Poleroviruses are phytoviruses strictly transmitted by phloem-feeding aphids in a circulative and nonpropagative mode. During ingestion, aphids sample virions in sieve tubes along with sap. Therefore, any sap protein bound to virions will be acquired by the insects and could potentially be involved in the transmission process. By developing in vitro virus-overlay assays on sap proteins collected from cucumber, we observed that approximately 20 proteins were able to bind to purified particles of Cucurbit aphid borne yellows virus (CABYV). Among them, eight proteins were identified by mass spectrometry. The role of two candidates belonging to the PP2-like family (predominant lectins found in cucurbit sap) in aphid transmission was further pursued by using purified orthologous PP2 proteins from Arabidopsis. Addition of these proteins to the virus suspension in the aphid artificial diet greatly increased virus transmission rate. This shift was correlated with an increase in the number of viral genomes in insect cells and with an increase of virion stability in vitro. Surprisingly, increase of the virus transmission rate was also monitored after addition of unrelated proteins in the aphid diet, suggesting that any soluble protein at sufficiently high concentration in the diet and acquired together with virions could stimulate virus transmission.
In addition to being essential for translation of eukaryotic mRNA, translation initiation factors are also key components of plant-virus interactions. In order to address the involvement of these factors in the infectious cycle of poleroviruses (aphid-transmitted, phloem-limited viruses), the accumulation of three poleroviruses was followed in Arabidopsis thaliana mutant lines impaired in the synthesis of translation initiation factors in the eIF4E and eIF4G families. We found that efficient accumulation of Turnip yellows virus (TuYV) in A. thaliana relies on the presence of eIF (iso)4G1, whereas Beet mild yellowing virus (BMYV) and Beet western yellows virus-USA (BWYV-USA) rely, instead, on eIF4E1. A role for these factors in the infectious processes of TuYV and BMYV was confirmed by direct interaction in yeast between these specific factors and the 5' viral genome-linked protein of the related virus. Although the underlying molecular mechanism is still unknown, this study reveals a totally unforeseen situation in which closely related viruses belonging to the same genus use different translation initiation factors for efficient infection of A. thaliana.
Poleroviruses are strictly transmitted by aphids. Glycosylation of Turnip yellows virus (TuYV) was previously reported and this modification was supposed to be required for aphid transmission. Using different approaches based on (i) a lectin-binding assay, (ii) use of specific complex glycan antibodies and (iii) mass spectrometry, we found no evidence that the structural proteins of TuYV and Cucurbit aphid-borne yellow virus (CABYV) carry glycan residues. Moreover, mutation of each of the four potential N-glycosylation sites of the structural protein sequences of CABYV indicated that, unless more than one site on the structural protein is glycosylated, N-glycosylation is not involved in aphid transmission. These results did not corroborate the previous hypothesis for the role of glycosylation in aphid transmission. They, however, revealed the presence of a glycosylated plant protein in purified polerovirus suspensions, whose function in aphid transmission should be further investigated.
Soft scales nymphs are suspected to propagate grapevine leafroll viruses in the vineyard, while moving from vine to vine, but results of long distance dispersion, especially those due to wind, are poorly known. Thus, a net of sticky cylindrical traps was placed in a newly planted grapevine plot to evaluate aerial dispersal of Parthenolecanium corni (Hemiptera Coccidae) nymphs. The plot was surrounded by vine plots infested by this species. Number of crawlers collected on traps was generally higher on the trap side exposed downwind and depended on population density of neighbouring vines. Coloured glitters of size and weight similar to that of crawlers were used to simulate wind transport. The highest number of glitters was trapped at the prevailing wind sector. A majority of glitters was found on traps close to dispersion cups, and the most remote were collected up to 165 m away from glitter source. Wind dispersal of crawlers was confirmed by colonisation of the new vine plot by P. corni, 2 years after plantation. The settling of P. corni on the young plot displayed no significant structure, as expected from a wind dispersal. Although grapevine leafroll viruses and Grapevine virus A were detected by reverse transcription-polymerase chain reaction in several batches of trapped nymphs, these viruses were not found in vines colonised by P. corni in the new plot within the 4 years following plantation.
Distribution patterns of the European fruit lecanium Parthenolecanium corni (Bouché) and of grapevine leafroll-associated virus-1 (GLRaV-1) and grapevine virus A (GVA) were monitored from 2003 to 2015 in a Riesling vine plot in the northeast of France. Virus spread was compared between two periods: 2003–2008 and 2009–2014. The percentage of infected vines increased from 54 to 78% for GLRaV-1 and from 14 to 26% for GVA. The spatial distribution of viruses and of P. corni was analysed using permutation tests and revealed an aggregative pattern. Virus distribution was not associated with the density of P. corni population on grapevines. However, GLRaV-1 and GVA spread mainly from initially infected vines. New GLRaV-1 and GVA infections were more frequent on vines near primarily infected vines, first anisotropically along the row, then between neighbouring rows. Virus spread was similar to those described in literature with grapevine mealybug species. This slow vine-to-vine progression suggests that P. corni was responsible for the virus spread, in accordance with the low mobility and low transmission capacities of its local population.
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