The RLCK-VND6 module coordinates secondary cell wall formation and adaptive growth in rice.

The orderly deposition of secondary cell wall (SCW) in plants is implicated in various biological programs and is precisely controlled. Although many positive and negative regulators of SCW have been documented, the molecular mechanisms underlying SCW formation coordinated with distinct cellular physiological processes during plant adaptive growth remain largely unclear. Here, we report the identification of Cellulose Synthase co-expressed Kinase1 (CSK1), which encodes a receptor-like cytoplasmic kinase, as a negative regulator of SCW formation and its signaling cascade in rice. Transcriptome deep sequencing of developing internodes and genome-wide co-expression assays revealed that CSK1 is co-expressed with cellulose synthase genes and is responsive to various stress stimuli. The increased SCW thickness and vigorous vessel transport in csk1 indicate that CSK1 functions as a negative regulator of SCW biosynthesis. Through observation of green fluorescent protein-tagged CSK1 in rice protoplasts and stable transgenic plants, we found that CSK1 is localized in the nucleus and cytoplasm adjacent to the plasma membrane. Biochemical and molecular assays demonstrated that CSK1 phosphorylates VASCULAR-RELATED NAC-DOMAIN 6 (VND6), a master SCW-associated transcription factor, in the nucleus, which reduces the transcription of a suite of SCW-related genes, thereby attenuating SCW accumulation. Consistently, genetic analyses show that CSK1 functions upstream of VND6 in regulating SCW formation. Interestingly, our physiological analyses revealed that CSK1 and VND6 are involved in abscisic acid-mediated regulation of cell growth and SCW deposition. Taken together, these results indicate that the CSK1-VND6 module is an important component of the SCW biosynthesis machinery, which coordinates SCW accumulation and adaptive growth in rice. Our study not only identifies a new regulator of SCW biosynthesis but also reveals a fine-tuned mechanism for precise control of SCW deposition, offering tools for rationally tailoring agronomic traits.

Excellent Catalytic Performance of ISOBAM Stabilized Co/Fe Colloidal Catalysts toward KBH4 Hydrolysis.

Recently, developing a cost-effective and high-performance catalyst is regarded as an urgent priority for hydrogen generation technology. In this work, ISOBAM-104 stabilized Co/Fe colloidal catalysts were prepared via a co-reduction method and used for the hydrogen generation from KBH4 hydrolysis. The obtained ISOBAM-104 stabilized Co10Fe90 colloidal catalysts exhibit an outstanding catalytic activity of 37,900 mL-H2 min-1 g-Co-1, which is far higher than that of Fe or Co monometallic nanoparticles (MNPs). The apparent activation energy (Ea) of the as-prepared Co10Fe90 colloidal catalysts is only 14.6 ± 0.7 kJ mol-1, which is much lower than that of previous reported noble metal-based catalysts. The X-ray photoelectron spectroscopy results and density functional theory calculations demonstrate that the electron transfer between Fe and Co atoms is beneficial for the catalytic hydrolysis of KBH4.

Research Progress on Coating Structure of Silicon Anode Materials for Lithium-Ion Batteries.

Silicon, which has been widely studied by virtue of its extremely high theoretical capacity and abundance, is recognized as one of the most promising anode materials for the next generation of lithium-ion batteries. However, silicon undergoes tremendous volume change during cycling, which leads to the destruction of the electrode structure and irreversible capacity loss, so the promotion of silicon materials in commercial applications is greatly hampered. In recent years, many strategies have been proposed to address these shortcomings of silicon. This Review focused on different coatings materials (e. g., carbon-based materials, metals, oxides, conducting polymers, etc.) for silicon materials. The role of different types of materials in the modification of silicon-based material encapsulation structure was reviewed to confirm the feasibility of the protective layer strategy. Finally, the future research direction of the silicon-based material coating structure design for the next-generation lithium-ion battery was summarized.

The receptor-like cytoplasmic kinase RIPK regulates broad-spectrum ROS signaling in multiple layers of plant immune system.

Production of reactive oxygen species (ROS) via the activity of respiratory burst oxidase homologs (RBOHs) plays a vital role in multiple layers of the plant immune system, including pathogen-associated molecular pattern-triggered immunity (PTI), damage-associated molecular pattern-triggered immunity (DTI), effector-triggered immunity (ETI), and systemic acquired resistance (SAR). It is generally established that RBOHD is activated by different receptor-like cytoplasmic kinases (RLCKs) in response to various immune elicitors. In this study, we showed that RPM1-INDUCED PROTEIN KINASE (RIPK), an RLCK VII subfamily member, contributes to ROS production in multiple layers of plant immune system. The ripk mutants showed reduced ROS production in response to treatment with all examined immune elicitors that trigger PTI, DTI, ETI, and SAR. We found that RIPK can directly phosphorylate the N-terminal region of RBOHD in vitro, and the levels of phosphorylated S343/S347 residues of RBOHD are sigfniciantly lower in ripk mutants compared with the wild type upon treatment with all tested immune elicitors. We further demonstrated that phosphorylation of RIPK is required for its function in regulating RBOHD-mediated ROS production. Similar to rbohd, ripk mutants showed reduced stomatal closure and impaired SAR, and were susceptible to the necrotrophic bacterium Pectobacterium carotovorum. Collectively, our results indicate that RIPK regulates broad-spectrum RBOHD-mediated ROS signaling during PTI, DTI, ETI, and SAR, leading to subsequent RBOHD-dependent immune responses.

Enhanced Diffusion Kinetics of Li Ions in Double-Shell Hollow Carbon Fibers.

The rational design and preparation of hierarchical hollow structures have promising potential in electrochemical energy storage systems. In this paper, double-shell hollow carbon fibers (DSHCFs) with tunable thickness and shell spacing are prepared using hollow electrospun polystyrene fibers as the hard template and in situ coated polypyrrole as the carbon source. The as-prepared DSHCFs with an optimized structure exhibit a submicrometer shell spacing and a nanoscaled shell thickness, which guarantees sufficient contact area with the electrolyte and provides abundant electrochemical active sites for Li+ storage. Owing to the unique structural advantages, a DSHCF-based anode shows favorable transport kinetics for both Li+ ions and electrons during the lithiation/delithiation process, and a high reversible capacity of 348 mAh g-1 at 5.0 A g-1 is well maintained even after 500 cycles with no obvious capacity attenuation. Particular emphasis is given to kinetic Li+ storage mechanisms in DSHCFs that are discussed in detail, providing a new avenue for developing high-performance carbon materials for the practical application of energy storage devices.

Molten salt synthesis of carbon-doped boron nitride nanosheets with enhanced adsorption performance.

Owing to their large specific areas, high thermal stability and chemical inertness, two-dimensional boron carbon nitride nanosheets (BCNNs) have captured much attention in recent years in the field of adsorption of pollutants. The formation of BCNNs via incorporating carbon into boron nitride (BN) can effectively improve the photoelectric and adsorption properties of the latter. In this work, carbon-doped BN (BCN) nanosheets were prepared at 1100 °C via a molten salt route using boric acid, melamine and glucose as the main starting materials. The effects of molten salt type and carbon doping level on the formation of BCN were investigated, and their isothermal adsorption properties in a methylene blue (MB) aqueous solution were evaluated based on the Langmuir and Freundlich models. The results indicated that using molten LiCl-KCl as a liquid medium was more favorable than NaCl-KCl to the formation of BCNNs. As-prepared BC0.4N sample possessed a sheet-like structure of about 10 nm thick and a specific surface area as high as 484 m2 g-1. Moreover, the adsorption test of MB demonstrated a high adsorption capacity of 249.04 mg g-1, which was about 14 times higher than that in the case of the pristine BN, and the kinetic rate constant value in the case of using BC0.4N is about ten times as high as that of BN following a pseudo-second-order model, suggesting that the as-formed BC0.4N nanosheets could be potentially used as a value-added effective adsorbent for future wastewater remediation.

Design and Application of a Rotatory Device for Detecting Transient Ca2+ Signals in Response to Mechanical Stimulation Using an Aequorin-Based Ca2+ Imaging System.

Elevation of the cytosolic free calcium ion (Ca2+ ) concentration ([Ca2+ ]cyt ) is one of the earliest responses to biotic and abiotic stress in plant cells. Among the various Ca2+ detection systems available, aequorin-based luminescence Ca2+ imaging systems provide a relatively amenable and robust method that facilitates large-scale genetic-mutant screening based on [Ca2+ ]cyt responses. Compared to that mediated by chemical elicitors, mechanical stimulation-induced elevation of [Ca2+ ]cyt is considerably more rapid, occurring within 10 s following stimulation. Therefore, its assessment using aequorin-based Ca2+ imaging systems represents a notable challenge, given that a time interval of ≥1 min is required to reduce the background light before operating the photon imaging detector. In this context, we designed a device that can rotate automatically within the confines of an enclosed dark box, and using this, we can record [Ca2+ ]cyt dynamics immediately after plants had been rotated to induce mechanical stimulation. This tool can facilitate the study of perception and early signal transduction in response to mechanical stimulation on a large scale based on [Ca2+ ]cyt responses. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Detection of background luminance signals in aequorin-transformed Arabidopsis seedlings using a photon imaging detector Basic Protocol 2: Construction of the rotatory device Basic Protocol 3: Calcium measurement in Arabidopsis seedlings after rotatory stimulation Basic Protocol 4: Data analysis and processing.

Lipopolysaccharides Trigger Two Successive Bursts of Reactive Oxygen Species at Distinct Cellular Locations.

Lipopolysaccharides (LPS) are major components of the outer membrane of gram-negative bacteria and are an important microbe-associated molecular pattern (MAMP) that triggers immune responses in plants and animals. A previous genetic screen in Arabidopsis (Arabidopsis thaliana) identified LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (LORE), a B-type lectin S-domain receptor kinase, as a sensor of LPS. However, the LPS-activated LORE signaling pathway and associated immune responses remain largely unknown. In this study, we found that LPS trigger biphasic production of reactive oxygen species (ROS) in Arabidopsis. The first transient ROS burst was similar to that induced by another MAMP, flagellin, whereas the second long-lasting burst was induced only by LPS. The LPS-triggered second ROS burst was found to be conserved in a variety of plant species. Microscopic observation of the generation of ROS revealed that the LPS-triggered second ROS burst was largely associated with chloroplasts, and functional chloroplasts were indispensable for this response. The lipid A moiety, the most conserved portion of LPS, appears to be responsible for the second ROS burst. Surprisingly, the LPS- and lipid A-triggered second ROS burst was only partially dependent on LORE. Together, our findings provide insight on the LPS-triggered ROS production and the associated signaling pathway.

A Gibberellin-Mediated DELLA-NAC Signaling Cascade Regulates Cellulose Synthesis in Rice.

Cellulose, which can be converted into numerous industrial products, has important impacts on the global economy. It has long been known that cellulose synthesis in plants is tightly regulated by various phytohormones. However, the underlying mechanism of cellulose synthesis regulation remains elusive. Here, we show that in rice (Oryza sativa), gibberellin (GA) signals promote cellulose synthesis by relieving the interaction between SLENDER RICE1 (SLR1), a DELLA repressor of GA signaling, and NACs, the top-layer transcription factors for secondary wall formation. Mutations in GA-related genes and physiological treatments altered the transcription of CELLULOSE SYNTHASE genes (CESAs) and the cellulose level. Multiple experiments demonstrated that transcription factors NAC29/31 and MYB61 are CESA regulators in rice; NAC29/31 directly regulates MYB61, which in turn activates CESA expression. This hierarchical regulation pathway is blocked by SLR1-NAC29/31 interactions. Based on the results of anatomical analysis and GA content examination in developing rice internodes, this signaling cascade was found to be modulated by varied endogenous GA levels and to be required for internode development. Genetic and gene expression analyses were further performed in Arabidopsis thaliana GA-related mutants. Altogether, our findings reveal a conserved mechanism by which GA regulates secondary wall cellulose synthesis in land plants and provide a strategy for manipulating cellulose production and plant growth.

Brittle Culm1, a COBRA-like protein, functions in cellulose assembly through binding cellulose microfibrils.

Cellulose represents the most abundant biopolymer in nature and has great economic importance. Cellulose chains pack laterally into crystalline forms, stacking into a complicated crystallographic structure. However, the mechanism of cellulose crystallization is poorly understood. Here, via functional characterization, we report that Brittle Culm1 (BC1), a COBRA-like protein in rice, modifies cellulose crystallinity. BC1 was demonstrated to be a glycosylphosphatidylinositol (GPI) anchored protein and can be released into cell walls by removal of the GPI anchor. BC1 possesses a carbohydrate-binding module (CBM) at its N-terminus. In vitro binding assays showed that this CBM interacts specifically with crystalline cellulose, and several aromatic residues in this domain are essential for binding. It was further demonstrated that cell wall-localized BC1 via the CBM and GPI anchor is one functional form of BC1. X-ray diffraction (XRD) assays revealed that mutations in BC1 and knockdown of BC1 expression decrease the crystallite width of cellulose; overexpression of BC1 and the CBM-mutated BC1s caused varied crystallinity with results that were consistent with the in vitro binding assay. Moreover, interaction between the CBM and cellulose microfibrils was largely repressed when the cell wall residues were pre-stained with two cellulose dyes. Treating wild-type and bc1 seedlings with the dyes resulted in insensitive root growth responses in bc1 plants. Combined with the evidence that BC1 and three secondary wall cellulose synthases (CESAs) function in different steps of cellulose production as revealed by genetic analysis, we conclude that BC1 modulates cellulose assembly by interacting with cellulose and affecting microfibril crystallinity.

Brittle culm15 encodes a membrane-associated chitinase-like protein required for cellulose biosynthesis in rice.

Plant chitinases, a class of glycosyl hydrolases, participate in various aspects of normal plant growth and development, including cell wall metabolism and disease resistance. The rice (Oryza sativa) genome encodes 37 putative chitinases and chitinase-like proteins. However, none of them has been characterized at the genetic level. In this study, we report the isolation of a brittle culm mutant, bc15, and the map-based cloning of the BC15/OsCTL1 (for chitinase-like1) gene affected in the mutant. The gene encodes the rice chitinase-like protein BC15/OsCTL1. Mutation of BC15/OsCTL1 causes reduced cellulose content and mechanical strength without obvious alterations in plant growth. Bioinformatic analyses indicated that BC15/OsCTL1 is a class II chitinase-like protein that is devoid of both an amino-terminal cysteine-rich domain and the chitinase activity motif H-E-T-T but possesses an amino-terminal transmembrane domain. Biochemical assays demonstrated that BC15/OsCTL1 is a Golgi-localized type II membrane protein that lacks classical chitinase activity. Quantitative real-time polymerase chain reaction and ß-glucuronidase activity analyses indicated that BC15/OsCTL1 is ubiquitously expressed. Investigation of the global expression profile of wild-type and bc15 plants, using Illumina RNA sequencing, further suggested a possible mechanism by which BC15/OsCTL1 mediates cellulose biosynthesis and cell wall remodeling. Our findings provide genetic evidence of a role for plant chitinases in cellulose biosynthesis in rice, which appears to differ from their roles as revealed by analysis of Arabidopsis (Arabidopsis thaliana).