scholarly journals Production of plum pox virus HC-Pro functionally active for aphid transmission in a transient-expression system

2006 ◽  
Vol 87 (11) ◽  
pp. 3413-3423 ◽  
Author(s):  
Elisa Goytia ◽  
Lourdes Fernández-Calvino ◽  
Belén Martínez-García ◽  
Dionisio López-Abella ◽  
Juan José López-Moya
Keyword(s):  
Expression System ◽  
Aphid Transmission ◽  
Plum Pox Virus ◽  
Infection Cycle ◽  
Feeding Assay ◽  
Multifunctional Protein ◽  
Virus Particles ◽  
Accessory Factor ◽  
Aphid Vectors ◽  
Transient System

Potyviruses are non-persistently transmitted by aphid vectors with the assistance of a viral accessory factor known as helper component (HC-Pro), a multifunctional protein that is also involved in many other essential processes during the virus infection cycle. A transient Agrobacterium-mediated expression system was used to produce Plum pox virus (PPV) HC-Pro in Nicotiana benthamiana leaves from constructs that incorporated the 5′ region of the genome, yielding high levels of HC-Pro in agroinfiltrated leaves. The expressed PPV HC-Pro was able to assist aphid transmission of purified virus particles in a sequential feeding assay, and to complement transmission-defective variants of the virus. Also, HC-Pro of a second potyvirus, Tobacco etch virus (TEV), was expressed and found to be functional for aphid transmission. These results show that this transient system can be useful for production of functionally active HC-Pro in potyviruses, and the possible uses of this approach to study the mechanism of transmission are discussed.

scholarly journals Small Bottleneck Size in a Highly Multipartite Virus during a Complete Infection Cycle

2018 ◽  
Vol 92 (14) ◽  
Author(s):  
Romain Gallet ◽  
Frédéric Fabre ◽  
Gaël Thébaud ◽  
Mircea T. Sofonea ◽  
Anne Sicard ◽  
...  
Keyword(s):  
Genomic Structure ◽  
Aphid Transmission ◽  
Infection Cycle ◽  
Experimental Conditions ◽  
Entire Genome ◽  
Content Type ◽  
Viral Particles ◽  
Theoretical Predictions ◽  
Aphid Vectors ◽  
Very High

ABSTRACT Multipartite viruses package their genomic segments independently and thus incur the risk of being unable to transmit their entire genome during host-to-host transmission if they undergo severe bottlenecks. In this paper, we estimated the bottleneck size during one infection cycle of Faba bean necrotic stunt virus (FBNSV), an octopartite nanovirus whose segments have been previously shown to converge to particular and unequal relative frequencies within host plants and aphid vectors. Two methods were used to derive this estimate, one based on the probability of transmission of the virus and the other based on the temporal evolution of the relative frequency of markers for two genomic segments, one frequent and one rare (segment N and S, respectively), both in plants and vectors. Our results show that FBNSV undergoes severe bottlenecks during aphid transmission. Further, even though the bottlenecks are always narrow under our experimental conditions, they slightly widen with the number of transmitting aphids. In particular, when several aphids are used for transmission, the bottleneck size of the segments is also affected by within-plant processes and, importantly, significantly differs across segments. These results indicate that genetic drift not only must be an important process affecting the evolution of these viruses but also that these effects vary across genomic segments and, thus, across viral genes, a rather unique and intriguing situation. We further discuss the potential consequences of our findings for the transmission of multipartite viruses. IMPORTANCE Multipartite viruses package their genomic segments in independent capsids. The most obvious cost of such genomic structure is the risk of losing at least one segment during host-to-host transmission. A theoretical study has shown that for nanoviruses, composed of 6 to 8 segments, hundreds of copies of each segment need to be transmitted to ensure that at least one copy of each segment was present in the host. These estimations seem to be very high compared to the size of the bottlenecks measured with other viruses. Here, we estimated the bottleneck size during one infection cycle of FBNSV, an octopartite nanovirus. We show that these bottlenecks are always narrow (few viral particles) and slightly widen with the number of transmitting aphids. These results contrast with theoretical predictions and illustrate the fact that a new conceptual framework is probably needed to understand the transmission of highly multipartite viruses.

scholarly journals Aphid Transmission of Potyvirus: The Largest Plant-Infecting RNA Virus Genus

Viruses ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 773
Author(s):  
Kiran R. Gadhave ◽  
Saurabh Gautam ◽  
David A. Rasmussen ◽  
Rajagopalbabu Srinivasan
Keyword(s):  
Rna Virus ◽  
Management Strategies ◽  
Aphid Transmission ◽  
Species Variation ◽  
Plant Biochemistry ◽  
Potyvirus Species ◽  
Wide Range ◽  
Aphid Vectors ◽  
Global Agriculture ◽  
Aphid Behavior

Potyviruses are the largest group of plant infecting RNA viruses that cause significant losses in a wide range of crops across the globe. The majority of viruses in the genus Potyvirus are transmitted by aphids in a non-persistent, non-circulative manner and have been extensively studied vis-à-vis their structure, taxonomy, evolution, diagnosis, transmission, and molecular interactions with hosts. This comprehensive review exclusively discusses potyviruses and their transmission by aphid vectors, specifically in the light of several virus, aphid and plant factors, and how their interplay influences potyviral binding in aphids, aphid behavior and fitness, host plant biochemistry, virus epidemics, and transmission bottlenecks. We present the heatmap of the global distribution of potyvirus species, variation in the potyviral coat protein gene, and top aphid vectors of potyviruses. Lastly, we examine how the fundamental understanding of these multi-partite interactions through multi-omics approaches is already contributing to, and can have future implications for, devising effective and sustainable management strategies against aphid-transmitted potyviruses to global agriculture.

scholarly journals Interaction between potyvirus helper component-proteinase and capsid protein in infected plants

2002 ◽  
Vol 83 (7) ◽  
pp. 1765-1770 ◽  
Author(s):  
Geneviève Roudet-Tavert ◽  
Sylvie German-Retana ◽  
Thierry Delaunay ◽  
Brigitte Delécolle ◽  
Thierry Candresse ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Aphid Transmission ◽  
Plum Pox Virus ◽  
Viral Coat Protein ◽  
Helper Component ◽  
Transmission Electron ◽  
Sandwich Elisa Assay ◽  
Viral Coat ◽  
Helper Component Proteinase

Monoclonal antibodies were raised against helper component-proteinase (HcPro) purified from plants infected with the potyvirus Lettuce mosaic virus (LMV). These antibodies were used in a two-site triple antibody sandwich ELISA assay together with polyclonal antibodies directed against purified virions. An interaction between HcPro and the viral coat protein (CP) was demonstrated in extracts of LMV-infected leaves, as well as for two other potyviruses, Plum pox virus and Potato virus Y. The CP–HcPro interaction was not abolished in LMV derivatives with an HcPro GFP N-terminal fusion, or with a deletion from the CP of the amino acids involved in aphid transmission. Electron microscopy indicated that HcPro probably does not interact with the CP in the form of assembled virions or virus-like particles. Together, these results suggest that the interaction detected between CP and HcPro might be involved in a process of the potyvirus cycle different from aphid transmission.

scholarly journals Electron microscopy of structures present in embryonic cells of plants infected with Plum pox virus

2008 ◽  
Vol 44 (No. 3) ◽  
pp. 81-84
Author(s):  
J. Polák ◽  
M. Jokeš ◽  
M. Ducháčová ◽  
A. Hauptmanová ◽  
P. Komínek
Keyword(s):  
Electron Microscopy ◽  
Plum Pox Virus ◽  
Embryonic Cells ◽  
Virus Particles ◽  
Ultrathin Sections ◽  
Peach Trees

Electron microscopy was used to detect the presence of virus particles or inclusions in growth tips and parenchymatic cells of leaves of plum, apricot and peach trees artificially infected with <I>Plum pox virus</I> (PPV). Typical pinwheels were found in ultrathin sections of leaves of PPV infected plums, apricots and peaches. Filamentous particles or their aggregates approximately 750 nm long were found in ultrathin sections of growth tips of plum, apricot, and peach shoots with a diameter of 0.5 mm. Pinwheels were never present in embryonic cells. No virus particles were found in ultrathin sections of growth tips of PPV infected plum, apricot and peach shoots of 0.2 mm in diameter. Embryonic cells of growth tips up to 0.2 mm in diameter are PPV free. PPV particles are present in growth tips at a distance 0.2–0.5 mm from the top; the virus is probably multiplied in this part of the growth tips.

scholarly journals Cauliflower mosaic virus is preferentially acquired from the phloem by its aphid vectors

2002 ◽  
Vol 83 (12) ◽  
pp. 3163-3171 ◽  
Author(s):  
Itziar Palacios ◽  
Martin Drucker ◽  
Stéphane Blanc ◽  
Silvia Leite ◽  
Aranzazu Moreno ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Electronic Monitoring ◽  
Total Duration ◽  
Aphid Transmission ◽  
Cauliflower Mosaic Virus ◽  
Transmission Factor ◽  
Stepwise Logistic Regression ◽  
Mesophyll Cells ◽  
Aphid Vectors ◽  
Stepwise Logistic Regression Model

Cauliflower mosaic virus (CaMV) is transmitted in a non-circulative manner by aphids following the helper strategy. Helper proteins P2 and P3 act as a bridge between virions and the aphid cuticle. Electronic monitoring of aphid stylet activities (EPG technique), transmission tests and electron microscopy showed that CaMV is preferentially acquired from the phloem by its most common aphid vectors, Brevycorine brassicae and Myzus persicae. We also found that CaMV is semipersistently transmitted and that the rate of acquisition does not follow a typical bimodal curve. Instead, the virus could be acquired from non-phloem tissues at a low and fairly constant rate after one or more intracellular punctures within a few minutes, but the probability of acquisition rose significantly when aphids reached the phase of committed ingestion from the phloem. The acquisition rate of CaMV did not increase with increasing number of intracellular punctures, but the total duration of intracellular puncture was one of the variables selected by the stepwise logistic regression model used to fit the data that best explained acquisition of CaMV. Furthermore, aphids reaching the phloem faster had a higher probability of acquiring the virus. Our results support the hypothesis that multiple intracellular punctures of epidermal and mesophyll cells result in loading aphids with the CaMV-encoded aphid transmission factor (P2), and that aphids, in most cases, subsequently acquire CaMV particles during phloem sap ingestion. Consistently, immunoelectron microscopy showed that P3–virions are frequently found in the sieve element lumen, whereas P2 could not be detected.

scholarly journals Prunus Host Range of Plum pox virus (PPV) in the United States by Aphid and Graft Inoculation

Plant Disease ◽  
2007 ◽  
Vol 91 (1) ◽  
pp. 18-23 ◽  
Author(s):  
V. D. Damsteegt ◽  
R. Scorza ◽  
A. L. Stone ◽  
W. L. Schneider ◽  
K. Webb ◽  
...  
Keyword(s):  
Virus Disease ◽  
Systemic Infection ◽  
Sour Cherry ◽  
Aphid Transmission ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Prunus Species ◽  
Wide Range

Plum pox (Sharka) is a serious virus disease of stone fruits caused by the Plum pox virus (PPV). To determine which species could function as potential hosts and virus reservoirs, we used aphid transmission and bud or chip grafting to evaluate the susceptibility of commercial, ornamental, and wild Prunus species to isolates of PPV found in Pennsylvania, USA. Following inoculation, test trees were observed for symptoms, analyzed by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR), back-assayed to healthy peach, and followed through at least four cold-induced dormancy (CID) cycles over 4 years. Thirty-one of 33 Prunus species and cultivars were systemically infected following aphid transmission. Systemic infection could not be detected in P. cerasus (sour cherry) and P. × ‘Snofozam’ (Snow Fountains) despite repeated aphid inoculation attempts. Following grafting of PPV-infected budwood, all 40 species and varieties became infected, although species differed in their susceptibility. Within most species, some individual plants remained PPV negative throughout the study despite repeated inoculations. Infection in some species could be detected only through quantitative reverse transcription (RT)-PCR. Most species displayed clear symptoms, were highly positive by ELISA and RT-PCR, and could be back-inoculated into peach seedlings following CID. Our results indicate that a wide range of native and ornamental Prunus species are susceptible to U.S. isolates of PPV-D.

scholarly journals First Report of Occurrence of Papaya ring spot virus Infecting Papaya in Bangladesh

Plant Disease ◽  
2004 ◽  
Vol 88 (2) ◽  
pp. 221-221 ◽  
Author(s):  
R. K. Jain ◽  
K. M. Nasiruddin ◽  
Jyoti Sharma ◽  
R. P. Pant ◽  
A. Varma
Keyword(s):  
Polyclonal Antibodies ◽  
Electron Microscopic Examination ◽  
Aphid Transmission ◽  
Electron Microscopic ◽  
Spot Virus ◽  
First Report ◽  
Virus Particles ◽  
Cp Gene ◽  
Papaya Ring Spot Virus ◽  
Ring Spot

Papaya (Carica papaya L.) is an important fruit crop in Bangladesh. During surveys conducted in Dhaka and Mymensingh regions from April to June 2003, >50% of papaya plants were observed to have leaf mottling, mosaic and mild distortion, and water-soaked streaks on petioles and stem, which are typical symptoms of Papaya ring spot virus (PRSV) infection. Electron-microscopic examination of negatively stained leaf-dip preparations from 10 symptomatic samples revealed the association of flexuous virus particles that were decorated with polyclonal antibodies raised to an isolate from India (PRSV-D). The identity of PRSV associated with the papaya disease in Bangladesh was further confirmed by reverse transcription polymerase chain reaction and sequence analysis (2). By using PRSV specific primers (2), the 3′-terminal region comprising a part of the nuclear inclusion b gene, the coat protein (CP) gene, and the untranslated region were amplified and sequenced (GenBank Accession No. AY423557). The CP gene consisted of 286 amino acids and the conserved regions common to the genus Potyvirus, such as WCIEN and QMKAA, were present. Like all known PRSV sequences (1), a stretch of glutamic acid and lysine repeats (EK region) after the aphid transmission motif (DAG) also was present. Comparative CP amino acid sequence analyses revealed that the virus infecting papaya in Bangladesh, designated as PRSV-Bd, shared 89 to 92% identity with PRSV isolates from India and 88 to 93% identity with isolates from other parts of the world. To our knowledge, this is the first report of occurrence of PRSV infecting papaya in Bangladesh. References: (1) M. F. Bateson et al. J. Gen. Virol. 83:2575, 2002. (2) R. K. Jain et al. Ann Appl. Biol. 132:413, 1998.

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