A Cork Cell Wall Approach To Swelling And Boiling

The bark of cork oak (Quercus suber L.) is mostly used for cork stopper production, whereas bark is undergoing a series of industrial procedures, boiling usually leading to changes in the characteristics of the tissue. Trees are traditionally grown under natural conditions; however, irrigation is now being used in plantations. These permanent water availability affects cork-oak development, while its effects on industrial procedures is unknown. This study provides a first insight into the behaviour of the cell walls of cork during the process of swelling and boiling when trees have been grown under irrigation, subject to a specific water regime. Cork tissue was analysed using environmental and scanning electron microscopy under three regimes: raw conditions; following immersion in water; and after boiling. Additionally, the radial expansion of samples was determined. The results showed greater cell-wall expansion in cork from the irrigated site than cork from the traditional rainfed plot, when hydrated for 24h. After boiling, the cell walls of the rainfed site were thinner than in the raw stage, in contrast to the irrigated cork. This study suggests that irrigation during cork-oak growth produces a higher capacity for adsorption, increasing cell-wall thickness from the raw stage to the boiling stage.

Heavy isotope labeling and mass spectrometry reveal unexpected remodeling of bacterial cell wall expansion in response to drugs

Antibiotics of the β-lactam (penicillin) family inactivate target enzymes called D,D-transpeptidases or penicillin-binding proteins (PBPs) that catalyze the last cross-linking step of peptidoglycan synthesis. The resulting net-like macromolecule is the essential component of bacterial cell walls that sustains the osmotic pressure of the cytoplasm. In Escherichia coli, bypass of PBPs by the YcbB L,D-transpeptidase leads to resistance to these drugs. We developed a new method based on heavy isotope labeling and mass spectrometry to elucidate PBP- and YcbB-mediated peptidoglycan polymerization. PBPs and YcbB similarly participated in single-strand insertion of glycan chains into the expanding bacterial side wall. This absence of any transpeptidase-specific signature suggests that the peptidoglycan expansion mode is determined by other components of polymerization complexes. YcbB did mediate β-lactam resistance by insertion of multiple strands that were exclusively cross-linked to existing tripeptide-containing acceptors. We propose that this unprecedented mode of polymerization depends upon accumulation of linear glycan chains due to PBP inactivation, formation of tripeptides due to cleavage of existing cross-links by a β-lactam-insensitive endopeptidase, and concerted cross-linking by YcbB.

Genome-Wide Analysis of the COBRA-Like Gene Family Supports Gene Expansion through Whole-Genome Duplication in Soybean (Glycine max)

The COBRA-like (COBL) gene family has been associated with the regulation of cell wall expansion and cellulose deposition. COBL mutants result in reduced levels and disorganized deposition of cellulose causing defects in the cell wall and inhibiting plant development. In this study, we report the identification of 24 COBL genes (GmCOBL) in the soybean genome. Phylogenetic analysis revealed that the COBL proteins are divided into two groups, which differ by about 170 amino acids in the N-terminal region. The GmCOBL genes were heterogeneously distributed in 14 of the 20 soybean chromosomes. This study showed that segmental duplication has contributed significantly to the expansion of the COBL family in soybean during all Glycine-specific whole-genome duplication events. The expression profile revealed that the expression of the paralogous genes is highly variable between organs and tissues of the plant. Only 20% of the paralogous gene pairs showed similar expression patterns. The high expression levels of some GmCOBLs suggest they are likely essential for regulating cell expansion during the whole soybean life cycle. Our comprehensive overview of the COBL gene family in soybean provides useful information for further understanding the evolution and diversification of COBL genes in soybean.

Molecular investigation of Tuscan sweet cherries sampled over three years: gene expression analysis coupled to metabolomics and proteomics

Sweet cherry (Prunus avium L.) is a stone fruit widely consumed and appreciated for its organoleptic properties, as well as its nutraceutical potential. We here investigated the characteristics of six non-commercial Tuscan varieties of sweet cherry maintained at the Regional Germplasm Bank of the CNR-IBE in Follonica (Italy) and sampled ca. 60 days post-anthesis over three consecutive years (2016-2017-2018). We adopted an approach merging genotyping and targeted gene expression profiling with metabolomics. To complement the data, a study of the soluble proteomes was also performed on two varieties showing the highest content of flavonoids. Metabolomics identified the presence of flavanols and proanthocyanidins in highest abundance in the varieties Morellona and Crognola, while gene expression revealed that some differences were present in genes involved in the phenylpropanoid pathway during the 3 years and among the varieties. Finally, proteomics on Morellona and Crognola showed variations in proteins involved in stress response, primary metabolism and cell wall expansion. To the best of our knowledge, this is the first multi-pronged study focused on Tuscan sweet cherry varieties providing insights into the differential abundance of genes, proteins and metabolites.

Hydrogen sulfide promotes hypocotyl elongation via increasing cellulose content and changing the arrangement of cellulose fibrils in alfalfa

Hydrogen sulfide (H2S) is known to have positive physiological functions in plant growth, but limited data are available on its influence on cell walls. Here, we demonstrate a novel mechanism by which H2S regulates the biosynthesis and deposition of cell wall cellulose in alfalfa (Medicago sativa). Treatment with NaHS was found to increase the length of epidermal cells in the hypocotyl, and transcriptome analysis indicated that it caused the differential expression of numerous of cell wall-related genes. These differentially expressed genes were directly associated with the biosynthesis of cellulose and hemicellulose, and with the degradation of pectin. Analysis of cell wall composition showed that NaHS treatment increased the contents of cellulose and hemicellulose, but decreased the pectin content. Atomic force microscopy revealed that treatment with NaHS decreased the diameter of cellulose fibrils, altered the arrangement of the fibrillar bundles, and increased the spacing between the bundles. The dynamics of cellulose synthase complexes (CSCs) were closely related to cellulose synthesis, and NaHS increased the rate of mobility of the particles. Overall, our results suggest that the H2S signal enhances the plasticity of the cell wall by regulating the deposition of cellulose fibrils and by decreasing the pectin content. The resulting increases in cellulose and hemicellulose contents lead to cell wall expansion and cell elongation.

A mechanical cusp catastrophe imposes a universal developmental constraint on the shapes of tip-growing cells

Understanding the mechanistic basis for cell morphology is a central problem in biology. Evolution has converged on tip growth many times, yielding filamentous cells, yet tip-growing cells display a range of apical morphologies. To understand this variability, we measured the spatial profiles of cell-wall expansion for three species that spanned the phylogeny and morphology of tip-growth. Profiles were consistent with a mechanical model whereby the wall was stratified and stretched by turgor pressure during cell growth. We calculated the spatial profiles of wall mechanical properties, which could be accurately fit with an empirical two-parameter function. Combined with the mechanical model, this function yielded a “morphospace” that accounted for the shapes of diverse tip-growing species. However, natural shapes were bounded by a cusp bifurcation in the morphospace that separated thin, fast-growing cells from (nonexistent) wide, slow-growing cells. This constraint has important implications for our understanding of the evolution of tip-growing cells.

High post-anthesis temperature effects on bread wheat (Triticum aestivum L.) grain transcriptome during early grain-filling

Background: High post-anthesis (p.a) temperatures reduce mature grain weight in wheat. However, the causes of this reduction are not entirely known. Control of grain expansion by the maternally derived pericarp of the grain has previously been suggested, although this interaction has not been investigated under high p.a. temperatures. Down-regulation of pericarp localised genes that regulate cell wall expansion under high p.a. temperatures may limit expansion of the encapsulated endosperm due to a loss of plasticity in the pericarp, reducing mature grain weight. Here the effect of high p.a. temperatures on the transcriptome of the pericarp and endosperm of the wheat grain during early grain-filling was investigated via RNA-Seq and is discussed alongside grain moisture dynamics during early grain development and mature grain weight. Results: High p.a. temperatures applied from 6-days after anthesis (daa) and until 18daa reduced the grain’s ability to accumulate water, with total grain moisture and percentage grain moisture content being significantly reduced from 14daa onwards. Mature grain weight was also significantly reduced by the same high p.a. temperatures applied from 6daa for 4-days or more, in a separate experiment. Comparison of our RNA-Seq data from whole grains, with existing data sets from isolated pericarp and endosperm tissues enabled the identification of subsets of genes whose expression was significantly affected by high p.a. temperature and predominantly expressed in either tissue. Hierarchical clustering and gene ontology analysis resulted in the identification of a number of genes implicated in the regulation of cell wall expansion, predominantly expressed in the pericarp and significantly down-regulated under high p.a. temperatures, including endoglucanase, xyloglucan endotransglycosylases and a β-expansin. An over-representation of genes involved in the ‘cuticle development’ functional pathway that were expressed in the pericarp and affected by high p.a. temperatures was also observed. Conclusions: High p.a. temperature induced down-regulation of genes involved in regulating pericarp cell wall expansion. This concomitant down-regulation with a reduction in total grain moisture content and grain weight following the same treatment period, adds support to the theory that high p.a. temperatures may cause a reduction in mature grain weight as result of decreased pericarp cell wall expansion.

High post-anthesis temperature effects on bread wheat (Triticum aestivum L.) grain transcriptome during early grain-filling

Background: High post-anthesis (p.a) temperatures reduce mature grain weight in wheat. However, the causes of this reduction are not entirely known. Control of grain expansion by the maternally derived pericarp of the grain has previously been suggested, although this interaction has not been investigated under high p.a. temperatures. Down-regulation of pericarp localised genes that regulate cell wall expansion under high p.a. temperatures may limit expansion of the encapsulated endosperm due to a loss of plasticity in the pericarp, reducing mature grain weight. Here the effect of high p.a. temperatures on the transcriptome of the pericarp and endosperm of the wheat grain during early grain-filling was investigated via RNA-Seq and is discussed alongside grain moisture dynamics during early grain development and mature grain weight. Results: High p.a. temperatures applied from 6-days after anthesis (daa) and until 18daa reduced the grain’s ability to accumulate water, with total grain moisture and percentage grain moisture content being significantly reduced from 14daa onwards. Mature grain weight was also significantly reduced by the same high p.a. temperatures applied from 6daa for 4-days or more, in a separate experiment. Comparison of our RNA-Seq data from whole grains, with existing data sets from isolated pericarp and endosperm tissues enabled the identification of subsets of genes whose expression was significantly affected by high p.a. temperature and predominantly expressed in either tissue. Hierarchical clustering and gene ontology analysis resulted in the identification of a number of genes implicated in the regulation of cell wall expansion, predominantly expressed in the pericarp and significantly down-regulated under high p.a. temperatures, including endoglucanase, xyloglucan endotransglycosylases and a β-expansin. An over-representation of genes involved in the ‘cuticle development’ functional pathway that were expressed in the pericarp and affected by high p.a. temperatures was also observed. Conclusions: High p.a. temperature induced down-regulation of genes involved in regulating pericarp cell wall expansion. This concomitant down-regulation with a reduction in total grain moisture content and grain weight following the same treatment period, adds support to the theory that high p.a. temperatures may cause a reduction in mature grain weight as result of decreased pericarp cell wall expansion.