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The calcium-sensing receptor (CAS), as a chloroplast thylakoid membrane protein, is involved in the process of external Ca

Hyperosmolality-gated calcium-permeable channels (OSCA) are characterized as an osmosensor in plants; they are able to recognize and respond to exogenous and endogenous osmotic changes, and play a vital role in plant growth and adaptability to environmental stress. To explore the potential biological functions of OSCAs in maize, we performed a bioinformatics and expression analysis of the ZmOSCA gene family. Using bioinformatics methods, we identified twelve OSCA genes from the genome database of maize. According to their sequence composition and phylogenetic relationship, the maize OSCA family was classified into four groups (Ⅰ, Ⅱ, Ⅲ, and Ⅳ). Multiple sequence alignment analysis revealed a conserved DUF221 domain in these members. We modeled the calcium binding sites of four OSCA families using the autodocking technique. The expression profiles of ZmOSCA genes were analyzed in different tissues and under diverse abiotic stresses such as drought, salt, high temperature, and chilling using quantitative real-time PCR (qRT-PCR). We found that the expression of twelve ZmOSCA genes is variant in different tissues of maize. Furthermore, abiotic stresses such as drought, salt, high temperature, and chilling differentially induced the expression of twelve ZmOSCA genes. We chose ZmOSCA2.2 and ZmOSCA2.3, which responded most strongly to temperature stress, for prediction of protein interactions. We modeled the calcium binding sites of four OSCA families using autodocking tools, obtaining a number of new results. These results are helpful in understanding the function of the plant OSCA gene family for study of the molecular mechanism of plant osmotic stress and response, as well as exploration of the interaction between osmotic stress, high-temperature stress, and low-temperature stress signal transduction mechanisms. As such, they can provide a theoretical basis for crop breeding.

SUMMARYFruit ripening evolved to be attractive to frugivores that derive energy and nutrition from the fruits in exchange for assisting seed dispersal, which is accompanied by the dramatically change of fruit characteristics, including color, aroma, and texture. The plant hormone ethylene plays a key role in climacteric fruit ripening, while the role of other phytohormones as well as their cross talk with ethylene in modulating fruit ripening remains elusive. Here, we report growth-promoting phytohormone brassinosteroids promote fruit ripening in tomato through regulation of ethylene biosynthesis. Exogenous BR treatment and the increase of endogenous BR content in SlCYP90B3-OE promoted ethylene production and fruit ripening. SlBZR1, a central component and positive regulator of BR signaling pathway, promotes ethylene production and carotenoid accumulation through direct transcriptional regulation of SlACO1, SlACO3 and SlPSY1. Furthermore, SlBIN2, a negative regulator of BR signaling upstream of SlBZR1, decreases ethylene production and carotenoid accumulation. Together, our results demonstrate that BR signaling integrates ethylene and carotenoid biosynthetic pathway to regulate fruit ripening.

Summary: Fruit softening indicated by firmness determines the texture, transportability, and shelf life of tomato products. However, the regulatory mechanism underlying firmness formation in tomato fruit is poorly understood. Here, we report the regulatory role of SlBES1, an essential component of brassinosteroid hormone signaling, in tomato fruit softening. We found that SlBES1 promotes fruit softening during tomato fruit ripening and postharvest storage. RNA-seq analysis suggested that PMEU1, which encodes a pectin methylesterase, might participate in SlBES1-mediated softening. Biochemical and immunofluorescence assays indicated that SlBES1 inhibited PMEU1-related pectin de-methylesterification. Further molecular and genetic evidence verified that SlBES1 directly binds to the E-box of PMEU1 to repress its expression, leading to fruits softening. Loss-of-function SlBES1 mutant generated by CRISPR-Cas9 showed firmer fruits and longer shelf life during postharvest storage without other quality alteration. Collectively, our results indicated the potential of manipulating SlBES1 to regulate firmness without negative consequence on visual and nutrition quality.

Solidago is a genus of more than 120 species of flowering plants in the Asteraceae family and mainly distributed in Americas. To explore the differences of volatile components and analyze their correlations as well variability among the five species (Solidago caesia, Solidago tortifolia, Solidago glomerata, Solidago nemoralis and Solidago rugosa) of this genus, their volatile components were determined by gas chromatography-mass spectrometry (GC-MS) and then the principal component analysis (PCA) was carried out with PAST (PAST2.08). The results indicated that Solidago caesia almost characterized by the same chemotype with Solidago glomerata due to their common essential oil compositions b-cubebene, (1R)-(+)-α-pinene and (−)-a-cadinol while Solidago nemoralis has little variability with Solidago rugosa and with the presence of germacrene D, limonene and sabenene as the representative compounds. But there is chemical variability between Solidago tortifolia and other four species. Sabenene and πcalacorene are the major volatile compounds of Solidago tortifolia.

Synthetic fungicides have been widely used to protect crops from fungal diseases. However, excessive use of synthetic fungicides leads to the generation of fungicide resistance in fungal pathogens. Recently, smart cargo delivery systems have been introduced for the construction of a pesticide delivery nanoplatform, benefiting from their controlled release performance. Herein, a fungal pathogen microenvironment-responsive supramolecular fungicide nanoplatform has been designed and constructed, using quaternary ammonium salt (Q)-modified mesoporous silica nanoparticles (MSN-Q NPs) as nanocarriers loaded with berberine hydrochloride (BH) and carboxylatopillar[5]arene (CP[5]A) as nanogates to form BH-loaded CP[5]A@MSN-Q NPs for effective inhibition of Botrytis cinerea. CP[5]A as nanogates can endow the fungicide nanoplatform with pH stimuli-responsive release features for the control of fungicide release. The loaded BH, as a natural plant fungicide, provides an ecofriendly alternative to synthetic fungicides for controlling B. cinerea. Interestingly, we use oxalic acid (OA) secreted by B. cinerea as a trigger so that BH can be released from the fungicide nanoplatform on demand under pathogen microenvironments for controlling B. cinerea. The experimental results indicate that the fabricated fungicide nanoplatform could effectively inhibit the mycelial growth and spore germination, providing a new way for the management of B. cinerea in actual application.

Due to the unique environmental conditions and vast territory, marine habitat breeds more abundant biological resources than terrestrial environment. Massive marine biological species provide valuable resources for obtaining a large number of natural products with diverse structure and excellent activity. In recent years, new breakthroughs have been made in the application of marine natural products in drug development. In addition, the use of marine natural products to develop insecticides and other pesticide products has also been widely concerned. Targeting marine plants, animals, and microorganisms, we have collected information on marine natural products with insecticidal activity for nearly decade, including alkaloids, terpenes, flavonoids and phenols fatty acids, peptides, and proteins, et al. In addition, some active crude extracts are also included. This review describes the insecticidal activities of marine natural products and their broad applications for future research in agriculture and health.

Firmness is one of the most important factors that affect postharvest properties of tomato fruit. However, the regulatory mechanism underlying firmness formation in tomato fruit is poorly understood. Here, we report a novel role of SlBES1, a transcriptional factor (TF) mediating brassinosteroid (BR) signaling, in tomato fruit softening. We first found that SlBES1 promotes fruit softening during tomato fruit ripening and postharvest storage. RNA-seq analysis suggested that PMEU1, which encodes a pectin de-methylesterification protein, might participate in SlBES1-mediated fruit softening. Biochemical and immunofluorescence assays in SlBES1 transgenic fruits indicated that SlBES1 inhibited PMEU1-related pectin de-methylesterification. Further molecular and genetic evidence verified that SlBES1 directly binds to the E-box in the promoter of PMEU1 to repress its expression, leading to the softening of the tomato fruits. Loss-of-function SlBES1 mutant generated by CRISPR/cas9 showed firmer fruits and longer shelf life during postharvest storage without the color, size and nutritional quality alteration. Collectively, our results indicated the potential of manipulating SlBES1 to regulate fruit firmness via transcriptional inhibition of PMEU1 without negative consequence on visual and nutrition quality.

Brassinosteroids (BRs) play critical roles in plant growth and development, as well as in responses to abiotic stresses. The BRASSINAZOLE RESISTANT 1 (BZR1) and BRI1-EMS-SUPPRESSOR 1 (BES1) families of transcription factors have been elucidated largely in Arabidopsis and rice but not in other plant species. Here, we studied the functional characterization of a tomato (Solanum lycopersicum) BZR homolog gene, SlBZR1, in BR-regulated plant growth and tolerance to salt stress. SlBZR1 was highly expressed in the flowers and developing fruits of tomato. Both SlBZR1 and SlBZR1D (proline to leucine mutation at the 239th amino acid of SlBZR1) were transcriptional repressors and localized in the nucleus. SlBZR1 or SlBZR1D could interact with SlMYB30, SlMYBL2, SlPIF4, SlHAT1, SlIWS1 and SlREF6 in tomato. Overexpression of SlBZR1D enhanced the BR response and improved tolerance to salt stress in Arabidopsis, consistent with the phenotype of the Arabidopsis bes1-D mutant. Moreover, SlBZR1D-overexpressing tomato lines showed a short plant height, smaller and curly leaves, and delayed flowering. Additionally, SlBZR1D positively regulated salt tolerance in tomato and upregulated the expression of multiple stress-related genes. Our study provides new insights for understanding the function and mechanism of BZR transcription factors in BR-regulated plant growth and abiotic stress responses.

Nanotechnology in agriculture and food industry has attracted significant attention over the past few decades. Benefiting from the introduction of controlled release capability, drug/cargo delivery systems possess great advantages for plants in delivering plant genes, hormones, pesticides, and fertilizers to mediate their growth. Herein, we construct a new smart multi-stimuli responsive hormone delivery system to regulate the release of gibberellin acid (GA3) for plant growth. Hollow mesoporous silica nanoparticles have been utilized as nanocarriers, upon which water soluble carboxylatopillar[5]arene ammonium (WP[5]A) functionalized Fe3O4 nanoparticles (WP[5]A-Fe3O4) are installed as nanovalves via host–guest interactions (WP[5]A-Fe3O4 capped HMSNs are denoted as HMSN/Fe3O4). Bidirectional pH-responsive supramolecular nanovalves have been constructed for the first time, and under acidic or alkaline conditions hormones can be released out from GA3-loaded HMSN/Fe3O4 (GA3-HMSN/Fe3O4) to promote the growth of plants. Interestingly, the GA3-HMSN/Fe3O4 nanoparticles can also respond to other stimuli including 1,4-butanediamine (BDA) and ultrasound. This smart delivery system has been successfully applied in plants like Arabidopsis thaliana (A. thaliana) and cabbages, and our experimental results suggested that the GA3-HMSN/Fe3O4 hybrid nanomaterials can obviously promote the growth of both plants. Therefore, we envision that this magnetic multi-stimuli responsive system will hopefully play an important role in promoting agricultural industry development.

Controlled fungicide delivery in response to the specific microenvironment produced by fungal pathogens is an advisable strategy to improve the efficacy of fungicides. Herein, the authors construct a smart fungicide nanoplatform, using mesoporous silica nanoparticles (MSNs) as nanocarriers loaded with eugenol (EU) and Ag

Vaccinium corymbosum (blueberry) is touted as a superfood with numerous health benefits due to its high levels of flavonoids. Flavanone 3-hydroxylase (F3H) is a key regulatory enzyme of the flavonoid pathway. In this study, we cloned the full-length cDNA of F3H (designated VcF3H) from young blueberry leaves using rapid amplification of cDNA ends (RACE). The cDNA contained a 1080-bp open reading frame that encoded a 359-amino acid protein. The deduced VcF3H protein showed high similarities to other plant F3Hs. Conserved amino acid motifs required for ferrous iron binding (HXD) and 2-oxoglutarate binding (RXS) were identified in VcF3H, VcFLS (flavonol synthase), and VcANS (anthocyanidin synthase). Quantitative RT-PCR analysis demonstrated that VcF3H was expressed in all tissues tested, with particularly high expression in young leaves, fruits (pink and blue), and stems. Anthocyanins accumulated mainly in fruits, whereas flavonols were found mainly in leaves and stems. Furthermore, the expression pat...

VcFLS from Vaccinium corymbosum promoted myricetin biosynthesis in Arabidopsis thaliana and VcFLS expression was induced by salicylic acid. Flavonoids are polyphenols with important functions in pigmentation, UV filtration, and symbiotic nitrogen fixation. Flavonols are a class of flavonoids that are produced by the desaturation of dihydroflavanols in a reaction that is catalyzed by flavonol synthase (FLS). In the study reported here, we cloned the full-length cDNA of FLS (designated as VcFLS) from Vaccinium corymbosum (blueberry) using rapid amplification of cDNA ends (RACE). The cDNA contained a 1005-bp open reading frame that encoded a 334-amino acid protein. Phylogenetic analysis showed that VcFLS was closely related to FaFLS, a flavonol synthase that catalyzed the formation of kaempferol and had little effect on the formation of quercetin. Quantitative RT-PCR analysis demonstrated that VcFLS was expressed in all of the tissues tested, with particularly high expression in the petals and young leaves (both green and red). The flavanols myricetin and quercetin also occurred in all of these tested tissues, with the highest levels detected in mature leaves. The expression of VcFLS was not consistent with the accumulation of quercetin and myricetin in different tissues, nor were the expressions of VcFLS, VcPAL, VcCHS, VcF3H, and VcF3′5′H consistent with the accumulation of the quercetin during fruit development. However, the change in the trend of VcCHS and VcF3H expression was similar with myricetin accumulation during fruit development. Expression profiling analysis revealed that VcFLS expression was induced by salicylic acid, a phytohormone involved in plant defense against pathogens, and was suppressed by gibberellic acid, a phytohormone involved in various aspects of plant development. Heterologous expression of VcFLS in Arabidopsis thaliana increased the content of myricetin, but did not affect quercetin content. Thus, we conclude that VcFLS is a key enzyme in the flavonol biosynthetic pathway and would appear to be involved in the plant defense response.

The dehydration-responsive element-binding (DREB) transcription factors play important roles in regulation of plant responses to abiotic stresses. In the present study, ZmDBF3, a novel DREB transcription factor gene from maize (Zea mays L.), was cloned and characterized. Sequence analyses revealed that ZmDBF3 is classified into A-4 group. It was demonstrated that ZmDBF3 was induced in by salt, drought, cold, and high temperature, as well as by signaling molecules abscisic acid (ABA), but no significant changes were observed under salicylic acid (SA) and methyl jasmonate (MeJA) conditions. The results of transient expression assays and transcriptional activity analysis revealed that ZmDBF3 is a nuclear protein with transcriptional activity. Overexpression of ZmDBF3 in yeast (Saccharomyces cerevisiae) exhibited increased survival rate under NaCl, KCl, Na2CO3, NaHCO3, PEG6000, freezing, and sorbitol treatment, compared with the control. Furthermore, ectopic expression of ZmDBF3 in Arabidopsis significantly enhanced tolerance to salt, drought, and freezing tolerance. Taken together, the findings indicated that the ZmDBF3 is a novel member of DREB transcription factor which may act as a regulatory factor involved in multiple stress response pathways.

The ZmCBF3 gene is a member of AP2/ERF transcription factor family, which is a large family of plant-specific transcription factors that share a well-conserved DNA-binding domain. To understand the regulatory mechanism of ZmCBF3 gene expression, we isolated and characterized the ZmCBF3 promoter (PZmCBF3). Three deletion fragments of PZmCBF3 were generated, C1–C3, from the translation start codon at position −1079, −638, and −234, and fused to the GUS reporter gene. Each deletion construct was analyzed by Agrobacterium-mediated stable transformation and expression in Arabidopsis thaliana. GUS expression assays indicated that the PZmCBF3 exhibited root-specific expression activity. A 234-bp fragment upstream of the ZmCBF3 gene conferred a high level of GUS activity in Arabidopsis. Some cis-acting elements involved in the down-regulation of gene expression were detected in the promoter, encompassing positions −1079 to −234. PZmCBF3 was activated by cold stress. The MYCCONSENSUSAT elements (CANNTG) were responsible for the ability of PZmCBF3 to respond to cold stress. The results of the present study suggest that PZmCBF3 might play a role in cold tolerance in maize.

The ZmRXO1 gene is a nucleotide-binding site leucine-rich repeat (NBS–LRR) type of R gene in maize (Zea mays). To understand the regulatory mechanism of ZmRXO1 gene expression, we isolated and characterized the ZmRXO1 promoter (PZmRXO1)—the 5′ flanking region of ZmRXO1. A series of PZmRXO1 deletion derivatives, R1–R4, from the translation start code (−1,576, −934, −829, and −582) were fused to the GUS reporter gene, and each deletion construct was analyzed by Agrobacterium-mediated transformation into tobacco. Sequence analysis showed that several cis-acting elements (MBS, Box-I, TGA-element and CCAAT-box) were located within the promoter. Deletion analysis of the promoter suggested that the 1,576-bp fragment upstream of ZmRXO1 gene showed a high level of GUS expression in tobacco. The promoter sequence (−582 to −1) was sufficient to improve transcription of GUS gene under hormones (MeJA, GA, ABA), drought and low temperature. Moreover, there might be repressor elements in the region (−1,576 to −934 bp) to repress ZmRXO1 gene expression under treatment with salicylic acid.

The human traffic in each bus station is estimated in real-time by means of an embedded ARM(Acorn RISC Machine) platform and the API(Application Programming Interface) in OpenCV(Open Source Computer Vision). For high-density human traffic situations, this paper proposed a method in which the Hough Circle Transform, centroid coordinates and color are used as the features for detecting, tracking and counting the moving targets. Experimental results show that the proposed method can solve the problem of counting the moving targets from the video sequences in the case of high-density human traffic.

Previous studies have shown that an ethylene (ET)-dependent pathway is involved in the cell death signalling triggered by Alternaria alternata f. sp. lycopersici (AAL) toxin in detached tomato (Solanum lycopersicum) leaves. In this study, the role of jasmonic acid (JA) signalling in programmed cell death (PCD) induced by AAL toxin was analysed using a 35S::prosystemin transgenic line (35S::prosys), a JA-deficient mutant spr2, and a JA-insensitive mutant jai1. The results indicated that JA biosynthesis and signalling play a positive role in the AAL toxin-induced PCD process. In addition, treatment with the exogenous ET action inhibitor silver thiosulphate (STS) greatly suppressed necrotic lesions in 35S::prosys leaves, although 35S::prosys leaflets co-treated with AAL toxin and STS still have a significant high relative conductivity. Application of 1-aminocyclopropane-1-carboxylic acid (ACC) markedly enhanced the sensitivity of spr2 and jai1 mutants to the toxin. However, compared with AAL toxin treatment alone, exogenous application of JA to the ET-insensitive mutant Never ripe (Nr) did not alter AAL toxin-induced cell death. In addition, the reduced ET-mediated gene expression in jai1 leaves was restored by co-treatment with ACC and AAL toxin. Furthermore, JA treatment restored the decreased expression of ET biosynthetic genes but not ET-responsive genes in the Nr mutant compared with the toxin treatment alone. Based on these results, it is proposed that both JA and ET promote the AAL toxin-induced cell death alone, and the JAI1 receptor-dependent JA pathway also acts upstream of ET biosynthesis in AAL toxin-triggered PCD.

The effect of brassinosteroid (BR) on relative water content (RWC), stomatal conductance (g s ), net photosynthetic rate ( P N), intercellular CO 2 concentration (Ci), lipid peroxidation level, activities of antioxidant enzymes and abscisic acid concentration (ABA) in tomato ( Lycopersicon esculentum ) seedlings under water stress was investigated. Two tomato genotypes, Mill. cv. Ailsa Craig (AC) and its ABA-deficient mutant notabilis ( not ), were used. Water stress was achieved by withholding water and both the AC and not plants were treated with 1 μM 24-epibrassinolide (EBR) or distilled water as a control. The RWC, g s , Ci and P N were significantly decreased under water stress. However, EBR treatment significantly alleviated water stress and increased the RWC and P N. EBR application also markedly increased the activities of antioxidant enzymes (catalase, ascorbate peroxidase and superoxide dismutase) while it decreased g s , Ci and the contents of H 2 O 2 and malondialdehyde (MDA). Interestingly, ABA concentration in AC and not plants was markedly elevated after EBR treatment although the increasing rate and amplitude of ABA in not plants treated by EBR was significantly lower than those in AC plants. Our study suggested that amelioration of the drought stress of tomato seedlings may be caused by EBR-induced elevation of endogenous ABA concentration and/or the activities of antioxidant enzymes.

Broccoli (Brassica oleracea var. italica) florets were packaged in polyethylene bags with no holes (M0), two microholes (M1), and four macroholes (M2), and then stored at 4 or 20 °C. The effects of modified atmosphere packaging (MAP) treatments on visual quality and glucosinolate contents were determined by comparing with non-wrapped florets. The results showed that MAP treatments, especially with M0 and M1, extended the shelf life and reduced the postharvest deterioration of broccoli florets stored at 4 and 20 °C. All three MAP treatments reduced the decreasing concentration rates of individual, total aliphatic and indole glucosinolates in broccoli florets when compared to those in the control, with M0 being the most significant, followed by M1 and M2 during 23 days of storage at 4 °C or 5 days of storage at 20 °C. Broccoli florets with M0 treatment maintained the visual quality and glucosinolate contents for 13 days at 4 °C and 3 days at 20 °C.

Montmorillonite (MMT) supported Sn catalyst was prepared by ion-exchange technique. Ketones are oxidized by 30% hydrogen peroxide in n -butanol and transformed into the corresponding lactones with very high selectivity catalyzed by Sn–montmorillonite. The catalyst can be prepared in large scale in a simple manner without the utilization of any expensive materials and can be recycled.

The intermolecular reaction of phenols with propiolic esters in TFA in the presence of a Pd(OAc)2 catalyst, affording coumarin derivatives, is described. An exclusive formation of 5,6,7-trimethoxy-...

The mycotoxin fumonisin B1 (FB1) causes the accumulation of reactive oxygen species (ROS) which then leads to programmed cell death (PCD) in Arabidopsis. In the process of studying FB1-induced biosynthesis of glucosinolates, we found that indole glucosinolate (IGS) is involved in attenuating FB1-induced PCD. Treatment with FB1 elevates the expression of genes related to the biosynthesis of camalexin and IGS. Mutants deficient in aliphatic glucosinolate (AGS) or camalexin biosynthesis display similar lesions to Col-0 upon FB1 infiltration; however, the cyp79B2 cyp79B3 double mutant, which lacks induction of both IGS and camalexin, displays more severe lesions. Based on the fact that the classic myrosinase β-thioglucoside glucohydrolase (TGG)-deficient double mutant tgg1 tgg2, rather than atypical myrosinase-deficient mutant pen2-2, is more sensitive to FB1 than Col-0, and the elevated expression of TGG1, but not of PEN2, correlates with the decrease in IGS, we conclude that TGG-dependent IGS hydrolysis is involved in FB1-induced PCD. Indole-3-acetonitrile (IAN) and indole-3-carbinol (I3C), the common derivatives of IGS, were used in feeding experiments, and this rescued the severe cell death phenotype, which is associated with reduced accumulation of ROS as well as increased activity of antioxidant enzymes and ROS-scavenging ability. Despite the involvement of indole-3-acetic acid (IAA) in restricting FB1-induced PCD, feeding of IAN and I3C attenuated FB1-induced PCD in the IAA receptor mutant tir1-1 just as in Col-0. Taken together, our results indicate that TGG-catalyzed breakdown products of IGS decrease the accumulation of ROS by their antioxidant behavior, and attenuate FB1 induced PCD in an IAA-independent way.

Simple, efficient, and economical recombinant plant binary expression vectors for deciphering large-scale functional genomic research in plants and promoting crop improvement by genetically engineering and biotechnology is in great demand. In this research, using the pCHF3, pCAMBIA1301, pCAMBIA3300, pCAMBIA3301 vectors, we successfully constructed general plant binary expression vectors carrying CaMV35S and Arabidopsis rd29A promoters mediating multiple cloning sites (SacI, KpnI, SmaI, BamHI, XbaI, SalI, and PstI). Meanwhile, a series of applicative binary expression vectors that can be utilized for subcellular localization were constructed by fusion of the MCS and eGFP. Subsequently, the recombinant vectors were successfully transferred into Arabidopsis thaliana and Nicotiana benthamiana for further investigation of functional elements in these plant binary expression vectors. Our results demonstrated that this system was a convenient and versatile vector system for phenotypic, functional, subcellular localization, and promoter activity analysis, and it provided a relatively high-efficiency and reliable platform for researchers in vector construction and may facilitate large-scale functional genomics analysis in plants.

Many heavy metals are essential for metabolic processes, but are toxic at elevated levels. Metal tolerance proteins provide resistance to this toxicity. In this study, we identified and characterized a heavy metal-associated protein, AcHMA1, from the halophyte, Atriplex canescens. Sequence analysis has revealed that AcHMA1 contains two heavy metal binding domains. Treatments with metals (Fe, Cu, Ni, Cd or Pb), PEG6000 and NaHCO3 highly induced AcHMA1 expression in A. canescens, whereas NaCl and low temperature decreased its expression. The role of AcHMA1 in metal stress tolerance was examined using a yeast expression system. Expression of the AcHMA1 gene significantly increased the ability of yeast cells to adapt to and recover from exposure to excess iron. AcHMA1 expression also provided salt, alkaline, osmotic and oxidant stress tolerance in yeast cells. Finally, subcellular localization of an AcHMA1/GFP fusion protein expressed in tobacco cells showed that AcHMA1 was localized in the plasma membrane. Thus, our results suggest that AcHMA1 encodes a membrane-localized metal tolerance protein that mediates the detoxification of iron in eukaryotes. Furthermore, AcHMA1 also participates in the response to abiotic stress.

Little information is available on gene expression profiling of halophyte A. canescens. To elucidate the molecular mechanism for stress tolerance in A. canescens, a full-length complementary DNA library was generated from A. canescens exposed to 400 mM NaCl, and provided 343 high-quality ESTs. In an evaluation of 343 valid EST sequences in the cDNA library, 197 unigenes were assembled, among which 190 unigenes (83.1% ESTs) were identified according to their significant similarities with proteins of known functions. All the 343 EST sequences have been deposited in the dbEST GenBank under accession numbers JZ535802 to JZ536144. According to Arabidopsis MIPS functional category and GO classifications, we identified 193 unigenes of the 311 annotations EST, representing 72 non-redundant unigenes sharing similarities with genes related to the defense response. The sets of ESTs obtained provide a rich genetic resource and 17 up-regulated genes related to salt stress resistance were identified by qRT-PCR. Six of these genes may contribute crucially to earlier and later stage salt stress resistance. Additionally, among the 343 unigenes sequences, 22 simple sequence repeats (SSRs) were also identified contributing to the study of A. canescens resources.

A new catalytic system, Pd–Mn-HPA (heteropolyacid), has been found to be very efficient in the oxidative carbonylation of phenol to diphenyl carbonate (DPC) with CO and oxygen at low pressure in the absence of solvent. Synergistic effect between W–Mo–heteropolyacid and Mn(OAc)2 was observed for regenerating the Pd(II) species from Pd(0). The ratio of W–Mo in heteropolyacid, efficiency of the heteropolyacid salts and the roles of tetrabutylammonium bromide and molecular sieves have been investigated in detail. Using suitable components of the catalyst system, 53% yield of DPC with a Pd turnover number of 827 can be attained.

Several novel and useful reactions involving transition metal-catalyzed C-H bond activation discovered in our laboratory have been summarized here, which includes olefin arylation, hydroarylation of alkynes, carboxylation of arenes and alkanes, and acetoxylation and aminomethylation of alkanes. Possible mechanisms of these reactions have been suggested.

A new and general method has been developed for preparation of coumarins and quinolinones by intramolecular hydroarylation of alkynes. Various aryl alkynoates and alkynanilides undergo fast intramolecular reaction at room temperature in the presence of a catalytic amount of Pd(OAc)(2) in a mixed solvent containing trifluoroacetic acid (TFA), affording coumarins and quinolinones in moderate to excellent yields with more than 1000 turnover numbers (TON) to Pd. The methodology proved to tolerate a number of functional groups such as Br and CHO. On the basis of isotope experiments, a possible mechanism involving ethynyl chelation-assisted electrophilic metalation of aromatic C-H bonds by in-situ generated cationic Pd(II) species has been discussed. Also the involvement of vinylcationic species has been suggested.

Simple heteroarenes such as pyrroles and indoles undergo addition reactions to C-C triple bonds in the presence of a catalytic amount of Pd(OAc)(2) under very mild conditions, affording cis-heteroarylalkenes in most cases. The cleavage of aromatic C-H bonds is the possible rate-determining step in CH(2)Cl(2), and the addition of heteroaromatic C-H bonds to C-C triple bonds is in a trans-fashion.

Efficient trans-hydroarylation of alkynes by simple arenes has been realized regio- and stereoselectively at room temperature in the presence of Pd(II) or Pt(II) catalysts and a mixed solvent containing trifluoroacetic acid (TFA). Various arenes undergo trans-hydroarylation selectively across terminal and internal C−C triple bondsincluding those conjugated to CHO, COMe, CO2H, and CO2Et groups, affording kinetically controlled cis-arylalkenes predominantly in most cases, especially, in good yields for electron-rich arenes and activated alkynes. The formation of arene/alkyne 1/2 or 2/1 adducts as side products is dependent on the arenes' and alkynes' substituents, which can be suppressed in some cases by changing the catalyst, catalyst concentration, and reaction time. The Pt(II) system, PtCl2/2AgOAc/TFA, shows lower catalytic activity than Pd(OAc)2/TFA, but higher selectivity, giving higher yields of adducts at the same conversion. On the basis of several isotope experiments and control reactions, a possib...

Efficient electrophilic metalation of aromatic C–H bonds leading to new C–C bond formation through regio- and stereoselective addition to alkynes and alkenes has been realized by a catalytic amount (0.02 to 5 mole percent) of palladium(II) or platinum(II) compounds in a mixed solvent containing trifluoroacetic acid at room temperature. Various arenes undergo unexpected selective trans hydroarylation to terminal or internal C≡C bonds inter- and intramolecularly with high efficiency (up to a turnover number of 4500 for palladium), especially for electron-rich arenes, giving thermodynamically unfavorable cis -alkenes, and the oxygen- and nitrogen-containing heterocycles. The simplicity, generality, and efficiency of this process should be very attractive to the possible industrial application for the functionalization of arenes.

The brassinosteroid (BR) response transcription factor Brassinazole resistant 1 (BZR1)-mediated BR signalling regulates many specific developmental processes including fruit ripening. Here, we report the effect of 2,4-epibrassinolide (EBR) and BZR1-1D overexpression on carotenoid accumulation and quality attributes of tomato (Solanum lycopersicum) fruit. EBR-treated pericarp discs of ethylene-insensitive mutant, Never ripe, accumulated significantly more carotenoid than those of the control. The results suggest that BR seems to be involved in modulating pigments accumulation. When three independent transgenic lines overexpressing the Arabidopsis BZR1-1D were used to evaluate the role of BZR1 in regulating tomato fruit carotenoid accumulation and quality attributes, fruits of all three transgenic lines exhibited enhanced carotenoid accumulation and increased soluble solid, soluble sugar and ascorbic acid contents during fruit ripening. In addition, the fruits of two transgenic lines showed dark green shoulder at mature green stage, in accordance with the up-regulated expression level of SlGLK2, which is involved in chloroplast development. Our results indicate the importance of BZR1-centred BR signalling in regulating carotenoid accumulation and quality attributes of tomato fruit and the potential application of the BZR1-like(s) for improvement of nutritional quality and flavour of tomato through genetic engineering.

A new and efficient system for the allylic acetoxylation of olefins has been developed, which consists of a palladium (II) catalyst and hydrogen peroxide as oxidant in acetic acid. The acetoxylation reaction competes with the epoxidation by the peracetic acid generated in situ in this system. Increase of the concentration of palladium catalyst and addition of benzoquinone (BQ), enhance the acetoxylation considerably. For the Pd(OAc)2BQH2O2 system, more than 1000 turnover number could be achieved in the acetoxylation of cyclohexene. Most internal and cyclic olefins tested gave the corresponding allylic acetates in fair to high yields. 1,3-Cyclohexadiene gave 1,4-diacetoxy-cyclohex-2-ene while 1,5-hexadiene afforded 3-acetoxy-methylenecyclopentane; 1,7-octadiene produced 1-octen-7-one and octan-2,7-dione. Terminal olefins gave methyl ketones in good yield with high turnover number both in the presence and absence of BQ. Possible mechanisms and involvement of palladium peroxidic species in both the catalytic acetoxylation and ketonization of olefins are discussed.

Several novel and useful reactions involving transition metal-catalyzed C-H bond activation discovered in our laboratory have been summarized here, which includes olefin arylation, hydroarylation of alkynes, carboxylation of arenes and alkanes, and acetoxylation and aminomethylation of alkanes. Possible mechanisms of these reactions have been sug- gested.

Several novel synthetic reactions of arenes and alkanes discovered and investigated in our laboratory are summarized here. These include olefin arylation, hydroarylation of alkynes, hydroxylation of arenes, carboxylation of arenes and alkanes, and aminomethylation and acetoxylation of alkanes. Most of these reactions are catalyzed by highly electrophilic transition metal cationic species generated in situ in an acid medium, involving electrophilic metalation of C−H bonds of arenes and alkanes which lead to the formation of aryl−metal and alkyl−metal σ-complexes.

Efficient inter- and intramolecular hydroarylations of C-C triple bonds by simple electron-rich arenes have been realized in the presence of Pd (II) or Pt (II) catalysts and an acid such as trifluoroacetic acid and acetic acid. The reaction of heteroarenes such as pyrroles and indoles occurs under very mild conditions (e.g. at room temperature in neutral solvents such as CH2Cl2). The reaction provides a very good strategy for functionalization of arenes as well as heteroarenes, affording arylalkenes. The intramolecular reaction offers a concise and straightforward route to biologically interesting heterocycles such as coumarins, quinolinones and thiocoumarins. The possible mechanism involving electrophilic metalation of aromatic C-H bonds by a cationic metallic species such as [Pd (O2CCF3)] + and the formation of a vinyl cationic species is discussed.

β-Alkenylpyrroles were prepared directly by Pd(II)-catalyzed addition of pyrroles to alkynoates at room temperature. This procedure was applied to direct synthesis of a β-alkenylpyrrole, the pyrrole fragment of haemin.

Reaction of alkoxyphenols and alkynoates in the presence of a catalytic amount of Pd(OAc)2 in trifluoroacetic acid at room temperature gave coumarin derivatives in high yields. This procedure provides a convenient method for direct synthesis of coumarin derivatives under very mild conditions.

A new catalytic system, which consists of Pd(OAc)2 as the catalyst and W–Mo-containing heteropolyacids (HPAs)/Mn(OAc)2 as the bicomponent cocatalysts, has been found to be very efficient in the oxidative carbonylation of phenol to diphenyl carbonate at low pressure in the absence of solvent. The efficiency of the catalytic system is dependent on the ratio of W/Mo in the HPAs, being best in a ratio of 6/6. The synergistic effect between W–Mo-containing HPAs and Mn(OAc)2 has been observed in the catalysis.

An environmentally benign and selective Baeyer–Villiger oxidation system is introduced. Palygorskite-supported Sn complexes were prepared by a simple procedure. Cyclic ketones and acyclic ketones were oxidized by hydrogen peroxide in a reaction catalyzed by palygorskite-supported Sn complexes, affording corresponding lactones or esters with selectivity for the product of 90–99%. The influence of the solvents, reaction temperature, the amount of catalyst used and the reaction time on the catalytic activity and product selectivity were investigated in detail. The catalyst is cheap, easy to be prepared in large scale and can be recycled.

Steroids function as signaling molecules in both animals and plants. While animal steroid hormones are perceived by nuclear receptor family of transcription factors, brassinosteroids (BR) in plants are perceived by a cell surface receptor kinase, BRI1. Recent studies have demonstrated that BR binding to the extracellular domain of BRI1 induces kinase activation and dimerization with another receptor kinase, BAK1. Activated BRI1 or BAK1 then regulate, possibly indirectly, the activities of BIN2 kinase and/or BSU1 phosphatase, which directly regulate the phosphorylation status and nuclear accumulation of two homologous transcription factors, BZR1 and BES1. BZR1 and BES1 directly bind to promoters of BR responsive genes to regulate their expression. The BR signaling pathway has become a paradigm for both receptor kinase signaling in plants and steroid signaling by cell surface receptors in general.