C. Desbiez

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.

Association of an atypical alphasatellite with a bipartite New World begomovirus

The complete sequence of a watermelon-infecting Venezuelan begomovirus was obtained; it corresponds to that of the partially characterized melon chlorotic mosaic virus (MeCMV). MeCMV is a typical bipartite New World begomovirus. A putative alphasatellite, MeCMVa1, was found associated with MeCMV. This is the first satellite detected in the New World, and the first natural association of an alphasatellite with a bipartite begomovirus. The sequence of MeCMVa1 diverged highly from those of other alphasatellites, except for two atypical ones, which, like MeCMVa1, had a putative ORF2 embedded in the Rep-encoding ORF1. These findings raise new questions about the origin and evolution of subviral agents associated with begomoviruses.

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.

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.

Biological and Molecular Characterization of Moroccan watermelon mosaic virus and a Potyvirus Isolate from Eastern Sudan

A potyvirus (Su-94-54) was isolated from a naturally infected snake cucumber (Cucumis melo var. flexuosus) plant with severe mosaic and leaf deformation symptoms collected in Eastern Sudan. This isolate has a host range limited to cucurbits and is serologically distantly related to Moroccan watermelon mosaic virus (MWMV) and to Papaya ringspot virus (PRSV). Coat protein sequence analysis of Su-94-54 and MWMV and comparison with other potyviruses indicate that Su-94-54 is more closely related to MWMV than to any other potyvirus. Based on the amino acid sequence identity in the core part of the coat protein with MWMV (86%), this isolate could be regarded as a distinct species. However, because of biological, cytological, and serological affinities with MWMV, we propose that this isolate be considered as a strain of MWMV, possibly an evolutionary intermediate between MWMV and PRSV, until more is known on the structure of the PRSV subgroup within the genus Potyvirus.

Molecular epidemiology of Zucchini yellow mosaic virus in France: an historical overview.

Cucurbit viruses are involved in complex and changing pathosystems in France, with new virus strains or species regularly reported. Zucchini yellow mosaic virus (ZYMV) is an archetypal emerging virus that was reported in France in 1979. It has since caused sporadic but occasionally very severe economic losses and its epidemiology still remains poorly understood. Partial sequencing of the viral genome has been used to characterize ZYMV isolates that occurred over a 29-year period in experimental plots at Montfavet, France (n=227), or that were received through a national survey for cucurbit viruses conducted in France from 2004 to 2007 (n=198). A total of 34 haplotypes were differentiated belonging to five molecular groups, three including isolates already described in France and two corresponding to isolates that emerged in France within the last 5 years. Comparison of haplotypes found at one location during successive years revealed contrasting situations. When they were either the same or closely related haplotypes, this suggested the availability of overwintering hosts, whereas when they belonged to different molecular groups this indicated shifts in viral populations with possible new introductions. The contribution of molecular epidemiology in tracing the origin of ZYMV in the French West Indies is also reviewed.

Cloning of the Arabidopsis rwm1 gene for resistance to Watermelon mosaic virus points to a new function for natural virus resistance genes

Arabidopsis thaliana represents a valuable and efficient model to understand mechanisms underlying plant susceptibility to viral diseases. Here, we describe the identification and molecular cloning of a new gene responsible for recessive resistance to several isolates of Watermelon mosaic virus (WMV, genus Potyvirus) in the Arabidopsis Cvi-0 accession. rwm1 acts at an early stage of infection by impairing viral accumulation in initially infected leaf tissues. Map-based cloning delimited rwm1 on chromosome 1 in a 114-kb region containing 30 annotated genes. Positional and functional candidate gene analysis suggested that rwm1 encodes cPGK2 (At1g56190), an evolutionary conserved nucleus-encoded chloroplast phosphoglycerate kinase with a key role in cell metabolism. Comparative sequence analysis indicates that a single amino acid substitution (S78G) in the N-terminal domain of cPGK2 is involved in rwm1-mediated resistance. This mutation may have functional consequences because it targets a highly conserved residue, affects a putative phosphorylation site and occurs within a predicted nuclear localization signal. Transgenic complementation in Arabidopsis together with virus-induced gene silencing in Nicotiana benthamiana confirmed that cPGK2 corresponds to rwm1 and that the protein is required for efficient WMV infection. This work uncovers new insight into natural plant resistance mechanisms that may provide interesting opportunities for the genetic control of plant virus diseases.

First report of cucurbit yellow stunting disorder virus in Morocco.

In October, 1999, severe yellowing symptoms were observed in a melon (Cucumis melo L.) crop grown under plastic tunnels in the region of Agadir, Morocco. Large populations of whiteflies (Bemisia tabaci) were noticed during the early stages of the crop. At harvest, leaf samples were collected from two symptomatic plants and one symptomless plant. A mature yellow leaf was assayed from each symptomatic plant and for one of these two plants a younger leaf exhibiting only yellow spots. Cucurbit aphid-borne yellows virus, which causes similar symptoms in melons, was not detected by double-antibody sandwich enzyme-linked immunosorbent assay tests. Total RNA was extracted from fresh leaf tissues and submitted to reverse transcription and polymerase chain reaction with primers specific to two whitefly-transmissible viruses: Beet pseudo-yellows virus (BPYV) and Cucurbit yellow stunting disorder virus (CYSDV) (2). No amplification was obtained with BPYV-specific primers. In contrast, an expected 465-bp product was amplified in all samples from symptomatic plants with CYSDV-specific primers. No amplification was detected in samples from the symptomless plant nor from healthy control plants. B. tabaci-transmitted CYSDV has been reported in the Middle East, southwestern Europe, and North America (1,4). This is the first report of CYSDV in Morocco, and it follows the first report of another B. tabaci-transmitted virus, Tomato yellow leaf curl virus, in tomato (3), suggesting an important change in the viral pathosystem affecting vegetable crops in Morocco. References: (1) Kao et al. Plant Dis. 84:101, 2000. (2) Livieratos et al. Plant Pathol. 47:362, 1998. (3) Peterschmitt et al. Plant Dis. 83:1074, 1999. (4) Wisler et al. Plant Dis. 82:270, 1998.