Cell Wall Extensibility Research Articles

Nitric oxide induces S-nitrosylation of CESA1 and CESA9 and increases cellulose content in Arabidopsis hypocotyls

Nitric oxide (NO), a small signaling gas molecule, participates in several growth and developmental processes in plants. However, how NO regulates cell wall biosynthesis remains unclear. Here, we demonstrate a positive effect of NO on cellulose content that may be related to S-nitrosylation of cellulose synthase 1 (CESA1) and CESA9. Two S-nitrosylated cysteine (Cys) residues, Cys562 and Cys641, which are exposed on the surface of CESA1 and CESA9 and located in the cellulose synthase catalytic domain, were identified to be S-nitrosylated. Meanwhile, Cys641 was located on the binding surface of CESA1 and CESA9, and Cys562 was very close to the binding surface. Cellulose synthase complexes (CSCs) dynamics are closely associated with cellulose content. S-nitrosylation of CESA1 and CESA9 improved particles mobility and thus increased the accumulation of cellulose in Arabidopsis hypocotyl cells. An increase in hemicellulose content as well as an alteration in pectin content facilitated cell wall extension and contributed to cell growth, finally promoting elongation of Arabidopsis hypocotyls. Overall, our work provides a path to investigate the way NO affects the cellulose content of plants.

Modification of Xyloglucan Metabolism during a Decrease in Cell Wall Extensibility in 1-Aminocyclopropane-1-Carboxylic Acid-Treated Azuki Bean Epicotyls.

The exogenous application of ethylene or 1-aminocyclopropane-1-carboxylic acid (ACC), the biosynthetic precursor for ethylene, to plants decreases the capacity of the cell wall to extend, thereby inhibiting stem elongation. In this study, the mechanism by which the extensibility of cell walls decreases in ACC-treated azuki bean epicotyls was studied. ACC decreased the total extensibility of cell walls, and such a decrease was due to the decrease in irreversible extensibility. ACC increased the molecular mass of xyloglucans but decreased the activity of xyloglucan-degrading enzymes. The expression of VaXTHS4, which only exhibits hydrolase activity toward xyloglucans, was downregulated by ACC treatment, whereas that of VaXTH1 or VaXTH2, which exhibits only transglucosylase activity toward xyloglucans, was not affected by ACC treatment. The suppression of xyloglucan-degrading activity by downregulating VaXTHS4 expression may be responsible for the increase in the molecular mass of xyloglucan. Our results suggest that the modification of xyloglucan metabolism is necessary to decrease cell wall extensibility, thereby inhibiting the elongation growth of epicotyls in ACC-treated azuki bean seedlings.

Influence of Low-Molar-Mass Xyloglucans on the Rheological Behavior of Concentrated Cellulose Nanocrystal Suspensions.

Hydrogels were prepared at high solid contents (70-100 g/L) with cellulose nanocrystals (CNC) and very short xyloglucans (XGs). At 70 g/L, CNCs form cholesteric liquid crystals regularly spaced by a distance of 30 nm. This structure was preserved after adsorption of XG with a molar mass (Mw) of 20,000 g/mol (XG20) but was lost at 40,000 g/mol (XG40). Rheological measurements discriminated domains where an increasing Mw from XG20 to XG40 gave rise to drastic changes in storage moduli (on 3 orders of magnitude). At 40,000 g/mol, transient systems were obtained and a re-entrant glass-gel-glass transition was observed with increasing XG concentrations. This was interpreted in terms of the length and stiffness of the chain in relation to the inter-CNC distance. Liquid-to-glass-to-gel transitions were attributed to an XG adsorption type according to train or trail conformations or interconnected structures. Such tunable properties may further have implications on the in vivo role of XG during cell wall extension.

Tomato STEROL GLYCOSYLTRANSFERASE 1 silencing unveils a major role of steryl glycosides in plant and fruit development

Free and glycosylated sterols localize in the plant cell plasma membrane, where in combination with other lipids regulate its structure and function. The role of glycosylated sterols in regulating membrane-associated biological processes is more relevant in plants like tomato (Solanum lycopersicum), in which glycosylated sterols are the predominant sterols. A proper ratio of free sterols versus glycosylated sterols has proven to be essential for proper plant performance in several species, but almost nothing is known in tomato. To assess the role of glycosylated sterols in tomato plant and fruit development, we generated transgenic lines of tomato cultivar Micro-Tom expressing two different amiRNAs devised to silence STEROL GLYCOSYLTRANSFERASE 1, the most actively expressed of the four genes encoding sterol glycosyltransferases in this plant. STEROL GLYCOSYLTRANSFERASE 1 gene silencing caused moderate plant dwarfism and reduced fruit size. Analysis of the profile of glycosylated sterols throughout fruit development demonstrated that the maintenance of proper levels of these compounds during the early stages of fruit development is essential for normal fruit growth, since reduced levels of glycosylated sterols trigger a transcriptional downregulatory response that affects genes involved in processes that are critical for proper fruit development, such as seed filling, cell wall extension and auxin signaling.

Pectin Characteristics Affect Root Growth in Spinach under Salinity.

In understanding the role of root cell wall mechanisms in plant tolerance to salinity, it is important to elucidate the changes in the pectin composition and physical properties of the cell wall. Two salt-sensitive (Helan 3 and Prius β) and one salt-tolerant (R7) spinach cultivars were used to investigate the pectin polysaccharides, the characteristics of pectin, including the degree of pectin methy-lesterification, the HG:RG-I ratio, neutral side chains (galactan/arabinangalactan), and elasticity and viscosity parameters in the root elongation zone under salinity. Root growth was inhibited by salinity, whereas the root diameter was thickened in all cultivars. Salinity significantly reduced cell wall extensibility in all cultivars, and increased cell wall viscosity in Helan 3 and R7 relative to Prius β. Pectin was significantly increased under salinity stress. Cell wall viscosity was affected by pectin due to the molar proportion of uronic acid and/or pectin characteristics (HG:RG-I ratio). The molar proportion of uronic acid in pectin was reduced in Helan 3 and R7 compared with Prius β. The length and degree of pectin methy-lesterification of neutral side chains were significantly decreased in the R7 cultivar, with no significant changes in the other two cultivars. Demethylation of pectin could alter root growth and boost salt tolerance in the R7 cultivar. In this study, it is shown that cell wall pectin played important roles in regulating the root growth of Spinacia oleracea L. under salinity stress.

The Arabidopsis Receptor-like Kinase CAP1 Promotes Shoot Growth under Ammonium Stress.

Simple SummaryUnderstanding the underlying mechanisms of NH4+ toxicity is essential for improving nitrogen use efficiency. Although numerous genes and factors have been identified to function in modulating the response to NH4+ stress, NH4+ toxicity remains poorly characterized. Our work reported here demonstrated a new role for CAP1 in shoot growth in response to NH4+ stress. The enhanced sensitivity of the cap1-1 mutant to NH4+ stress is linked with the role of CAP1 in regulation cell wall loosening and ROS accumulation.High levels of ammonium (NH4+) in soils inhibit plant growth and nitrogen utilization efficiency. Elucidating the underlying mechanisms of NH4+ toxicity is essential for alleviating the growth inhibition caused by high NH4+. Our previous work showed that [Ca2+]cyt-associated protein kinase 1 (CAP1) regulates root hair growth in response to NH4+ in Arabidopsis thaliana, and the cap1-1 mutant produces short root hairs under NH4+ stress conditions. However, it is unclear whether CAP1 functions in other physiological processes in response to NH4+. In the present study, we found that CAP1 also plays a role in attenuating NH4+ toxicity to promote shoot growth. The cap1-1 mutant produced smaller shoots with smaller epidermal cells compared with the wild type in response to NH4+ stress. Disruption of CAP1 enhanced the NH4+-mediated inhibition of the expression of cell enlargement-related genes. The cap1-1 mutant showed elevated reactive oxygen species (ROS) levels under NH4+ stress, as well as increased expression of respiratory burst oxidase homologue genes and decreased expression of catalase genes compared with the wild type. Our data reveal that CAP1 attenuates NH4+-induced shoot growth inhibition by promoting cell wall extensibility and ROS homeostasis, thereby highlighting the role of CAP1 in the NH4+ signal transduction pathway.

Genome-wide association, RNA-seq and iTRAQ analyses identify candidate genes controlling radicle length of wheat.

The radicle, present in the embryo of a seed, is the first root to emerge at germination, and its rapid growth is essential for establishment and survival of the seedling. However, there are few studies on the critical mechanisms underlying radicle and then radicle length in wheat seedlings, despite its importance as a food crop throughout the world. In the present study, 196 wheat accessions from the Huanghuai Wheat Region were screened to measure radicle length under 4 hydroponic culture environments over 3 years. Different expression genes and proteins (DEGs/DEPs) between accessions with extremely long [Yunong 949 (WRL1), Zhongyu 9,302 (WRL2)] and short roots [Yunong 201 (WRS1), Beijing 841 (WRS2)] were identified in 12 sets of root tissue samples by RNA-seq and iTRAQ (Isobaric tags for relative and absolute quantification). Phenotypic results showed that the elongation zone was significantly longer in root accessions with long roots compared to the short-rooted accessions. A genome-wide association study (GWAS) identified four stable chromosomal regions significantly associated with radicle length, among which 1A, 4A, and 7A chromosomes regions explained 7.17% to12.93% of the phenotypic variation. The omics studies identified the expression patterns of 24 DEGs/DEPs changed at both the transcriptional and protein levels. These DEGs/DEPs were mainly involved in carbon fixation in photosynthetic organisms, photosynthesis and phenylpropanoid biosynthesis pathways. TraesCS1A02G104100 and TraesCS2B02G519100 were involved in the biosynthesis of tricin-lignins in cell walls and may affect the extension of cell walls in the radicle elongation zone. A combination of GWAS and RNA-seq analyses revealed 19 DEGs with expression changes in the four accessions, among which, TraesCS1A02G422700 (a cysteine-rich receptor-like protein kinase 6, CRK6) also showed upregulation in the comparison group by RNA-seq, iTRAQ, and qRT-PCR. BSMV-mediated gene silencing also showed that TaCRK6 improves root development in wheat. Our data suggest that TaCRK6 is a candidate gene regulating radicle length in wheat.

ROP GTPase-dependent polarity establishment during tip growth in plants.

Polar cell growth in plants requires a cell peripheral region that undergoes membrane extension and cell wall remodeling. Since the 1990s, RHO-RELATED GTPASES FROM PLANTS (ROPs) have been identified as master regulators that determine the site of cell growth. ROPs function to regulate actin and microtubule cytoskeletons, calcium gradients, and exocytosis, thus directing the delivery of materials for membrane and cell wall extension. In recent years, our understanding of the regulatory mechanisms underlying polar localization and the activation of ROPs has greatly advanced. Evidence points to the crucial roles of membrane lipids, receptor-like kinases, and cell wall components. In this review, we provide updates on the mechanisms underlying polarity control in tip-growing cells, with a focus on ROP effectors and membrane-associated signals. By integrating knowledge from pollen tubes, root hairs, and findings in bryophyte protonema cells and rhizoids, we hope to offer important insights into a common conceptual framework on polarity establishment governed by intercellular and extracellular signals.

Boron-mediated lignin metabolism in response to aluminum toxicity in citrus (Poncirus trifoliata (L.) Raf.) root

Aluminum (Al) toxicity has conspicuous detrimental effects on citrus production whereas boron (B) has been shown to alleviate its toxicity. Lignin plays a critical role in the cell wall extensibility and root elongation under stressed conditions. Hence, the interaction between B and Al on cell wall structure and lignin-related metabolic pathway was investigated in root of trifoliate orange (Poncirus trifoliata (L.) Raf.) seedlings. The results showed B supply considerably decreased the Al content in root, particularly in cell wall, and reduced Al-induced damage on growth-related parameters and thickness of cell wall. Boron application decreased the hydrogen peroxide (H2O2), malondialdehyde (MDA), and lignin contents in the Al-treated root, which prevents the inhibitory effects of Al on the root length. Moreover, metabonomics results showed that B addition resulted in the reduction of metabolites involved in the lignin biosynthesis pathways (phenylpropanoid metabolic) i.e., shikimic acid, tyrosine, caffeic acid, chlorogenic acid, coniferyl alcohol, sinapinic acid, sinapaldehyde, and sinapyl alcohol, as well as distinctively restrain the activities of lignin biosynthesis-related enzymes (4-coumarate-CoA ligase (4CL), cinnamyl-alcohol dehydrogenase (CAD)) under Al toxicity. Collectively, our findings suggest that the positive effects of B on the resistance of Al toxicity may be it reduces Al accumulation in the cell wall, lignin biosynthesis, and cell wall thickness, thereby increasing the extensibility and elasticity of cell wall and thus promoting root elongation.

MsPG4-mediated hydrolysis of pectins increases the cell wall extensibility and aluminum resistance of alfalfa

MsPG4-mediated hydrolysis of pectins increases the cell wall extensibility and aluminum resistance of alfalfa

Cell Wall Components and Extensibility Regulate Root Growth in Suaeda salsa and Spinacia oleracea under Salinity.

Understanding the role of root cell walls in the mechanism of plant tolerance to salinity requires elucidation of the changes caused by salinity in the interactions between the mechanical properties of the cell walls and root growth, and between the chemical composition of the cell walls and root growth. Here, we investigated cell wall composition and extensibility of roots by growing a halophyte (Suaeda salsa) and a glycophyte (Spinacia oleracea) species under an NaCl concentration gradient. Root growth was inhibited by increased salinity in both species. However, root growth was more strongly reduced in S. oleracea than in S. salsa. Salinity reduced cell wall extensibility in S. oleracea significantly, whereas treatment with up to 200 mM NaCl increased it in S. salsa. Meanwhile, S. salsa root cell walls exhibited relatively high cell wall stiffness under 300 mM NaCl treatment, which resist wall deformation under such stress conditions. There was no decrease in pectin content with salinity treatment in the cell walls of the elongation zone of S. salsa roots. Conversely, a decrease in pectin content was noted with increasing salinity in S. oleracea, which might be due to Na+ accumulation. Cellulose content and uronic acid proportions in pectin increased with salinity in both species. Our results suggest that (1) cell wall pectin plays important roles in cell wall extension in both species under salinity, and that the salt tolerance of glycophyte S. oleracea is affected by the pectin; (2) cellulose limits root elongation under saline conditions in both species, but in halophytes, a high cell wall content and the proportion of cellulose in cell walls may be a salt tolerance mechanism that protects the stability of cell structure under salt stress; and (3) the role of the cell wall in root growth under salinity is more prominent in the glycophyte than in the halophyte.

Silicon confers aluminium tolerance in rice via cell wall modification in the root transition zone.

The root-apex transition zone (TZ), the major perception site for aluminium (Al) toxicity, is crucial for the Al-induced root-growth inhibition, while the mechanism underlying silicon-mediated alleviation of Al toxicity in the TZ is largely unknown. In this study, the role of silicon (Si) in alleviating Al-induced damage in the TZ and root-growth inhibition of rice was investigated. We found that Si had direct alleviative effect on Al toxicity as revealed by less root growth-inhibition, Al accumulation, and callose formation. Si reversed Al-induced decreases of the cell wall elongation and extensibility, and reduced Al-induced increments of cell wall polysaccharides in the TZ. The similar distribution patterns of Al and Si in the cell wall indicated that Si might detoxify Al by forming hydroxyaluminumsilicates in the apoplast of the root-apex TZ. Moreover, the wall-bound form of Si reduced Al binding sites, thereby reducing the capability of Al bound to the cell wall. These results suggest that Si-mediated cell wall modification in the TZ alleviates Al-induced root-growth inhibition in rice involving the promotion of cell wall extensibility and the decrease of Al accumulation in the cell wall.

Exogenous gibberellin treatment improves fruit quality in self-pollinated apple

Although a few apple (Malus × ×domestica) varieties are self-compatible, little is known about the differences in fruit quality between self- and cross-pollinated apple. In our current study, we compared the fruit quality of self-pollinated apple plants (cultivar ‘Hanfu’) in self-pollination or cross-pollinated by another cultivar ‘Qinguan’. Analysis of fruit quality revealed substantial differences in the external qualities between self- and cross-pollinated apple fruit, but not in the internal qualities. Fruits harvested from self-pollinated ‘Hanfu’ were smaller and more asymmetrical than those harvested from the cross-pollinated plants. We developed a mathematical model describing how seed number and distribution affect fruit growth. According to this model, the fewer the seeds, the greater the force released from the seeds and the more asymmetrical the fruit. Detection of endogenous hormone and the associated gene expression revealed that gibberellin (GA) levels and GA transporter gene expression on the seedless side were significantly lower than those on the seeded side. Analysis of fruit pectin methylesterase activity and demethylated pectin levels indicated that the lack of GA limits fruit cell wall extension. Additionally, spraying the self-pollinating plants with gibberellic acid increased the fruit weight and lowered the proportion of asymmetrical fruit, recovering the exterior fruit quality to that of the cross-pollinated fruit. Furthermore, exogenous GA treatment increased the wax layer thickness and reduced the fruit water loss rate, leading to a dramatic improvement in fruit storage capacity. Therefore, exogenous GA treatment could be used to ensure regular fruit production of self-pollinated ‘Hanfu’.

ZmXTH, a xyloglucan endotransglucosylase/hydrolase gene of maize, conferred aluminum tolerance in Arabidopsis

Aluminum (Al) toxicity is one of the primary factors limiting crop production in acid soils worldwide. The cell wall is the major target of Al toxicity owing to the presence of many Al binding sites. Previous studies have found that XTH, encoding xyloglucan endohydrolase (XEH) and xyloglucan endotransglucosylase (XET), could participate in cell wall extension and affect the binding ability of the cell wall to Al by impeding the activities of these two enzymes. In this study, we found that ZmXTH, an XTH gene in maize, was involved in Al detoxification. The Al-induced up-regulation of ZmXTH occurred in the roots, prominently in the root tips. Additionally, the expression of ZmXTH was specifically induced by Al3+ but no other divalent or trivalent cations. Compared with the wild-type Arabidopsis, ZmXTH overexpressing plants grew more healthy and had decreased Al content in their root and root cell wall after Al stress. Overall, the results suggest that ZmXTH could confer the Al tolerance of transgenic Arabidopsis plants by reducing the Al accumulation in their roots and cell walls.

Apical-root apoplastic acidification affects cell wall extensibility in wheat under salinity stress.

Plant salt tolerance is associated with a high rate of root growth. Although root growth is governed by cell wall and apoplastic pH, the relationship between these factors in the root elongation zone under salinity stress remains unclear. Herein, we assess apoplastic pH, pH- and expansin-dependent cell wall extensibility, and expansin expression in the root elongation zone of salt-sensitive (Yongliang-15) and -tolerant (JS-7) cultivars under salinity stress. A six-day 80 mM NaCl treatment significantly reduced apical root apoplastic pH in both cultivars. Using a pH-dependent cell wall extensibility experiment, we found that, under 0 mM NaCl treatment, the optimal pH for cell wall loosening was 6.0 in the salinity-tolerant cultivar and 4.6 in the salinity-sensitive cultivar. Under 80 mM treatment, a pH of 5.0 mitigated the cell wall stiffness caused by salinity stress in the salinity-tolerant cultivar but promoted cell wall stiffening in the salinity-sensitive cultivar. Salinity stress altered expansin expression and differentially affecting cell wall extensibility under pH5.0 and 6.0. TaEXPA8 might be relative to cell wall loosening at pH5.0, whereas TaEXPA5 relative to cell wall loosening at pH6.0. These results elucidate the relationship between expansins and cell wall extensibility in the root elongation zone, with important implications for enhancing plant growth under salinity stress.

Plasmodiophora brassicae-Triggered Cell Enlargement and Loss of Cellular Integrity in Root Systems Are Mediated by Pectin Demethylation.

Gall formation on the belowground parts of plants infected with Plasmodiophora brassicae is the result of extensive host cellular reprogramming. The development of these structures is a consequence of increased cell proliferation followed by massive enlargement of cells colonized with the pathogen. Drastic changes in cellular growth patterns create local deformities in the roots and hypocotyl giving rise to mechanical tensions within the tissue of these organs. Host cell wall extensibility and recomposition accompany the growth of the gall and influence pathogen spread and also pathogen life cycle progression. Demethylation of pectin within the extracellular matrix may play an important role in P. brassicae-driven hypertrophy of host underground organs. Through proteomic analysis of the cell wall, we identified proteins accumulating in the galls developing on the underground parts of Arabidopsis thaliana plants infected with P. brassicae. One of the key proteins identified was the pectin methylesterase (PME18); we further characterized its expression and conducted functional and anatomic studies in the knockout mutant and used Raman spectroscopy to study the status of pectin in P. brassicae-infected galls. We found that late stages of gall formation are accompanied with increased levels of PME18. We have also shown that the massive enlargement of cells colonized with P. brassicae coincides with decreases in pectin methylation. In pme18-2 knockout mutants, P. brassicae could still induce demethylation; however, the galls in this line were smaller and cellular expansion was less pronounced. Alteration in pectin demethylation in the host resulted in changes in pathogen distribution and slowed down disease progression. To conclude, P. brassicae-driven host organ hypertrophy observed during clubroot disease is accompanied by pectin demethylation in the extracellular matrix. The pathogen hijacks endogenous host mechanisms involved in cell wall loosening to create an optimal cellular environment for completion of its life cycle and eventual release of resting spores facilitated by degradation of demethylated pectin polymers.

Raman imaging of Micrasterias: new insights into shape formation

The algae Micrasterias with its star-shaped cell pattern is a perfect unicellular model system to study morphogenesis. How the indentations are formed in the primary cell wall at exactly defined areas puzzled scientists for decades, and they searched for chemical differences in the primary wall of the extending tips compared to the resting indents. We now tackled the question by Raman imaging and scanned in situ Micrasterias cells at different stages of development. Thousands of Raman spectra were acquired from the mother cell and the developing semicell to calculate chemical images based on an algorithm finding the most different Raman spectra. Each of those spectra had characteristic Raman bands, which were assigned to molecular vibrations of BaSO4, proteins, lipids, starch, and plant cell wall carbohydrates. Visualizing the cell wall carbohydrates revealed a cell wall thickening at the indentations of the primary cell wall of the growing semicell and uniplanar orientation of the cellulose microfibrils to the cell surface in the secondary cell wall. Crystalline cellulose dominated in the secondary cell wall spectra, while in the primary cell wall spectra, also xyloglucan and pectin were reflected. Spectral differences between the indent and tip region of the primary cell wall were scarce, but a spectral mixing approach pointed to more cellulose fibrils deposited in the indent region. Therefore, we suggest that cell wall thickening together with a denser network of cellulose microfibrils stiffens the cell wall at the indent and induces different cell wall extensibility to shape the lobes.

Differential responses of roots for varying tolerance to salinity stress in wheat with special reference to elasticity

Two salt-sensitive (Yongliang-15, GS-6058) and two salt-tolerant (JS-7, Xinchun-31) wheat cultivars were used to investigate the extension, extensibility (viscoelastic parameters), and chemical composition of the cell walls in their root elongation regions (apical 10 mm-long root segments), under salinity stress. The elasticity of the root cell wall, indicated by E0, significantly decreased in the salt-sensitive cultivars, whereas the E0 in the salt-tolerant cultivars was maintained at the same level as that in the non-saline condition. Root extension and the differences among cultivars were largely dependent on elastic extension, which accounts for one-half to two-thirds of the total extension. Viscosity, indicated by η0, and the plastic extension of the root cell walls did not change across the treatments and cultivars. The significant decrease in cell wall elasticity in the root elongation region was one of the factors that depressed root growth in salt-sensitive cultivars under the saline condition. The well-maintained elasticity of salt-tolerant cultivars alleviated the depression of root growth by NaCl. Cell wall elasticity was positively correlated with the relative pectin and hemicellulose I contents and negatively correlated with the relative cellulose content. Under saline conditions, the relative hemicellulose II content did not change in the salt-sensitive cultivars; however, it decreased in the salt-tolerant ones. Thus, changes in chemical composition of the cell wall were correspond with the cell wall extensibility and root growth in wheat cultivars with different degrees of salt tolerance.

Over-Expression of AtEXPB2 Alleviates Salt Stress Damage in Transgenic Tobacco Plants

Expansin protein is a component of the cell wall that is generally accepted as a key regulator of the cell wall extension during plant growth and stress response. Expansins loosen the cell wall in a pH-dependent manner. To investigate the role of AtEXPB2, an Arabidopsis expansin gene, its mRNA expression level was analyzed in response to salt stress in overexpressed tobacco (Nicotiana tabacum L.). This over-expression of AtEXPB2 in transgenic lines was studied under salt stress in order to evaluate their resistance to salt stress conditions. In the findings of the current study, however, transgenic plants’ cellular characteristics were probably altered because of the changes in some morphological traits like capsule seed weight, capsule weight, and root length; this alteration did not have any unfavorable effects on transgenic plant phenotypes. Transgenic lines were better than wild-type plant in terms of some morphological parameters, such as capsule number, capsule weight, total seed weight, and capsule seed number. In other words, seed yield was higher in transgenic lines than those in the control. Overexpression of AtEXPB2 conferred tolerance to salt stress by enhancing the activity of antioxidant enzymes, proline content, RWC, and chlorophyll, as well as reducing electrolyte leakage index (ELI). In addition, relative expression of AtEXPB2 and H+-ATPase gene increased in transgenic plants under salinity stress. Overall, these results confirmed the role of expansin proteins in plant stress tolerance.

Accumulation and cross-linkage of β-1,3/1,6-glucan lead to loss of basal stipe cell wall extensibility in mushroom Coprinopsis cinerea

The mature basal stipe of mushroom Coprinopsis cinerea loses wall extensibility. We found that an endo-β-1,3-glucanase ENG from C. cinerea could restore mature basal stipe wall extensibility via pretreatment such that the ENG-pretreated basal stipe walls could be induced to extend by chitinase ChiIII. ENG pretreatment released glucose, laminaribiose, and 3-O-D-gentiobiose-D-glucose from the basal stipe walls, consistent with ENG-digested products of β-1,6-branched β-1,3-glucan. Different effects of endo-β-1,3-glucanase ENG and exo-β-1,3-glucanase EXG pretreatment on the structure, amount and ratio (β-1,3-glucoside bonds to β-1,6-glucoside bonds) of products from the basal stipe and the apical stipe cell walls, respectively, and on the cell wall extensibility and the cell wall ultra-architecture of the basal stipes were analyzed. All results demonstrate that the more accumulation and cross-linkage of β-1,6-branched β-1,3-glucan with wall maturation lead to loss of wall extensibility of the basal stipe regions compared to the apical stipe cell walls.