Genome-wide identification and evolutionary analysis of MLO gene family in Rosaceae plants

Mutants lacking wild-type MLO(Mildew resistance Locus O)proteins show broad-spectrum resistance to the powdery mildew fungus,and dysregulated cell death control,with spontaneous cell death in response to developmental or abiotic stimuli.In order to understand the evolution and divergence patterns of the MLO gene family in Rosaceae plants,we analysed systematically genome-wide data from Fragaria vesca,Prunus persica,Prunus mume,Malus domestica,Pyrus bretschneideri and Rubus occidentalis based on bioinformatics methods.Using three phylogenetic methods(the neighbour-joining,maximum likelihood,and Bayesian methods),we identified 117 MLO genes from 6 Rosaceae species.The results of all three phylogenetic analysis methods supported that these genes were divided into six clades.Conserved motif analysis found that only motif 2 was present in all MLO proteins and had 3 nearly invariant amino acid residues.The findings indicated that motif 2 might be shared by the MLO gene family.The structural features of these genes showed large variations in sequence length among different species,although the lengths and the numbers of exons exhibited high degrees of similarity.Selective pressure analysis showed extremely significant differences in all 6 clades,with 2,1,and 1 site(s)under significant positive selection detected in clades III,IV,and VI,respectively.These positive selection sites were important driving forces for the promotion of the functional differentiation of the MLO genes.Functional divergence analysis showed that the significantly divergent sites were located within the domains of the MLO genes.Functional distance analysis showed that the clade V had more conservative functions and might have retained more original functions during the evolutionary process.However,clade I may have undergone extensive altered functional constraints as a specialised functional role.Moreover,the most original function of the MLO genes in Rosaceae could be related to the evolution of their resistance to powdery mildew,which then gradually evolved into functions such as the regulation of flower development,the control of root morphology,and seed evolution due to the different evolutionary rates after gene duplication.These results provide a theoretical basis for further studies of the molecular evolutionary patterns of the plant MLO gene family.

Comparative transcriptomic and metabonomic analysis revealed the relationships between biosynthesis of volatiles and flavonoid metabolites in Rosa rugosa

Rosa rugosa is not only cultivated as a landscaping plant,but also used in cosmetics,the medical and food industries.However,little information is currently available on the gene regulatory networks involved in its scent and color biosynthesis and metabolism.In this study,R.rugosa Thunb.f.rosea Rehd with red petals(RR)and its white petal variant(WR),were used to study the molecular mechanisms in flower color and scent.Sixty-five differential flavonoid metabolites and 15 volatiles were found to have significant differences between RR and WR.Correspondingly,the key regulators(MYB-bHLH-WD40)of anthocyanin synthesis pathway and their structural genes involved in flavonoid biosynthesis,benzenoid/phenylpropanoid biosynthesis,terpenoid biosynthesis pathways were also found to be differentially expressed by comparative transcriptome.Further,qPCR permitted the identification of some transcripts encoding proteins that were putatively associated with scent and color biosynthesis in roses.Particularly,the results showed that the ACT gene(encoding CoA geraniol/citronellol acetyltransferase,GeneID:112190420),which expressed lower in WR,was involved in three pathways:flavonoid biosynthesis,phenylpropanoid biosynthesis and terpenoid biosynthesis,however,GT5(anthocyanin glycosylation gene,GeneID:112186660),expressed higher in WR,was involved in both flavonoid and phenylpropanoid biosynthesis pathways.These results suggested that ACT and GT5 might play important roles in regulating the relationship of color pigmentation and volatile emission.

CATase-immobilized hydrogel platform molded by photo-enzyme coupling polymerization for effectively preventing postoperative abdominal adhesion

Redox homeostasis,which is regulated by enzymes acting as regulatory valves,is crucial for maintaining the proper functioning of biomolecules and a stable microenvironment for physiological processes by modulating the homeostasis of reactive oxygen species(ROS).Antioxidant enzymes in biocatalysis are used in the prevention or treatment of oxidative stress-related disease by counteracting the harmful effects of ROS.However,designing a system that can efficiently immobilize antioxidant enzymes with high catalytic activity and stability is still challenging.Bioinspired by photo-biocatalysis,a novel and effective catalase(CATase)-immobilized hydrogel platform has been developed by the proposed photo-enzymatic coupled radical polymerization strategy of the visible light coupling with the porphyrin-centered CATase.The higher catalytic stability and activity can therefore be achieved due to the preferential polymerization of CATase-immobilized hydrogel platform with a favorable three-dimensional network of enhanced coupling efficacy between light and enzymes.The mechanisms of free radical-initiated polymerization as well as the antioxidant cycle in the photo-CATase coupling system have been explored.Intriguingly,the CATase-immobilized hydrogel platform affords an unprecedented antioxidant ability to scavenge ROS and provide an effective cellular protection mechanism against external oxidative stress.Additionally,the CATase-immobilized hydrogel platform can effectively prevent peritoneal adhesion by reducing the expression of inflammatory cytokines.Therefore,the novel CATase-immobilized hydrogel platform is a potential candidate for physical barriers that effectively prevent postoperative adhesion formation,offering a new anti-adhesion strategy for clinical applications.

Multi-Functional Supramolecular-Polymeric Hybrid Hydrogel for Promoting Cutaneous Wound Repair

Supramolecular-polymeric hybrid hydrogels combining advantages of the assembled supramolecular networks and polymeric networks have attracted considerable attention recently.However,they are rarely used as multi-functional wound dressing materials.In this work,we report a supramolecular-polymeric hybrid hydrogel consisting of a co-assembled DBS-COOH/Naproxen network(DBS/NAP)and a polymeric gelatin(G)/quaternary ammonium group(Q)and phenylboronic acid groups(PBA)-grafted chitosan(QCS-PBA)network(G/QCS)for accelerating skin wound healing.The polymeric networks could obviously enhance the mechanical properties of the drug-derived supramolecular networks.Q groups endowed the hydrogel with enhanced antibacterial properties.Borate ester crosslinking between PBA groups and diol groups in DBS-COOH could effectively scavenge reactive radicals.The hybrid hydrogel exhibited tunable mechanical and adhesive properties due to the temperature-responsive gelatin networks.NAP could gradually release from hybrid hydrogel networks thus continuously relieving inflammation.Finally,the biocompatible,adhesive,antibacterial and antioxidant hybrid G/QCS-DBS/NAP gel could promote the in vivo healing stages in a full-thickness wounds with increased collagen deposition,upregulated CD31 expression and down-regulated TNF-αexpression.Overall,this multi-functional supramolecular-polymeric hybrid hydrogel exhibited great potential for treating skin defects as promising wound dressing materials.

Graft-accelerated virus-induced gene silencing facilitates functional genomics in rose flowers

Rose has emerged as a model ornamental plant for studies of flower development, senescence, and morphology, as well as the metabolism of floral fragrances and colors.Virus-induced gene silencing(VIGS) has long been used in functional genomics studies of rose by vacuum infiltration of cuttings or seedlings with an Agrobacterium suspension carrying TRV-derived vectors. However, VIGS in rose flowers remains a challenge because of its low efficiency and long time to establish silencing. Here we present a novel and rapid VIGS method that can be used to analyze gene function in rose,called ‘graft-accelerated VIGS’, where axil ary sprouts are cut from the rose plant and vacuum infiltrated with Agrobacterium. The inoculated scions are then grafted back onto the plants to flower and silencing phenotypes can be observed within 5 weeks, post-infiltration. Using this new method, we successfully silenced expression of the RhDFR, RhA G, and RhNUDXin rose flowers, and affected their color, petal number, as well as fragrance, respectively. This grafting method will facilitate high-throughput functional analysis of genes in rose flowers. Importantly, it may also be applied to other woody species that are not currently amenable to VIGS by conventional leaf or plantlet/seedling infiltration methods.

Supramolecular protein glue to boost enzyme activity

Proteins possess many biological functions.However, they can easily degrade or aggregate, thus losing their bioactivity. Therefore, it is very important to develop materials capable of interacting with proteins and forming nanostructures for protein storage and delivery. In this study,we serendipitously found a novel peptide-based supramolecular protein glue(Nap-GFFYK(γE)2-NH2, compound 1) that could co-assemble with proteins into nanofibers and hydrogels. We found that compound 1 rapidly folded into a β-sheet conformation upon contact with many proteins but not with polymers. Total internal reflection fluorescence microscopy(TIRFM) images clearly show the formation of co-assembled nanofibers by proteins and the peptide. The supramolecular protein glue could improve the dispersion of enzymes(lipase and lysozyme) and therefore enhance their catalytic activity,especially at high temperatures. More importantly, the supramolecular protein glue could co-assemble with two enzymes, glucose oxidase/horseradish peroxidase(GOx/HRP)and GOx/cytochrome c(cyt c), to form nanofibers that significantly enhanced the catalytic activity of tandem enzymatic reactions. We envisioned the great potential of our supramolecular protein glue for protein storage, delivery, and bioactivity manipulation.

Regenerated hydrogel electrolyte towards an all-gel supercapacitor

Electrolyte regeneration is an important goal for environmental protection and sustainable development efforts.Herein,we report a facile strategy inspired by the transformation of edible dough from flour to regenerate hydrogel electrolytes from their dehydrated copolymer granules(CGs)via direct addition of water or salt solution.With the aid of heating,this procedure is efficient,relatively quick,and easily implemented.The dehydrated CGs are lightweight,reusable and stable under long-term storage.Even after 5 cycles of dehydration and regeneration,the regeneration efficiency of the hydrogel electrolytes,as evaluated based on retention of mechanical strength,is over 60%.The regenerated electrolytes possess considerable ionic conductivity,reprocessability,and 3D-printability.Furthermore,an all-gel supercapacitor assembled from the regenerated hydrogel electrolyte and activated carbon electrode with CGs as binder demonstrates excellent interfacial compatibility.The assembled all-gel supercapacitor can maintain 98.7% of its original specific capacitance after 100 bending tests,and can operate in a wide temperature range spanning from-15 to 60°C.This work may provide a new access to the development of renewable materials for various applications in the fields of intelligent devices,wearable electronics and soft robotics.

Dynamic assembly and biocatalysis-selected gelation endow self-compartmentalized multienzyme superactivity

Cellular metabolism in multiple organelles utilizes compartmentalized multienzyme efficient catalysis to realize substance metabolism, energy conversion and immune defenses. The convenient and biomimetic design of artificial multienzymes has become an emerging research topic. Herein, we employ a facile enzyme-initiated radical polymerization to self-anchor multienzyme in cell-like hydrogels with mesoscale compartments. The dynamic assembly of glucose oxidase/cytochrome c(GOx/Cyt c) with methacrylate-modified hyaluronic acid can form nanoaggregates, where only the bound enzyme pairs with the adjacent position can catalyze the polymerization to compartmentalize multienzymes within hydrogel. Consequently, the cascade enzymes within hydrogel display 33.9 times higher activity compared to free enzymes, as well as excellent thermostability and multiple recyclability. The mechanism study indicates that the compartmental effect of the hydrogel and the anchoring effect of Cyt c synergistically enhance GOx/Cyt c activity. According to the density functional theory(DFT) calculation, Cyt c activity increment originates from its ligand changes of Fe(Ⅲ) porphyrin, which has a smaller energy barrier of the catalytic reaction.This study provides a promising strategy for autonomous colocalization of multienzyme in biocompatible hydrogels which can be potentially applied in cascade enzyme induced catalysis applications.

Spatiotemporally-regulated multienzymatic polymerization endows hydrogel continuous gradient and spontaneous actuation

There are several natural materials which have evolved functional gradients,ingeniously attaining maximal efficacy from limited components.Herein,we utilized the spatiotemporal distribution of initiator acetylacetone to regulate the multienzyme polymerization and fabricate a chitosan-polymer hydrogel.The temporal priority order of acetylacetone was higher than phenolmodified chitosan by density functional theory calculation.The acetylacetone within the gelatin could gradually diffuse spatially into the chitosan hydrogel to fabricate the composite hydrogel with gradient network structure.The gradient hydrogel possessed a transferring topography from the two-dimensional pattern.A continuously decreased modulus along with acetylacetone diffusion was confirmed by atomic force microscope-based force mapping experiment.The water-retaining ability of various regions was confirmed by low-field nuclear magnetic resonance(NMR)and thermogravimetric analysis(TG)analysis,which led to the spontaneous actuation of gradient hydrogel with maximum 1821°/h curling speed and 227°curling angle.Consequently,the promising gradient hydrogels could be applied as intelligent actuators and flexible robots.

Construction of self-assembled nanogel as mulitenzyme mimics for bioresponsive tandem-catalysis imaging

The self-assembled phospholipid-or cytosolassociated multienzyme complexes constitute necessary components of the foundation of life.As a proof of concept,metalcoordinated supramolecular nanogels (MCSGs) have been designed,with the self-assembly of di-lysine coordinated iron(Fe(Lys)_(2))-functionalized peptide gelators on the interface by an in situ amidation-induced protonation process.The monoatomic and highly dispersed active centers of Fe(Lys)_(2) offered the nanogel mimics with excellent reaction rates due to the high density and nano compartmental structure similar to the natural matrix-associated multienzyme complex.SiO_(2)@MCSGs show both superoxide dismutase (SOD) activity and peroxidase (POD) activity,and the higher activities compared with the activity of free Fe(Lys);molecules can be detected.After loading the substrate 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS),SiO_(2)@MCSGs;can responsively convert O^(-)_(2) in the tumor microenvironment into H_(2)O_(2) intermediates and then tandem catalyze the oxidization of ABTS for contrast photoacoustic (PA) imaging of tumor by the SOD-POD mimic activity,showing their great potential as the efficient enzymatic agents for pathological theranostics.