scholarly journals Components of Arabidopsis Defense- and Ethylene-Signaling Pathways Regulate Susceptibility to Cauliflower mosaic virus by Restricting Long-Distance Movement

2007 ◽  
Vol 20 (6) ◽  
pp. 659-670 ◽  
Author(s):  
Andrew J. Love ◽  
Valérie Laval ◽  
Chiara Geri ◽  
Janet Laird ◽  
A. Deri Tomos ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Systemic Acquired Resistance ◽  
Fluorescent Protein ◽  
Acquired Resistance ◽  
Systemic Infection ◽  
Cauliflower Mosaic Virus ◽  
Virus Movement ◽  
Wild Type ◽  
Long Distance ◽  
Systemic Spread

We analyzed the susceptibility of Arabidopsis mutants with defects in salicylic acid (SA) and jasmonic acid (JA)/ethylene (ET) signaling to infection by Cauliflower mosaic virus (CaMV). Mutants cpr1-1 and cpr5-2, in which SA-dependent defense signaling is activated constitutively, were substantially more resistant than the wild type to systemic infection, implicating SA signaling in defense against CaMV. However, SA-deficient NahG, sid2-2, eds5-1, and pad4-1 did not show enhanced susceptibility. A cpr5 eds5 double mutant also was resistant, suggesting that resistance in cpr5 may function partially independently of SA. Treatment of cpr5 and cpr5 eds5, but not cpr1, with salicyl-hydroxamic acid, an inhibitor of alternative oxidase, partially restored susceptibility to wild-type levels. Mutants etr1-1, etr1-3, and ein2-1, and two mutants with lesions in ET/JA-mediated defense, eds4 and eds8, also showed reduced virus susceptibility, demonstrating that ET-dependent responses also play a role in susceptibility. We used a green fluorescent protein (GFP)-expressing CaMV recombinant to monitor virus movement. In mutants with reduced susceptibility, cpr1-1, cpr5-2, and etr1-1, CaMV-GFP formed local lesions similar to the wild type, but systemic spread was almost completely absent in cpr1 and cpr5 and was substantially reduced in etr1-1. Thus, mutations with enhanced systemic acquired resistance or compromised ET signaling show diminished long-distance virus movement.

Limitations on the use of fused green fluorescent protein to investigate structure–function relationships for the cauliflower mosaic virus movement protein

Microbiology ◽  
2000 ◽  
Vol 81 (7) ◽  
pp. 1851-1855 ◽  
Author(s):  
Carole L. Thomas ◽  
Andrew J. Maule
Keyword(s):  
Green Fluorescent Protein ◽  
Mosaic Virus ◽  
Fluorescent Protein ◽  
Movement Protein ◽  
Cauliflower Mosaic Virus ◽  
Virus Movement ◽  
Wild Type ◽  
Tubule Formation ◽  
Mouth Disease Virus ◽  
Green Fluorescent

To investigate the process of tubule formation for the cauliflower mosaic virus movement protein (CaMV MP), the green fluorescent protein (GFP) was fused to the MP to provide a vital marker for MP location after expression in insect cells. In contrast to the long tubular structures seen previously following baculovirus-based expression of the wild-type MP, the fusion protein produced only aggregates of fluorescing material in the cytoplasm. However, by co-expressing wild-type MP and GFP–MP, or by engineering their co-accumulation by introducing a foot-and-mouth disease virus 2A cleavage sequence between GFP and MP, long GFP-fluorescing tubules were formed. The experiments suggest that the presence of GFP at the N or C terminus of the tubule-forming domain of the CaMV MP places steric constraints upon the aggregation of the MP into a tubule but that this can be overcome by providing wild-type protein for inclusion in the aggregate.

scholarly journals Requirement of the Movement Protein for Long Distance Spread of Tobacco Mosaic Virus in Grafted Plants

1997 ◽  
Vol 10 (6) ◽  
pp. 691-699 ◽  
Author(s):  
Patricio Arce-Johnson ◽  
Ulrich Reimann-Philipp ◽  
Hal S. Padgett ◽  
Rafael Rivera-Bustamante ◽  
Roger N. Beachy
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Movement Protein ◽  
Systemic Infection ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Long Distance ◽  
Gene Encoding ◽  
Systemic Spread ◽  
Root Stock

Systemic spread of tobacco mosaic virus (TMV) that lacks a functional movement protein (TMVΔMP) was investigated in grafted tobacco (Nicotiana tabacum) plants. Transgenic plants that express the 30-kDa movement protein (MP) gene (MP) under the control of the rolC (phloem-specific) or pal2 (xylem-specific) promoters were unable to support systemic infection by the mutant virus, while plants that express the MP gene from the cauliflower mosaic virus 35S promoter (35S:MP) led to systemic infection. Doubly grafted plants were constructed in which plants containing the 35S:MP gene were used as root stock and plants carrying various MP constructs constituted the middle scion. The upper scion contained the 35S:MP gene in plants that produce a hypersensitive response when systemically infected by TMV. TMVΔMP moved systemically and produced complete necrosis in the upper scion when expression of MP in the middle scion was under the control of the rolC or 35S promoter, but not when the pal2 promoter was used. When plants expressing a gene encoding a defective MP were used as the middle scion, there was no systemic infection by TMVΔMP, and a delay in systemic infection by wild-type TMV. In grafted plants with middle scions that expressed the TMV 54 kDa gene sequence there was no apparent systemic infection by TMVΔMP in the upper scion. The results obtained indicate that the MP has a role in long distance movement, and support the suggestion that replication is necessary for systemic infection of these grafted plants.

scholarly journals Effects of Movement Protein Mutations on the Formation of Tubules in Plant Protoplasts Expressing a Fusion Between the Green Fluorescent Protein and Cauliflower mosaic virus Movement Protein

2001 ◽  
Vol 14 (8) ◽  
pp. 1026-1031 ◽  
Author(s):  
Zhong Huang ◽  
Yu Han ◽  
Stephen H. Howell
Keyword(s):  
Green Fluorescent Protein ◽  
Mosaic Virus ◽  
Fluorescent Protein ◽  
Movement Protein ◽  
Cauliflower Mosaic Virus ◽  
Virus Movement ◽  
Plant Protoplasts ◽  
Tubule Formation ◽  
Leaf Mesophyll ◽  
Green Fluorescent

Fusions between the green fluorescent protein (GFP) and the Cauliflower mosaic virus (CaMV) movement protein (MP) induce the formation of fluorescent foci and surface tubules in Arabidopsis thaliana leaf mesophyll protoplasts. Tubules elongate coordinately and progressively in an assembly process approximately 6 to 12 h following transfection of protoplasts with GFP-MP constructs. Tubules are not formed in protoplasts transfected by GFP-MPER2A, a MP mutation that renders CaMV noninfectious. A small number of short tubules are formed on protoplasts transfected by GFP-MPN6 and GFP-MPN13, two second-site revertants of ER2A that partially restore infectivity. Protoplasts cotransfected with cyan fluorescent protein (CFP)-MPWT and GFP-MPER2A form tubules containing both MP fusions, indicating that although the GFP-MPER2A cannot induce tubule formation, GFP-MPER2A can coassemble or colocalize with CFP-MPWT in tubules. Thus, CaMV MP-induced tubule formation in protoplasts correlates closely with the infectivity of mutation ER2A and its revertants, suggesting that tubule-forming capacity in plant protoplasts reflects a process required for virus infection or movement.

scholarly journals Arabidopsis thaliana FLOWERING LOCUS D Is Required for Systemic Acquired Resistance

2013 ◽  
Vol 26 (9) ◽  
pp. 1079-1088 ◽  
Author(s):  
Vijayata Singh ◽  
Shweta Roy ◽  
Mrunmay Kumar Giri ◽  
Ratnesh Chaturvedi ◽  
Zulkarnain Chowdhury ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Salicylic Acid ◽  
Systemic Acquired Resistance ◽  
Acquired Resistance ◽  
Wild Type ◽  
Long Distance ◽  
Epistasis Analysis ◽  
Flowering Locus ◽  
Systemic Accumulation

Localized infection in plants often induces systemic acquired resistance (SAR), which provides long-term protection against subsequent infections. A signal originating in the SAR-inducing organ is transported to the distal organs, where it stimulates salicylic acid (SA) accumulation and priming, a mechanism that results in more robust activation of defenses in response to subsequent pathogen infection. In recent years, several metabolites that promote long-distance SAR signaling have been identified. However, the mechanism or mechanisms by which plants perceive and respond to the SAR signals are largely obscure. Here, we show that, in Arabidopsis thaliana, the FLOWERING LOCUS D (FLD) is required for responding to the SAR signals leading to the systemic accumulation of SA and enhancement of disease resistance. Although the fld mutant was competent in accumulating the SAR-inducing signal, it was unable to respond to the SAR signal that accumulates in petiole exudates of wild-type leaves inoculated with a SAR-inducing pathogen. Supporting FLD's role in systemic SAR signaling, we observed that dehydroabietinal and azelaic acid, two metabolites that, in wild-type plants, promote SAR-associated systemic accumulation of SA and priming, respectively, were unable to promote SAR in the fld mutant. FLD also participates in flowering, where it functions to repress expression of the flowering repressor FLOWERING LOCUS C (FLC). However, epistasis analysis indicates that FLD's function in SAR is independent of FLC.

scholarly journals Virulence and Differential Local and Systemic Spread of Cucumber mosaic virus in Tobacco are Affected by the CMV 2b Protein

2002 ◽  
Vol 15 (7) ◽  
pp. 647-653 ◽  
Author(s):  
Avril J. Soards ◽  
Alex M. Murphy ◽  
Peter Palukaitis ◽  
John P. Carr
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Laser Scanning ◽  
Fluorescent Protein ◽  
Wild Type Virus ◽  
Virus Spread ◽  
Wild Type ◽  
Mesophyll Cells ◽  
Systemic Spread ◽  
2B Protein

A mutant of the Cucumber mosaic virus subgroup IA strain Fny (Fny-CMV) lacking the gene encoding the 2b protein (Fny-CMVΔ2b) induced a symptomless systemic infection in tobacco. Both the accumulation of Fny-CMVΔ2b in inoculated tissue and the systemic movement of the virus appeared to proceed more slowly than for wild-type Fny-CMV. The influence of the 2b protein on virus movement in the inoculated leaf was examined using viral constructs derived from Fny-CMV and Fny-CMVΔ2b expressing the green fluorescent protein. Laser scanning confocal microscopy was used to visualize the movement of these viruses. Whereas the wild-type virus spread between the epidermal cells as well as the mesophyll cells, the mutant virus spread less efficiently through the epidermal layer and moved preferentially through the mesophyll. Thus, the 2b protein of Fny-CMV influences the dynamics of movement of the virus both within the inoculated leaf and through the whole plant. We propose that this altered movement profile of Fny-CMVΔ2b results in the absence of disease symptoms in tobacco.

scholarly journals Are Tubules Generated by the 3a Protein Necessary for Cucumber Mosaic Virus Movement?

1999 ◽  
Vol 12 (11) ◽  
pp. 985-993 ◽  
Author(s):  
Tomas Canto ◽  
Peter Palukaitis
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Movement Protein ◽  
Virus Movement ◽  
Wild Type ◽  
Gene Encoding ◽  
Green Fluorescent ◽  
Infected Tobacco

The 3a movement protein of cucumber mosaic virus (CMV), fused to the jellyfish green fluorescent protein (3a-GFP) generated surface punctate aggregates as well as tubules protruding from infected tobacco and Nicotiana benthamiana protoplasts. Fluorescent tubules also appeared on the surface of protoplasts prepared from transgenic tobacco plants expressing 3a-GFP, indicating that the 3a protein is the only viral component required for the formation of the tubules. CMV with a mutation in the gene encoding the 3a protein, M8 CMV, could infect tobacco systemically, but tubules were not detected protruding from infected protoplasts when the mutated 3a protein was fused to the GFP [(M8)3a-GFP]. This indicates that the ability of the 3a protein to generate tubules in the surface of protoplasts is not a function required for the spread of CMV in tobacco. On the other hand, the (M8)3a-GFP did not traffic through plasmodesmata interconnecting tobacco epidermal cells, in contrast to the wild-type 3a-GFP. This suggests that there may be a correlation between the ability of the 3a protein to assemble tubules in protoplasts and its ability to promote movement within certain tissues.

scholarly journals Cucumber mosaic virus-Plant Interactions: Identification of 3a Protein Sequences Affecting Infectivity, Cell-to-Cell Movement, and Long-distance Movement

2001 ◽  
Vol 14 (3) ◽  
pp. 378-385 ◽  
Author(s):  
Qiubo Li ◽  
Ki Hyun Ryu ◽  
Peter Palukaitis
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Virus Movement ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Plant Interactions ◽  
Wild Type ◽  
Long Distance ◽  
In Trans ◽  
Long Distance Movement

Mutants of the Cucumber mosaic virus (CMV) movement protein (MP) were generated and analyzed for their effects on virus movement and pathogenicity in vivo. Similar to the wild-type MP, mutants M1, M2, and M3, promoted virus movement in eight plant species. Mutant M3 showed some differences in pathogenicity in one host species. Mutant M8 showed some host-specific alterations in movement in two hypersensitive hosts of CMV. Mutant M9 showed altered pathogenicity on three hosts and was temperature sensitive for long-distance movement, demonstrating that cell-to-cell and long-distance movement are distinct movement functions for CMV. Four mutants (M4, M5, M6, and M7) were debilitated from movement in all hosts tested. Mutants M4, M5, and M6 could be complemented in trans by the wild-type MP expressed transgenically, although not by each other or by mutant M9 (at the restrictive temperature). Mutant M7 showed an inability to be complemented in trans. From these mutants, different aspects of the CMV movement process could be defined and specific roles for particular sequence domains assigned. The broader implications of these functions are discussed.

scholarly journals Asymmetry in Synergistic Interaction Between Wheat streak mosaic virus and Triticum mosaic virus in Wheat

2019 ◽  
Vol 32 (3) ◽  
pp. 336-350 ◽  
Author(s):  
Satyanarayana Tatineni ◽  
Jeff Alexander ◽  
Adarsh K. Gupta ◽  
Roy French
Keyword(s):  
Mosaic Virus ◽  
Great Plains ◽  
Fluorescent Protein ◽  
Systemic Infection ◽  
Wheat Streak Mosaic Virus ◽  
Synergistic Interaction ◽  
Long Distance ◽  
Genomic Rnas ◽  
Long Distance Movement ◽  
Prior Infection

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV), distinct members in the family Potyviridae, are economically important wheat-infecting viruses in the Great Plains region. Previously, we reported that coinfection of wheat by WSMV and TriMV caused disease synergism with increased concentration of both viruses. The mechanisms of synergistic interaction between WSMV and TriMV and the effects of prior infection of wheat by either of these “synergistically interacting partner” (SIP) viruses on the establishment of local and systemic infection by the other SIP virus are not known. In this study, using fluorescent protein-tagged viruses, we found that prior infection of wheat by WSMV or TriMV negatively affected the onset and size of local foci elicited by subsequent SIP virus infection compared with those in buffer-inoculated wheat. These data revealed that prior infection of wheat by an SIP virus has no measurable advantage for another SIP virus on the initiation of infection and cell-to-cell movement. In TriMV-infected wheat, WSMV exhibited accelerated long-distance movement and increased accumulation of genomic RNAs compared with those in buffer-inoculated wheat, indicating that TriMV-encoded proteins complemented WSMV for efficient systemic infection. In contrast, TriMV displayed delayed systemic infection in WSMV-infected wheat, with fewer genomic RNA copies in early stages of infection compared with those in buffer-inoculated wheat. However, during late stages of infection, TriMV accumulation in WSMV-infected wheat increased rapidly with accelerated long-distance movement compared with those in buffer-inoculated wheat. Taken together, these data suggest that interactions between synergistically interacting WSMV and TriMV are asymmetrical; thus, successful establishment of synergistic interaction between unrelated viruses will depend on the order of infection of plants by SIP viruses.

scholarly journals Cauliflower Mosaic Virus Isolate NY8153 Breaks Resistance in Arabidopsis Ecotype En-2

1997 ◽  
Vol 87 (8) ◽  
pp. 792-798 ◽  
Author(s):  
W. Tang ◽  
S. M. Leisner
Keyword(s):  
Arabidopsis Thaliana ◽  
Mosaic Virus ◽  
Virus Isolate ◽  
Systemic Infection ◽  
Cauliflower Mosaic Virus ◽  
Limited Extent ◽  
Long Distance ◽  
Mechanism Of Resistance ◽  
Arabidopsis Ecotype ◽  
Long Distance Movement

Arabidopsis thaliana ecotype En-2 was previously shown to be resistant to cauliflower mosaic caulimovirus (CaMV) isolate CM4-184. In this study, En-2 plants were screened with eight other isolates of CaMV to identify viruses capable of overcoming resistance and to determine if the mechanism of resistance was the same for each virus. En-2 resistance to most CaMV isolates was mediated by the same mechanism, i.e., preventing virus long-distance movement. One CaMV isolate, NY8153, was found that produced a severe systemic infection on En-2 plants. In addition, the CM1841 isolate was able to spread systemically through En-2 plants, to a limited extent, without producing visible symptoms. These data indicate that the resistance shown by En-2 plants is not an all-or-none phenomenon. En-2 plants were susceptible to turnip mosaic potyvirus, suggesting that resistance is specific to CaMV.

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