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In this study, a water transfer method was developed to fabricate conducive thin-film patterns on 3D curvilinear surfaces. Crystalline silver nanoplates (AgNPLs) with a dimension of 700 nm and a thickness of 35 nm were suspended in ethanol with an anionic surfactant, sodium dodecyl sulfate, to improve the suspension stability. The prepared AgNPL suspension was then spread over the water surface via the Langmuir-Blodgett approach to generate a self-assembled thin film. By dipping an accepting object with a robotic arm, the floating AgNPL thin film with nanometer thickness can be effectively transferred to the object surfaces and exhibited a superior conductivity up to 15% of bulk silver without thermal sintering. Besides good conductivity, the AgNPL conductive thin films can also be transferred efficiently on any curvilinear (concave and convex) surface. Moreover, with the help of masks, conductive patterns can be produced on water surfaces and transferred to curvilinear surfaces for electronic applications. As a proof of concept, several examples were demonstrated to display the capability of this approach for radiofrequency identification and other printed circuit applications.
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Begomoviruses frequently inflict upward or downward leaf curling symptoms on infected plants, leading to severe economic damages. Knowledge of the underlying mechanism controlling the leaf curling severity may facilitate the development of alternative disease management strategies. In this study, through genomic recombination between Ageratum yellow vein virus Nan-Tou strain (AYVV-NT) and Tomato leaf curl virus Tai-Chung Strain (TLCV-TC), which caused upward and downward leaf curling on Nicotiana benthamiana, respectively, it was found that the coding region of C4 protein might be involved in the determination of leaf curling directions. Sequence comparison and mutational analysis revealed that the cysteine and glycine at position 8 and 14 of AYVV-TC C4 protein, respectively, are involved in the modulation of leaf curling symptoms. Cross-protection assays further demonstrated that N. benthamiana inoculated with AYVV-carrying mutations of the aforementioned amino acids exhibited attenuated leaf curling symptoms under the challenge of wild-type AYVV-NT. Together, these findings revealed a new function of begomovirus C4 proteins involved in the modulation of leaf curling severity during symptom formation and suggested potential applications for managing viral diseases through manipulating the symptoms.
Assuntos
Begomovirus , Solanum lycopersicum , Aminoácidos , Begomovirus/genética , Doenças das PlantasRESUMO
In this work, a simple and rapid synthesis method was developed to prepare silver nanoplates (AgNPLs) with a high aspect ratio. A microwave heating process with a high heating rate and uniform heating was used to promote the silver reduction reaction. Silver nitrate (AgNO3) was used as the precursor of AgNPLs, and N,N-dimethylformamide (DMF) played the role of a solvent and reducing agent. Poly(vinylpyrrolidone) (PVP) with a molecular weight of 29,000 and a PVP/AgNO3 ratio of 10 were used to control the shape of synthesized AgNPLs. By adjusting the optimal microwave heating parameters, temperature ramping rate, reaction time, and reaction temperature, triangular AgNPLs with high aspect ratios could be produced. The synthesized AgNPLs had an edge length up to 700 nm and a thickness of 35 nm with aspect ratios up to 20. The AgNPLs were also used to produce conductive patterns via pen writing with a conductivity of 2 × 106 S/m to demonstrate the feasibility of applying the synthesized nanomaterials for electronic applications.
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In this study, an ink formulation was developed to prepare conductive copper thin films with compact structure by using intense pulsed light (IPL) sintering. To improve inter-particle connections in the sintering process, a cuprous oxide shell was synthesized over copper nanoparticles (CuNP). This cuprous oxide shell can be reduced by IPL with the presence of a reductant and fused to form connection between large copper particles. However, the thermal yield stress after strong IPL sintering resulted in cracks of conductive copper film. Thus, a multiple pulse sintering with an off time of 2 s was needed to reach a low resistivity of 10-5 Ω·cm. To increase the light absorption efficiency and to further decrease voids between CuNPs in the copper film, cupric oxide nanoparticles (CuONP) of 50 nm, were also added into ink. The results showed that these CuONPs can be reduced to copper with a single pulse IPL and fused with the surrounding CuNPs. With an optimal CuNP/CuONP weight ratio of 1/80, the copper film showed a lowest resistivity of 7 × 10-5 Ω·cm, ~25% conductivity of bulk copper, with a single sintering energy at 3.08 J/cm2. The ink can be printed on flexible substrates as conductive tracks and the resistance remained nearly the same after 10,000 bending cycles.
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Serological methods are relatively convenient and simple for the detection of pathogens for front-line workers. On-site visualization of the test results plays a pivotal role in the process. However, an efficient, universal labeling agent for antibodies is needed for the development of efficient serological detection tools. In this study, a Bamboo mosaic virus (BaMV)-based viral vector was employed to express recombinant proteins, collectively designated GfED, consisting of Staphylococcus aureus Protein A domain ED (SpaED) fused to either the N- or C-terminal of an improved green florescent protein (GFP) with or without the coat protein (CP) of BaMV, efficiently in Chenopodium quinoa. The GfED in crude leaf extracts could specifically attach to IgG molecules of rabbits and mice, effectively labeling IgG with GFP, emitting green light at 506 nm when excited at 450 nm using simple, handheld equipment. To demonstrate the applicability of GfED in serological assays, we have developed a fluorescent dot blot assay for the rapid detection of Acidovorax citrulli (Ac), a bacterial pathogen of cucurbits, and BaMV, a viral pathogen of bamboos. By using the crude extracts of inoculated C. quinoa leaves expressing GfED as an IgG-labeling agent, the pathogens were easily and quickly detected through uncomplicated operations using simple equipment, with results observable by the naked eye. Examination using fluorescent microscopy and transmission electron microscopy revealed that the GfED subunits may assemble into virus-like particles, which were further involved in the formation of aggregates of GfED-antibody-antigen complexes with the potential for fluorescence signal enhancement. The results suggested that plant-expressed GfED may serve as a promising alternative of IgG-labeling agent for current serological assays.
Assuntos
Chenopodium/metabolismo , Comamonadaceae/isolamento & purificação , Imunofluorescência/métodos , Vetores Genéticos , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Potexvirus/isolamento & purificação , Western Blotting , Chenopodium/virologia , Comamonadaceae/genética , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Folhas de Planta/metabolismo , Reação em Cadeia da Polimerase , Potexvirus/genéticaRESUMO
In this study, we report a facile and green process to synthesize high-quality and few-layer graphene (FLG) derived from graphite via a liquid exfoliation process. The corresponding characterizations of FLG, such as scanning electron microscopy (SEM), transmission electron microscope (TEM), atomic force microscopy (AFM) and Raman spectroscopy, were carried out. The results of SEM show that the lateral size of as-synthesized FLG is 1â»5 µm. The results of TEM and AFM indicate more than 80% of graphene layers is <10 layers. The most surprising thing is that D/G ratio of graphite and FLG are 0.15 and 0.19, respectively. The result of the similar D/G ratio demonstrates that little structural defects were created via the liquid exfoliation process. Electronic conductivity tests and resistance of composite film, in terms of different contents of graphite/polyvinylidene difluoride (PVDF) and FLG/PVDF, were carried out. Dramatically, the FLG/PVDF composite demonstrates superior performance compared to the graphite/PVDF composite at the same ratio. In addition, the post-sintering process plays an important role in improving electronic conductivity by 85%. The composition-optimized FLG/PVDF thin film exhibits 81.9 S·cm-1. These results indicate that the developed FLG/PVDF composite adhesives could be a potential candidate for conductive adhesive applications.
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Bamboo mosaic virus (BaMV), a member of the genus Potexvirus, is the major threat to bamboo cultivation. Similar to most potexviruses, the transmission of BaMV by insect vectors has not been documented previously. However, field observations of BaMV disease incidences suggested that insect vectors might be involved. In this study, we aimed to investigate the possibility of insect-mediated transmission of BaMV among bamboo clumps, in order to provide further insights into the infection cycles of BaMV for the development of effective disease management measures. From the major insects collected from infected bamboo plantations, BaMV genomic RNAs were detected inside the bodies of two dipteran insects, Gastrozona fasciventris and Atherigona orientalis, but not in thrips (Scirtothrips dorsalis). Artificial feeding assays using green fluorescent protein-tagged BaMV virions revealed that BaMV could enter the digestive systems and survive in the regurgitant and excretion of the dipterans. BaMV RNA could be retained in the dipterans for up to 4 weeks. Insect-mediated transmission assays indicated that both dipterans could transmit BaMV to bamboo seedlings through artificially created wounds with low infection efficiency (14 - 41%), suggesting that the dipterans may mediate the transmission in a mechanical-like manner. These results demonstrated that dipterans with sponge-like mouthparts may also serve as vectors for at least one potexvirus, BaMV, among bamboo plants. The finding suggested that dipteran insect control should be integrated into the disease management measures against BaMV infections.
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A cryptic prokaryotic promoter, designated AV3 promoter, has been previously identified in certain begomovirus genus, including ageratum yellow vein virus isolate NT (AYVV-NT). In this study, we demonstrated that the core nucleotides in the putative -10 and -35 boxes are necessary but not sufficient for promoter activity in Escherichia coli, and showed that AYVV-NT AV3 promoter could specifically interact with single-stranded DNA-binding protein and sigma 70 of E. coli involved in transcription. Several AYVV-NT-encoded proteins were found to increase the activity of AV3 promoter. The transcription start sites downstream to AV3 promoter were mapped to nucleotide positions 803 or 805 in E. coli, and 856 in Nicotiana benthamiana. The eukaryotic activity of AV3 promoter and the translatability of a short downstream open reading frame were further confirmed by using a green fluorescent protein reporter construct in yeast (Saccharomyces cerevisiae) cells. These results suggested that AV3 promoter might be a remnant of evolution that retained cryptic activity at present.
Assuntos
Ageratum/virologia , Begomovirus/genética , Nicotiana/virologia , Fases de Leitura Aberta , Regiões Promotoras Genéticas , Proteínas Virais/genética , Agrobacterium/genética , Sequência de Bases , Begomovirus/metabolismo , Evolução Biológica , Escherichia coli/genética , Escherichia coli/virologia , Genoma Viral , Dados de Sequência Molecular , Doenças das Plantas/virologia , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/virologia , Transcrição Gênica , Proteínas Virais/química , Proteínas Virais/metabolismoRESUMO
Geminiviruses are known to exhibit both prokaryotic and eukaryotic features in their genomes, with the ability to express their genes and even replicate in bacterial cells. We have demonstrated previously the existence of unit-length single-stranded circular DNAs of Ageratum yellow vein virus (AYVV, a species in the genus Begomovirus, family Geminiviridae) in Escherichia coli cells, which prompted our search for unknown prokaryotic functions in the begomovirus genomes. By using a promoter trapping strategy, we identified a novel prokaryotic promoter, designated AV3 promoter, in nts 762-831 of the AYVV genome. Activity assays revealed that the AV3 promoter is strong, unidirectional, and constitutive, with an endogenous downstream ribosome binding site and a translatable short open reading frame of eight amino acids. Sequence analyses suggested that the AV3 promoter might be a remnant of prokaryotic ancestors that could be related to certain promoters of bacteria from marine or freshwater environments. The discovery of the prokaryotic AV3 promoter provided further evidence for the prokaryotic origin in the evolutionary history of geminiviruses.
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Begomovirus/genética , DNA Viral/genética , Genoma Viral , Regiões Promotoras Genéticas , Sequência de Aminoácidos , Sequência de Bases , Begomovirus/química , Begomovirus/classificação , Evolução Biológica , DNA Viral/química , Escherichia coli/virologia , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Plantas/virologiaRESUMO
Most infectious clones of geminiviruses consist of (partial) tandem repeats of viral genomes in the vectors, which usually involve tedious, multi-step assemblies of genomic fragments in the construction process. A simplified procedure was devised to circumvent these problems, which employs limited restriction digestion of multimeric viral genomes produced by rolling circle amplification (RCA), followed by direct cloning into appropriate vectors. The efficiency of the procedure, and infectivity of the dimeric constructs it produced, were demonstrated using three different geminiviruses, namely ageratum yellow vein virus, tomato leaf curl virus, and squash leaf curl virus.
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Begomovirus/genética , Clonagem Molecular/métodos , Genoma Viral , Doenças das Plantas/virologia , Enzimas de Restrição do DNA/metabolismoRESUMO
Replication of genomic DNAs of plant-pathogenic begomoviruses has been demonstrated in prokaryotes, which supported the possibility of analyzing DNA replication process of begomoviruses in bacteria. However, previous studies indicated that the replication of begomovirus DNAs in prokaryotes requires tandem constructs of viral genomes with at least two copies of the origin of replication (ori). In this study, phage M13 vector harboring the unit-length genome with only a single copy of ori of a mono-partite begomovirus, Ageratum yellow vein virus PD isolate (AYVV-[PD]), was constructed and used to investigate the replication of AYVV-[PD] DNAs in Escherichia coli. The generation of single-stranded, circular DNAs (sscDNAs) corresponding to the unit-length AYVV-[PD] genome of both polarity was observed and verified. Replication-associated (Rep) protein of AYVV-[PD] was detected only in bacteria generating the corresponding sscDNAs, whereas disruption of the Rep gene abolished the phenomenon. The results suggested that a single copy of ori is sufficient for the prokaryotes to support the generation of unit-length, genomic sscDNAs of begomoviruses, which requires the presence of functional Rep protein.