Pectin Chemistry and Cellulose Crystallinity Govern Pavement Cell Morphogenesis in a Multi-Step Mechanism.

Simple plant cell morphologies, such as cylindrical shoot cells, are determined by the extensibility pattern of the primary cell wall, which is thought to be largely dominated by cellulose microfibrils, but the mechanism leading to more complex shapes, such as the interdigitated patterns in the epidermis of many eudicotyledon leaves, is much less well understood. Details about the manner in which cell wall polymers at the periclinal wall regulate the morphogenetic process in epidermal pavement cells and mechanistic information about the initial steps leading to the characteristic undulations in the cell borders are elusive. Here, we used genetics and recently developed cell mechanical and imaging methods to study the impact of the spatio-temporal dynamics of cellulose and homogalacturonan pectin distribution during lobe formation in the epidermal pavement cells of Arabidopsis (Arabidopsis thaliana) cotyledons. We show that nonuniform distribution of cellulose microfibrils and demethylated pectin coincides with spatial differences in cell wall stiffness but may intervene at different developmental stages. We also show that lobe period can be reduced when demethyl-esterification of pectins increases under conditions of reduced cellulose crystallinity. Our data suggest that lobe initiation involves a modulation of cell wall stiffness through local enrichment in demethylated pectin, whereas subsequent increase in lobe amplitude is mediated by the stress-induced deposition of aligned cellulose microfibrils. Our results reveal a key role of noncellulosic polymers in the biomechanical regulation of cell morphogenesis.

Honey vesicle-like nanoparticles protect aged liver from non-alcoholic steatohepatitis.

Non-alcoholic steatohepatitis (NASH), an advanced form of non-alcoholic fatty liver disease (NAFLD), has emerged as the leading cause of liver failure and related death. Currently, no medication is specifically approved to treat NAFLD or NASH. Here we report that oral administration of honey vesicle-like nanoparticles (H-VLNs) to naturally aged mice protects the liver from NASH development. H-VLNs are dominantly taken up by Kupffer cells in the liver and suppress hepatic chronic inflammation and further development of fibrosis and nodule formation in aged mice. Besides their reported anti-inflammasome function, H-VLNs are found to inhibit the transcriptional activities of C-JUN and nuclear factor-kappa B (NF-κB). MicroRNAs miR5119 and miR5108 and phenolic compound luteolin in H-VLNs are identified in suppressing both the C-JUN and NF-κB pathways. Collectively, oral intake of H-VLNs represents a promising new user-friendly modality to prevent the development of NASH.

3D Visualization of Microtubules in Epidermal Pavement Cells.

The plant cytoskeleton is instrumental in cellular processes such as cell growth, differentiation, and immune response. Microtubules, in particular, play a crucial role in morphogenesis by governing the deposition of plant cell wall polysaccharides and, in consequence, the cell wall mechanics and cell shape. Scrutinizing the microtubule dynamics is therefore integral to understanding the spatiotemporal regulation of cellular activities. In this chapter, we outline steps to acquire 3D images of microtubules in epidermal pavement cells of Arabidopsis thaliana cotyledons using a confocal microscope. We introduce the steps to assess the microtubule distribution and organization using image processing software Bitplane Imaris and ImageJ. We also demonstrate how the interpretation of image material can be facilitated by post-processing with general-purpose image enhancement software using methods trained by artificial intelligence-based algorithms.

TaNAC032 transcription factor regulates lignin-biosynthetic genes to combat Fusarium head blight in wheat.

Fusarium head blight (FHB) is a destructive disease affecting cereal crops globally due to mycotoxin contamination of grains that reduce yield and quality. Among hundreds of QTLs identified for resistance, the QTL-Fhb1 is of significant interest even today, for its major contribution to FHB resistance. Previously, QTL-Fhb1 dissection based on a combined metabolo-genomics approach, identified a few potential resistance genes, including a NAC like transcription factor for FHB resistance. Sequencing and phylogenetic analysis confirmed NAC to be the wheat TaNAC032. Also, the quantitative RT-PCR studies revealed a greater induced expression of TaNAC032 in resistant NIL in comparison to susceptible NIL upon Fusarium graminearum (Fg) infection. The virus-induced gene silencing (VIGS) based functional validation of TaNAC032 in resistant NIL confirmed increased disease severity and fungal biomass. Metabolic profiling revealed low abundances of resistance-related (RR) metabolites in TaNAC032 silenced NIL-R compared to non-silenced. Silenced plants showed decreased transcript abundances of RR metabolite biosynthetic genes associated with a reduction in total lignin content in rachis, confirming the regulatory role of TaNAC032 in wheat in response to Fg infection. If TaNA032 is mutated in an FHB susceptible cultivar, it can be edited to enhance FHB resistance.

Mechanical Stress Initiates and Sustains the Morphogenesis of Wavy Leaf Epidermal Cells.

Pavement cells form wavy interlocking patterns in the leaf epidermis of many plants. We use computational mechanics to simulate the morphogenetic process based on microtubule organization and cell wall chemistry. Based on the in silico simulations and experimental evidence, we suggest that a multistep process underlies the morphogenesis of pavement cells. The in silico model predicts alternatingly located, feedback-augmented mechanical heterogeneity of the periclinal and anticlinal walls. It suggests that the emergence of waves is created by a stiffening of the emerging indented sides, an effect that matches cellulose and de-esterified pectin patterns in the cell wall. Further, conceptual evidence for mechanical buckling of the cell walls is provided, a mechanism that has the potential to initiate wavy patterns de novo and may precede chemical and geometrical symmetry breaking.

Understanding plant cell morphogenesis requires real-time monitoring of cell wall polymers.

Plant cell development and growth are determined by the expansion pattern of the cell wall, a matrix of mixed polysaccharide polymers and proteins. To understand the different roles of these polymers in the regulation of the morphogenetic process, their spatial dynamics need to be monitored over time. Recent developments in the live cell labeling of polysaccharides include specific dyes whose insertion into the wall does not interfere with wall properties and growth, as well as metabolically inserted labeling. The present review explains the motivation and necessity for novel polysaccharide labeling techniques and provides an overview of the insight gained with these strategies.