Comparative Transcriptome Analysis Provides Insights into the Effect of Epicuticular Wax Accumulation on Salt Stress in Coconuts.

The coconut is an important tropical economical crop and exhibits high tolerance to various types of salinity stress. However, little is known about the molecular mechanism underlying its salt tolerance. In this study, RNA-Seq was applied to examine the different genes expressed in four coconut varieties when exposed to a salt environment, resulting in the generation of data for 48 transcriptomes. Comparative transcriptome analysis showed that some genes involved in cutin and wax biosynthesis were significantly upregulated in salt treatment compared to the control, including CYP86A4, HTH, CER1, CER2, CER3, DCR, GPAT4, LTP3, LTP4, and LTP5. In particular, the expression of CER2 was induced more than sixfold, with an RPKM value of up to 205 ten days after salt treatment in Hainan Tall coconut, demonstrating superior capacity in salt tolerance compared to dwarf coconut varieties. However, for yellow dwarf and red dwarf coconut varieties, the expression level of the CER2 gene was low at four different time points after exposure to salt treatment, suggesting that this gene may contribute to the divergence in salt tolerance between tall and dwarf coconut varieties. Cytological evidence showed a higher abundance of cuticle accumulation in tall coconut and severe damage to cuticular wax in dwarf coconut.

The small peptide VISP1 acts as a selective autophagy receptor regulating plant-virus interactions.

Selective macroautophagy/autophagy is tightly regulated by cargo receptors that recruit specific substrates to the ATG8-family proteins for autophagic degradation. Therefore, identification of selective receptors and their new cargoes will improve our understanding of selective autophagy functions in plant development and stress responses. We have recently demonstrated that the small peptide VISP1 acts as a selective autophagy receptor to mediate degradation of suppressors of RNA silencing (VSRs) of several RNA and DNA viruses. Moreover, VISP1 induces symptom recovery through fine-tuning the balance of plant immunity and virus pathogenicity. Our findings provide new insights into the double-edged sword roles of selective autophagy in plant-virus interactions.

A selective autophagy receptor VISP1 induces symptom recovery by targeting viral silencing suppressors.

Selective autophagy is a double-edged sword in antiviral immunity and regulated by various autophagy receptors. However, it remains unclear how to balance the opposite roles by one autophagy receptor. We previously identified a virus-induced small peptide called VISP1 as a selective autophagy receptor that facilitates virus infections by targeting components of antiviral RNA silencing. However, we show here that VISP1 can also inhibit virus infections by mediating autophagic degradation of viral suppressors of RNA silencing (VSRs). VISP1 targets the cucumber mosaic virus (CMV) 2b protein for degradation and attenuates its suppression activity on RNA silencing. Knockout and overexpression of VISP1 exhibit compromised and enhanced resistance against late infection of CMV, respectively. Consequently, VISP1 induces symptom recovery from CMV infection by triggering 2b turnover. VISP1 also targets the C2/AC2 VSRs of two geminiviruses and enhances antiviral immunity. Together, VISP1 induces symptom recovery from severe infections of plant viruses through controlling VSR accumulation.

Metabolomics and transcriptomics provide insights into the molecular mechanisms of anthocyanin accumulation in the seed coat of differently colored mung bean (Vigna radiata L.).

Black mung bean is rich in anthocyanin, however, the accumulation and the molecular mechanism of anthocyanin synthesis in black mung bean are unclear. In this study, anthocyanin metabolomics and transcriptomics on the seed coats of two different colors of mung bean were performed to clarify the composition of anthocyanins, and identify transcription factors involved in regulating anthocyanin biosynthesis. In the mature stage, 23 kinds of anthocyanin compounds were identified. All anthocyanin components contents were significantly higher in seed coat of black mung bean compare with green mung bean. Transcriptome analysis suggested that most of the structural genes for anthocyanin biosynthesis and some potential regulatory genes were significantly differentially expressed. WGCNA suggested VrMYB90 was an important regulatory gene in anthocyanin biosynthesis. Arabidopsis thaliana overexpressing VrMYB90 showed significant accumulation of anthocyanins. PAL, 4CL, DFR, F3'5'H, LDOX, F3'H and UFGT were up-regulated in 35S:VrMYB90 Arabidopsis thaliana. These findings provide valuable information for understanding the synthesis mechanism of anthocyanins in black mung bean seed coats.

Trityl Quinodimethane Derivatives as Unimolecular Triple-Function Extracellular EPR Probes for Redox, pH, and Oxygen.

Electron paramagnetic resonance (EPR) spectroscopy and imaging coupled with the use of suitable probes is a promising tool for assessment of the tumor microenvironment (TME). Measurement of multiple TME parameters by EPR is very desirable but challenging. Herein, we designed and synthesized a class of negative-charged trityl quinodimethane MTPs as unimolecular triple-function extracellular probes for redox, pH, and oxygen (O2) levels. Using the deuterated analogue, dMTP5, which has an optimal pKa as well as high sensitivity to bioreduction and O2, we reasonably evaluated pH effects on efflux of reducing agents from HepG2 cells and cellular O2 consumption.

Integrative Physiological, Transcriptome, and Proteome Analyses Provide Insights into the Photosynthetic Changes in Maize in a Maize-Peanut Intercropping System.

Intercropping is a traditional and sustainable planting method that can make rational use of natural resources such as light, temperature, fertilizer, water, and CO2. Due to its efficient resource utilization, intercropping, in particular, maize and legume intercropping, is widespread around the world. However, the molecular details of these pathways remain largely unknown. In this study, physiological, transcriptome, and proteome analyses were compared between maize monocropping and maize-peanut intercropping. The results show that an intercropping system enhanced the ability of carbon fixation and carboxylation of maize leaves. Apparent quantum yield (AQY), the light-saturated net photosynthetic rate (LSPn), the light saturation point (LSP), and the light compensation point (LCP) were increased by 11.6%, 9.4%, 8.9%, and 32.1% in the intercropping system, respectively; carboxylation efficiency (CE), the CO2 saturation point (Cisat), the Rubisco maximum carboxylation rate (Vcmax), the maximum electron transfer rate (Jmax), and the triose phosphate utilization rate (TPU) were increased by 28.5%, 7.3%, 18.7%, 29.2%, and 17.0%, respectively; meanwhile, the CO2 compensation point (Γ) decreased by 22.6%. Moreover, the transcriptome analysis confirmed the presence of 588 differentially expressed genes (DEGs), and the numbers of up-regulated and down-regulated genes were 383 and 205, respectively. The DEGs were primarily concerned with ribosomes, plant hormone signal transduction, and photosynthesis. Furthermore, 549 differentially expressed proteins (DEPs) were identified in the maize leaves in both the maize monocropping and maize-peanut intercropping systems. Bioinformatics analysis revealed that 186 DEPs were related to 37 specific KEGG pathways in each of the two treatment groups. Based on the physiological, transcriptome, and proteome analyses, it was demonstrated that the photosynthetic characteristics in maize leaves can be improved by maize-peanut intercropping. This may be related to PS I, PS II, cytochrome b6f complex, ATP synthase, and photosynthetic CO2 fixation, which is caused by the improved CO2 carboxylation efficiency. Our results provide a more in-depth understanding of the high yield and high-efficiency mechanism in maize and peanut intercropping.

Photocatalytic nitrogen fixation: Oxygen vacancy modified novel micro-nanosheet structure Bi2O2CO3 with band gap engineering.

As a promising ammonia synthesis approach to replace the industrial Harber method, the biggest problem restricting photocatalytic nitrogen fixation is the suboptimal efficiency. Herein, novel surface oxygen vacancies modified micro-nanosheet structure Bi2O2CO3 (namely BOC/OV) were successfully synthesized via facile formation under room temperature. These defects-rich nanosheets exhibit outstanding performance for photocatalytic nitrogen fixation under visible light. The surface oxygen vacancies provide abundant active sites for molecular N2 activation, and the effect of scattered nanometer-size could facilitate the separation of photo-generated charges. Moreover, the energy band can be consecutively tuned with the accumulation of surface oxygen vacancies by lowering the conduction band position. Among all as-prepared samples, BOC/OV3 exhibited the highest NH4+ yield, reaching 1178 µmol·L-1·g-1·h-1, which is 10 times than that of pristine Bi2O2CO3. In this work, all samples synthesis and defects formation were conducted without requiring any secondary energy, which is of great significance for realizing green and efficient artificial ammonia synthesis.

Enantioselective Synthesis of Atropisomeric Anilides via Pd(II)-Catalyzed Asymmetric C-H Olefination.

Atropisomeric anilides have received tremendous attention as a novel class of chiral compounds possessing restricted rotation around an N-aryl chiral axis. However, in sharp contrast to the well-studied synthesis of biaryl atropisomers, the catalytic asymmetric synthesis of chiral anilides remains a daunting challenge, largely due to the higher degree of rotational freedom compared to their biaryl counterparts. Here we describe a highly efficient catalytic asymmetric synthesis of atropisomeric anilides via Pd(II)-catalyzed atroposelective C-H olefination using readily available L-pyroglutamic acid as a chiral ligand. A broad range of atropisomeric anilides were prepared in high yields (up to 99% yield) and excellent stereoinduction (up to >99% ee) under mild conditions. Experimental studies indicated that the atropostability of those anilide atropisomers toward racemization relies on both steric and electronic effects. Experimental and computational studies were conducted to elucidate the reaction mechanism and rate-determining step. DFT calculations revealed that the amino acid ligand distortion is responsible for the enantioselectivity in the C-H bond activation step. The potent applications of the anilide atropisomers as a new type of chiral ligand in Rh(III)-catalyzed asymmetric conjugate addition and Lewis base catalysts in enantioselective allylation of aldehydes have been demonstrated. This strategy could provide a straightforward route to access atropisomeric anilides, one of the most challenging types of axially chiral compounds.

Pd(II)-Catalyzed Tandem Enantioselective Methylene C(sp3 )-H Alkenylation-Aza-Wacker Cyclization to Access ß-Stereogenic γ-Lactams.

Herein, we describe an unprecedented cascade reaction to ß-stereogenic γ-lactams involving Pd(II)-catalyzed enantioselective aliphatic methylene C(sp3 )-H alkenylation-aza-Wacker cyclization through syn-aminopalladation. Readily available 3,3'-substituted BINOLs are used as chiral ligands, providing the corresponding γ-lactams with broad scope and high enantioselectivities (up to 98 % ee).

Detosylative (Deutero)alkylation of Indoles and Phenols with (Deutero)alkoxides.

An efficient strategy for N/O-(deutero)alkylation of indoles and phenols with alkoxides/alcohols as the alkylation reagents is described. The consecutive detosylation/alkylation transformations feature mild reaction conditions, high ipso-selectivity, and good functional group tolerance (>50 examples). A one-pot selective N-alkylation of unprotected indoles with alcohols and TsCl is also realized. The application of this method is demonstrated by the introduction of isotope-labeled (CD3 and 13CH3) groups using the readily accessible labeled alcohols and the synthesis of pharmaceuticals.

Iron-Catalyzed Intramolecular Amination of Aliphatic C-H Bonds of Sulfamate Esters with High Reactivity and Chemoselectivity.

It is challenging to develop simple and low cost catalytic systems while maintaining high reactivity and selectivity. An iron-catalyzed intramolecular C-H amination of sulfamate esters using simple and cheap ligands is reported with general substrate scope (31 examples, up to 95% yield). The addition of second ligand, bipyridine, is able to accelerate the reaction and increase the yield. The ready availability of these iron catalysts provides a promising approach to selective introduction of nitrogen into hydrocarbon feedstock.

Chimeric Japanese Encephalitis Virus SA14/SA14-14-2 Was Virulence Attenuated and Protected the Challenge of Wild-Type Strain SA14.

The attenuated Japanese encephalitis virus (JEV) live vaccine SA14-14-2 prepared from wild-type (WT) strain SA14 was licensed to prevent Japanese encephalitis (JE) in 1989 in China. Many studies showed that the premembrane (prM) and envelope (E) protein were the crucial determinant of virulence and immunogenicity of JEV. So we are interested in whether the substitution of prM/E of JEV WT SA14 with those of vaccine strain SA14-14-2 could decrease neurovirulence and prevent the challenge of JEV WT SA14. Molecular clone technique was used to replace the prM/E gene of JEV WT strain SA14 with those of vaccine strain SA14-14-2 to construct the infectious clone of chimeric virus (designated JEV SA14/SA14-14-2), the chimeric virus recovered from BHK21 cells upon electrotransfection of RNA into BHK21 cells. The results showed that the recovered chimeric virus was highly attenuated in mice, and a single immunization elicited strong protective immunity in a dose-dependent manner. This study increases our understanding of the molecular mechanisms of neurovirulence attenuation and immunogenicity of JEV.

Effective inhibition of different Japanese encephalitis virus genotypes by RNA interference targeting two conserved viral gene sequences in vitro and in vivo.

Japanese encephalitis is a zoonotic, mosquito-borne, infectious disease caused by Japanese encephalitis virus (JEV), which is prevalent in China. At present, there are no specific drugs or therapies for JEV infection, which can only be treated symptomatically. Lentivirus-mediated RNA interference (RNAi) is a highly efficient method to silence target genes. In this study, two lentiviral shRNA, LV-C and LV-NS5, targeting the conserved viral gene sequences were used to inhibit different JEV genotypes strains in BHK21 cells and mice. The results showed that LV-C significantly inhibited JEV genotype I and genotype III strains in cells and mice. Quantitative RT-PCR analysis showed that JEV mRNA were reduced by 83.2-90.9% in cells by LV-C and that flow cytometry analysis confirmed the inhibitory activity of LV-C. The viral titers were reduced by about 1000-fold in cells and the brains of suckling mice by LV-C, and the pretreatment of LV-C protected 60-80% of mice against JEV-induced lethality. The inhibitory activities of LV-NS5 in cells and mice were weaker than those of LV-C. These results indicate that RNAi targeting of the two conserved viral gene sequences had significantly suppressed the replication of different JEV genotypes strains in vitro and in vivo, highlighting the feasibility of RNAi targeting of conserved viral gene sequences for controlling JEV infection.

Diastereoisomers of l-proline-linked trityl-nitroxide biradicals: synthesis and effect of chiral configurations on exchange interactions.

The exchange (J) interaction of organic biradicals is a crucial factor controlling their physiochemical properties and potential applications and can be modulated by changing the nature of the linker. In the present work, we for the first time demonstrate the effect of chiral configurations of radical parts on the J values of trityl-nitroxide (TN) biradicals. Four diastereoisomers (TNT1, TNT2, TNL1 and TNL2) of TN biradicals were synthesized and purified by the conjugation of a racemic (R/S) nitroxide with the racemic (M/P) trityl radical vial-proline. The absolute configurations of these diastereoisomers were assigned by comparing experimental and calculated electronic circular dichroism (ECD) spectra as (M, S, S) for TNT1, (P, S, S) for TNT2, (M, S, R) for TNL1 and (P, S, R) for TNL2. Electron paramagnetic resonance (EPR) results showed that the configuration of the nitroxide part instead of the trityl part is dominant in controlling the exchange interactions and the order of the J values at room temperature is TNT1 (252 G) > TNT2 (127 G) ≫ TNL2 (33 G) > TNL1 (14 G). Moreover, the J values of TNL1/TNL2 with the S configuration in the nitroxide part vary with temperature and the polarity of solvents due to their flexible linker, whereas the J values of TNT1/TNT2 are almost insensitive to these two factors due to the rigidity of their linkers. The distinct exchange interactions between TNT1,2 and TNL1,2 in the frozen state led to strongly different high-field dynamic nuclear polarization (DNP) enhancements with ε = 7 for TNT1,2 and 40 for TNL1,2 under 800 MHz DNP conditions.

Proteomic Analysis of Vernalization Responsive Proteins in Winter Wheat Jing841.

BACKGROUND: Vernalization is one of the pivotal ways for plants to flower. The twodimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-offlight/ time-of-flight mass spectrometry (MALDI-TOF/TOF MS) were applied to analyze the changes in protein expression profiles in responding to vernalization in leaves of wheat seedling before (0d) and after (30d) of vernalization. OBJECTIVE: The main objective of this study was to analyze the vernalization-responsive proteins in winter wheat after vernalization. METHODS: Winter wheat seedling leaf proteins were extracted by phenol extraction coupled with ammonium acetate in methanol. 2-DE was conducted according to procedures described in the manual given by the GE manufacture. The selected protein spots were identified by MALDITOF/ TOF MS. Gene ontology (GO) classification was applied to classify the functions of the differentially expressed proteins. Pathway enrichment analysis identified significantly enriched metabolic pathways or signal transduction pathways relative to the whole proteins background. RESULTS: The results of 2-DE and MALDI-TOF/TOF MS showed that among the 65 differentially expressed proteins that were successfully identified under vernalization, 30 were up-regulated whereas 35 were down-regulated after vernalization, respectively. These vernalization-responsive proteins were found to play roles in carbohydrate metabolism, protein metabolism, photosynthesis, defense and stress-resistance and may therefore participate in many biological processes in responding to vernalization. The enhanced accumulation of proteins after vernalization, such as thiamine thiazole synthase, late embryogenesis abundant protein, and glutathione-S-transferase, probably play vital roles in the mechanisms underlying vernalization response in wheat. CONCLUSION: Our results indicated these vernalization-responsive proteins were found to be involved in protein metabolism, carbohydrate metabolism, photosynthesis, and stress resistance/ defense. The responses of plants to low temperature were very complex, involving in a wide range of cellular pathways for signal transduction, gene regulation, protein modifications, and metabolic regulation. Studying on wheat proteomic profiles in response to vernalization can improve our understanding the molecular mechanisms underlying vernalization in cereals. The results obtained in this study have provided a novel insight into the mechanisms underlying vernalization in cereal crops.

Identification of vernalization responsive genes in the winter wheat cultivar Jing841 by transcriptome sequencing.

This study aimed to identify vernalization responsive genes in the winter wheat cultivar Jing841 by comparing the transcriptome data with that of a spring wheat cultivar Liaochun10. For each cultivar, seedlings before and after the vernalization treatment were sequenced by Solexa/Illumina sequencing. Genes differentially expressed after and before vernalization were identified as differentially expressed genes (DEGs) using false discovery rate (FDR) ≤ 0.001 and |log2 (fold change)|>1 as cutoffs. The Jing841-specific DEGs were screened and subjected to functional annotation using gene ontology (GO) database. Vernalization responsive genes among the specific genes were selected for validation by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and the expression change over the time was investigated for the top 11 genes with the most significant expression differences. A total of 138,062 unigenes were obtained. Overall, 636 DEGs were identified as vernalization responsive genes including some known genes such as VRN-1 and COR14a, and some unknown contigs. The qRT-PCR validated changes in the expression of 18 DEGs that were detected by RNA-seq. Among them, 11 genes displayed four different types of expression patterns over time during the 30-day-long vernalization treatment. Genes or contigs such as VRN-A1, COR14a, IRIP, unigene1806 and Cl18953. Contig2 probably have critical roles in vernalization.

Virus-induced gene silencing-based functional verification of six genes associated with vernalization in wheat.

Vernalization requirement is an important characteristic in crop breeding. Wheat is a widely grown crop in the world that possesses enormous economic significance. To better understand the gene networks in vernalization process, we performed a high-throughput RNA sequencing analysis comparing the transcriptomes of spring and winter wheat cultivars, with and without vernalization (unpublished data). In this study, we selected six unigenes (CL14010, CL12788, CL176, Unigene 16777, CL8746 and Unigene10196) from our transcriptome analysis based on their expression differences to further characterize their function. Transient silencing of the six unigenes individually were achieved through virus-induced gene silencing (VIGS) using BSMV vector. The period from germination to spike differentiation were recorded and compared between plants underwent VIGS silencing and the control. Our result showed that VIGS of the six unigenes significantly shortened the period from seedling to double ridge (DR) stage. Resulting in SD period ranging from 59.8 ± 0.60 to 65.8 ± 0.48 days, compared to 85.0 ± 0.73 days in the control. The results indicated that these six unigenes function as suppressors in vernalization process and silence or down-regulation of these genes promoted flower development in wheat. Further characterization of these six unigenes and their function in vernalization and flowering control is needed.

[Effects of jiawei huzhang san decoction on the expressions of inflammatory factors MCP-1 and PDGF-BB in rat models of experimental autoimmune prostatitis].

OBJECTIVE: To study the effects of Jiawei Huzhang San (JWHZS) decoction on the expressions of the inflammatory factors monocyte chemoattractant protein-1 (MCP-1) and platelet-derived growth factor-BB (PDGF-BB) on experimental autoimmune prostatitis in rats. METHODS: Twelve male Wistar rats were taken as normal controls, and models of experimental autoimmune prostatitis were established in another 60 by injection of SC purified prostate protein with FCA, and then divided into five groups to be treated with normal saline, indomethacin, high-dose JWHZS (0.445 g/kg), medium-dose JWHZS (0.223 g/kg) and low-dose JWHZS (0.089 g/kg), respectively. All the rats were sacrificed at 30 days after the treatment for detection of the mRNA and protein expressions of inflammatory factors by immunohistochemistry and fluorescent quantitative RT-PCR. RESULTS: In the high-, medium- and low-dose JWHZS groups, the mRNA expressions of MCP-1 (0.31 +/- 0.14, 0.49 +/- 0.21 and 0.62 +/- 0.28) and PDGF-BB (0.50 +/- 0.22, 0.54 +/- 0.17 and 0.71 +/- 0.29), and the protein expressions of MCP-1 (677 +/- 208, 725 +/- 311 and 1302 +/- 884) and PDGF-BB (1265 +/- 698, 1347 +/- 827 and 1655 +/- 812) were significantly lower than in the model control group (MCP-1 mRNA: 1.12 +/- 0.43; MCP-1 protein: 2201 +/- 934; PDGF-BB mRNA: 1.14 +/- 0.51; PDGF-BB protein: 2754 +/- 852) (P < 0.05). And JWHZS exhibited a significantly better activity at high and medium doses than at a low dose (P < 0.05). In the indomethacin control group, both the mRNA and protein expressions of MCP-1 (0.71 +/- 0.34 and 1824 +/- 1157) and PDGF-BB (1.08 +/- 0.37 and 2493 +/- 924) were markedly higher than in the JWHZS groups (P < 0.01). CONCLUSION: Down-regulation of the inflammatory factors MCP-1 and PDGF-BB may be the important molecular mechanism of JWHZS acting on experimental autoimmune prostatitis.