriacous pathogens, the genomes of two N. sichuanensis isolates exhibited fewer secondary metabolite enzymes, carbohydrate-active enzymes, plant cell wall degrading enzymes, secreted proteins, and effectors. Comparative genomics analysis suggested that N. sichuanensis shares more similar characteristics with hemibiotrophic pathogens. Based on single carbon source tests, N. sichuanensis strains demonstrated a higher potential for xylan decomposition than pectin and cellulose. The proportion of cell wall-degrading enzyme effectors occupied a high proportion of the total effectors of the N. sichuanensis genomes. These findings provide valuable insights into uncovering the pathogenesis of N. sichuanensis toward the efficient management of rhombic-spot disease of fishscale bamboo.

Early and late responses to Fusarium Head blight in durum wheat: Focus on phenylpropanoid biosynthetic pathway

Durum wheat is among the cereal crops most susceptible to Fusarium Head Blight (FHB), a fungal disease that can lead to significant yield losses. Despite this, only limited research efforts have been directed towards understanding FHB resistance in durum wheat. Wheat grains naturally contain phenolic compounds, and anthocyanins are particularly present in the so-called pigmented wheat genotypes, such as purple pericarp ones. In this study the effects of the biotic stress caused by Fusarium graminearum infection on phenylpropanoid biosynthetic pathway in durum wheat spikes were explored, considering three genotypes with different susceptibility (including a purple pericarp genotype), and two time points (an early stage time point: 2 days post infection, and a late stage time point: 21 days post infection). At early infection stage, the F. graminearum infection triggered upregulation of all the considered genes involved in the phenylpropanoid pathway in the resistant genotype, while, in the purple pericarp genotype, the infection caused an increase in quercetin accumulation in the soluble fraction of spike extract. At late infection stage, the infection caused (in all the genotypes) a degradation of secondary cell wall and the release of the hydroxycinnamic acids esterified with arabinoxylans (ferulic acid and p-coumaric acid) and lignin-derived monomers (vanillic acid). Furthermore, chalcone synthase gene (CHS) and the transcription factor Ppm1 (Purple pericarp MYB 1) were boosted in the pigmented genotype due to infection at late infection stage. These findings contribute to the understanding of host-pathogen interactions for future breeding programs focused on improving FHB resistance in durum wheat varieties, with a particular focus on pigmented genotypes.

Constrained Catalytic Itinerary of a Retaining 3,6‐Anhydro‐D‐Galactosidase, a Key Enzyme in Red Algal Cell Wall Degradation

AbstractThe marine Bacteroidota Zobellia galactanivorans has a polysaccharide utilization locus dedicated to the catabolism of the red algal cell wall galactan carrageenan and its unique and industrially important α‐3,6‐anhydro‐D‐galactose (ADG) monosaccharide. Here we present the first analysis of the specific molecular interactions that the exo‐(α‐1,3)‐3,6‐anhydro‐D‐galactosidase ZgGH129 uses to cope with the strict steric restrictions imposed by its bicyclic ADG substrate — which is ring flipped relative to D‐galactose. Crystallographic snapshots of key catalytic states obtained with the natural substrate and novel chemical tools designed to mimic species along the reaction coordinate, together with quantum mechanics/molecular mechanics (QM/MM) metadynamics methods and kinetic studies, demonstrate a retaining mechanism where the second step is rate limiting. The conformational landscape of the constrained 3,6‐anhydro‐D‐galactopyranose ring proceeds through enzyme glycosylation B1,4→[E4]≠→E4/1C4 and deglycosylation E4/1C4→[E4]≠→B1,4 itineraries limited to the Southern Hemisphere of the Cremer–Pople sphere. These results demonstrate the conformational changes throughout catalysis in a non‐standard, sterically restrained, bicyclic monosaccharide, and provide a molecular framework for mechanism‐based inhibitor design for anhydro‐type carbohydrate‐processing enzymes and for future applications involving carrageenan degradation. In addition, our study provides a rare example of distinct niche‐based conformational itineraries within the same carbohydrate‐active enzyme family.

Constrained Catalytic Itinerary of a Retaining 3,6-Anhydro-D-Galactosidase, a Key Enzyme in Red Algal Cell Wall Degradation.

The marine Bacteroidota Zobellia galactanivorans has a polysaccharide utilization locus dedicated to the catabolism of the red algal cell wall galactan carrageenan and its unique and industrially important α-3,6-anhydro-D-galactose (ADG) monosaccharide. Here we present the first analysis of the specific molecular interactions that the exo-(α-1,3)-3,6-anhydro-D-galactosidase ZgGH129 uses to cope with the strict steric restrictions imposed by its bicyclic ADG substrate - which is ring flipped relative to D-galactose. Crystallographic snapshots of key catalytic states obtained with the natural substrate and novel chemical tools designed to mimic species along the reaction coordinate, together with quantum mechanics/molecular mechanics (QM/MM) metadynamics methods and kinetic studies, demonstrate a retaining mechanism where the second step is rate limiting. The conformational landscape of the constrained 3,6-anhydro-D-galactopyranose ring proceeds through enzyme glycosylation B1,4→[E4]≠→E4/1C4 and deglycosylation E4/1C4→[E4]≠→B1,4 itineraries limited to the Southern Hemisphere of the Cremer-Pople sphere. These results demonstrate the conformational changes throughout catalysis in a non-standard, sterically restrained, bicyclic monosaccharide, and provide a molecular framework for mechanism-based inhibitor design for anhydro-type carbohydrate-processing enzymes and for future applications involving carrageenan degradation. In addition, our study provides a rare example of distinct niche-based conformational itineraries within the same carbohydrate-active enzyme family.

Characterization and Pathogenicity of Colletotrichum truncatum Causing Hylocereus undatus Anthracnose through the Changes of Cell Wall-Degrading Enzymes and Components in Fruits.

Anthracnose is one of the destructive diseases of pitaya that seriously affects the plant growth and fruit quality and causes significant yield and economic losses worldwide. However, information regarding the species of pathogens that cause anthracnose in pitaya (Hylocereus undatus) fruits in Gansu Province, China, and its pathogenic mechanism is unknown. Thus, the purposes of our present study were to identify the species of pathogens causing H. undatus fruits anthracnose based on the morphological and molecular characteristics and determine its pathogenic mechanism by physiological and biochemical methods. In our present study, forty-six isolates were isolated from the collected samples of diseased H. undatus fruits and classified as three types (named as H-1, H-2, and H-3), according to the colony and conidium morphological characteristics. The isolation frequencies of H-1, H-2, and H-3 types were 63.04%, 21.74%, and 15.22%, respectively. The representative single-spore isolate of HLGTJ-1 in H-1 type has significant pathogenicity, and finally we identified Colletotrichum truncatum as the pathogen based on the morphological characteristics as well as multi-locus sequence analysis. Moreover, the H. undatus fruits inoculated with C. truncatum had a significantly increased activity of cell wall-degrading enzymes (CWDEs) cellulase (Cx), β-glucosidase (β-Glu), polygalacturonase (PG), and pectin methylgalacturonase (PMG), while having a decreased level of cell wall components of original pectin and cellulose in comparison to control. The average increased activities of Cx, β-Glu, PG, and PMG were 30.73%, 40.40%, 51.55%, and 32.23% from day 0 to 6 after inoculation, respectively. In contrast, the average decreased contents of original pectin and cellulose were 1.82% and 16.47%, respectively, whereas the average increased soluble pectin content was 38.31% in comparison to control. Our results indicate that C. truncatum infection increased the activities of CWDEs in H. undatus fruits to disassemble their cell wall components, finally leading to the fruits' decay and deterioration. Thus, our findings will provide significant evidence in the controlling of pitaya anthracnose in the future.

Effect of alkali treatment and fungal degradation on the nanostructure and cellulose arrangement in Scots pine cell walls – A neutron and X-ray scattering study

Research on new conservation treatments for historical wood requires considerable amounts of appropriate wood material, which is hard to acquire. Thus, we produced biologically and chemically degraded model wood that could be used as a representative material in future research on consolidating agents. Using chemical composition determinations, we found that fungal decay targeted mainly polysaccharides, while alkaline treatment mostly reduced hemicelluloses and lignin content. X-ray and neutron scattering showed that all decayed samples had increased disorder in microfibril alignment and larger elementary fibril cross-sections, and alkaline-treated samples had much larger elementary fibril spacing compared to those decayed by fungi. These nanoscale and chemical differences correlate with physical property changes. For example, decreased cellulose crystallinity and increased disorder of the microfibrils in degraded cell walls likely contribute to the lower elastic moduli measured for these cell walls. The obtained data improves understanding of how degradation alters wood structures and properties across length scales and will be valuable for future studies focusing on archeological wood. Moreover, it leads to the conclusion that it is more appropriate to develop treatments that consider not only spatial variability and degree of wood degradation but also the corresponding molecular and nanoscale changes in the cell walls.

Mass Transfer Resistance and Reaction Rate Kinetics for Carbohydrate Digestion with Cell Wall Degradation by Cellulase.

This paper introduces an enzymatic approach to estimate internal mass-transfer resistances during food digestion studies. Cellulase has been used to degrade starch cell walls (where cellulose is a significant component) and reduce the internal mass-transfer resistance, so that the starch granules are released and hydrolysed by amylase, increasing the starch hydrolysis rates, as a technique for measuring the internal mass-transfer resistance of cell walls. The estimated internal mass-transfer resistances for granular starch hydrolysis in a beaker and stirrer system for simulating the food digestion range from 2.2 × 107 m-1 s at a stirrer speed of 100 rpm to 6.6 × 107 m-1 s at 200 rpm. The reaction rate constants for cellulase-treated starch are about three to eight times as great as those for starch powder. The beaker and stirrer system provides an in vitro model to quantitatively understand external mass-transfer resistance and compare mass-transfer and reaction rate kinetics in starch hydrolysis during food digestion. Particle size analysis indicates that starch cell wall degradation reduces starch granule adhesion (compared with soaked starch samples), though the primary particle sizes are similar, and increases the interfacial surface area, reducing internal mass-transfer resistance and overall mass-transfer resistance. Dimensional analysis (such as the Damköhler numbers, Da, 0.3-0.5) from this in vitro system shows that mass-transfer rates are greater than reaction rates. At the same time, SEM (scanning electron microscopy) images of starch particles indicate significant morphology changes due to the cell wall degradation.

Mutation of tomato xyloglucan transglucosylase/hydrolase5 increases fruit firmness and contributes to prolonged shelf life

Fruit ripening in tomato is a highly coordinated developmental process accompanied with fruit softening, which is closely associated with cell wall degradation and remodeling. Xyloglucan endotransglucosylase/hydrolases (XTHs) are known to play an essential role in cell wall xyloglucan metabolism. Tomato XTH5 exhibits xyloglucan endotransglucosylase (XET) activity in vitro, but the understanding of its biological role in fruit ripening remains unclear. In this study, we revealed that SlXTH5 is highly expressed in mature fruits. Knockout mutant plants of SlXTH5 were generated by CRISPR/Cas9 gene editing strategy in tomato cultivar Micro-Tom. The mutant fruits showed accelerated transition from unripe to ripe process and earlier ethylene accumulation compared to wild type fruits. Although the mutation of SlXTH5 did not affect the size, weight and number of fruits, it indeed increased fruit firmness and extended shelf life, which is probably attributed to the increased cell layer and cell wall thickness of pericarp tissue. Pathogen infection experiment showed the enhanced resistance of mutant fruits to Botrytis cinerea. These results revealed the role of SlXTH5 in fruit ripening process, and provide new insight into how cell wall metabolism and remodeling regulate fruit softening and shelf life.

Dual RNA sequencing during Trichoderma harzianum-Phytophthora capsici interaction reveals multiple biological processes involved in the inhibition and highlights the cell wall as a potential target.

Phytophthora capsici is a destructive oomycete pathogen, causing huge economic losses for agricultural production. The genus Trichoderma represents one of the most extensively researched categories of biocontrol agents, encompassing a diverse array of effective strains. The commercial biocontrol agent Trichoderma harzianum strain T-22 exhibits pronounced biocontrol effects against many plant pathogens, but its activity against P. capsici is not known. T. harzianum T-22 significantly inhibited the growth of P. capsici mycelia and the culture filtrate of T-22 induced lysis of P. capsici zoospores. Electron microscopic analyses indicated that T-22 significantly modulated the ultrastructural composition of P. capsici, with a severe impact on the cell wall integrity. Dual RNA sequencing revealed multiple biological processes involved in the inhibition during the interaction between these two microorganisms. In particular, a marked upregulation of genes was identified in T. harzianum that are implicated in cell wall degradation or disruption. Concurrently, the presence of T. harzianum appeared to potentiate the susceptibility of P. capsici to cell wall biosynthesis inhibitors such as mandipropamid and dimethomorph. Further investigations showed that mandipropamid and dimethomorph could strongly inhibit the growth and development of P. capsici but had no impact on T. harzianum even at high concentrations, demonstrating the feasibility of combining T. harzianum and these cell wall synthesis inhibitors to combat P. capsici. These findings provided enhanced insights into the biocontrol mechanisms against P. capsici with T. harzianum and evidenced compatibility between specific biological and chemical control strategies. © 2024 Society of Chemical Industry.

Transcriptome analysis reveals the potential mechanism of methyl jasmonate alleviated ripening disorder in mango fruit at low temperature

High susceptibility of mangoes to low temperature leads to ripening failure that restricts the marketability of products. This study investigated the effect of methyl jasmonate (MeJA) on ripening disorder and mechanism involved in mangoes during refrigeration. Results showed that 50 μM MeJA ameliorated ripening disorder, as indicated by accelerated advancement of ripening-related parameters. Transcriptome analysis revealed that 17,414 significantly differentially expressed genes were mainly enriched in ethylene synthesis, cell wall degradation, starch degradation and sugar transport. Moreover, 8 AP2/ERF transcription factors and 12 ripening-related genes were characterized via qRT-PCR. Afterwards, through the analysis of transcription factor binding sites and cis-acting elements, a regulatory network of ERFs mediated alleviation of ripening disorder conferred by MeJA was constructed. Finally, the interactions between MiERFs and the promoters of target genes were verified by yeast one-hybrid assay. Our findings provide a theoretical basis for improving cold tolerance via counteracting ripening disorder in mangoes.

Preharvest methyl jasmonate application delays cell wall degradation and upregulates phenolic metabolism and antioxidant activities in cold stored raspberries

The effects of preharvest methyl jasmonate (MeJA) spray application on the physicochemical quality, metabolism of phenolics, and cell wall components in raspberries were investigated during a 10-day cold storage period. MeJA spray reduced firmness loss, decay incidence, and weight loss, while maintained higher levels of soluble solids content, ascorbic acid, anthocyanins and flavonoids in raspberries. Furthermore, MeJA application resulted in increased total pectin and protopectin levels, as well as lowered water-soluble pectin, and activities of pectin methyl esterase, polygalacturonase and cellulase enzymes. Additionally, MeJA treatment upregulated the phenylpropanoid pathway, leading to higher endogenous phenolics and activities of phenylalanine-ammonia lyase and shikimate dehydrogenase. In conclusion, preharvest MeJA spray application could be adopted to enhance the storage potential of cold-stored raspberries for 10 days by maintaining higher firmness, assuring better physicochemical quality, and increasing phenolic metabolism, while reducing cell wall hydrolysis.

Foliar spraying of boron prolongs preservation period of strawberry fruits by altering boron form and boron distribution in cell.

Boron (B), an essential micronutrient for fruit development, also plays a crucial role in maintaining the shelf life of strawberries (Fragaria ananassa Duch.) by affecting cell wall structure and components. We investigated the distribution pattern of B within cells and cell walls in strawberry fruits under different B levels and revealed the relationship between the B distribution in cell walls and fruit firmness after harvesting. Foliar spraying of 0.1% H3BO3 promoted the growth of strawberry seedlings and improved fruit yield and flesh firmness by 45.7% and 25.6%. During the fruit softening and decay process, the content of bound B and cell wall-B decreased while more B was allocated to the protoplast and apoplast. The changes in B distribution in cells were attributed to cell damage during fruit decay, and B extended the freshness period of the fruits by alleviating the decrease of B distribution in cell walls. After leaving the fruits at room temperature for 10 h, the B content in different cell wall components significantly decreased, while foliar spraying of B alleviated the reduction of B content in covalently bound pectin (CBP), cellulose, and hemicellulose. Meanwhile, B spraying on fruit decreased the activity of cell wall degradation enzymes, including polygalacturonase (PG) and pectin lyase (PL), by 20.2% and 38.1%, while enhancing the demethylation of pectin by increasing pectin methylesterase (PME) activity from 21.6 U/g to 25.7 U/g. Thus, foliar spraying of 0.1% H3BO3 enhances the cross-linking of B with cell wall components and maintains cell wall structure, thereby prolonging the shelf life of strawberry fruits.

Genomic Sequencing and Functional Analysis of the Ex-Type Strain of Malbranchea zuffiana.

Malbranchea is a genus within the order Onygenales (phylum Ascomycota) that includes predominantly saprobic cosmopolitan species. Despite its ability to produce diverse secondary metabolites, no genomic data for Malbranchea spp. are currently available in databases. Therefore, in this study, we obtained, assembled, and annotated the genomic sequence of the ex-type strain of Malbranchea zuffiana (CBS 219.58). For the genomic sequencing, we employed both the Illumina and PacBio platforms, followed by hybrid assembly using MaSuRCA. Quality assessment of the assembly was performed using QUAST and BUSCO tools. Annotation was conducted using BRAKER2, and functional annotation was completed with InterProScan. The resulting genome was of high quality, with a size of 26.46 Mbp distributed across 38 contigs and a BUSCO completion rate of 95.7%, indicating excellent contiguity and assembly completeness. A total of 8248 protein-encoding genes were predicted, with functional annotations assigned to 73.9% of them. Moreover, 82 genes displayed homology with entries in the Pathogen Host Interactions (PHI) database, while 494 genes exhibited similarity to entries in the Carbohydrate-Active Enzymes (CAZymes) database. Furthermore, 30 biosynthetic gene clusters (BGCs) were identified, suggesting significant potential for the biosynthesis of diverse secondary metabolites. Comparative functional analysis with closely related species unveiled a considerable abundance of domains linked to enzymes involved in keratin degradation, alongside a restricted number of domains associated with enzymes engaged in plant cell wall degradation in all studied species of the Onygenales. This genome-based elucidation not only enhances our comprehension of the biological characteristics of M. zuffiana but also furnishes valuable insights for subsequent investigations concerning Malbranchea species and the order Onygenales.

Varying concentrations of ethephon induce antioxidant defences and cell wall degradation to regulate storage quality of fresh-cut lotus root

This study examined the impact of varying concentrations of ethephon (ET) immersion on the storage quality of fresh-cut lotus root. Compared to the 0.1%, the 0.8% and 1.6% ET groups inhibited phenolic oxidation and showed stronger antioxidant and reactive oxygen species balancing capacity during early storage, which led to better browning inhibition. During the early storage period, varying concentrations of ET treatments were inversely associated with microbial growth, and Pseudomonas as the major microorganism was significantly inhibited. During the post-storage period, the ET group exacerbated cell wall metabolism, contributed to the softening of lotus root slices. Additionally, ET stimulated enhanced respiration and reduced total soluble solids content as ET concentration increased. The heightened expression of NnERF in the late storage stage suggested a potential link to the regulation of root quality deterioration. Both browning inhibition and quality deterioration promotion exhibited gradient shifts in accordance with ET concentration. Combined with DNA affinity purification sequencing technology to identify the presence of motifs that bind specifically to ERF1B in lotus root, and screening for the possible target genes NnPL18 and NnACO that may be regulated by NnERF1B-like (104610030), it further suggests that NnERF1B-like (104610030) may mediate the process of lotus root flake softening and ethylene biosynthesis.

THE USE OF PURPLE EGGPLANT FRUIT CALYX STEM (Solanum melongena L.) AS AN ALTERNATIVE SOURCE OF LACTASE-PRODUCING LACTIC ACID BACTERIA

Lactose intolerance is a condition where body is unable to digest lactose due to an enzyme lactase deficiency in digestive system, leading to various health problems ranging from bloating, diarrhea, and abdominal pain to severe problems that can result in stunting in children. One attempt to prevent stunting due to lactose intolerance is administration of low-lactose dairy products, such as yogurt. The production of yogurt requires a starter known as lactic acid bacteria (LAB). Starter bacteria are often derived from yeast or commercially available bacteria, which are relatively expensive. This research aims to utilize waste products, namely the purple eggplant (Solanum melongena L.) stalks, as a new source of LAB for lactase enzyme production. Fermenting cow's milk using purple eggplant stalks results in yogurt. Inoculating this yogurt onto MRS Broth and MRS Agar media confirms the presence of LAB from the milk fermentation. To test the lactase enzyme, bacterial cell wall degradation is achieved using sonication, allowing for lactase enzyme testing. Characterization test for lactase enzyme is conducted at various temperatures and pH levels to determine the optimum conditions for the lactase enzyme. The optimum temperature for lactase enzyme is at 70°C with a specific enzyme activity of 0.0061 U/mg, optimum pH is 7 with a specific enzyme activity of 0.0044 U/mg. This study demonstrates that purple eggplant stalks can be utilized as a new source of LAB to produce lactase enzyme as an alternative starter bacterium for yogurt production, particularly for lactose intolerance therapy in the future.