Arabinosylation of cell wall extensin is required for the directional response to salinity in roots.

Soil salinity is a major contributor to crop yield losses. To improve our understanding of root responses to salinity, we developed and exploited a real-time salt-induced tilting assay. This assay follows root growth upon both gravitropic and salt challenges, revealing that root bending upon tilting is modulated by Na+ ions, but not by osmotic stress. Next, we measured this salt-specific response in 345 natural Arabidopsis (Arabidopsis thaliana) accessions and discovered a genetic locus, encoding the cell wall-modifying enzyme EXTENSIN ARABINOSE DEFICIENT TRANSFERASE (ExAD) that is associated with root bending in the presence of NaCl (hereafter salt). Extensins are a class of structural cell wall glycoproteins known as hydroxyproline (Hyp)-rich glycoproteins, which are posttranslationally modified by O-glycosylation, mostly involving Hyp-arabinosylation. We show that salt-induced ExAD-dependent Hyp-arabinosylation influences root bending responses and cell wall thickness. Roots of exad1 mutant seedlings, which lack Hyp-arabinosylation of extensin, displayed increased thickness of root epidermal cell walls and greater cell wall porosity. They also showed altered gravitropic root bending in salt conditions and a reduced salt-avoidance response. Our results suggest that extensin modification via Hyp-arabinosylation is a unique salt-specific cellular process required for the directional response of roots exposed to salinity.

Revisiting an ecophysiological oddity: Hydathode-mediated foliar water uptake in Crassula species from southern Africa.

Hydathodes are usually associated with water exudation in plants. However, foliar water uptake (FWU) through the hydathodes has long been suspected in the leaf-succulent genus Crassula (Crassulaceae), a highly diverse group in southern Africa, and, to our knowledge, no empirical observations exist in the literature that unequivocally link FWU to hydathodes in this genus. FWU is expected to be particularly beneficial on the arid western side of southern Africa, where up to 50% of Crassula species occur and where periodically high air humidity leads to fog and/or dew formation. To investigate if hydathode-mediated FWU is operational in different Crassula species, we used the apoplastic fluorescent tracer Lucifer Yellow in combination with different imaging techniques. Our images of dye-treated leaves confirm that hydathode-mediated FWU does indeed occur in Crassula and that it might be widespread across the genus. Hydathodes in Crassula serve as moisture-harvesting structures, besides their more common purpose of guttation, an adaptation that has likely played an important role in the evolutionary history of the genus. Our observations suggest that ability for FWU is independent of geographical distribution and not restricted to arid environments under fog influence, as FWU is also operational in Crassula species from the rather humid eastern side of southern Africa. Our observations point towards no apparent link between FWU ability and overall leaf surface wettability in Crassula. Instead, the hierarchically sculptured leaf surfaces of several Crassula species may facilitate FWU due to hydrophilic leaf surface microdomains, even in seemingly hydrophobic species. Overall, these results confirm the ecophysiological relevance of hydathode-mediated FWU in Crassula and reassert the importance of atmospheric humidity for some arid-adapted plant groups.

Cell wall polysaccharide and glycoprotein content tracks growth-form diversity and an aridity gradient in the leaf-succulent genus Crassula.

Cell wall traits are believed to be a key component of the succulent syndrome, an adaptive syndrome to drought, yet the variability of such traits remains largely unknown. In this study, we surveyed the leaf polysaccharide and glycoprotein composition in a wide sampling of Crassula species that occur naturally along an aridity gradient in southern Africa, and we interpreted its adaptive significance in relation to growth form and arid adaptation. To study the glycomic diversity, we sampled leaf material from 56 Crassula taxa and performed comprehensive microarray polymer profiling to obtain the relative content of cell wall polysaccharides and glycoproteins. This analysis was complemented by the determination of monosaccharide composition and immunolocalization in leaf sections using glycan-targeting antibodies. We found that compact and non-compact Crassula species occupy distinct phenotypic spaces in terms of leaf glycomics, particularly in regard to rhamnogalacturonan I, its arabinan side chains, and arabinogalactan proteins (AGPs). Moreover, these cell wall components also correlated positively with increasing aridity, which suggests that they are likely advantageous in terms of arid adaptation. These differences point to compact Crassula species having more elastic cell walls with plasticizing properties, which can be interpreted as an adaptation toward increased drought resistance. Furthermore, we report an intracellular pool of AGPs associated with oil bodies and calcium oxalate crystals, which could be a peculiarity of Crassula and could be linked to increased drought resistance. Our results indicate that glycomics may be underlying arid adaptation and drought resistance in succulent plants.

Computer habits and digital literacy in geriatric patients: A survey.

Objective: Among hospitalised geriatric patients, only half are computer users. However, many of them refrain from using telehealth solutions. This study aimed to investigate geriatric patients' computer and Internet habits and digital literacy and their associations with stress levels and frequency of Internet use. Methods: Inpatients and outpatients aged 65 years or older, all computer users, were consecutively surveyed. Besides information about computer and Internet habits, computer support, and computer stress, the survey also collected information about digital literacy using the electronic Health Literacy Assessment toolkit. Results: A total of 124 computer users with a mean age of 80.6 ± 7.4 years participated in the study from 1 October to 1 December 2019. Most patients received computer support from their children and grandchildren, whereas 6% did not seek support. They found themselves 'most familiar with using a keyboard' (79%), 59% 'were unfamiliar with the Copy Paste function', and only one-third 'were open to new ways of using computers'. Digital literacy was associated with the frequency of Internet use (P = 0.001), and higher digital literacy was associated with less computer stress (P = 0.01). Conclusions: Geriatric computer users are challenged by their basic computer skills, which may influence their choice of participation in telehealth solutions. If telehealth solutions are to succeed among geriatric patients, individualised computer support based on their basic computer skills and user-friendly computer devices are a prerequisite. For ongoing support, it is also necessary to introduce people close to the patient to telehealth solutions.

Focus on the relationships between the cell wall composition in the extraction of anthocyanins and tannins from grape berries.

Concentrations of anthocyanins and tannins after extraction from berries in wines and from skin macerations in model solutions have been studied for two grape varieties, two maturation levels and two vintages berries. Characterization of the cell wall polysaccharides has also been performed, the classical method based on the analysis of the neutral sugars after depolymerization being completed by a comprehensive microarray polymer profiling (CoMPP). Extraction was lower in model solutions than in wines, with the same ranking: non acylated anthocyanins> tannins > p-coumaroylated anthocyanins. The polysaccharidic composition suggested a role of homogalacturonans, rhamnogalacturonans and extensins in the extraction process. A global explanation of the interactions between anthocyanins, tannins and polysaccharides is proposed.

Combined high-throughput and fractionation approaches reveal changes of polysaccharides in blueberry skin cell walls during fermentation for wine production.

There have been rare reports about the structure/composition of polymers in blueberry skin and their changes during fermentation for wine production. In this study, the compositional changes occurring in blueberry skin during fermentation were tracked by a combination of cell wall analysis techniques including infra-red spectroscopy, monosaccharide analysis, and comprehensive microarray polymer profiling (CoMPP). The cross-corroborating data revealed that blueberry skin cell wall is particularly rich in xyloglucan. Chemical fractionation analysis indicated that the KOH soluble fraction is a dominant fraction in fermented blueberry skin. Interestingly, the KOH soluble fraction contained abundant epitopes associated with pectin branch chains, indicating tight binding of some enzyme-resistant pectin polymers to hemicellulose. This study provides important implications for the development of effective strategies to extract beneficial substances (such as aromatics, tannins and pigments) from berry tissues during processing.

Are cell wall traits a component of the succulent syndrome?

Succulence is an adaptation to low water availability characterised by the presence of water-storage tissues that alleviate water stress under low water availability. The succulent syndrome has evolved convergently in over 80 plant families and is associated with anatomical, physiological and biochemical traits. Despite the alleged importance of cell wall traits in drought responses, their significance in the succulent syndrome has long been overlooked. Here, by analyzing published pressure-volume curves, we show that elastic adjustment, whereby plants change cell wall elasticity, is uniquely beneficial to succulents for avoiding turgor loss. In addition, we used comprehensive microarray polymer profiling (CoMPP) to assess the biochemical composition of cell walls in leaves. Across phylogenetically diverse species, we uncover several differences in cell wall biochemistry between succulent and non-succulent leaves, pointing to the existence of a 'succulent glycome'. We also highlight the glycomic diversity among succulent plants, with some glycomic features being restricted to certain succulent lineages. In conclusion, we suggest that cell wall biomechanics and biochemistry should be considered among the characteristic traits that make up the succulent syndrome.

Corrigendum: Strategies for Efficient Gene Editing in Protoplasts of Solanum tuberosum Theme: Determining gRNA Efficiency Design by Utilizing Protoplast (Research).

[This corrects the article DOI: 10.3389/fgeed.2021.795644.].

Tele-Rehabilitation in Community-Dwelling Older People with Reduced Functional Capacity: A 4-Patient Case Report.

Tele-rehabilitation (TR) can limit physical contact in older patients if long transportation times or physical attendance challenges their health. Digital literacy decreases with age, which might interfere with geriatric patients' ability to benefit from TR. The purpose was to describe the TR intervention, adherence to training sessions, and level of digital literacy. TR via videoconferencing was delivered both individually and in groups where more were challenged by joining training outside their homes. Improvements in functional capacity were found. Combining individual and group exercises, high adherence to the TR programme was achieved. Digital literacy was on par with younger adult computer users. Individual-and group TR sessions might secure high adherence to the exercise programme. Digital literacy at a certain level might be a prerequisite when supplied with both oral and written material Suggestions on how to raise older people's digital literacy to use telehealth solutions in the future are provided.

Prevalence of Computer Use among Geriatric In- and Outpatients.

Objective: Despite its many benefits, tele-rehabilitation is not widely used by the older generations. This study aimed to investigate the opportunity to offer tele-rehabilitation in a geriatric population by determining the prevalence of computer use and to examine whether the patients' characteristics affect computer use. Design: Cross-sectional study. Patients: Cognitive well-functioning in- and outpatients aged 65 years or older. Methods: Patients were consecutively included and surveyed. Results: A total of 249 patients participated in the survey. Among them, 124 were computer users. Four of these never went online. Compared to non-users, computer users were younger OR: 0.91 (95% CI: 0.87; 0.94) p = 0.001, less frail OR: 0.37 (95% CI: 0.25;0.55) p = 0.001, had a higher functional capacity OR: 1.02 (95% CI 1.01; 1.03) p = 0.001 and more often had an education at high school level or higher OR: 1.7 (95% CI: 1.41; 2.40). Conclusion: Only half of the geriatric patients are computer users. If tele-rehabilitation is to be adopted by a wider geriatric population challenged by reduced mobility, long transportation times, or frailty, computer training, user friendly devices and computer support should be considered beforehand.

Immune-instructive copolymer scaffolds using plant-derived nanoparticles to promote bone regeneration.

BACKGROUND: Age-driven immune signals cause a state of chronic low-grade inflammation and in consequence affect bone healing and cause challenges for clinicians when repairing critical-sized bone defects in elderly patients. METHODS: Poly(L-lactide-co-ɛ-caprolactone) (PLCA) scaffolds are functionalized with plant-derived nanoparticles from potato, rhamnogalacturonan-I (RG-I), to investigate their ability to modulate inflammation in vitro in neutrophils and macrophages at gene and protein levels. The scaffolds' early and late host response at gene, protein and histological levels is tested in vivo in a subcutaneous rat model and their potential to promote bone regeneration in an aged rodent was tested in a critical-sized calvaria bone defect. Significant differences were tested using one-way ANOVA, followed by a multiple-comparison Tukey's test with a p value ≤ 0.05 considered significant. RESULTS: Gene expressions revealed PLCA scaffold functionalized with plant-derived RG-I with a relatively higher amount of galactose than arabinose (potato dearabinated (PA)) to reduce the inflammatory state stimulated by bacterial LPS in neutrophils and macrophages in vitro. LPS-stimulated neutrophils show a significantly decreased intracellular accumulation of galectin-3 in the presence of PA functionalization compared to Control (unmodified PLCA scaffolds). The in vivo gene and protein expressions revealed comparable results to in vitro. The host response is modulated towards anti-inflammatory/ healing at early and late time points at gene and protein levels. A reduced foreign body reaction and fibrous capsule formation is observed when PLCA scaffolds functionalized with PA were implanted in vivo subcutaneously. PLCA scaffolds functionalized with PA modulated the cytokine and chemokine expressions in vivo during early and late inflammatory phases. PLCA scaffolds functionalized with PA implanted in calvaria defects of aged rats downregulating pro-inflammatory gene markers while promoting osteogenic markers after 2 weeks in vivo. CONCLUSION: We have shown that PLCA scaffolds functionalized with plant-derived RG-I with a relatively higher amount of galactose play a role in the modulation of inflammatory responses both in vitro and in vivo subcutaneously and promote the initiation of bone formation in a critical-sized bone defect of an aged rodent. Our study addresses the increasing demand in bone tissue engineering for immunomodulatory 3D scaffolds that promote osteogenesis and modulate immune responses.

The effect of enzyme treatment on polyphenol and cell wall polysaccharide extraction from the grape berry and subsequent sensory attributes in Cabernet Sauvignon wines.

Pectolytic enzyme maceration is common for producing red wines, but the effects on bitterness and astringency are not well understood. Glycan microarrays assessed polysaccharide diversity and with polyphenol analysis was correlated with sensory data on descriptors of astringency and their perceived levels in enzyme-crafted Cabernet Sauvignon wines. Enzyme use is shown to have no effect on bitterness, but enzyme-macerated wines are more astringent. The data suggests that pectolytic enzymes are much more pronounced in their effect on the cell wall matrix than the ripeness of the berries at harvest and subsequent sensory perception. Enzyme-macerated red wines showed higher levels of polyphenol which were more polymerized and galloylated. The polyphenol-rich wines were described as hard, chalky, grippy, grainy and dry. The non-enzyme wines had elevated levels of arabinogalactan protein and pectin epitopes (notably biomarker mAbs JIM8 and JIM13) with the wines being characterized as soft, fine and velvety.

Elastic and collapsible: current understanding of cell walls in succulent plants.

Succulent plants represent a large functional group of drought-resistant plants that store water in specialized tissues. Several co-adaptive traits accompany this water-storage capacity to constitute the succulent syndrome. A widely reported anatomical adaptation of cell walls in succulent tissues allows them to fold in a regular fashion during extended drought, thus preventing irreversible damage and permitting reversible volume changes. Although ongoing research on crop and model species continuously reports the importance of cell walls and their dynamics in drought resistance, the cell walls of succulent plants have received relatively little attention to date, despite the potential of succulents as natural capital to mitigate the effects of climate change. In this review, we summarize current knowledge of cell walls in drought-avoiding succulents and their effects on tissue biomechanics, water relations, and photosynthesis. We also highlight the existing knowledge gaps and propose a hypothetical model for regulated cell wall folding in succulent tissues upon dehydration. Future perspectives of methodological development in succulent cell wall characterization, including the latest technological advances in molecular and imaging techniques, are also presented.

Impact of the variety on the adsorption of anthocyanins and tannins on grape flesh cell walls.

BACKGROUND: During winemaking, after extraction from the skins, anthocyanins and tannins adsorb onto the pulp flesh cell walls. The present study aimed to quantify the amounts adsorbed and their impact on wine composition, the impact of variety and ethanol on adsorption, and whether the presence of anthocyanins plays a role and impacts tannin adsorption. RESULTS: Anthocyanin and tannin fractions obtained by mimicking winemaking conditions were mixed with fresh flesh cell walls of two varieties: Carignan and Grenache. Adsorption isotherms were measured. Adsorption of tannins was higher with Carignan than with Grenache and decreased when the ethanol content increased. In comparison, anthocyanins were adsorbed in small amounts, and their mixing with tannins had no impact on their adsorption. The differences were related to differences in pulp cell wall composition, particularly in terms of extensins and arabinans. CONCLUSION: Adsorption of tannins, which can reach 50% of the initial amount, depends on the pulp cell wall composition. This needs to be investigated further. © 2021 Society of Chemical Industry.

A diverse member of the fungal Avr4 effector family interacts with de-esterified pectin in plant cell walls to disrupt their integrity.

Effectors are small, secreted proteins that promote pathogen virulence. Although key to microbial infections, unlocking the intrinsic function of effectors remains a challenge. We have previously shown that members of the fungal Avr4 effector family use a carbohydrate-binding module of family 14 (CBM14) to bind chitin in fungal cell walls and protect them from host chitinases during infection. Here, we show that gene duplication in the Avr4 family produced an Avr4-2 paralog with a previously unknown effector function. Specifically, we functionally characterize PfAvr4-2, a paralog of PfAvr4 in the tomato pathogen Pseudocercospora fuligena, and show that although it contains a CBM14 domain, it does not bind chitin or protect fungi against chitinases. Instead, PfAvr4-2 interacts with highly de-esterified pectin in the plant's middle lamellae or primary cell walls and interferes with Ca2+-mediated cross-linking at cell-cell junction zones, thus loosening the plant cell wall structure and synergizing the activity of pathogen secreted endo-polygalacturonases.

Comprehensive Leaf Cell Wall Analysis Using Carbohydrate Microarrays Reveals Polysaccharide-Level Variation between Vitis Species with Differing Resistance to Downy Mildew.

The cell wall acts as one of the first barriers of the plant against various biotic stressors. Previous studies have shown that alterations in wall polysaccharides may influence crop disease resistance. In the grapevine family, several native species (e.g., Chinese wild grapevine) show a naturally higher resistance to microbial pathogens than cultivated species (e.g., Vitis vinifera), and this trait could be inherited through breeding. Despite the importance of the cell wall in plant immunity, there are currently no comprehensive cell wall profiles of grapevine leaves displaying differing resistance phenotypes, due to the complex nature of the cell wall and the limitations of analytical techniques available. In this study, the cutting-edge comprehensive carbohydrate microarray technology was applied to profile uninfected leaves of the susceptible cultivar (Vitis vinifera cv. "Cabernet Sauvignon"), a resistant cultivar (Vitis amurensis cv. "Shuanghong") and a hybrid offspring cross displaying moderate resistance. The microarray approach uses monoclonal antibodies, which recognize polysaccharides epitopes, and found that epitope abundances of highly esterified homogalacturonan (HG), xyloglucan (with XXXG motif), (galacto)(gluco)mannan and arabinogalactan protein (AGP) appeared to be positively correlated with the high resistance of Vitis amurensis cv. "Shuanghong" to mildew. The quantification work by gas chromatography did not reveal any significant differences for the monosaccharide constituents, suggesting that polysaccharide structural alterations may contribute more crucially to the resistance observed; this is again supported by the contact infrared spectroscopy of cell wall residues, revealing chemical functional group changes (e.g., esterification of pectin). The identification of certain wall polysaccharides that showed alterations could be further correlated with resistance to mildew. Data from the use of the hybrid material in this study have preliminarily suggested that these traits could be inherited and may be applied as potential structural biomarkers in future breeding work.

A group-based real-time videoconferencing telerehabilitation programme in recently discharged geriatric patients: a feasibility study.

PURPOSE: Exercise at home and improvement in the ability to undertake daily tasks are highly valued by older people after hospitalisation. New telerehabilitation (TR) technologies make it possible to supervise and communicate with exercising participants through videoconferencing equipment. This technology has been shown to be both feasible and effective in Danish chronic obstructive pulmonary disease patients in terms of basic mobility, safety, social interactions and patient perception. This study sought to examine whether it was feasible to carry out TR through home exercises in groups. METHODS: Both medical and hip-fracture home-dwelling patients aged 65 years and older admitted to the Emergency Department (ED) and Department of Geriatrics for acute reasons were asked to participate in the study just before their discharge. The inclusion criteria were normal cognitive function, being dependent on a walking aid and computer users before hospitalisation. RESULTS: At discharge, 333 patients were consecutively screened for participation. Of those, 300 patients were excluded. Thirty-three patients met the inclusion criteria. They had a mean age of 82.3 years (± 7.8) and 76% were women. Nine patients agreed to participate, but seven withdrew. The most frequent explanation was exhaustion in the continuation of hospitalisation. CONCLUSION: It was not possible to conduct a videoconference TR study in a geriatric population, as many were excluded and those who were eligible withdrew after inclusion. During the COVID-19 pandemic, TR may be an important tool for isolated older persons to hinder functional decline. Aspects such as recruitment procedures and IT solutions designed for older people must be considered.

Strategies for Efficient Gene Editing in Protoplasts of Solanum tuberosum Theme: Determining gRNA Efficiency Design by Utilizing Protoplast (Research).

Potato, Solanum tuberosum is a highly diverse tetraploid crop. Elite cultivars are extremely heterozygous with a high prevalence of small length polymorphisms (indels) and single nucleotide polymorphisms (SNPs) within and between cultivars, which must be considered in CRISPR/Cas gene editing strategies and designs to obtain successful gene editing. In the present study, in-depth sequencing of the gene encoding glucan water dikinase (GWD) 1 and the downy mildew resistant 6 (DMR6-1) genes in the potato cultivars Saturna and Wotan, respectively, revealed both indels and a 1.3-2.8 higher SNP prevalence when compared to the heterozygous diploid RH genome sequence as expected for a tetraploid compared to a diploid. This complicates guide RNA (gRNA) and diagnostic PCR designs. At the same time, high editing efficiencies at the cell pool (protoplast) level are pivotal for achieving full allelic knock-out in tetraploids. Furthermore, high editing efficiencies reduce the downstream cumbersome and delicate ex-plant regeneration. Here, CRISPR/Cas ribonucleoprotein particles (RNPs) were delivered transiently to protoplasts by polyethylene glycol (PEG) mediated transformation. For each of GWD1 and the DMR6-1, 6-10 gRNAs were designed to target regions comprising the 5' and the 3' end of the two genes. Similar to other studies including several organisms, editing efficiency of the individual RNPs varied significantly, and some generated specific indel patterns. RNP's targeting the 5' end of GWD1 yielded significantly higher editing efficiency as compared to targeting the 3' end. For DMR6-1, such an effect was not seen. Simultaneously targeting each of the two target regions with two RNPs (multiplexing) yielded a clear positive synergistic effect on the total editing when targeting the 3' end of the GWD1 gene only. Multiplexing of the two genes, residing on different chromosomes, yielded no or a slightly negative effect on editing from the single or combined gRNA/RNPs. These initial findings may instigate much larger studies needed for facilitating and optimizing precision breeding in plants.

Amylose/cellulose nanofiber composites for all-natural, fully biodegradable and flexible bioplastics.

Thermoplastic, polysaccharide-based plastics are environmentally friendly. However, typical shortcomings include lack of water resistance and poor mechanical properties. Nanocomposite manufacturing using pure, highly linear, polysaccharides can overcome such limitations. Cast nanocomposites were fabricated with plant engineered pure amylose (AM), produced in bulk quantity in transgenic barley grain, and cellulose nanofibers (CNF), extracted from agrowaste sugar beet pulp. Morphology, crystallinity, chemical heterogeneity, mechanics, dynamic mechanical, gas and water permeability, and contact angle of the films were investigated. Blending CNF into the AM matrix significantly enhanced the crystallinity, mechanical properties and permeability, whereas glycerol increased elongation at break, mainly by plasticizing the AM. There was significant phase separation between AM and CNF. Dynamic plasticizing and anti-plasticizing effects of both CNF and glycerol were demonstrated by NMR demonstrating high molecular order, but also non-crystalline, and evenly distributed 20 nm-sized glycerol domains. This study demonstrates a new lead in functional polysaccharide-based bioplastic systems.

Genome Wide Identification and Comparative Analysis of the Serpin Gene Family in Brachypodium and Barley.

Serpins (serine protease inhibitors) constitute one of the largest and most widely distributed superfamilies of protease inhibitors and have been identified in nearly all organisms. To gain significant insights, a comprehensive in silico analysis of the serpin gene family was carried out in the model plant for temperate grasses Brachypodium distachyon and barley Hordeum vulgare using bioinformatic tools at the genome level for the first time. We identified a total of 27 BdSRPs and 25 HvSRP genes in Brachypodium and barley, respectively, showing an unexpectedly high gene number in these model plants. Gene structure, conserved motifs and phylogenetic comparisons of serpin genes supported the role of duplication events in the expansion and evolution of serpin gene family. Further, purifying selection pressure was found to be a main driving force in the evolution of serpin genes. Genome synteny analysis indicated that BdSRP genes were present in syntenic regions of barley, rice, sorghum and maize, suggesting that they evolved before the divergence of these species from common ancestor. The distinct expression pattern in specific tissues further suggested a specialization of functions during development and in plant defense. These results suggest that the LR serpins (serpins with Leu-Arg residues at P2-P1') identified here can be utilized as candidates for exploitation in disease resistance, pest control and preventing stress-induced cell death. Additionally, serpins were identified that could lead to further research aimed at validating and functionally characterizing the role of potential serpin genes from other plants.