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1.
Curr Genet ; 70(1): 6, 2024 May 11.
Article En | MEDLINE | ID: mdl-38733432

The gene products of PRS1-PRS5 in Saccharomyces cerevisiae are responsible for the production of PRPP (5-phospho-D-ribosyl-α-1-pyrophosphate). However, it has been demonstrated that they are also involved in the cell wall integrity (CWI) signalling pathway as shown by protein-protein interactions (PPIs) with, for example Slt2, the MAP kinase of the CWI pathway. The following databases: SGD, BioGRID and Hit Predict, which collate PPIs from various research papers, have been scrutinized for evidence of PPIs between Prs1-Prs5 and components of the CWI pathway. The level of certainty in PPIs was verified by interaction scores available in the Hit Predict database revealing that well-documented interactions correspond with higher interaction scores and can be graded as high confidence interactions based on a score > 0.28, an annotation score ≥ 0.5 and a method-based high confidence score level of ≥ 0.485. Each of the Prs1-Prs5 polypeptides shows some degree of interaction with the CWI pathway. However, Prs5 has a vital role in the expression of FKS2 and Rlm1, previously only documented by reporter assay studies. This report emphasizes the importance of investigating interactions using more than one approach since every method has its limitations and the use of different methods, as described herein, provides complementary experimental and statistical data, thereby corroborating PPIs. Since the experimental data described so far are consistent with a link between PRPP synthetase and the CWI pathway, our aim was to demonstrate that these data are also supported by high-throughput bioinformatic analyses promoting our hypothesis that two of the five PRS-encoding genes contain information required for the maintenance of CWI by combining data from our targeted approach with relevant, unbiased data from high-throughput analyses.


Cell Wall , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae , Cell Wall/metabolism , Cell Wall/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Signal Transduction , Protein Interaction Maps , Protein Interaction Mapping
2.
Proc Natl Acad Sci U S A ; 121(21): e2319707121, 2024 May 21.
Article En | MEDLINE | ID: mdl-38743622

Glycogen is a glucose storage molecule composed of branched α-1,4-glucan chains, best known as an energy reserve that can be broken down to fuel central metabolism. Because fungal cells have a specialized need for glucose in building cell wall glucans, we investigated whether glycogen is used for this process. For these studies, we focused on the pathogenic yeast Cryptococcus neoformans, which causes ~150,000 deaths per year worldwide. We identified two proteins that influence formation of both glycogen and the cell wall: glycogenin (Glg1), which initiates glycogen synthesis, and a protein that we call Glucan organizing enzyme 1 (Goe1). We found that cells missing Glg1 lack α-1,4-glucan in their walls, indicating that this material is derived from glycogen. Without Goe1, glycogen rosettes are mislocalized and ß-1,3-glucan in the cell wall is reduced. Altogether, our results provide mechanisms for a close association between glycogen and cell wall.


Cell Wall , Cryptococcus neoformans , Fungal Proteins , Glucans , Glycogen , Cell Wall/metabolism , Glycogen/metabolism , Glucans/metabolism , Fungal Proteins/metabolism , Cryptococcus neoformans/metabolism , Glucosyltransferases/metabolism , beta-Glucans/metabolism
3.
Carbohydr Polym ; 337: 122149, 2024 Aug 01.
Article En | MEDLINE | ID: mdl-38710571

Phytopathogen cell wall polysaccharides have important physiological functions. In this study, we isolated and characterized the alkali-insoluble residue on the inner layers of the Rhizoctonia solani AG1 IA cell wall (RsCW-AIR). Through chemical composition and structural analysis, RsCW-AIR was mainly identified as a complex of chitin/chitosan and glucan (ChCsGC), with glucose and glucosamine were present in a molar ratio of 2.7:1.0. The predominant glycosidic bond linkage of glucan in ChCsGC was ß-1,3-linked Glcp, both the α and ß-polymorphic forms of chitin were presented in it by IR, XRD, and solid-state NMR, and the ChCsGC exhibited a degree of deacetylation measuring 67.08 %. RsCW-AIR pretreatment effectively reduced the incidence of rice sheath blight, and its induced resistance activity in rice was evaluated, such as inducing a reactive oxygen species (ROS) burst, leading to the accumulation of salicylic acid (SA) and the up-regulation of SA-related gene expression. The recognition of RsCW-AIR in rice is partially dependent on CERK1.


Cell Wall , Chitin , Chitosan , Glucans , Oryza , Plant Diseases , Rhizoctonia , Rhizoctonia/drug effects , Oryza/microbiology , Oryza/chemistry , Cell Wall/chemistry , Chitosan/chemistry , Chitosan/pharmacology , Chitin/chemistry , Chitin/pharmacology , Glucans/chemistry , Glucans/pharmacology , Plant Diseases/microbiology , Disease Resistance , Reactive Oxygen Species/metabolism
4.
J R Soc Interface ; 21(214): 20240008, 2024 May.
Article En | MEDLINE | ID: mdl-38715319

Multicellular organisms grow and acquire their shapes through the differential expansion and deformation of their cells. Recent research has addressed the role of cell and tissue mechanical properties in these processes. In plants, it is believed that growth rate is a function of the mechanical stress exerted on the cell wall, the thin polymeric layer surrounding cells, involving an effective viscosity. Nevertheless, recent studies have questioned this view, suggesting that cell wall elasticity sets the growth rate or that uptake of water is limiting for plant growth. To assess these issues, we developed a microfluidic device to quantify the growth rates, elastic properties and hydraulic conductivity of individual Marchantia polymorpha plants in a controlled environment with a high throughput. We characterized the effect of osmotic treatment and abscisic acid on growth and hydromechanical properties. Overall, the instantaneous growth rate of individuals is correlated with both bulk elastic modulus and hydraulic conductivity. Our results are consistent with a framework in which the growth rate is determined primarily by the elasticity of the wall and its remodelling, and secondarily by hydraulic conductivity. Accordingly, the coupling between the chemistry of the cell wall and the hydromechanics of the cell appears as key to set growth patterns during morphogenesis.


Cell Wall , Cell Wall/physiology , Marchantia/growth & development , Marchantia/physiology , Abscisic Acid/metabolism , Models, Biological , Biomechanical Phenomena , Plant Development/physiology
5.
Appl Microbiol Biotechnol ; 108(1): 324, 2024 May 07.
Article En | MEDLINE | ID: mdl-38713211

Laccase, a copper-containing polyphenol oxidase, is an important green biocatalyst. In this study, Laccase Lcc5 was homologous recombinantly expressed in Coprinopsis cinerea and a novel strategy of silencing chitinase gene expression was used to enhance recombinant Lcc5 extracellular yield. Two critical chitinase genes, ChiEn1 and ChiE2, were selected by analyzing the transcriptome data of C. cinerea FA2222, and their silent expression was performed by RNA interference (RNAi). It was found that silencing either ChiEn1 or ChiE2 reduced sporulation and growth rate, and increased cell wall sensitivity, but had no significant effect on mycelial branching. Among them, the extracellular laccase activity of the ChiE2-silenced engineered strain Cclcc5-antiChiE2-5 and the control Cclcc5-13 reached the highest values (38.2 and 25.5 U/mL, respectively) at 250 and 150 rpm agitation speeds, corresponding to productivity of 0.35 and 0.19 U/mL·h, respectively, in a 3-L fermenter culture. Moreover, since Cclcc5-antiChiE2-5 could withstand greater shear forces, its extracellular laccase activity was 2.6-fold higher than that of Cclcc5-13 when the agitation speed was all at 250 rpm. To our knowledge, this is the first report of enhanced recombinant laccase production in C. cinerea by silencing the chitinase gene. This study will pave the way for laccase industrial production and accelerate the development of a C. cinerea high-expression system. KEY POINTS: • ChiEn1 and ChiE2 are critical chitinase genes in C. cinerea FA2222 genome. • Chitinase gene silencing enhanced the tolerance of C. cinerea to shear forces. • High homologous production of Lcc5 is achieved by fermentation in a 3-L fermenter.


Chitinases , Gene Silencing , Laccase , Chitinases/genetics , Chitinases/metabolism , Chitinases/biosynthesis , Laccase/genetics , Laccase/metabolism , Laccase/biosynthesis , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Agaricales/genetics , Agaricales/enzymology , Fermentation , RNA Interference , Fungal Proteins/genetics , Fungal Proteins/metabolism , Mycelium/genetics , Mycelium/growth & development , Mycelium/enzymology , Cell Wall/metabolism , Cell Wall/genetics
6.
Sci Rep ; 14(1): 10237, 2024 05 03.
Article En | MEDLINE | ID: mdl-38702505

Enzymatic degradation of algae cell wall carbohydrates by microorganisms is under increasing investigation as marine organic matter gains more value as a sustainable resource. The fate of carbon in the marine ecosystem is in part driven by these degradation processes. In this study, we observe the microbiome dynamics of the macroalga Fucus vesiculosus in 25-day-enrichment cultures resulting in partial degradation of the brown algae. Microbial community analyses revealed the phylum Pseudomonadota as the main bacterial fraction dominated by the genera Marinomonas and Vibrio. More importantly, a metagenome-based Hidden Markov model for specific glycosyl hydrolyses and sulphatases identified Bacteroidota as the phylum with the highest potential for cell wall degradation, contrary to their low abundance. For experimental verification, we cloned, expressed, and biochemically characterised two α-L-fucosidases, FUJM18 and FUJM20. While protein structure predictions suggest the highest similarity to a Bacillota origin, protein-protein blasts solely showed weak similarities to defined Bacteroidota proteins. Both enzymes were remarkably active at elevated temperatures and are the basis for a potential synthetic enzyme cocktail for large-scale algal destruction.


Cell Wall , Fucus , Metagenomics , Cell Wall/metabolism , Fucus/metabolism , Fucus/genetics , Fucus/microbiology , Metagenomics/methods , Bacteroidetes/genetics , Bacteroidetes/enzymology , Metagenome , Microbiota , Phylogeny
7.
Nat Commun ; 15(1): 3792, 2024 May 06.
Article En | MEDLINE | ID: mdl-38710711

Infection with the apicomplexan protozoan Toxoplasma gondii can be life-threatening in immunocompromised hosts. Transmission frequently occurs through the oral ingestion of T. gondii bradyzoite cysts, which transition to tachyzoites, disseminate, and then form cysts containing bradyzoites in the central nervous system, resulting in latent infection. Encapsulation of bradyzoites by a cyst wall is critical for immune evasion, survival, and transmission. O-glycosylation of the protein CST1 by the mucin-type O-glycosyltransferase T. gondii (Txg) GalNAc-T3 influences cyst wall rigidity and stability. Here, we report X-ray crystal structures of TxgGalNAc-T3, revealing multiple features that are strictly conserved among its apicomplexan homologues. This includes a unique 2nd metal that is coupled to substrate binding and enzymatic activity in vitro and cyst wall O-glycosylation in T. gondii. The study illustrates the divergence of pathogenic protozoan GalNAc-Ts from their host homologues and lays the groundwork for studying apicomplexan GalNAc-Ts as therapeutic targets in disease.


Protozoan Proteins , Toxoplasma , Toxoplasma/enzymology , Toxoplasma/genetics , Glycosylation , Protozoan Proteins/metabolism , Protozoan Proteins/genetics , Protozoan Proteins/chemistry , Humans , Crystallography, X-Ray , Glycosyltransferases/metabolism , Glycosyltransferases/genetics , Cell Wall/metabolism , Animals
8.
Glycobiology ; 34(6)2024 Apr 24.
Article En | MEDLINE | ID: mdl-38690785

Cellulose is an abundant component of plant cell wall matrices, and this para-crystalline polysaccharide is synthesized at the plasma membrane by motile Cellulose Synthase Complexes (CSCs). However, the factors that control CSC activity and motility are not fully resolved. In a targeted chemical screen, we identified the alkylated nojirimycin analog N-Dodecyl Deoxynojirimycin (ND-DNJ) as a small molecule that severely impacts Arabidopsis seedling growth. Previous work suggests that ND-DNJ-related compounds inhibit the biosynthesis of glucosylceramides (GlcCers), a class of glycosphingolipid associated with plant membranes. Our work uncovered major changes in the sphingolipidome of plants treated with ND-DNJ, including reductions in GlcCer abundance and altered acyl chain length distributions. Crystalline cellulose content was also reduced in ND-DNJ-treated plants as well as plants treated with the known GlcCer biosynthesis inhibitor N-[2-hydroxy-1-(4-morpholinylmethyl)-2-phenyl ethyl]-decanamide (PDMP) or plants containing a genetic disruption in GLUCOSYLCERAMIDE SYNTHASE (GCS), the enzyme responsible for sphingolipid glucosylation that results in GlcCer synthesis. Live-cell imaging revealed that CSC speed distributions were reduced upon treatment with ND-DNJ or PDMP, further suggesting an important relationship between glycosylated sphingolipid composition and CSC motility across the plasma membrane. These results indicate that multiple interventions compromising GlcCer biosynthesis disrupt cellulose deposition and CSC motility, suggesting that GlcCers regulate cellulose biosynthesis in plants.


Arabidopsis , Cellulose , Glucosylceramides , Glucosyltransferases , Arabidopsis/metabolism , Glucosyltransferases/metabolism , Glucosyltransferases/genetics , Cellulose/metabolism , Cellulose/biosynthesis , Glucosylceramides/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , 1-Deoxynojirimycin/pharmacology , 1-Deoxynojirimycin/analogs & derivatives , Cell Wall/metabolism
9.
Physiol Plant ; 176(3): e14330, 2024.
Article En | MEDLINE | ID: mdl-38698648

Wall-associated kinases (WAKs) have been determined to recognize pathogenic signals and initiate plant immune responses. However, the roles of the family members in host resistance against Valsa canker, a serious fungal disease of apples and pears, are largely unknown. Here, we identified MbWAK1 in Malus baccata, a resistant germplasm differentially expressed during infection by Valsa mali (Vm). Over-expression of MbWAK1 enhanced the Valsa canker resistance of apple and pear fruits and 'Duli-G03' (Pyrus betulifolia) suspension cells. A large number of phloem, cell wall, and lipid metabolic process-related genes were differentially expressed in overexpressed suspension cell lines in response to Valsa pyri (Vp) signals. Among these, the expression of xyloglucan endotransglucosylase/hydrolase (XTH) gene PbeXTH1 and sieve element occlusion B-like (SEOB) gene PbeSEOB1 were significantly inhibited. Transient expression of PbeXTH1 or PbeSEOB1 compromised the expressional induction of MbWAK1 and the resistance contributed by MbWAK1. In addition, PbeXTH1 and PbeSEOB1 suppressed the immune response induced by MbWAK1. Our results enriched the molecular mechanisms for MbWAK1 against Valsa canker and resistant breeding.


Disease Resistance , Gene Expression Regulation, Plant , Malus , Plant Diseases , Plant Proteins , Pyrus , Plant Diseases/microbiology , Plant Diseases/genetics , Plant Diseases/immunology , Disease Resistance/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Pyrus/genetics , Pyrus/microbiology , Malus/genetics , Malus/microbiology , Malus/immunology , Malus/enzymology , Cell Wall/metabolism
10.
Proc Natl Acad Sci U S A ; 121(21): e2314570121, 2024 May 21.
Article En | MEDLINE | ID: mdl-38739804

Lipid polymers such as cutin and suberin strengthen the diffusion barrier properties of the cell wall in specific cell types and are essential for water relations, mineral nutrition, and stress protection in plants. Land plant-specific glycerol-3-phosphate acyltransferases (GPATs) of different clades are central players in cutin and suberin monomer biosynthesis. Here, we show that the GPAT4/6/8 clade in Arabidopsis thaliana, which is known to mediate cutin formation, is also required for developmentally regulated root suberization, in addition to the established roles of GPAT5/7 in suberization. The GPAT5/7 clade is mainly required for abscisic acid-regulated suberization. In addition, the GPAT5/7 clade is crucial for the formation of the typical lamellated suberin ultrastructure observed by transmission electron microscopy, as distinct amorphous globular polyester structures were deposited in the apoplast of the gpat5 gpat7 double mutant, in contrast to the thinner but still lamellated suberin deposition in the gpat4 gpat6 gpat8 triple mutant. Site-directed mutagenesis revealed that the intrinsic phosphatase activity of GPAT4, GPAT6, and GPAT8, which leads to monoacylglycerol biosynthesis, contributes to suberin formation. GPAT5/7 lack an active phosphatase domain and the amorphous globular polyester structure observed in the gpat5 gpat7 double mutant was partially reverted by treatment with a phosphatase inhibitor or the expression of phosphatase-dead variants of GPAT4/6/8. Thus, GPATs that lack an active phosphatase domain synthetize lysophosphatidic acids that might play a role in the formation of the lamellated structure of suberin. GPATs with active and nonactive phosphatase domains appear to have nonredundant functions and must cooperate to achieve the efficient biosynthesis of correctly structured suberin.


Arabidopsis Proteins , Arabidopsis , Lipids , Plant Roots , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis/growth & development , Plant Roots/metabolism , Plant Roots/growth & development , Plant Roots/genetics , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Lipids/chemistry , Gene Expression Regulation, Plant , Glycerol-3-Phosphate O-Acyltransferase/metabolism , Glycerol-3-Phosphate O-Acyltransferase/genetics , Membrane Lipids/metabolism , Abscisic Acid/metabolism , Cell Wall/metabolism , 1-Acylglycerol-3-Phosphate O-Acyltransferase
11.
Int J Mol Sci ; 25(9)2024 Apr 30.
Article En | MEDLINE | ID: mdl-38732136

In the context of sustainable agriculture and biomaterial development, understanding and enhancing plant secondary cell wall formation are crucial for improving crop fiber quality and biomass conversion efficiency. This is especially critical for economically important crops like upland cotton (Gossypium hirsutum L.), for which fiber quality and its processing properties are essential. Through comprehensive genome-wide screening and analysis of expression patterns, we identified a particularly high expression of an R2R3 MYB transcription factor, GhMYB52 Like, in the development of the secondary cell wall in cotton fiber cells. Utilizing gene-editing technology to generate a loss-of-function mutant to clarify the role of GhMYB52 Like, we revealed that GhMYB52 Like does not directly contribute to cellulose synthesis in cotton fibers but instead represses a subset of lignin biosynthesis genes, establishing it as a lignin biosynthesis inhibitor. Concurrently, a substantial decrease in the lint index, a critical measure of cotton yield, was noted in parallel with an elevation in lignin levels. This study not only deepens our understanding of the molecular mechanisms underlying cotton fiber development but also offers new perspectives for the molecular improvement of other economically important crops and the enhancement of biomass energy utilization.


Cotton Fiber , Gene Expression Regulation, Plant , Gossypium , Lignin , Plant Proteins , Lignin/biosynthesis , Gossypium/genetics , Gossypium/metabolism , Gossypium/growth & development , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Cell Wall/metabolism , Cell Wall/genetics , Cellulose/biosynthesis , Cellulose/metabolism , Biosynthetic Pathways
12.
ACS Biomater Sci Eng ; 10(5): 2956-2966, 2024 May 13.
Article En | MEDLINE | ID: mdl-38593061

Bacteria experience substantial physical forces in their natural environment, including forces caused by osmotic pressure, growth in constrained spaces, and fluid shear. The cell envelope is the primary load-carrying structure of bacteria, but the mechanical properties of the cell envelope are poorly understood; reports of Young's modulus of the cell envelope of Escherichia coli range from 2 to 18 MPa. We developed a microfluidic system to apply mechanical loads to hundreds of bacteria at once and demonstrated the utility of the approach for evaluating whole-cell stiffness. Here, we extend this technique to determine Young's modulus of the cell envelope of E. coli and of the pathogens Vibrio cholerae and Staphylococcus aureus. An optimization-based inverse finite element analysis was used to determine the cell envelope Young's modulus from observed deformations. The Young's modulus values of the cell envelope were 2.06 ± 0.04 MPa for E. coli, 0.84 ± 0.02 MPa for E. coli treated with a chemical (A22) known to reduce cell stiffness, 0.12 ± 0.03 MPa for V. cholerae, and 1.52 ± 0.06 MPa for S. aureus (mean ± SD). The microfluidic approach allows examination of hundreds of cells at once and is readily applied to Gram-negative and Gram-positive organisms as well as rod-shaped and cocci cells, allowing further examination of the structural causes behind differences in cell envelope Young's modulus among bacterial species and strains.


Elastic Modulus , Escherichia coli , Staphylococcus aureus , Vibrio cholerae , Staphylococcus aureus/physiology , Staphylococcus aureus/drug effects , Vibrio cholerae/physiology , Escherichia coli/physiology , Escherichia coli/drug effects , Finite Element Analysis , Cell Membrane/physiology , Cell Membrane/drug effects , Cell Wall/drug effects
13.
J Mol Biol ; 436(10): 168570, 2024 May 15.
Article En | MEDLINE | ID: mdl-38604529

Cellular mRNA levels, particularly under stress conditions, can be finely regulated by the coordinated action of transcription and degradation processes. Elements of the 5'-3' mRNA degradation pathway, functionally associated with the exonuclease Xrn1, can bind to nuclear chromatin and modulate gene transcription. Within this group are the so-called decapping activators, including Pat1, Dhh1, and Lsm1. In this work, we have investigated the role of Pat1 in the yeast adaptive transcriptional response to cell wall stress. Thus, we demonstrated that in the absence of Pat1, the transcriptional induction of genes regulated by the Cell Wall Integrity MAPK pathway was significantly affected, with no effect on the stability of these transcripts. Furthermore, under cell wall stress conditions, Pat1 is recruited to Cell Wall Integrity-responsive genes in parallel with the RNA Pol II complex, participating both in pre-initiation complex assembly and transcriptional elongation. Indeed, strains lacking Pat1 showed lower recruitment of the transcription factor Rlm1, less histone H3 displacement at Cell Wall Integrity gene promoters, and impaired recruitment and progression of RNA Pol II. Moreover, Pat1 and the MAPK Slt2 occupied the coding regions interdependently. Our results support the idea that Pat1 and presumably other decay factors behave as transcriptional regulators of Cell Wall Integrity-responsive genes under cell wall stress conditions.


Cell Wall , Gene Expression Regulation, Fungal , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae , Cell Wall/metabolism , Cell Wall/genetics , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , MAP Kinase Signaling System , Transcription, Genetic , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA Polymerase II/metabolism , RNA Polymerase II/genetics , RNA Stability , Transcription Factors/metabolism , Transcription Factors/genetics , Endoribonucleases/metabolism , Endoribonucleases/genetics , RNA-Binding Proteins , MADS Domain Proteins
14.
J Hazard Mater ; 471: 134313, 2024 Jun 05.
Article En | MEDLINE | ID: mdl-38669927

Secondary lignification of the root exodermis of Kandelia obovata is crucial for its response to adversity such as high salinity and anaerobic environment, and this lignification is also effective in blocking cadmium transport to the roots. However, how the differences in lignification of root exodermis at different developmental stages respond to Cd stress and its regulatory mechanisms have not been revealed. In this study, after analyzing the root structure and cell wall thickness using a Phenom scanning electron microscope as well as measuring cadmium content in the root cell wall, we found that the exodermis of young and mature roots of K. obovata responded to Cd stress through the polymerization of different lignin monomers, forming two different mechanisms: chelation and blocking. Through small RNA sequencing, RLM-5'-RACE and dual luciferase transient expression system, we found that miR397 targets and regulates KoLAC4/17/7 expression. The expression of KoLAC4/17 promoted the accumulation of guaiacyl lignin during lignification and enhanced the binding of cadmium to the cell wall. Meanwhile, KoLAC7 expression promotes the accumulation of syringyl lignin during lignification, which enhances the obstruction of cadmium and improves the tolerance to cadmium. These findings enhance our understanding of the molecular mechanisms underlying the differential lignification of the root exodermis of K. obovata in response to cadmium stress, and provide scientific guidance for the conservation of mangrove forests under heavy metal pollution.


Cadmium , Lignin , MicroRNAs , Plant Roots , Lignin/chemistry , Cadmium/toxicity , Plant Roots/drug effects , Plant Roots/metabolism , Plant Roots/growth & development , MicroRNAs/metabolism , MicroRNAs/genetics , Stress, Physiological/drug effects , Gene Expression Regulation, Plant/drug effects , Polymerization/drug effects , Cell Wall/drug effects , Cell Wall/metabolism , Araceae/drug effects , Araceae/metabolism , Plant Proteins/metabolism , Plant Proteins/genetics
15.
Plant Physiol Biochem ; 210: 108568, 2024 May.
Article En | MEDLINE | ID: mdl-38581806

Postharvest physiological deterioration (PPD) reduces the availability and economic value of fresh produces, resulting in the waste of agricultural products and becoming a worldwide problem. Therefore, many studies have been carried out at the anatomical structural, physiological and biochemical levels and molecular levels of PPD of fresh produces to seek ways to manage the postharvest quality of fresh produce. The cell wall is the outermost structure of a plant cell and as such represents the first barrier to prevent external microorganisms and other injuries. Many studies on postharvest quality of crop storage organs relate to changes in plant cell wall-related components. Indeed, these studies evidence the non-negligible role of the plant cell wall in postharvest storage ability. However, the relationship between cell wall metabolism and postharvest deterioration of fresh produces has not been well summarized. In this review, we summarize the structural changes of cell walls in different types of PPD, metabolic changes, and the possible molecular mechanism regulating cell wall metabolism in PPD of fresh produce. This review provides a basis for further research on delaying the occurrence of PPD of fresh produce.


Cell Wall , Cell Wall/metabolism , Fruit/metabolism , Fruit/physiology
16.
Plant Physiol Biochem ; 210: 108621, 2024 May.
Article En | MEDLINE | ID: mdl-38604012

To enhance the postharvest quality of avocado (Persea americana Mill.) fruit, this study investigates alterations in cell wall metabolism and reactive oxygen species (ROS) metabolism during near-freezing temperature (NFT) storage, and explores their impact on fruit softening. The fruit was stored at 25 °C, 5 °C, 2 °C, and NFT, respectively. NFT storage retarded firmness loss and chilling injury in comparison with 25 °C, 5 °C, and 2 °C. NFT storage delayed the decrease of ionic-soluble pectin (ISP) and cellulose (CLL) contents by suppressing cell wall degradation enzyme activities. Correlation analysis showed that cell wall degradation enzyme activities were positively correlated to rates of ethylene release and respiration. Moreover, NFT storage maintained higher levels of DPPH and ABTS scavenging abilities, activities of superoxide dismutase, peroxidase, and catalase, as well as ascorbate-glutathione cycle (ascorbic acid, glutathione, glutathione disulfide, ascorbate peroxidase, cycle-related enzymes), thereby inhibited the increase of ROS content, malondialdehyde content, and cell membrane permeability. Fruit firmness and chilling injury were correlated with the contents of hydrogen (H2O2), superoxide anion (O2.-), ISP, and CLL. These results suggested that NFT could suppress fruit softening and chilling injury by inhibiting cell wall degradation through delaying respiration and ethylene production and suppressing ROS production via activation of antioxidant systems, thereby maintaining quality and prolonged storage life during avocado fruit storage.


Cell Wall , Fruit , Persea , Reactive Oxygen Species , Persea/metabolism , Cell Wall/metabolism , Reactive Oxygen Species/metabolism , Fruit/metabolism , Food Storage/methods , Cold Temperature , Freezing , Ethylenes/metabolism , Pectins/metabolism , Cellulose/metabolism
17.
Int J Biol Macromol ; 267(Pt 1): 131420, 2024 May.
Article En | MEDLINE | ID: mdl-38583835

Natural product bulgecin A potentiates the activity of ß-lactam antibiotics by inhibition of three lytic transglycosylases in Pseudomonas aeruginosa, of which MltD is one. MltD exhibits both endolytic and exolytic reactions in the turnover of the cell-wall peptidoglycan and tolerates the presence or absence of stem peptides in its substrates. The present study reveals structural features of the multimodular MltD, presenting a catalytic module and four cell-wall-binding LysM modules that account for these attributes. Three X-ray structures are reported herein for MltD that disclose one unpredicted LysM module tightly attached to the catalytic domain, whereas the other LysM modules are mobile, and connected to the catalytic domain through long flexible linkers. The formation of crystals depended on the presence of bulgecin A. The expansive active-site cleft is highlighted by the insertion of a helical region, a hallmark of the family 1D of lytic transglycosylases, which was mapped out in a ternary complex of MltD:bulgecinA:chitotetraose, revealing at the minimum the presence of eight subsites (from -4 to +4, with the seat of reaction at subsites -1 and + 1) for binding of sugars of the substrate for the endolytic reaction. The mechanism of the exolytic reaction is revealed in one of the structures, showing how the substrate's terminal anhydro-NAM moiety could be sequestered at subsite +2. Our results provide the structural insight for both the endolytic and exolytic activities of MltD during cell-wall-turnover events.


Catalytic Domain , Pseudomonas aeruginosa , Pseudomonas aeruginosa/enzymology , Models, Molecular , Glycosyltransferases/chemistry , Glycosyltransferases/metabolism , Biological Products/chemistry , Biological Products/pharmacology , Crystallography, X-Ray , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Cell Wall , Substrate Specificity
18.
Methods Mol Biol ; 2788: 81-95, 2024.
Article En | MEDLINE | ID: mdl-38656510

Atomic force microscopy (AFM) has broken boundaries in the characterization of the supramolecular architecture of cell wall assemblies and single cell wall polysaccharides at the nanoscale level. Moreover, AFM provides an opportunity to evaluate the mechanical properties of cell wall material which is not possible with any other method. However, in the case of plant tissue, the critical step is a smart sample preparation that should not affect the polysaccharide structure or assembly and on the other hand should consider device limitations, especially scanner ranges. In this chapter, the protocols from the sample preparation, including isolation of cell wall material and extraction of cell wall polysaccharide fractions, through AFM imaging of polysaccharide assemblies and single molecules until an image analysis to obtain quantitative data characterizing the biopolymers are presented.


Cell Wall , Microscopy, Atomic Force , Microscopy, Atomic Force/methods , Cell Wall/ultrastructure , Cell Wall/chemistry , Polysaccharides/chemistry , Polysaccharides/analysis
19.
Proc Natl Acad Sci U S A ; 121(15): e2321759121, 2024 Apr 09.
Article En | MEDLINE | ID: mdl-38579009

Adjacent plant cells are connected by specialized cell wall regions, called middle lamellae, which influence critical agricultural characteristics, including fruit ripening and organ abscission. Middle lamellae are enriched in pectin polysaccharides, specifically homogalacturonan (HG). Here, we identify a plant-specific Arabidopsis DUF1068 protein, called NKS1/ELMO4, that is required for middle lamellae integrity and cell adhesion. NKS1 localizes to the Golgi apparatus and loss of NKS1 results in changes to Golgi structure and function. The nks1 mutants also display HG deficient phenotypes, including reduced seedling growth, changes to cell wall composition, and tissue integrity defects. These phenotypes are comparable to qua1 and qua2 mutants, which are defective in HG biosynthesis. Notably, genetic interactions indicate that NKS1 and the QUAs work in a common pathway. Protein interaction analyses and modeling corroborate that they work together in a stable protein complex with other pectin-related proteins. We propose that NKS1 is an integral part of a large pectin synthesis protein complex and that proper function of this complex is important to support Golgi structure and function.


Arabidopsis Proteins , Arabidopsis , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cell Adhesion/genetics , Pectins/metabolism , Golgi Apparatus/genetics , Golgi Apparatus/metabolism , Cell Wall/metabolism
20.
Plant Mol Biol ; 114(3): 38, 2024 Apr 11.
Article En | MEDLINE | ID: mdl-38605193

The cell wall (CW) is the dynamic structure of a plant cell, acting as a barrier against biotic and abiotic stresses. In grape berries, the modifications of pulp and skin CW during softening ensure flexibility during cell expansion and determine the final berry texture. In addition, the CW of grape berry skin is of fundamental importance for winemaking, controlling secondary metabolite extractability. Grapevine varieties with contrasting CW characteristics generally respond differently to biotic and abiotic stresses. In the context of climate change, it is important to investigate the CW dynamics occurring upon different stresses, to define new adaptation strategies. This review summarizes the molecular mechanisms underlying CW modifications during grapevine berry fruit ripening, plant-pathogen interaction, or in response to environmental stresses, also considering the most recently published transcriptomic data. Furthermore, perspectives of new biotechnological approaches aiming at modifying the CW properties based on other crops' examples are also presented.


Fruit , Vitis , Fruit/genetics , Fruit/metabolism , Vitis/genetics , Vitis/metabolism , Gene Expression Profiling , Cell Wall/metabolism , Stress, Physiological
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