scholarly journals The P25 Protein of Potato Virus X (PVX) Is the Main Pathogenicity Determinant Responsible for Systemic Necrosis in PVX-Associated Synergisms

2014 ◽  
Vol 89 (4) ◽  
pp. 2090-2103 ◽  
Author(s):  
Emmanuel Aguilar ◽  
David Almendral ◽  
Lucía Allende ◽  
Remedios Pacheco ◽  
Bong Nam Chung ◽  
...  
Keyword(s):  
Potato Virus ◽  
Rna Silencing ◽  
Potato Virus X ◽  
Threshold Level ◽  
Plant Viruses ◽  
Plum Pox Virus ◽  
Systemic Necrosis ◽  
Suppressor Activity ◽  
Content Type ◽  
Reading Frame

ABSTRACTMost plant viruses counter the RNA silencing-based antiviral defense by expressing viral suppressors of RNA silencing (VSRs). In this sense, VSRs may be regarded as virulence effectors that can be recognized by the host as avirulence (avr) factors to induceR-mediated resistance. We made use ofAgrobacterium-mediated transient coexpression of VSRs in combination withPotato virus X(PVX) to recapitulate in local tissues the systemic necrosis (SN) caused by PVX-potyvirus synergistic infections inNicotiana benthamiana. The hypersensitive response (HR)-like response was associated with an enhanced accumulation of PVX subgenomic RNAs. We further show that expression of P25, the VSR of PVX, in the presence of VSR from different viruses elicited an HR-like response inNicotianaspp. Furthermore, the expression of P25 by aPlum pox virus(PPV) vector was sufficient to induce an increase of PPV pathogenicity that led to necrotic mottling. A frameshift mutation in the P25 open reading frame (ORF) of PVX did not lead to necrosis when coexpressed with VSRs. These findings indicate that P25 is the main PVX determinant involved in eliciting a systemic HR-like response in PVX-associated synergisms. Moreover, we show that silencing ofSGT1andRAR1attenuated cell death in both PVX-potyvirus synergistic infection and the HR-like response elicited by P25. Our study underscores that P25 variants that have impaired ability to suppress RNA silencing cannot act as elicitors when synergized by the presence of other VSRs. These findings highlight the importance of RNA silencing suppression activity in the HR-like response elicited by VSRs in certain hosts.IMPORTANCEThe work presented here describes how the activity of the PVX suppressor P25 elicits an HR-like response inNicotianaspp. when overexpressed with other VSR proteins. This finding suggests that the SN response caused by PVX-associated synergisms is a delayed immune response triggered by P25, once it reaches a threshold level by the action of other VSRs. Moreover, this work supports the contention that the silencing suppressor activity of PVX P25 protein is a prerequisite for HR elicitation. We propose that unidentified avr determinants could be involved in other cases of viral synergisms in which heterologous “helper” viruses encoding strong VSRs exacerbate the accumulation of the avr-encoding virus.

scholarly journals Characterization of the Recombinant Forms Arising from a Potato virus X Chimeric Virus Infection under RNA Silencing Pressure

2006 ◽  
Vol 19 (8) ◽  
pp. 904-913 ◽  
Author(s):  
D. Barajas ◽  
F. Tenllado ◽  
J. R. Díaz-Ruíz
Keyword(s):  
Transgenic Plants ◽  
Potato Virus ◽  
Rna Silencing ◽  
Defense Mechanism ◽  
Selective Pressure ◽  
Significant Proportion ◽  
Potato Virus X ◽  
Plum Pox Virus ◽  
Recombination Processes

Recombination is a frequent phenomenon in RNA viruses whose net result is largely influenced by selective pressures. RNA silencing in plants acts as a defense mechanism against viruses and can be used to engineer virus resistance. Here, we have investigated the influence of RNA silencing as a selective pressure to favor recombinants of PVX-HCT, a chimeric Potato virus X (PVX) vector carrying the helper-component proteinase (HC-Pro) gene from Plum pox virus (PPV). All the plants from two lines expressing a silenced HC-Pro transgene were completely resistant to PPV. However a significant proportion became infected with PVX-HCT. Analysis of viral RNAs accumulating in silenced plants revealed that PVX-HCT escaped silencing-based resistance by removal of the HC-Pro sequences that represented preferential targets for transgene-promoted silencing. The virus vector also tended to lose the HC-Pro insert when infecting transgenic plants containing a nonsilenced HC-Pro transgene or wild-type (wt) Nicotiana benthamiana plants. Nevertheless, loss of HC-Pro sequences was faster in nonsilenced transgenic plants than in wt plants, suggesting the transgene plays a role in promoting a higher selective pressure in favor of recombinant virus versions. These results indicate that the outcome of recombination processes depends on the strength of selection pressures applied to the virus.

scholarly journals Inhibition of RNA silencing by the coat protein of Pelargonium flower break virus: distinctions from closely related suppressors

2009 ◽  
Vol 90 (2) ◽  
pp. 519-525 ◽  
Author(s):  
Sandra Martínez-Turiño ◽  
Carmen Hernández
Keyword(s):  
Coat Protein ◽  
Potato Virus ◽  
Rna Silencing ◽  
Potato Virus X ◽  
Plant Viruses ◽  
Small Interfering Rnas ◽  
Silencing Suppression ◽  
Viral Suppressors

Viral-derived double-stranded RNAs (dsRNAs) activate RNA silencing, generating small interfering RNAs (siRNAs) which are incorporated into an RNA-induced silencing complex (RISC) that promotes homology-dependent degradation of cognate RNAs. To counteract this, plant viruses express RNA silencing suppressors. Here, we show that the coat protein (CP) of Pelargonium flower break virus (PFBV), a member of the genus Carmovirus, is able to efficiently inhibit RNA silencing. Interestingly, PFBV CP blocked both sense RNA- and dsRNA-triggered RNA silencing and did not preclude generation of siRNAs, which is in contrast with the abilities that have been reported for other carmoviral CPs. We have also found that PFBV CP can bind siRNAs and that this ability correlates with silencing suppression activity and enhancement of potato virus X pathogenicity. Collectively, the results indicate that PFBV CP inhibits RNA silencing by sequestering siRNAs and preventing their incorporation into a RISC, thus behaving similarly to unrelated viral suppressors but dissimilarly to orthologous ones.

scholarly journals Characterization of Pathogenicity-Associated V2 Protein of Tobacco Curly Shoot Virus

2021 ◽  
Vol 22 (2) ◽  
pp. 923
Author(s):  
Mingjun Li ◽  
Changchang Li ◽  
Kairong Jiang ◽  
Ke Li ◽  
Junlei Zhang ◽  
...  
Keyword(s):  
Rna Silencing ◽  
Viral Infections ◽  
Infectious Clone ◽  
Potato Virus X ◽  
Transient Expression Assay ◽  
Systemic Necrosis ◽  
Suppressor Activity ◽  
Amino Acid Region ◽  
Tobacco Curly Shoot Virus ◽  
Silencing Suppression

V2 proteins encoded by some whitefly-transmitted geminiviruses were reported to be functionally important proteins. However, the functions of the V2 protein of tobacco curly shoot virus (TbCSV), a monopartite begomovirus that causes leaf curl disease on tomato and tobacco in China, remains to be characterized. In our report, an Agrobacterium infiltration-mediated transient expression assay indicated that TbCSV V2 can suppress local and systemic RNA silencing and the deletion analyses demonstrated that the amino acid region 1–92 of V2, including the five predicted α-helices, are required for local RNA silencing suppression. Site-directed substitutions showed that the conserved basic and ring-structured amino acids in TbCSV V2 are critical for its suppressor activity. Potato virus X-mediated heteroexpression of TbCSV V2 in Nicotiana benthamiana induced hypersensitive response-like (HR-like) cell death and systemic necrosis in a manner independent of V2′s suppressor activity. Furthermore, TbCSV infectious clone mutant with untranslated V2 protein (TbCSV∆V2) could not induce visual symptoms, and coinfection with betasatellite (TbCSB) could obviously elevate the viral accumulation and symptom development. Interestingly, symptom recovery occurred at 15 days postinoculation (dpi) and onward in TbCSV∆V2/TbCSB-inoculated plants. The presented work contributes to understanding the RNA silencing suppression activity of TbCSV V2 and extends our knowledge of the multifunctional role of begomovirus-encoded V2 proteins during viral infections.

scholarly journals The cysteine-rich proteins of beet necrotic yellow vein virus and tobacco rattle virus contribute to efficient suppression of silencing in roots

2012 ◽  
Vol 93 (8) ◽  
pp. 1841-1850 ◽  
Author(s):  
Ida Bagus Andika ◽  
Hideki Kondo ◽  
Masamichi Nishiguchi ◽  
Tetsuo Tamada
Keyword(s):  
Mosaic Virus ◽  
Potato Virus ◽  
Rna Silencing ◽  
Tobacco Rattle Virus ◽  
Potato Virus X ◽  
Plant Viruses ◽  
Yellow Vein ◽  
Negative Strand ◽  
Silencing Suppression ◽  
Leaves And Roots

Many plant viruses encode proteins that suppress RNA silencing, but little is known about the activity of silencing suppressors in roots. This study examined differences in the silencing suppression activity of different viruses in leaves and roots of Nicotiana benthamiana plants. Infection by tobacco mosaic virus, potato virus Y and cucumber mosaic virus but not potato virus X (PVX) resulted in strong silencing suppression activity of a transgene in both leaves and roots, whereas infection by beet necrotic yellow vein virus (BNYVV) and tobacco rattle virus (TRV) showed transgene silencing suppression in roots but not in leaves. For most viruses tested, viral negative-strand RNA accumulated at a very low level in roots, compared with considerable levels of positive-strand genomic RNA. Co-inoculation of leaves with PVX and either BNYVV or TRV produced an increase in PVX negative-strand RNA and subgenomic RNA (sgRNA) accumulation in roots. The cysteine-rich proteins (CRPs) BNYVV p14 and TRV 16K showed weak silencing suppression activity in leaves. However, when either of these CRPs was expressed from a PVX vector, there was an enhancement of PVX negative-strand RNA and sgRNA accumulation in roots compared with PVX alone. Such enhancement of PVX sgRNAs was also observed by expression of CRPs of other viruses and the well-known suppressors HC-Pro and p19 but not of the potato mop-top virus p8 CRP. These results indicate that BNYVV- and TRV-encoded CRPs suppress RNA silencing more efficiently in roots than in leaves.

scholarly journals Identification of the Potential Virulence Factors and RNA Silencing Suppressors of Mulberry Mosaic Dwarf-Associated Geminivirus

Viruses ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 472 ◽  
Author(s):  
Xiuling Yang ◽  
Yanxiang Ren ◽  
Shaoshuang Sun ◽  
Dongxue Wang ◽  
Fanfan Zhang ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Rna Silencing ◽  
Fluorescent Protein ◽  
Potato Virus X ◽  
Defense Responses ◽  
Plant Viruses ◽  
Silencing Suppressor ◽  
Long Distance ◽  
Suppressor Activity ◽  
Rna Silencing Suppressor

Plant viruses encode virulence factors or RNA silencing suppressors to reprogram plant cellular processes or to fine-tune host RNA silencing-mediated defense responses. In a previous study, Mulberry mosaic dwarf-associated virus (MMDaV), a novel, highly divergent geminivirus, has been identified from a Chinese mulberry tree showing mosaic and dwarfing symptoms, but the functions of its encoded proteins are unknown. In this study, all seven proteins encoded by MMDaV were screened for potential virulence and RNA silencing suppressor activities. We found that V2, RepA, and Rep affect the pathogenicity of a heterologous potato virus X. We showed that V2 could inhibit local RNA silencing and long-distance movement of the RNA silencing signal, but not short-range spread of the green fluorescent protein (GFP) silencing signal in Nicotiana benthamiana 16c plants. In addition, V2 localized to both subnuclear foci and the cytoplasm. Deletion mutagenesis of V2 showed that the basic motif from amino acids 61 to 76 was crucial for V2 to form subnuclear foci and for suppression of RNA silencing. Although the V2 protein encoded by begomoviruses or a curtovirus has been shown to have silencing suppressor activity, this is the first identification of an RNA silencing suppressor from a woody plant-infecting geminivirus.

scholarly journals Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

2015 ◽  
Vol 81 (9) ◽  
pp. 3039-3048 ◽  
Author(s):  
Stefanie Rettcher ◽  
Felicitas Jungk ◽  
Christoph Kühn ◽  
Hans-Joachim Krause ◽  
Greta Nölke ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Plant Pathogens ◽  
High Sensitivity ◽  
Potato Virus X ◽  
Plant Viruses ◽  
Economic Losses ◽  
Virus Concentration ◽  
Content Type ◽  
Agricultural Industry ◽  
Sum Frequency

ABSTRACTPlant pathogens cause major economic losses in the agricultural industry because late detection delays the implementation of measures that can prevent their dissemination. Sensitive and robust procedures for the rapid detection of plant pathogens are therefore required to reduce yield losses and the use of expensive, environmentally damaging chemicals. Here we describe a simple and portable system for the rapid detection of viral pathogens in infected plants based on immunofiltration, subsequent magnetic detection, and the quantification of magnetically labeled virus particles.Grapevine fanleaf virus(GFLV) was chosen as a model pathogen. Monoclonal antibodies recognizing the GFLV capsid protein were immobilized onto immunofiltration columns, and the same antibodies were linked to magnetic nanoparticles. GFLV was quantified by immunofiltration with magnetic labeling in a double-antibody sandwich configuration. A magnetic frequency mixing technique, in which a two-frequency magnetic excitation field was used to induce a sum frequency signal in the resonant detection coil, corresponding to the virus concentration within the immunofiltration column, was used for high-sensitivity quantification. We were able to measure GFLV concentrations in the range of 6 ng/ml to 20 μg/ml in less than 30 min. The magnetic immunoassay could also be adapted to detect other plant viruses, includingPotato virus XandTobacco mosaic virus, with detection limits of 2 to 60 ng/ml.

scholarly journals Phenotypes and Functional Effects Caused by Various Viral RNA Silencing Suppressors in Transgenic Nicotiana benthamiana and N. tabacum

2008 ◽  
Vol 21 (2) ◽  
pp. 178-187 ◽  
Author(s):  
Shahid Aslam Siddiqui ◽  
Cecilia Sarmiento ◽  
Erkki Truve ◽  
Harry Lehto ◽  
Kirsi Lehto
Keyword(s):  
Mosaic Virus ◽  
Potato Virus ◽  
Nicotiana Benthamiana ◽  
Rna Silencing ◽  
Fluorescent Protein ◽  
Potato Virus X ◽  
Mottle Virus ◽  
African Cassava Mosaic Virus ◽  
Rice Yellow Mottle Virus ◽  
Green Fluorescent

RNA silencing suppressor genes derived from six virus genera were transformed into Nicotiana benthamiana and N. tabacum plants. These suppressors were P1 of Rice yellow mottle virus (RYMV), P1 of Cocksfoot mottle virus, P19 of Tomato bushy stunt virus, P25 of Potato virus X, HcPro of Potato virus Y (strain N), 2b of Cucumber mosaic virus (strain Kin), and AC2 of African cassava mosaic virus (ACMV). HcPro caused the most severe phenotypes in both Nicotiana spp. AC2 also produced severe effects in N. tabacum but a much milder phenotype in N. benthamiana, although both HcPro and AC2 affected the leaf tissues of the two Nicotiana spp. in similar ways, causing hyperplasia and hypoplasia, respectively. P1-RYMV caused high lethality in the N. benthamiana plants but only mild effects in the N. tabacum plants. Phenotypic alterations produced by the other transgenes were minor in both species. Interestingly, the suppressors had very different effects on crucifer-infecting Tobamovirus (crTMV) infections. AC2 enhanced both spread and brightness of the crTMV-green fluorescent protein (GFP) lesions, whereas 2b and both P1 suppressors enhanced spread but not brightness of these lesions. P19 promoted spread of the infection into new foci within the infiltrated leaf, whereas HcPro and P25 suppressed the spread of crTMV-GFP lesions.

scholarly journals The Cucumber vein yellowing virus Silencing Suppressor P1b Can Functionally Replace HCPro in Plum pox virus Infection in a Host-Specific Manner

2012 ◽  
Vol 25 (2) ◽  
pp. 151-164 ◽  
Author(s):  
Alberto Carbonell ◽  
Gabriela Dujovny ◽  
Juan Antonio García ◽  
Adrian Valli
Keyword(s):  
Rna Silencing ◽  
Deleterious Effect ◽  
Prunus Persica ◽  
Plant Viruses ◽  
Natural Host ◽  
Plum Pox Virus ◽  
Silencing Suppressor ◽  
Specific Manner ◽  
Silencing Suppression ◽  
Yellowing Virus

Plant viruses of the genera Potyvirus and Ipomovirus (Potyviridae family) use unrelated RNA silencing suppressors (RSS) to counteract antiviral RNA silencing responses. HCPro is the RSS of Potyvirus spp., and its activity is enhanced by the upstream P1 protein. Distinctively, the ipomovirus Cucumber vein yellowing virus (CVYV) lacks HCPro but contains two P1 copies in tandem (P1aP1b), the second of which functions as RSS. Using chimeras based on the potyvirus Plum pox virus (PPV), we found that P1b can functionally replace HCPro in potyviral infections of Nicotiana plants. Interestingly, P1a, the CVYV protein homologous to potyviral P1, disrupted the silencing suppression activity of P1b and reduced the infection efficiency of PPV in Nicotiana benthamiana. Testing the influence of RSS in host specificity, we found that a P1b-expressing chimera poorly infected PPV's natural host, Prunus persica. Conversely, P1b conferred on PPV chimeras the ability to replicate locally in cucumber, CVYV's natural host. The deleterious effect of P1a on PPV infection is host dependent, because the P1aP1b-expressing PPV chimera accumulated in cucumber to higher levels than PPV expressing P1b alone. These results demonstrate that a potyvirus can use different RSS, and that particular RSS and upstream P1-like proteins contribute to defining the virus host range.

scholarly journals Peptide display on Potato virus X: molecular features of the coat protein-fused peptide affecting cell-to-cell and phloem movement of chimeric virus particles

2006 ◽  
Vol 87 (10) ◽  
pp. 3103-3112 ◽  
Author(s):  
Chiara Lico ◽  
Floriana Capuano ◽  
Giovanni Renzone ◽  
Marcello Donini ◽  
Carla Marusic ◽  
...  
Keyword(s):  
Coat Protein ◽  
Potato Virus ◽  
Potato Virus X ◽  
Structural Features ◽  
Plant Viruses ◽  
Chimeric Virus ◽  
Expression Vectors ◽  
In Planta ◽  
Virus Particles ◽  
Molecular Features

The potexvirus Potato virus X (PVX) can be modified genetically to generate chimeric virus particles (CVPs) carrying heterologous peptides fused to coat protein (CP) subunits. A spontaneous PVX mutant expressing a truncated, but functional, form of the CP has been isolated. With the aim of exploiting this virus to display peptides useful for vaccine formulations, two novel viral expression vectors based on pPVX201 (bearing the wild-type PVX genome) were constructed encoding the truncated CP. Both vectors were able to produce infectious virus particles in planta and were used to insert a panel of sequences encoding peptides of biopharmaceutical interest as N-terminal fusions to the truncated cp gene. The analysis of infection progression induced by the different constructs enabled identification of two important structural features of the fused peptide, namely tryptophan content and isoelectric point, critically affecting the formation of PVX CVPs and virus movement through the plant. These results are discussed in view of the rising interest in engineered plant viruses for development of peptide-based epitope vaccines.

Sign in / Sign up

Export Citation Format

Share Document