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Abstract
BackgroundGrapevine Algerian latent virus (GALV) is a tombusvirus first isolated in 1989 from an Algerian grapevine (Vitis spp.) plant and more recently from water samples and commercial nipplefruit and statice plants. No further reports of natural GALV infections in grapevine have been published in the last two decades, and artificial inoculations of grapevine plants have not been reported. We developed and tested a synthetic GALV construct for the inoculation of Nicotiana benthamiana plants and different grapevine genotypes to investigate the ability of this virus to infect and spread systemically in different hosts.MethodsWe carried out a phylogenetic analysis of all known GALV sequences and an epidemiological survey of grapevine samples to detect the virus. A GALV-Nf clone under the control of the T7 promoter was chemically synthesized based on the full-length sequence of the nipplefruit isolate GALV-Nf, the only available sequence at the time the project was conceived, and the infectious transcripts were tested in N. benthamiana plants. A GALV-Nf-based binary vector was then developed for the agroinoculation of N. benthamiana and grapevine plants. Infections were confirmed by serological and molecular analysis and the resulting ultrastructural changes were investigated in both species.ResultsSequence analysis showed that the GALV coat protein is highly conserved among diverse isolates. The first epidemiological survey of cDNAs collected from 152 grapevine plants with virus-like symptoms did not reveal the presence of GALV in any of the samples. The agroinoculation of N. benthamiana and grapevine plants with the GALV-Nf binary vector promoted efficient infections, as revealed by serological and molecular analysis. The GALV-Nf infection of grapevine plants was characterized in more detail by inoculating different cultivars, revealing distinct patterns of symptom development. Ultrastructural changes induced by GALV-Nf in N. benthamiana were similar to those induced by tombusviruses in other hosts, but the cytopathological alterations in grapevine plants were less severe.ConclusionsThis is the first report describing the development of a synthetic GALV-Nf cDNA clone, its artificial transmission to grapevine plants and the resulting symptoms and cytopathological alterations.Electronic supplementary materialThe online version of this article (doi:10.1186/1743-422X-11-186) contains supplementary material, which is available to authorized users.
Highlights
Grapevine Algerian latent virus (GALV) is a tombusvirus first isolated in 1989 from an Algerian grapevine (Vitis spp.) plant and more recently from water samples and commercial nipplefruit and statice plants
Variability of GALV isolates We collected all available published GALV sequences by searching the NCBI Nucleotide Database, which yielded a single complete genome sequence derived from a nipplefruit isolate (GALV-Nf) plus seven further GALV coat protein mRNA sequences
A further complete genome sequence derived from a grapevine isolate [1] was deposited recently, which hereafter we describe as GALVVv.2 to distinguish it from the GALV-Nf sequence
Summary
Grapevine Algerian latent virus (GALV) is a tombusvirus first isolated in 1989 from an Algerian grapevine (Vitis spp.) plant and more recently from water samples and commercial nipplefruit and statice plants. We developed and tested a synthetic GALV construct for the inoculation of Nicotiana benthamiana plants and different grapevine genotypes to investigate the ability of this virus to infect and spread systemically in different hosts. Grapevine Algerian latent virus (GALV) was first isolated in Italy in 1989 from an Algerian vine infected by Grapevine fanleaf virus (GFLV) [1]. The potential economic consequences of this virus in the grapevine industry are unknown because it has been detected only once in a mixed infection [1] but it caused severe stunting, chlorotic spots and mosaic symptoms in nipplefruit and statice plants [6,7]. Several experimental hosts have been identified in plant families such as Solanaceae, Chenopodiaceae and Amaranthaceae [1,2,3,5,6,7]
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