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Abstract Virus‐induced gene silencing (VIGS) is a powerful tool for transient gene functional analysis in plants, especially for monocot species (e.g., grasses) that are recalcitrant to transformation. Despite various VIGS systems that have been developed in different plant species, none was previously available for the bioenergy crop Miscanthus. Here, we report the establishment of an efficient and robust VIGS system mediated by Tobacco Rattle Virus (TRV) in Miscanthus. We first investigated the impact of various factors that may affect gene silencing efficiency using the Miscanthus sinensis Phytoene Desaturase (MsPDS) gene as a visual indicator of photobleaching. Then, we optimized the TRV‐elicited VIGS procedure using an orthogonal experimental design with four factors (sprout size, Agrobacterium concentration, vacuum infiltration time, and co‐incubation time) each at three levels. The following led to the highest silencing efficiency (~76%): inoculation of germinating seedlings (1.0–2.0 mm), Agrobacterium tumefaciens culture grown to optical density at 600 nm (OD600) of 0.4, vacuum infiltration for 90 min, and co‐incubation for 5 h. The VIGS system established was applicable for both M. sinensis and M. lutarioriparius, with comparable gene silencing efficiency. We verified the efficacy of the VIGS system via the functional characterization of the role of a MYB transcription factor, MsMYB112, in salt stress tolerance. Expression of MsMYB112 was successfully knocked down using the VIGS system, and this led to compromised salt tolerance in the silenced Miscanthus plants. The TRV‐based VIGS system established may, therefore, substantially facilitate functional genomic studies in Miscanthus.

The homeodomain-leucine zipper (HD-ZIP) transcription factors, representing one of the largest plant-specific superfamilies, play important roles in the response to various abiotic stresses. However, the functional roles of HD-ZIPs in abiotic stress tolerance and the underlying mechanisms remain relatively limited in Miscanthus sinensis. In this study, we isolated an HD-ZIP TF gene, MsHDZ23, from Miscanthus and ectopically expressed it in Arabidopsis. Transcriptome and promoter analyses revealed that MsHDZ23 responded to salt, alkali, and drought treatments. The overexpression (OE) of MsHDZ23 in Arabidopsis conferred higher tolerance to salt and alkali stresses compared to wild-type (WT) plants. Moreover, MsHDZ23 was able to restore the hb7 mutant, the ortholog of MsHDZ23 in Arabidopsis, to the WT phenotype. Furthermore, MsHDZ23-OE lines exhibited significantly enhanced drought stress tolerance, as evidenced by higher survival rates and lower water loss rates compared to WT. The improved drought tolerance may be attributed to the significantly smaller stomatal aperture in MsHDZ23-OE lines compared to WT. Furthermore, the accumulation of the malondialdehyde (MDA) under abiotic stresses was significantly decreased, accompanied by dramatically enhanced activities in several antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in the transgenic plants. Collectively, these results demonstrate that MsHDZ23 functions as a multifunctional transcription factor in enhancing plant resistance to abiotic stresses.

Exsolution, as an effective approach to constructing particle-decorated interfaces, is still challenging to yield interfacial films rather than isolated particles. Inspired by in vivo near-infrared laser photothermal therapy, using 3 mol% Y _2 O _3 stabilized tetragonal zirconia polycrystals (3Y-TZP) as host oxide matrix and iron-oxide (Fe _3 O _4 / γ -Fe _2 O _3 / α -Fe _2 O _3 ) materials as photothermal modulator and exsolution resource, femtosecond laser ultrafast exsolution approach is presented enabling to conquer this challenge. The key is to trigger photothermal annealing behavior via femtosecond laser ablation to initialize phase transition from monoclinic zirconia (m-ZrO _2 ) to tetragonal zirconia (t-ZrO _2 ) and induce t-ZrO _2 columnar crystal growth. Fe-ions rapidly segregate along grain boundaries and diffuse towards the outmost surface, and become ‘frozen’, highlighting the potential to use photothermal materials and ultrafast heating/quenching behaviors of femtosecond laser ablation for interfacial exsolution. Triggering interfacial iron-oxide coloring exsolution is composition and concentration dependent. Photothermal materials themselves and corresponding photothermal transition capacity play a crucial role, initializing at 2 wt%, 3 wt%, and 5 wt% for Fe _3 O _4 / γ -Fe _2 O _3 / α -Fe _2 O _3 doped 3Y-TZP samples. Due to different photothermal effects, exsolution states of ablated 5 wt% Fe _3 O _4 / γ -Fe _2 O _3 / α -Fe _2 O _3 -doped 3Y-TZP samples are totally different, with whole coverage, exhaustion (ablated away) and partial exsolution (rich in the grain boundaries in subsurface), respectively. Femtosecond laser ultrafast photothermal exsolution is uniquely featured by up to now the deepest microscale (10 μ m from 5 wt%-Fe _3 O _4 -3Y-TZP sample) Fe-elemental deficient layer for exsolution and the whole coverage of exsolved materials rather than the formation of isolated exsolved particles by other methods. It is believed that this novel exsolution method may pave a good way to modulate interfacial properties for extensive applications in the fields of biology, optics/photonics, energy, catalysis, environment, etc.

Abstract For the first time, a configurable NbOx memristor is achieved that can be configured as an artificial synapse or neuron after fabrication by controlling the forming compliance current (FCC). When the FCC ≤ 2 mA, the memristors exhibit the resistive‐switching (RS) property, enabling multiple types of synaptic plasticity, including short‐term potentiation, paired‐pulse facilitation, short‐term memory, and long‐term memory. When the FCC ≥ 3 mA, the memristors can be electroformed and exhibit the threshold switching (TS) property with excellent endurance (>1012), thus achieving various biological neuron characteristics, such as threshold‐triggering, strength‐modulation of spike frequency, and leaky integrate‐and‐fire. This enables the successful implementation of a spiking Pavlov's dog that employs the spikes as information carrier by connecting an RS NbOx memristor as artificial synapse and a TS memristor as artificial neuron in series. Furthermore, a fully NbOx memristors‐based single‐layer spiking neural network is simulated. It is first found that, due to the forgetting property of synapse, the recognition accuracy for the Modified National Institute of Standards and Technology handwritten digits is increased from 85.49% to 91.45%. This study provides a solid foundation for the development of neuromorphic machines based on the principles of the human brain.

Biochar has been widely reported to improve soil conditions and affect plant growth. However, its effectiveness is limited by soil type and production technology. Considering the application effect of biochar in saline alkali soil, there is currently a lack of in-depth mechanism explanations in the research. Therefore, we designed an experiment to explore the effect of biochar on plant growth in saline alkali soil and conducted soil column experiments in a greenhouse environment using composite inorganic fertilizer (NPK). The results showed that biochar significantly affected the distribution of soil nutrient content at different depths, with a significant increase in fertility levels in the surface and middle layers and a decrease in fertility levels in deep soils. Compared to using fertilizers alone, the combined use of biochar and fertilizers further expands the enrichment effect and significantly reduces the leaching of fertilizers into deeper layers. At the same time, the application of biochar also improved soil properties, including an increase in electrical conductivity and organic matter content, as well as an increase in soil enzyme activity. On the other hand, the application of biochar also increases the activity of antioxidant enzymes and the content of osmoregulation substances in plants, reducing the environmental stress that plants are subjected to. Therefore, our results indicate that biochar can reduce the leaching of fertilizers into deep soil layers, improve soil properties, and promotes the growth of Miscanthus in saline alkali soils.

Abstract Duckweed is one kind of promising bioenergy plant with prominent advantages such as fast growth rate and high starch content. However, almost all previous studies focused on the natural duckweed germplasms. In this study, heavy‐ion irradiation was used to establish a mutant library of Lemna aequinoctialis 6002, and one mutant named submarine‐1 (sub‐1) was screened, which could accumulate more starch but with smaller granules. Unexpectedly, under proper external growth conditions such as poor nutritional status and insufficient growth space, sub‐1 mutant would sink underwater due to formation of dense tissue structure and large amount of fine starch particles with the extension of cultivation time. The starch content in the sinking sub‐1 increased to over 45% (dry weight) and was 12% higher than the floating sub‐1, highlighting that submergence can be considered as a spontaneous and efficient indicator for screening of high‐starch duckweed. Additionally, the saccharification efficiency of starch and ethanol yield had increased in sub‐1 mutant compared to the wild type. Based on the unique characteristics of sub‐1 mutant, a cultivation model of submerged duckweed in a simulated aquaculture pond was designed to get more starch‐rich biomass, enabling effective production of renewable bioenergy.

Abstract Miscanthus is a promising lignocellulosic bioenergy crop for the production of cellulosic ethanol. The recalcitrance of lignocellulose hampers the efficient conversion of Miscanthus biomass to fermentable sugars. Therefore, screening of Miscanthus germplasms with enhanced lignocellulose saccharification efficiency is of important significance to the cellulosic ethanol production from Miscanthus biomass. In this study, we utilize heavy‐ion irradiation mutagenesis to establish a mutant library of Miscanthus lutarioriparius, and screened a reddish stem (rs) mutant with improved lignocellulose saccharification efficiency. The rs mutant was characterized by a noticeable reddish coloration in various lignified cell walls in phloem and vascular bundle sheath, whereas the biomass yield was not compromised. The reddish coloration was attributed to the reduced incorporation of several phenolic compounds including 3‐caffeoylquinic acid, dihydroflavonol and proanthocyanin A, which shared common precursors with lignin monolignol biosynthesis via the phenylpropanoid pathway. Correspondingly, the lignin content was decreased by 10.2% and the cellulose content was concomitantly increased by 9.5% in rs mutant compared with the control plant. Meanwhile, the saccharification efficiency was substantially improved by 22.4%–25.6% in rs mutant compared to the control, when the lignocellulosic biomass was pretreated with or without 1.5% H2SO4. The rs mutant identified here holds a potential utilization as an ideal candidate feedstock in cellulosic bioethanol production.

The GRAS family genes encode plant-specific transcription factors that play important roles in a diverse range of developmental processes and abiotic stress responses. However, the information of GRAS gene family in the bioenergy crop Miscanthus has not been available. Here, we report the genome-wide identification of GRAS gene family in Micanthus sinensis. A total of 123 MsGRAS genes were identified, which were divided into ten subfamilies based on the phylogenetic analysis. The co-linearity analysis revealed that 59 MsGRAS genes experienced segmental duplication, forming 35 paralogous pairs. The expression of six MsGRAS genes in responding to salt, alkali, and mixed salt-alkali stresses was analyzed by transcriptome and real-time quantitative PCR (RT-qPCR) assays. Furthermore, the role of MsGRAS60 in salt and alkali stress response was characterized in transgenic Arabidopsis. The MsGRAS60 overexpression lines exhibited hyposensitivity to abscisic acid (ABA) treatment and resulted in compromised tolerance to salt and alkali stresses, suggesting that MsGRAS60 is a negative regulator of salt and alkali tolerance via an ABA-dependent signaling pathway. The salt and alkali stress-inducible MsGRAS genes identified serve as candidates for the improvement of abiotic stress tolerance in Miscanthus.

Brassinosteroid (BR) signaling has long been reported to have an effect on xylem development, but the detailed mechanism remains unclear, especially in tree species. In this study, we find PdC3H17, which was demonstrated to mediate xylem formation driven by auxin in our previous report, is also involved in BR-promoted xylem development. Y1H analysis, EMSA, and transcription activation assay confirmed that PdC3H17 was directly targeted by PdBES1, which is a key transcriptional regulator in BR signaling. Tissue specificity expression analysis and in situ assay revealed that PdC3H17 had an overlapping expression profile with PdBES1. Hormone treatment examinations verified that xylem phenotypes in PdC3H17 transgenic plants, which were readily apparent in normal condition, were attenuated by treatment with either brassinolide or the BR biosynthesis inhibitor propiconazole. The subsequent quantitative real-time polymerase chain reaction (qRT-PCR) analyses further revealed that BR converged with PdC3H17 to influence transcription of downstream xylem-related genes. Additionally, the enhancement of xylem differentiation by auxin in PdC3H17 overexpression plants was significantly attenuated compared with wild-type and dominant negative plants due to BR deficiency, which suggested that the BR- and auxin-responsive gene PdC3H17 acted as an mediation of these two hormones to facilitate xylem development. Taken together, our results demonstrate that BR signaling converges with auxin-mediated PdC3H17 to regulate xylem formation in Populus and thus provide insight into the regulation mechanism of BRs and the crosstalk with auxin signaling on xylem formation.

Obesity leads to multiple health problems, including diabetes, fatty liver, and even cancer. Here, we report that urolithin A (UA), a gut-microflora-derived metabolite of pomegranate ellagitannins (ETs), prevents diet-induced obesity and metabolic dysfunctions in mice without causing adverse effects. UA treatment increases energy expenditure (EE) by enhancing thermogenesis in brown adipose tissue (BAT) and inducing browning of white adipose tissue (WAT). Mechanistically, UA-mediated increased thermogenesis is caused by an elevation of triiodothyronine (T3) levels in BAT and inguinal fat depots. This is also confirmed in UA-treated white and brown adipocytes. Consistent with this mechanism, UA loses its beneficial effects on activation of BAT, browning of white fat, body weight control, and glucose homeostasis when thyroid hormone (TH) production is blocked by its inhibitor, propylthiouracil (PTU). Conversely, administration of exogenous tetraiodothyronine (T4) to PTU-treated mice restores UA-induced activation of BAT and browning of white fat and its preventive role on high-fat diet (HFD)-induced weight gain. Together, these results suggest that UA is a potent antiobesity agent with potential for human clinical applications.

Abstract Miscanthus is a promising lignocellulosic bioenergy crop for bioethanol production. To identify candidate genes and regulation networks involved in secondary cell wall (SCW) development in Miscanthus, we performed de novo transcriptome analysis of a developing internode. According to the histological and in-situ histochemical analysis, an elongating internode of M. lutarioriparius can be divided into three distinct segments, the upper internode (UI), middle internode (MI) and basal internode (BI), each representing a different stage of SCW development. The transcriptome analysis generated approximately 300 million clean reads, which were de novo assembled into 79,705 unigenes. Nearly 65% of unigenes was annotated in seven public databases. Comparative profiling among the UI, MI and BI revealed four distinct clusters. Moreover, detailed expression profiling was analyzed for gene families and transcription factors (TFs) involved in SCW biosynthesis, assembly and modification. Based on the co-expression patterns, putative regulatory networks between TFs and SCW-associated genes were constructed. The work provided the first transcriptome analysis of SCW development in M. lutarioriparius. The results obtained provide novel insights into the biosynthesis and regulation of SCW in Miscanthus. In addition, the genes identified represent good candidates for further functional studies to unravel their roles in SCW biosynthesis and modification.

Aiming at the support crushing disasters in proximity to coal seam beneath a room mining goaf caused by the different pressure time of the roof above pillars and the immediate roof，according to the nonlinear variation of the pillars'instability failure under the influence of the overlying load，a system mechanics model composed of roof-pillars-gangue was established.In addition，by using the cusp catastrophe theory，the instability mechanisms of the system under the linear loads were studied，the necessary sufficient condition for instability failure of the system and the formula of the jump value and the releasing energy of displacement of pillars were gained，and the main factors for instability failure of the system were analyzed.The results show that the unstable failure of pillars leads to the support crushing disasters.By using the energy method，the attacking dynamic factor and the working resistance of support were calculated.The rationality of the theoretical derivation has been verified by an engineering example.

The prevalence of obesity has increased dramatically worldwide in the past ~50 years. Searching for safe and effective anti-obesity strategies are urgently needed. Lactucin, a plant-derived natural small molecule, is known for anti-malaria and anti-hyperalgesia. The study is to investigate whether lactucin plays a key role in adipogenesis. To this end, in vivo male C57BL/6 mice fed a high-fat diet (HFD) were treated with 20 mg/kg/day of lactucin or vehicle by gavage for seven weeks. Compared with vehicle-treated controls, Lactucin-treated mice showed lower body mass and mass of adipose tissue. Consistently, in vitro 3T3-L1 cells were treated with 20 μM of lactucin. Compared to controls, lactucin-treated cells showed significantly less lipid accumulation during adipocyte differentiation and lower levels of lipid synthesis markers. Mechanistically, we showed the anti-adipogenic property of lactucin was largely limited to the early stage of adipogenesis. Lactucin-treated cells fail to undergo mitotic clonal expansion (MCE). Further studies demonstrate that lactucin-induced MCE arrests might result from reduced phosphorylation of JAK2 and STAT3. We then asked whether activation of JAK2/STAT3 would restore the inhibitory effect of lactucin on adipogenesis with pharmacological STAT3 activator colivelin. Our results revealed similar levels of lipid accumulation between lactucin-treated cells and controls in the presence of colivelin, indicating that inactivation of STAT3 is the limiting factor for the anti-adipogenesis of lactucin in these cells. Together, our results provide the indication that lactucin exerts an anti-adipogenesis effect, which may open new therapeutic options for obesity.

Fatty liver is a major health problem worldwide. People with hereditary deficiency of hormone-sensitive lipase (HSL) are reported to develop fatty liver. In this study, systemic and tissue-specific HSL-deficient mice were used as models to explore the underlying mechanism of this association. We found that systemic HSL deficient mice developed fatty liver in an age-dependent fashion between 3 and 8 months of age. To further explore the mechanism of fatty liver in HSL deficiency, liver-specific HSL knockout mice were created. Surprisingly, liver HSL deficiency did not influence liver fat content, suggesting that fatty liver in HSL deficiency is not liver autonomous. Given the importance of adipose tissue in systemic triglyceride metabolism, we created adipose-specific HSL knockout mice and found that adipose HSL deficiency, to a similar extent as systemic HSL deficiency, causes age-dependent fatty liver in mice. Mechanistic study revealed that deficiency of HSL in adipose tissue caused inflammatory macrophage infiltrates, progressive lipodystrophy, abnormal adipokine secretion and systemic insulin resistance. These changes in adipose tissue were associated with a constellation of changes in liver: low levels of fatty acid oxidation, of very low density lipoprotein secretion and of triglyceride hydrolase activity, each favoring the development of hepatic steatosis. In conclusion, HSL-deficient mice revealed a complex interorgan interaction between adipose tissue and liver: the role of HSL in the liver is minimal but adipose tissue deficiency of HSL can cause age-dependent hepatic steatosis. Adipose tissue is a potential target for treating the hepatic steatosis of HSL deficiency.

In this paper, we present two kinds of Hermite-type collocation methods for linear Volterra integral equations of the second kind with highly oscillatory Bessel kernels. One method is direct Hermite collocation method, which used direct two-points Hermite interpolation in the whole interval. The other one is piecewise Hermite collocation method, which used a two-points Hermite interpolation in each subinterval. These two methods can calculate the approximate value of function value and derivative value simultaneously. Both methods are constructed easily and implemented well by the fast computation of highly oscillatory integrals involving Bessel functions. Under some conditions, the asymptotic convergence order with respect to oscillatory factor of these two methods are established, which are higher than the existing results. Some numerical experiments are included to show efficiency of these two methods.

Crustal movement and incremental-movement data observed repeatedly at GPS stations during 1999–2009 were analyzed to study the effect of two earthquakes of Ms8.1 and Ms8.0 that occurred in 2001 and 2008, respectively, in Qinghai-Tibet sub-plate and its eastern margin. The result revealed certain anomalous pre-earthquake deformation and some large co-seismic changes. Prior to the 2008 Wenchuan Ms8.0 earthquake, the seismogenic Kunlunshan fault zone became a geographic boundary between different regional movements. At the time of the earthquake, there was an average cross-fault crustal shortening of −1.04 m and an average right-lateral strike slip of 0.76 m along the ruptured segment, as well as a strain-energy release of −62. 66 × 10.7.

Abstract We report herein a facile method for the preparation of sodium tungsten bronzes hollow nanospheres using hydrogen gas bubbles as reactant for chemical reduction of tungstate to tungsten and as template for the formation of hollow nanospheres at the same time. The chemical composition and the crystalline state of the as-prepared hollow Na0.15WO3nanospheres were characterized complementarily, and the hollow structure formation mechanism was proposed. The hollow Na0.15WO3nanospheres showed large Brunauer–Emment–Teller specific area (33.8 m2 g−1), strong resistance to acids, and excellent ability to remove organic molecules such as dye and proteins from aqueous solutions. These illustrate that the hollow nanospheres of Na0.15WO3should be a useful adsorbent.

Based upon the fast computation of the coefficients of the interpolation polynomials at Chebyshev-type points by FFT, together with the efficient evaluation of the modified moments by forward recursions or by Oliver’s algorithms, this paper presents fast and stable interpolating integration algorithms, by using the coefficients and modified moments, for Clenshaw-Curtis, Fejér’s first- and second-type rules for Jacobi weights or Jacobi weights multiplied by a logarithmic function. Numerical examples illustrate the stability, efficiency, and accuracy of these quadratures.

Titania nanotube-based local drug delivery is an attractive strategy for combating implant-associated infection. In our previous study, we demonstrated that the gentamicin-loaded nanotubes could dramatically inhibit bacterial adhesion and biofilm formation on implant surfaces. Considering the overuse of antibiotics may lead to the evolution of antibiotic-resistant bacteria, we synthesized a new quaternized chitosan derivative (hydroxypropyltrimethyl ammonium chloride chitosan, HACC) with a 27% degree of substitution (DS; referred to as 27% HACC) that had a strong antibacterial activity and simultaneously good biocompatibility with osteogenic cells. Titania nanotubes with various diameters (80, 120, 160, and 200 nm) and 200 nm length were loaded with 2 mg of HACC using a lyophilization method and vacuum drying. Two standard strain, methicillin-resistant Staphylococcus aureus (American Type Culture Collection 43300) and Staphylococcus epidermidis (American Type Culture Collection 35984), and two clinical isolates, S. aureus 376 and S. epidermidis 389, were selected to investigate the bacterial adhesion at 6 h and biofilm formation at 24, 48, and 72 h on the HACC-loaded nanotubes (NT-H) using the spread plate method, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM). Smooth titanium (Smooth Ti) was also investigated and compared. We found that NT-H could significantly inhibit bacterial adhesion and biofilm formation on its surface compared with Smooth Ti, and the NT-H with 160 nm and 200 nm diameters had stronger antibacterial activity because of the extended HACC release time of NT-H with larger diameters. Therefore, NT-H can significantly improve the antibacterial ability of orthopedic implants and provide a promising strategy to prevent implant-associated infections.

BACKGROUND: Homeodomain-leucine zipper (HD-ZIP) proteins are plant-specific transcriptional factors known to play crucial roles in plant development. Although sequence phylogeny analysis of Populus HD-ZIPs was carried out in a previous study, no systematic analysis incorporating genome organization, gene structure, and expression compendium has been conducted in model tree species Populus thus far. PRINCIPAL FINDINGS: In this study, a comprehensive analysis of Populus HD-ZIP gene family was performed. Sixty-three full-length HD-ZIP genes were found in Populus genome. These Populus HD-ZIP genes were phylogenetically clustered into four distinct subfamilies (HD-ZIP I-IV) and predominately distributed across 17 linkage groups (LG). Fifty genes from 25 Populus paralogous pairs were located in the duplicated blocks of Populus genome and then preferentially retained during the sequential evolutionary courses. Genomic organization analyses indicated that purifying selection has played a pivotal role in the retention and maintenance of Populus HD-ZIP gene family. Microarray analysis has shown that 21 Populus paralogous pairs have been differentially expressed across different tissues and under various stresses, with five paralogous pairs showing nearly identical expression patterns, 13 paralogous pairs being partially redundant and three paralogous pairs diversifying significantly. Quantitative real-time RT-PCR (qRT-PCR) analysis performed on 16 selected Populus HD-ZIP genes in different tissues and under both drought and salinity stresses confirms their tissue-specific and stress-inducible expression patterns. CONCLUSIONS: Genomic organizations indicated that segmental duplications contributed significantly to the expansion of Populus HD-ZIP gene family. Exon/intron organization and conserved motif composition of Populus HD-ZIPs are highly conservative in the same subfamily, suggesting the members in the same subfamilies may also have conservative functionalities. Microarray and qRT-PCR analyses showed that 89% (56 out of 63) of Populus HD-ZIPs were duplicate genes that might have been retained by substantial subfunctionalization. Taken together, these observations may lay the foundation for future functional analysis of Populus HD-ZIP genes to unravel their biological roles.