The nucleotide sequence of the putative coat protein open reading frame of seven previously uncharacterized AMV strains from Italy and France was determined and aligned with comparable sequences of other AMV strains (425 L, 425 M, YSMV, S, VRU, 15/64 and Da). The data set of AMV sequences was used to determine phylogenetic relationships by both a stochastic (stationary Markov model) and a deterministic method (maximum-parsimony) of analysis. The topology of the trees obtained with the two methods was essentially the same showing that all AMV strains clustered in two monophyletic groups. Close clustering of Italian strains in subgroup I and of French strains in subgroup II seems to suggests the effect of geographic distinctiveness of evolutionary dynamics of these AMV strains. This separation did not correlate with differences in host range or symptoms (necrotic or non necrotic) induced in tomato but rather it reflected variations in the amino acid sequence of their CP, which might be related to structural properties of virus particles. A simple and rapid procedure based on the reverse transcriptase-polymerase chain reaction (RT-PCR) followed by ezymatic digestion (RFLP) was developed to identify and classify AMV isolates into the two subgroups. The method applied to a number of other AMV isolates from Italy and France supported their division in two distinct subgroups. This RT-PCR RFLP method may be useful way to investigate the dynamics of AMV populations in nature.
SUMMARYThe RNA content of four different CMV strains was investigated. All RNA preparations always contained the four major RNA species described by Kaper & West (I972), while most preparations also contained several minor RNA species. The four largest RNAs were separated by sucrose density-gradient centrifuging and by electrophoresis on polyacrylamide gels. Combination experiments suggest, for all strains, that the genome consists of the three largest RNAs (1 + 2 + 3), while RNA 4 is not required for infectivity.The distribution of RNAs in the capsids could not be established unequivocally, and the situation possibly differs in different strains.
The dominant gene Am from Lycopersicon hirsutum f. sp. glabratum PI134417 confers resistance to most strains of Alfalfa mosaic virus, including the recently identified necrotic strains. The phenotypic response includes a lack of symptom development following mechanical inoculation of leaves. To study the resistance mechanism controlled by Am, biological (back-inoculation to susceptible hosts), serological (double-antibody sandwich, enzyme-linked immunosorbent assay), and molecular (reverse transcription-polymerase chain reaction and hybridization with specific riboprobes) methods of virus detection have been conducted on mechanically inoculated PI134417 leaves. The virus was never recovered, indicating that Am acts by an inhibition of viral accumulation during the early events of the virus life cycle. Am has been mapped genetically to the short arm of tomato chromosome 6 in the resistance hotspot, which includes the R-genes Mi and Cf-2/Cf-5 and the quantitative resistance factors Ty-1, Ol-1, and Bw-5.
In single inoculations, both PVY and PVMV replicated in inoculated leaves of Nicotiana tabacum cv. ‘Xanthi nc’ plants, but only PVY infected the tobacco plants systemically, whereas PVMV caused localized infection.
A mixed infection by the PVY‐To72 and PVMV‐type strains was experimentally realized in ‘Xanthi nc’ plants. In the presence of PVY, PVMV migrated systemically into the upper leaves of the tobacco plant, as was proved by back inoculation.
It would appear that in tobacco, PVY acts as a “helper” virus, providing PVMV with the necessary component factor for migration.
In extracts from the co–infected leaves. Immune Electron Microscopy (IEM) revealed phenotypic mixed particles which contained a mixture of coat proteins of PVY and PVMV.
The role of the structural and functional interactions between the two viruses, which enable PVMV to migrate systemically in tobacco plants, is discussed.
Characterization of tobamovirus isolates collected from pepper crops in South East France has revealed the existance of several strains belonging to different viruses of the group. The biological and serological properties of three isolates (Vi76, Adam and Eve), selected as representative strains, have been studied. They have been compared with TMV and ToMV strains already isolated in our region as well as with other reference strains of ToMV‐D and TMV‐U1. They were also compared with other tobamovirus strains P11, P8 and P14, from the Netherlands, and PMMV‐W from Italy also isolated from pepper genotypes possessing the “L” gene for resistance to TMV and ToMV strains.
According to biological and serological reactions, Vi76 is more related to ToMV than to TMV but it is not related to Adam and Eve which are more closely related to PMMV. On the basis of the interaction with the “L” gene for resistance in pepper genotypes we have found that the Adam and P8 strains belong to the P1‐2 pathotype and the Eve and P14 strains to the P1‐2‐3 pathotype and all the 4 strains belong to the PMMV‐W group.
In France, there have been no reports of the isolation of the P11 strain which is distantly related to the Adam strain.
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