The evolutionary history and global spatio-temporal dynamics of potato virus Y

Potato virus Y (PVY) is a destructive plant pathogen that causes considerable losses to global potato and tobacco production. Although the molecular structure of PVY is well characterized, the evolutionary and global transmission dynamics of this virus remain poorly understood. We investigated the phylodynamics of the virus by analysing 253 nucleotide sequences of the genes encoding the third protein (P3), cylindrical inclusion protein (CI), and the nuclear inclusion protein (NIb). Our Bayesian phylogenetic analyses showed that the mean substitution rates of different regions of the genome ranged from 8.50 × 10−5 to 1.34 × 10−4 substitutions/site/year, whereas the time to the most recent common ancestor of PVY varied with the length of the genomic regions and with the number of viral isolates being analysed. Our phylogeographic analysis showed that the PVY population originated in South America and was introduced into Europe in the 19th century, from where it spread around the globe. The migration pathways of PVY correlate well with the trade routes of potato tubers, suggesting that the global spread of PVY is associated with human activities.

Cylindrical Inclusion Protein of Wheat Yellow Mosaic Virus Is Involved in Differential Infection of Wheat Cultivars

Wheat yellow mosaic virus (WYMV) belongs to the genus Bymovirus in the family Potyviridae and has a bipartite genome (RNA1 and RNA2). WYMV in Japan is classified into three pathotypes (I to III) based on its pathogenicity to wheat cultivars. Among these three, pathotypes I and II are discriminated by their pathogenicity to the wheat cultivar Fukuho; pathotype I infects Fukuho but pathotype II does not. In the present study, the genomic regions that are involved in such pathogenicity were examined using infectious viral cDNA clones of pathotypes I and II. Reassortant experiments between viral RNA1 and RNA2 revealed the presence of a viral factor related to pathogenicity in RNA1. A chimeric pathotype II virus harboring a cylindrical inclusion (CI) cistron from pathotype I facilitated systemic infection of Fukuho, indicating that CI protein is involved in pathogenicity. Furthermore, analysis of chimeric and site-directed mutants revealed that three amino acids at the N-terminal region of CI protein were involved in pathogenicity to Fukuho. On the other hand, at the single-cell level, pathotype II replicated in protoplasts of Fukuho similar to that of pathotype I virus. These data suggest that differential pathogenicity between pathotypes I and II was considered to depend on the ability of cell-to-cell or long-distance viral movement, in which CI protein is involved. To the best of our knowledge, this is the first report to show the involvement of the bymoviral CI protein in pathogenicity.

Cylindrical inclusion protein of potato virus A is associated with a subpopulation of particles isolated from infected plants

Potato virus A (PVA) particles were purified by centrifugation through a 30 % sucrose cushion and the pellet (P1) was resuspended and sedimented through a 5–40 % sucrose gradient. The gradient separation resulted in two different virus particle populations: a virus fraction (F) that formed a band in the gradient and one that formed a pellet (P2) at the bottom of the gradient. All three preparations contained infectious particles that retained their integrity when visualized by electron microscopy (EM). Western blotting of the P1 particles revealed that the viral RNA helicase, cylindrical inclusion protein (CI), co-purified with virus particles. This result was confirmed with co-immunoprecipitation experiments. CI was detected in P2 particle preparations, whereas F particles were devoid of detectable amounts of CI. ATPase activity was detected in all three preparations with the greatest amount in P2. Results from immunogold-labelling EM experiments suggested that a fraction of the CI present in the preparations was localized to one end of the virion. Atomic force microscopy (AFM) studies showed that P1 and P2 contained intact particles, some of which had a protruding tip structure at one end, whilst F virions were less stable and mostly appeared as beaded structures under the conditions of AFM. The RNA of the particles in F was translated five to ten times more efficiently than RNA from P2 particles when these preparations were subjected to translation in wheat-germ extracts. The results are discussed in the context of a model for CI-mediated functions.

Mutations in Turnip mosaic virus genomes that have adapted to Raphanus sativus

The genetic basis for virulence in potyviruses is largely unknown. Earlier studies showed that there are two host types of Turnip mosaic virus (TuMV); the Brassica/Raphanus (BR)-host type infects both Brassica and Raphanus systemically, whereas the Brassica (B)-host type infects Brassica fully and systemically, but not Raphanus. The genetic basis of this difference has been explored by using the progeny of an infectious clone, p35Tunos; this clone is derived from the UK1 isolate, which is of the B-host type, but rarely infects Raphanus systemically and then only asymptomatically. Two inocula from one such infection were adapted to Raphanus by passaging, during which the infectivity and concentration of the virions of successive infections increased. The variant genomes in the samples, 16 in total, were sequenced fully. Four of the 39 nucleotide substitutions that were detected among the Raphanus sativus-adapted variant genomes were probably crucial for adaptation, as they were found in several variants with independent passage histories. These four were found in the protein 1 (P1), protein 3 (P3), cylindrical inclusion protein (CI) and genome-liked viral protein (VPg) genes. One of four ‘parallel evolution’ substitutions, 3430G→A, resulted in a 1100Met→Ile amino acid change in the C terminus of P3. It seems likely that this site is important in the initial stages of adaptation to R. sativus. Other independent substitutions were mostly found in the P3, CI and VPg genes.