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Phytohormone signalling pathways have an important role in defence against pathogens mediated by cell-surface pattern recognition receptors and intracellular nucleotide-binding leucine-rich repeat class immune receptors1,2 (NLR). Pathogens have evolved counter-defence strategies to manipulate phytohormone signalling pathways to dampen immunity and promote virulence3. However, little is known about the surveillance of pathogen interference of phytohormone signalling by the plant innate immune system. The pepper (Capsicum chinense) NLR Tsw, which recognizes the effector nonstructural protein NSs encoded by tomato spotted wilt orthotospovirus (TSWV), contains an unusually large leucine-rich repeat (LRR) domain. Structural modelling predicts similarity between the LRR domain of Tsw and those of the jasmonic acid receptor COI1, the auxin receptor TIR1 and the strigolactone receptor partner MAX2. This suggested that NSs could directly target hormone receptor signalling to promote infection, and that Tsw has evolved a LRR resembling those of phytohormone receptors LRR to induce immunity. Here we show that NSs associates with COI1, TIR1 and MAX2 through a common repressor-TCP21-which interacts directly with these phytohormone receptors. NSs enhances the interaction of COI1, TIR1 or MAX2 with TCP21 and blocks the degradation of corresponding transcriptional repressors to disable phytohormone-mediated host immunity to the virus. Tsw also interacts directly with TCP21 and this interaction is enhanced by viral NSs. Downregulation of TCP21 compromised Tsw-mediated defence against TSWV. Together, our findings reveal that a pathogen effector targets TCP21 to inhibit phytohormone receptor function, promoting virulence, and a plant NLR protein has evolved to recognize this interference as a counter-virulence strategy, thereby activating immunity.
Assuntos
Capsicum , Doenças das Plantas , Reguladores de Crescimento de Plantas , Imunidade Vegetal , Proteínas de Plantas , Receptores de Reconhecimento de Padrão , Leucina , Doenças das Plantas/imunologia , Doenças das Plantas/virologia , Reguladores de Crescimento de Plantas/metabolismo , Imunidade Vegetal/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo , Receptores de Reconhecimento de Padrão/química , Receptores de Reconhecimento de Padrão/imunologia , Receptores de Reconhecimento de Padrão/metabolismo , Reconhecimento da Imunidade Inata , Capsicum/imunologia , Capsicum/metabolismo , Capsicum/virologia , VirulênciaRESUMO
Orthotospoviruses, the plant-infecting bunyaviruses, cause serious diseases in agronomic crops and pose major threats to global food security. The family of Tospoviridae contains more than 30 members that are classified into two geographic groups, American-type and Euro/Asian-type orthotospovirus. However, the genetic interaction between different species and the possibility, during mixed infections, for transcomplementation of gene functions by orthotospoviruses from different geographic groups remains underexplored. In this study, minireplicon-based reverse genetics (RG) systems have been established for Impatiens necrotic spot virus (INSV) (an American-type orthotospovirus) and for Calla lily chlorotic spot virus and Tomato zonate spot virus (CCSV and TZSV) (two representative Euro/Asian orthotospoviruses). Together with the earlier established RG system for Tomato spotted wilt virus (TSWV), a type species of the Orthotospovirus American-clade, viral replicase/movement proteins were exchanged and analyzed on interspecies transcomplementation. Whereas the homologous RNA-dependent RNA polymerase (RdRp) and nucleocapsid (N) protein supported the replication of orthotospoviruses from both geographic groups, heterologous combinations of RdRp from one group and N from the other group were unable to support the replication of viruses from both groups. Furthermore, the NSm movement protein (MP), from both geographic groups of orthotospoviruses, was able to transcomplement heterologous orthotospoviruses or a positive-strand Cucumber mosaic virus (CMV) in their movement, albeit with varying efficiency. MP from Rice stripe tenuivirus (RSV), a plant-infecting bunyavirus that is distinct from orthotospoviruses, or MP from CMV also moves orthotospoviruses. Our findings gain insights into the genetic interaction/reassortant potentials for the segmented plant orthotospoviruses. IMPORTANCE Orthotospoviruses are agriculturally important negative-strand RNA viruses and cause severe yield-losses on many crops worldwide. Whereas the emergence of new animal-infecting bunyaviruses is frequently associated with genetic reassortants, this issue remains underexposed with the plant-infecting orthotospovirus. With the development of reverse genetics systems for orthotospoviruses from different geographic regions, the interspecies/intergroup replication/movement complementation between American- and Euro/Asian-type orthotospoviruses were investigated. Genomic RNAs from American orthotospoviruses can be replicated by the RdRp and N from those of Euro/Asia-group orthotospoviruses, and vice versa. However, their genomic RNAs cannot be replicated by a heterologous combination of RdRp from one geographic group and N from another geographic group. Cell-to-cell movement of viral entity is supported by NSm from both geographic groups, with highest efficiency by NSm from viruses belonging to the same group. Our findings provide important insights into the genetic interaction and exchange ability of viral gene functions between different species of orthotospovirus.
Assuntos
Genética Reversa , Tospovirus , Replicação Viral , Animais , Genética Reversa/métodos , RNA Polimerase Dependente de RNA , Tospovirus/genética , Estados Unidos , Replicação Viral/genética , RNA Viral/genética , Proteínas do Nucleocapsídeo/genéticaRESUMO
Many persistent transmitted plant viruses, including rice stripe virus (RSV), cause serious damage to crop production worldwide. Although many reports have indicated that a successful insect-mediated virus transmission depends on a proper interaction between the virus and its insect vector, the mechanism(s) controlling this interaction remained poorly understood. In this study, we used RSV and its small brown planthopper (SBPH) vector as a working model to elucidate the molecular mechanisms underlying the entrance of RSV virions into SBPH midgut cells for virus circulative and propagative transmission. We have determined that this non-enveloped tenuivirus uses its non-structural glycoprotein NSvc2 as a helper component to overcome the midgut barrier(s) for RSV replication and transmission. In the absence of this glycoprotein, purified RSV virions were unable to enter SBPH midgut cells. In the RSV-infected cells, this glycoprotein was processed into two mature proteins: an amino-terminal protein (NSvc2-N) and a carboxyl-terminal protein (NSvc2-C). Both NSvc2-N and NSvc2-C interact with RSV virions. Our results showed that the NSvc2-N could bind directly to the surface of midgut lumen via its N-glycosylation sites. Upon recognition, the midgut cells underwent endocytosis followed by compartmentalization of RSV virions and NSvc2 into early and then late endosomes. The NSvc2-C triggered cell membrane fusion via its highly conserved fusion loop motifs under the acidic condition inside the late endosomes, leading to the release of RSV virions from endosomes into cytosol. In summary, our results showed for the first time that a rice tenuivirus utilized its glycoprotein NSvc2 as a helper component to ensure a proper interaction between its virions and SBPH midgut cells for its circulative and propagative transmission.
Assuntos
Glicoproteínas/fisiologia , Hemípteros/genética , Tenuivirus/metabolismo , Animais , Sistema Digestório/metabolismo , Sistema Digestório/virologia , Glicoproteínas/metabolismo , Insetos Vetores/metabolismo , Insetos Vetores/virologia , Insetos , Doenças das Plantas/virologia , Tenuivirus/patogenicidade , Vírion , Replicação Viral/fisiologiaRESUMO
Plant intracellular nucleotide-binding leucine-rich repeat (NLR) receptors play critical roles in mediating host immunity to pathogen attack. We use tomato Sw-5b::tospovirus as a model system to study the specific role of the compartmentalized plant NLR in dictating host defenses against the virus at different infection steps. We demonstrated here that tomato NLR Sw-5b distributes to the cytoplasm and nucleus, respectively, to play different roles in inducing host resistances against tomato spotted wilt orthotospovirus (TSWV) infection. The cytoplasmic-enriched Sw-5b induces a strong cell death response to inhibit TSWV replication. This host response is, however, insufficient to block viral intercellular and long-distance movement. The nuclear-enriched Sw-5b triggers a host defense that weakly inhibits viral replication but strongly impedes virus intercellular and systemic movement. Furthermore, the cytoplasmic and nuclear Sw-5b act synergistically to dictate a full host defense of TSWV infection. We further demonstrated that the extended N-terminal Solanaceae domain (SD) of Sw-5b plays critical roles in cytoplasm/nucleus partitioning. Sw-5b NLR controls its cytoplasm localization. Strikingly, the SD but not coil-coil domain is crucial for Sw-5b receptor to import into the nucleus to trigger the immunity. The SD was found to interact with importins. Silencing both importin α and ß expression disrupted Sw-5b nucleus import and host immunity against TSWV systemic infection. Collectively, our findings suggest that Sw-5b bifurcates disease resistances by cytoplasm/nucleus partitioning to block different infection steps of TSWV. The findings also identified a new regulatory role of extra domain of a plant NLR in mediating host innate immunity.
Assuntos
Solanum lycopersicum , Tospovirus , Núcleo Celular , Resistência à Doença , Doenças das Plantas , Domínios ProteicosRESUMO
Large hysteresis and low energy density of pure AgNbO3 ceramics limit their further application in pulsed power techniques. Here, less-harmful Sm2O3-modified AgNbO3 antiferroelectric ceramics were synthesized by a rolling process, in order to improve the energy storage performance. All the Sm2O3-doped samples display reduced hysteresis due to disrupted long-range order, as certified by Raman spectra. As expected, due to the large dielectric breakdown strength and reduced hysteresis, an improved energy storage density, Wrec, of 5.85 J cm-3 and satisfactory energy efficiency, η, of 77% can be simultaneously achieved in the studied antiferroelectric ceramics, showing great superiority over other bulk electronic ceramics. Along with excellent temperature stability within the range of 20-150 °C at 320 kV cm-1 (Wrec > 4.3 J cm-3, with minimal variation of ≤4%) in the studied samples, this shows that AgNbO3-based environmentally friendly ceramics are promising materials for pulsed power techniques.
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Circulating tumor cells (CTCs) released from the primary tumor to peripheral blood are promising targets for liquid biopsies. Their biological information is vital for early cancer detection, efficacy assessment, and prognostic monitoring. Despite the tremendous clinical applications of CTCs, development of effective separation techniques are still demanding. Traditional separation methods usually use batch processing for enrichment, which inevitably destroy cell integrity and affect the complete information acquisition. Considering the rarity and heterogeneity of CTCs, it is urgent to develop effective separation methods. Microfluidic chips with precise fluid control at the micron level are promising devices for CTC separation. Their further combination with micro-/nanostructure arrays adds more biomolecule binding sites and exhibit unique fluid barrier effect, which significantly improve the CTC capture efficiency, purity, and sensitivity. This review summarized the recent advances in micro-/nanostructure array integrated microfluidic devices for CTC separation, including microrods, nanowires, and 3D micro-/nanostructures. The mechanisms by which these structures contribute to improved capture efficiency are discussed. Two major categories of separation methods, based on the physical and biological properties of CTCs, are discussed separately. Physical separation includes the design and preparation of micro-/nanostructure arrays, while chemical separation additionally involves the selection and modification of specific capture probes. These emerging technologies are expected to become powerful tools for disease diagnosis in the future.
RESUMO
Piezoelectric energy harvesting technology using the piezoelectric circular diaphragm (PCD) has drawn much attention because it has great application potential in replacing chemical batteries to power microelectronic devices. In this article, we have found a non-uniform strain distribution inside the PCD energy harvester. From the edge to the center of the ceramic disk, its output voltage first increases and then decreases. This uneven output voltage reduces the output power of the PCD energy harvester. Based on this phenomenon, we reduce the ceramic disk diameter and dig a hole in the center, analyzing the effect of removing the ceramic disk's low output voltage part on the PCD energy harvester. The experimental results show that removing the ceramic disk's low output voltage part can improve the output power, reduce the resonance frequency, and increase the optimal impedance of the PCD energy harvester. Under the conditions of 10 g proof mass, 9.8 m/s2 acceleration, the PCD energy harvester with a 19-mm diameter and a 6-mm hole can reach a maximum output power of 8.34 mW.
RESUMO
Fluorinated polyethylene propylene (FEP) bipolar ferroelectret films with a specifically designed concentric tunnel structure were prepared by means of rigid-template based thermoplastic molding and contact polarization. The properties of the fabricated films, including the piezoelectric response, mechanical property, and thermal stability, were characterized, and two kinds of energy harvesters based on such ferroelectret films, working in 33- and 31-modes respectively, were investigated. The results show that the FEP films exhibit significant longitudinal and radial piezoelectric activities, as well as superior thermal stability. A quasi-static piezoelectric d33 coefficient of up to 5300 pC/N was achieved for the FEP films, and a radial piezoelectric sensitivity of 40,000 pC/N was obtained in a circular film sample with a diameter of 30 mm. Such films were thermally stable at 120 °C after a reduction of 35%. Two types of vibrational energy harvesters working in 33-mode and 31-mode were subsequently designed. The results show that a power output of up to 1 mW was achieved in an energy harvester working in 33-mode at a resonance frequency of 210 Hz, referring to a seismic mass of 33.4 g and an acceleration of 1 g (g is the gravity of the earth). For a device working in 31-mode, a power output of 15 µW was obtained at a relatively low resonance frequency of 26 Hz and a light seismic mass of 1.9 g. Therefore, such concentric tunnel FEP ferroelectric films provide flexible options for designing vibrational energy harvesters working either in 33-mode or 31-mode to adapt to application environments.
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Tospovirus is a tripartite negative stranded RNA virus and is considered as one of the most devastating plant viruses. Successful virus infection in plant requires many host factors. To date, very few host factors have been identified as important in Tospovirus infection in plants. We reported earlier that NSm protein encoded by Tomato spotted wilt virus (TSWV), a type species of the genus Orthotospovirus, plays critical roles in viral cell-to-cell and long-distance movement. In this study, we determined that molecular co-chaperone NbSGT1 interacted with TSWV NSm in Nicotianabenthamiana. TSWV infection significantly upregulated the expression of NbSGT1 gene and transient overexpression of NbSGT1 in N.benthamiana leaves accelerated TSWV infection. In contrast, silencing the NbSGT1 gene expression using a virus-induced gene silencing (VIGS) approach strongly inhibited TSWV NSm cell-to-cell movement, as well as TSWV local and systemic infection in N.benthamiana plants. Furthermore, NbSGT1 was found to regulate the infection of both American and Euro/Asia type tospoviruses in N.benthamiana plant. Collectively, our findings presented in this paper and the results published previously indicated that molecular co-chaperone NbSGT1 plays important roles in modulating both positive stranded and tripartite negative stranded RNA virus infection in plants.
Assuntos
Glucosiltransferases/metabolismo , Interações Hospedeiro-Patógeno , Chaperonas Moleculares/metabolismo , Nicotiana/enzimologia , Tospovirus/fisiologia , Internalização do Vírus , Liberação de Vírus , Proteínas do Movimento Viral em Plantas/metabolismo , Ligação Proteica , Nicotiana/virologiaRESUMO
This paper presents a method for harvesting electric energy from mechanical vibration using a mechanically excited piezoelectric circular membrane array. The piezoelectric circular diaphragm array consists of four plates with series and parallel connection, and the electrical characteristics of the array are examined under dynamic conditions. With an optimal load resistor of 160 kΩ, an output power of 28 mW was generated from the array in series connection at 150 Hz under a prestress of 0.8 N and a vibration acceleration of 9.8 m/s(2), whereas a maximal output power of 27 mW can be obtained from the array in parallel connection through a resistive load of 11 kΩ under the same frequency, prestress, and acceleration conditions. The results show that using a piezoelectric circular diaphragm array can significantly increase the output of energy compared with the use of a single plate. By choosing an appropriate connection pattern (series or parallel connections) among the plates, the equivalent impedance of the energy harvesting devices can be tailored to meet the matched load of different applications for maximal power output.