H. Lecoq

The nucleotide sequence of Watermelon mosaic virus (WMV, Potyvirus) reveals interspecific recombination between two related potyviruses in the 5' part of the genome

Summary.Watermelon mosaic virus (WMV, Potyvirus) is a potyvirus with a worldwide distribution, mostly in temperate and mediterranean regions. According to the partial sequences that were available, WMV appeared to share high sequence similarity with Soybean mosaic virus (SMV), and it was almost considered as a strain of SMV in spite of its different and much broader host range. Like SMV, it was also related to legume-infecting potyviruses belonging to the “Bean common mosaic virus (BCMV) subgroup”. In this paper we obtained the full-length sequence of WMV, and we confirmed that this virus is very closely related to SMV in most of its genome; however, there is evidence for an interspecific recombination in the P1 protein, as the P1 of WMV was 135 amino-acids longer than that of SMV, and the N-terminal half of the P1 showed no relation to SMV but was 85% identical to BCMV. This suggests that WMV has emerged through an ancestral recombination event, and supports the distinction of WMV and SMV as separate taxonomic units.

Pepper mild mottle virus, a plant virus associated with specific immune responses, Fever, abdominal pains, and pruritus in humans.

Background Recently, metagenomic studies have identified viable Pepper mild mottle virus (PMMoV), a plant virus, in the stool of healthy subjects. However, its source and role as pathogen have not been determined. Methods and Findings 21 commercialized food products containing peppers, 357 stool samples from 304 adults and 208 stool samples from 137 children were tested for PMMoV using real-time PCR, sequencing, and electron microscopy. Anti-PMMoV IgM antibody testing was concurrently performed. A case-control study tested the association of biological and clinical symptoms with the presence of PMMoV in the stool. Twelve (57%) food products were positive for PMMoV RNA sequencing. Stool samples from twenty-two (7.2%) adults and one child (0.7%) were positive for PMMoV by real-time PCR. Positive cases were significantly more likely to have been sampled in Dermatology Units (p<10−6), to be seropositive for anti-PMMoV IgM antibodies (p = 0.026) and to be patients who exhibited fever, abdominal pains, and pruritus (p = 0.045, 0.038 and 0.046, respectively). Conclusions Our study identified a local source of PMMoV and linked the presence of PMMoV RNA in stool with a specific immune response and clinical symptoms. Although clinical symptoms may be imputable to another cofactor, including spicy food, our data suggest the possibility of a direct or indirect pathogenic role of plant viruses in humans.

Inheritance of zucchini yellow mosaic virus resistance in Cucumis melo L.

SummaryResistance to zucchini yellow mosaic virus has been found in the muskmelon line ‘PI 414723’ from India. This resistance is effective against the ZYMV strains E15 and 1318 belonging respectively to the NF and F pathotypes. Resistance to E15 (no vein clearing and yellowing symptoms) is governed by one dominant gene (symbol Zym) according to segregations observed in F1, F2 and BC1 progenies. This gene is epistatic dominant over Fn, which induces wilting and necrosis after inoculation with F pathotype. Linkage studies suggest that Zym inherits independently from Fom-1, Fom-2, Vat, Wmv and Fn but is linked with a (13.1 ±2.4 units).

Epidemiological risks from mixed virus infections and transgenic plants expressing viral genes.

Publisher Summary This chapter focuses on the epidemiological risks from mixed virus infections and transgenic plants expressing viral genes. Several types of interaction may occur between a virus infecting a transgenic plant and either the transgene transcript or its translation product. These include heterologous encapsidation or interaction with a transmission accessory factor, which have the potential to alter vector specificity; complementation from a protein involved in cell-to-cell or long-distance movement, with possible effects on tissue tropism or the ability to establish a systemic infection; synergism resulting in increased replication and/or increased symptom severity; and recombination resulting in gain of function or alteration of pathogenicity. The chapter reviews the general aspects of vector specificity in different virus groups, as this could be altered either by recombination or by virus–transgene interactions. Potential complementation in a transgenic plant would also be limited in epidemiological effect, as the complementing transgene would not be present in nontransgenic plants. The chapter also discusses various means of minimizing the perceived risks of interactions between viruses and transgenically expressed viral sequences and the potential benefits to be gained from virus-resistant transgenic plants in comparison to the perceived risks.

Cytological and molecular evidence that the whitefly-transmitted Cucumber vein yellowing virus is a tentative member of the family Potyviridae

Cucumber vein yellowing virus (CVYV) is widespread in cucurbits in the Middle East. CVYV has filamentous particles and is transmitted by Bemisia tabaci by the semi-persistent mode. It has not yet been assigned to a specific genus or family. Ultramicroscopic observations revealed numerous cylindrical cytoplasmic inclusions in melon and cucumber cells infected by CVYV isolates from Israel and Jordan. Depending on the section orientation, the inclusions appeared as pinwheels or as bundles. In addition, a 1.9 kb DNA fragment was amplified by RT-PCR from CVYV-infected plant extracts using primers designed to detect all potyvirids. Sequence comparisons with the amplified fragment indicated that CVYV is more closely related to Sweet potato mild mottle virus than to any other virus in the family Potyviridae: These results suggest that CVYV can be considered as a tentative new member of the genus Ipomovirus:, family Potyviridae:

Potential Involvement of Melon Fruit in the Long Distance Dissemination of Cucurbit Potyviruses

Papaya ringspot virus (PRSV) and Zucchini yellow mosaic virus(ZYMV) are potyviruses frequently reported in cucurbits in Mediterranean, subtropical, and tropical regions. Occasionally, epidemics are also observed in more temperate regions, but the ways these viruses are introduced into new areas are not yet fully determined. We investigated the possibility that infected imported melon fruit could be a route for the introduction of PRSV and ZYMV. Imported melon fruits of the yellow canary type infected by ZYMV and PRSV were exposed in the fields next to healthy melon or squash bait plants. During this period, aphids were observed landing and probing on the fruits. In four independent experiments using different fruits, 3.1 to 25% of bait plants were infected by ZYMV and/or PRSV. PRSV was more frequently transmitted to bait plants than ZYMV. Comparison of partial sequences of the isolates from fruits and from bait plants showed a very high, if not complete, identity within each experiment, confirming that a natural transmission did occur from the fruit to the bait plants. These results suggest that globalization of melon production and international trade may be a factor in the spread of cucurbit potyviruses between countries or continents.

Genetic diversity of plant virus populations: towards hypothesis testing in molecular epidemiology.

Publisher Summary This chapter presents results obtained on plant virus diversity and population genetics. It illustrates how the analysis of the patterns of genome variation could be a ‘window’ through which to measure the role of the different evolutionary forces that shape plant virus populations. It also emphasizes that analysis of the representativeness of samples could help compare the diversity in different plant virus populations. Simple approaches have been presented that can be used to assess statistically the role of different mechanisms affecting plant virus populations. The development of genome exploration techniques and analytical methods for studying population genetic should allow a more precise understanding of plant virus epidemiology through the study of genomic diversity within virus populations. This strategy has, however, several inconveniences: (1) it relies on regular epidemics and for many viruses and it might not be desirable to introduce laboratory virus strains in the field; (2) these experiments only explore a limited fraction of natural epidemiological situations. An easier and less costly approach is to collect virus samples in different epidemiological situations and most studies of plant virus diversity have been performed in such a context. Consequently, few environmental parameters are controlled and analysis is rendered more complex. Based on knowledge of plant virus diversity and structure, several guidelines are provided to optimize molecular epidemiology.

Use of modified plum pox virus coat protein genes developed to limit heteroencapsidation-associated risks in transgenic plants.

Aphid transmission of a non-aphid-transmissible strain of zucchini yellow mosaic virus (ZYMV-NAT) occurs in transgenic plants expressing the plum pox potyvirus (PPV) coat protein (CP) gene. Heteroencapsidation has been shown to be responsible for this modification in the epidemiological characteristics of the infecting virus. In order to prevent this biological risk, several modified PPV CP constructs were produced that were designed to interfere with heteroencapsidation itself or to block aphid transmission of heteroencapsidated virions. These constructs were first expressed in Escherichia coli in order to check for the accumulation of pseudo-particles by electron microscopy. Virus-like particles (VLPs) were found with the full-length CP and with a PPV CP lacking the DAG amino acid triplet involved in aphid transmission. However, no VLPs were observed with CP lacking R220, Q221 or D264, amino acids known to be essential for the assembly of other potyvirus CPs. Transgenic Nicotiana benthamiana lines expressing the different PPV CP constructs were infected with ZYMV-NAT. Aphid transmission assays performed with these plants demonstrated that the strategies developed here provide an effective means of minimizing the biological risks associated with heteroencapsidation.

Virus transmission by host-specific strains ofOlpidium bornovanus andOlpidium brassicae

Zoospores of 12 isolatesO. bornovanus from geographically diverse sites and representing the three host specific cucurbit strains were tested as vectors for seven viruses using watermelon bait plants and the in vitro acquisition method. All isolates of the cucumber, melon, and squash strains transmitted melon necrotic spot carmovirus (MNSV) and cucumber necrosis tombusvirus (CNV) but none transmitted petunia asteroid mosaic tombusvirus (PAMV) or tobacco necrosis necrovirus (TNV). The isolates varied as vectors of three other carmoviruses: cucumber leaf spot virus (CLSV); cucumber soil borne virus (CSBV); and squash necrosis virus (SqNV). All cucumber isolates transmitted CLSV and SqNV but not CSBV. Some of the melon isolates transmitted CLSV and SqNV but none transmitted CSBV. Two squash isolates transmitted CSBV and SqNV but not CLSV. Two isolates ofO. brassicae transmitted only TNV and a third did not transmit any of the viruses. The species of bait plant sometimes affected transmission. The most efficient vector strains ofO. bornovanus, as determined by reducing zoospores and virus in the inoculum, were the cucumber strain for CLSV; the cucumber strain for CNV if cucumber was the bait plant or melon strain if watermelon was the bait plant; and the squash strain for SqNV. The plurivorous strain ofO. brassicae was the most efficient vector of TNV.Olpidium bornovanus is the first vector reported for CSBV and is confirmed as a vector of SqNV. It is proposed that all carmoviruses may have fungal vectors.Ligniera sp. did not transmit any of the viruses in one attempt.

Serological and molecular variability of watermelon mosaic virus (genus Potyvirus)

SummaryWatermelon mosaic virus (WMV, genus Potyvirus) is very common in cucurbits worldwide, but its variability has been little studied. In France, where WMV has been known since 1974, unusually severe symptoms on zucchini squash have been found to be associated with WMV since 1999. We have developed serological and molecular tools to study WMV variability and the origin of severe strains. Eight monoclonal antibodies were obtained, characterized by epitope mapping, and used to assess the serological variability of 42 isolates from different countries. Sequence analysis based on the NIb-CP region revealed an important variability, with three distinct molecular groups. These analyses also suggested frequent intraspecific recombination in WMV.