- Volume 89, Issue 3, 2008
Volume 89, Issue 3, 2008
- Animal
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- DNA viruses
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The F protein of Helicoverpa armigera single nucleopolyhedrovirus can be substituted functionally with its homologue from Spodoptera exigua multiple nucleopolyhedrovirus
More LessF proteins of group II nucleopolyhedroviruses (NPVs) are envelope fusion proteins essential for virus entry and egress. An F-null Helicoverpa armigera single nucleocapsid NPV (HearNPV) bacmid, HaBacΔF, was constructed. This bacmid could not produce infectious budded virus (BV) when transfected into HzAM1 cells, showing that F protein is essential for cell-to-cell transmission of BVs. When HaBacΔF was pseudotyped with the homologous F protein (HaBacΔF-HaF, positive control) or with the heterologous F protein from Spodoptera exigua multinucleocapsid NPV (SeMNPV) (HaBacΔF-SeF), infectious BVs were produced with similar kinetics. In the late phase of infection, the BV titre of HaBacΔF-SeF virus was about ten times lower than that of HaBacΔF-HaF virus. Both pseudotyped viruses were able to fuse HzAM1 cells in a similar fashion. The F proteins of both HearNPV and SeMNPV were completely cleaved into F1 and F2 in the BVs of vHaBacΔF-HaF and vHaBacΔF-SeF, respectively, but the cleavage of SeF in vHaBacΔF-SeF-infected HzAM1 cells was incomplete, explaining the lower BV titre of vHaBacΔF-SeF. Polyclonal antisera against HaF1 and SeF1 specifically neutralized the infection of vHaBacΔF-HaF and vHaBacΔF-SeF, respectively. HaF1 antiserum showed some cross-neutralization with vHaBacΔF-SeF. These results demonstrate that group II NPV F proteins can be functionally replaced with a homologue of other group II NPVs, suggesting that the interaction of F with other viral or host proteins is not absolutely species-specific.
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- Plant
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Suppression of tobacco mosaic virus-induced hypersensitive-type necrotization in tobacco at high temperature is associated with downregulation of NADPH oxidase and superoxide and stimulation of dehydroascorbate reductase
More LessTissue necroses and resistance during the hypersensitive response (HR) of tobacco to tobacco mosaic virus (TMV) are overcome at temperatures above 28 °C and the virus multiplies to high levels in the originally resistant N-gene expressing plants. We have demonstrated that chemical compounds that generate reactive oxygen species (ROS) or directly applied hydrogen peroxide (H2O2) are able to induce HR-type necroses in TMV-inoculated Xanthi-nc tobacco even at high temperatures (e.g. 30 °C). The amount of superoxide (O2 •−) decreased, while H2O2 slightly increased in TMV- and mock-inoculated leaves at 30 °C, as compared with 20 °C. Activity of NADPH oxidase and mRNA levels of genes that encode NADPH oxidase and an alternative oxidase, respectively, were significantly lower, while activity of dehydroascorbate reductase was significantly higher at 30 °C, as compared with 20 °C. It was possible to reverse or suppress the chemically induced HR-type necrotization at 30 °C by the application of antioxidants, such as superoxide dismutase and catalase, demonstrating that the development of HR-type necroses indeed depends on a certain level of superoxide and other ROS. Importantly, high TMV levels at 30 °C were similar in infected plants, whether the HR-type necrotization developed or not. Suppression of virus multiplication in resistant, HR-producing tobacco at lower temperatures seems to be independent of the appearance of necroses but is associated with temperatures below 28 °C.
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Structure–function relationship between the tobamovirus TMV-Cg coat protein and the HR-like response
The tobamovirus TMV-Cg induces an HR-like response in Nicotiana tabacum cv. Xanthi nn sensitive plants lacking the N or N′ resistance genes. This response has been characterized by the appearance of necrotic lesions in the inoculated leaf and viral systemic spread, although the defence pathways are activated in the plant. A previous study demonstrated that the coat protein (CP) of TMV-Cg (CPCg) was the elicitor of this HR-like response. We examined the influence of four specific amino acid substitutions on the structure of CPCg, as well as on the development of the host response. To gain insights into the structural implications of these substitutions, a set of molecular dynamic experiments was performed using comparative models of wild-type and mutant CPCg as well as the CP of the U1 strain of TMV (CPU1), which is not recognized by the plants. A P21L mutation produces severe changes in the three-dimensional structure of CPCg and is more unstable when this subunit is laterally associated in silico. This result may explain the observed incapacity of this mutant to assemble virions. Two other CPCg mutations (R46G and S54K) overcome recognition by the plant and do not induce an HR-like response. A double CPCg mutant P21L-S54K recovered its capacity to form virions and to induce an HR-like response. Our results suggest that the structural integrity of the CP proteins is important for triggering the HR-like response.
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Supervirulent pseudorecombination and asymmetric synergism between genomic components of two distinct species of begomovirus associated with severe tomato leaf curl disease in India
More LessIsolates of two distinct begomovirus species, the severe strain of the species Tomato leaf curl New Delhi virus (tomato leaf curl New Delhi virus-[India:New Delhi:Severe:1992]; ToLCNDV-[IN:ND:Svr:92], bipartite) and the Varanasi strain of the species Tomato leaf curl Gujarat virus (tomato leaf curl Gujarat virus-[India:Varanasi:2001]; ToLCGV-[IN:Var:01], mono/bipartite) infect tomato (Lycopersicon esculentum) and cause severe yield losses in northern India. This study investigated the infectivity properties of genomic components of these two species. Both pseudorecombinants were infectious in Nicotiana benthamiana, Nicotiana tabacum and L. esculentum. Enhanced pathogenicity was observed when DNA-A of ToLCNDV-[IN:ND:Svr:92] was trans-complemented with ToLCGV-[IN:Var:01] DNA-B, and was consistently associated with an increase in accumulation of ToLCGV-[IN:Var:01] DNA-B. Mixed infection of ToLCNDV-[IN:ND:Svr:92] and ToLCGV-[IN:Var:01] always showed extremely severe symptoms, suggesting a synergistic interaction between these two viruses. Southern blot analysis of viral DNAs from infected plants showed a significantly higher level of accumulation of both ToLCNDV components and DNA-B of ToLCGV-[IN:Var:01] with no alteration to levels of DNA-A of ToLCGV-[IN:Var:01]. Symptom development and/or higher infectivity of the supervirulent pseudorecombinants correlated with the increased levels of DNA-B accumulation. Protoplast replication assays revealed that enhanced infectivity by the pseudorecombinant occurred at the level of replication, as DNA-A of ToLCNDV-[IN:ND:Svr:92] enhanced ToLCGV-[IN:Var:01] DNA-B replication, whose accumulation was in turn increased by ToLCGV-[IN:Var:01] DNA-A. This is the first report demonstrating a virulent pseudorecombinant between two distinct species of begomoviruses that infect tomato, and is the second report on synergism between begomoviruses. The results revealed that ToLCGV-[IN:Var:01] DNA-B is capable of associating with different DNA-A components, despite having different iteron sequences.
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Cylindrical inclusion protein of potato virus A is associated with a subpopulation of particles isolated from infected plants
Potato virus A (PVA) particles were purified by centrifugation through a 30 % sucrose cushion and the pellet (P1) was resuspended and sedimented through a 5–40 % sucrose gradient. The gradient separation resulted in two different virus particle populations: a virus fraction (F) that formed a band in the gradient and one that formed a pellet (P2) at the bottom of the gradient. All three preparations contained infectious particles that retained their integrity when visualized by electron microscopy (EM). Western blotting of the P1 particles revealed that the viral RNA helicase, cylindrical inclusion protein (CI), co-purified with virus particles. This result was confirmed with co-immunoprecipitation experiments. CI was detected in P2 particle preparations, whereas F particles were devoid of detectable amounts of CI. ATPase activity was detected in all three preparations with the greatest amount in P2. Results from immunogold-labelling EM experiments suggested that a fraction of the CI present in the preparations was localized to one end of the virion. Atomic force microscopy (AFM) studies showed that P1 and P2 contained intact particles, some of which had a protruding tip structure at one end, whilst F virions were less stable and mostly appeared as beaded structures under the conditions of AFM. The RNA of the particles in F was translated five to ten times more efficiently than RNA from P2 particles when these preparations were subjected to translation in wheat-germ extracts. The results are discussed in the context of a model for CI-mediated functions.
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- ERRATUM
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Volumes and issues
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Volume 105 (2024)
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Volume 2 (1968)
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Volume 1 (1967)