Strain-dependent mutational effects for Pepino mosaic virus in a natural host

[EN] Pepino mosaic virus (PepMV) is an emerging plant pathogen that infects tomatoes worldwide. Understanding the factors that influence its evolutionary success is essential for developing new control strategies that may be more robust against the evolution of new viral strains. One of these evolutionary factors is the distribution of mutational fitness effect (DMFE), that is, the fraction of mutations that are lethal, deleterious, neutral, and beneficial on a given viral strain and host species. The goal of this study was to characterize the DMFE of introduced nonsynonymous mutations on a mild isolate of PepMV from the Chilean 2 strain (PepMV-P22). Additionally, we also explored whether the fitness effect of a given mutation depends on the gene where it appears or on epistatic interactions with the genetic background. To address this latter possibility, a subset of mutations were also introduced in a mild isolate of the European strain (PepMV-P11) and the fitness of the resulting clones measured.This study was financially supported by grant 2011/01/D/NZ9/00279, from the Poland National Science Center, to B.H.J and by grants BFU2015-65037-P, from Spain Ministry of Economy and Competitiveness-FEDER, and PROMETEOII/2014/021, from Generalitat Valenciana, to S.F.E.Minicka, J.; Elena Fito, SF.; Borodynko-Filas, N.; Rubis, B.; Hasiów-Jaroszewska, B. (2017). Strain-dependent mutational effects for Pepino mosaic virus in a natural host. BMC Evolutionary Biology. 17:1-11. https://doi.org/10.1186/s12862-017-0920-4S11117Steinhauer DA, Domingo E, Holland JJ. Lack of evidence for proofreading mechanisms associated with an RNA virus polymerase. Gene. 1992;122:281–8.Sanjuán R, Nebot MR, Chirico N, Mansky LM, Belshaw R. Viral mutation rates. J Virol. 2010;84:9733–48.Domingo E. Viruses at the edge of adaptation. 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Partial characterization of Sunn-hemp mosaic tobamovirus [SHMV] isolated from bean plants [Phaseolus vulgaris L.]

This work presents some properties of Sunn-hemp mosaic tobamovirus (SHMV) orginally isolated from bean plants. Virus infected host range and induced symptoms that were typical for SHMV. Following plant species distinquished SHMV from tobacco mosaic tobamovirus (TMV): Phaseolus vulgaris, Pisum sativum, Lupinus albus and Lycopersicon esculentum. In immunoblotting the serum against SHMV did not react with TMV and Tomato mosaic tobamovirus (ToMV). The electrophoretical patterns of whole virions and capsid proteins were characteristic for SHMV and different from that of TMV and ToMV.W pracy przedstawiono charakterystykę wirusa mozaiki krotalarii (SHMV) wyizolowanego z siewek fasoli wyrosłych z zainfekowanych nasion. Stwierdzono, że zakres roślin gospodarzy badanego izolatu SHMV oraz wywoływane przez niego objawy chorobowe były typowe dla tego wirusa. Gatunki roślin takie jak: Phaseolus vulgaris, Pisum sativum, Lupinus albus i Lycopersicon esculentum różnicują SHMV od TMV. W teście immunoblotingu surowica przeciwko SHMV nie reagowała z TMV-U i ToMV-2., z kolei surowica przeciwko TMV reagowała jedynie z TMV-U₁ i ToMV-2. Obrazy elektroforetyczne zarówno całych virionów jak i białek otoczki wirusowej były charakterystyczne dla SHMV i różne od tych dla TMV-U₁ oraz ToMV-2

Biologiczna i molekularna charakterystyka polskich izolatów wirusa mozaiki cukinii (Zuchinii yellow mosaic virus)

The diversity of Zucchini yellow mosaic virus (ZYMV) isolates from cucumber and zucchini plants growing in different regions of Poland was analyzed using biological tests and molecular biology techniques. The isolates differed in their host range and symptoms induced by them on a series of plant species. In addition, the analysis of the genetic diversity of the coat protein (CP) gene revealed high level of nucleotide variability among the isolates. Comparison of the CP gene sequences of 70 isolates from different geographical regions worldwide showed that the Polish isolates belong to different groups and they do not form a monophyletic cluster with European isolates. Interestingly, among the central European ZYMV isolates lower variability has been observed previously. The ratio of nonsynonymous to synonymous polymorphic sites showed a dominant negative selection however codons which might undergo positive selection were also identified. Moreover, the evidences for recombination in analyzed sequences of the CP gene of the analyzed ZYMV isolates were provided.Wirus żółtej mozaiki cukinii (Zucchini yellow mosaic virus, ZYMV) charakteryzuje się wysokim stopniem zróżnicowania zarówno biologicznego, jak i genetycznego. W pracy analizowano zakres roślin gospodarzy i symptomy wywoływane przez polskie izolaty ZYMV. Ponadto przeprowadzono analizę filogenetyczną i przebiegu rekombinacji z wykorzystaniem sekwencji nukleotydów genu kodującego białko płaszcza polskich i 67 innych izolatów ZYMV, których sekwencje zdeponowano w Banku Genów. Badania wykazały, że polskie izolaty nie tylko różniły się od siebie w teście biologicznym, ale należą do różnych grup filogenetycznych, nie tworząc tym samym jednej grupy z innymi izolatami europejskimi. Analiza presji selekcyjnej działającej na populację izolatów ZYMV wykazać dominację presji negatywnej, aczkolwiek zidentyfikowano również kodony, które mogą podlegać wpływowi pozytywnej presji selekcyjnej

Rapid evolutionary dynamics of the Pepino mosaic virus - status and future perspectives

Pepino mosaic virus (PepMV) has emerged as an important pathogen of greenhouse tomato crops and is currently distributed worldwide. Population genetic studies have revealed a shift in the dominant PepMV genotype from European (EU) to Chilean 2 (CH2) in North America and several European countries. New genetic variants are constantly being created by mutation and recombination events. Single nucleotide substitutions in different parts of the genome were found to affect on development of symptoms resulting in new pathotypes and accumulation of viral RNA. The variability of the PepMV population has a great impact on designing specific diagnostic tools and developing efficient and durable strategies of disease control. In this paper we review the current knowledge about the PepMV population, the evolutionary dynamics of this highly infective virus, methods for its detection and plant protection strategies

Cloning and sequencing of full-length cDNAs of RNA1 and RNA2 of a Tomato black ring virus isolate from Poland

International audienceFull-length cDNA clones corresponding to the RNA1 and RNA2 of the Polish isolate MJ of Tomato black ring virus (TBRV, genus Nepovirus) were obtained using a direct recombination strategy in yeast, and their complete nucleotide sequences were established. RNA1 is 7358 nucleotides and RNA2 is 4633 nucleotides in length, excluding the poly(A) tails. Both RNAs contain a single open reading frame encoding polyproteins of 254thinspkDa and 149thinspkDa for RNA1 and RNA2 respectively. Putative cleavage sites were identified, and the relationships between TBRV and related nepoviruses were studied by sequence comparison

Ratio of mutated versus wildtype coat protein sequences in Pepino mosaic virus determines nature and severity of yellowing symptoms on tomato plants

Recently, Pepino mosaic virus (PepMV) infections causing severe yellowing symptoms in tomato plants have been reported in glasshouse tomato crops. When studying this phenomenon in commercial glasshouses, two different types of yellowing symptoms, occurring in adjacent plants, were distinguished: interveinal leaf yellowing and yellow mosaics. After several weeks, the interveinal leaf yellowing symptoms gradually disappeared and the plant heads became green again, with yellow mosaic patterns on the leaves as an intermediate stage. The sequencing of multiple isolates causing interveinal leaf yellowing identified two point mutations, occurring in positions 155 and 166 of the coat protein (CP), as unique to the yellowing pathotype. Site-directed mutagenesis of infectious clones confirmed that both CP mutations are determinants of the interveinal leaf yellowing symptoms. Sequencing of CP clones from plants or plant parts with the yellow mosaic symptoms resulted in a mixture of wild-type and mutated sequences, whereas sequencing of CP clones from the green heads of recovered plants resulted in only wild-type sequences. Yellow mosaic symptoms could be reproduced by inoculation of an artificial 1:1 mixture of RNA transcripts from the wild-type and mutated infectious clones. These results show that the ratio of mutated versus wild-type sequences can determine the nature and severity of symptom development. The gradual recovery of the plants, which coincides with the disappearance of the yellowing mutations, suggests that selection pressure acts to the advantage of the wild-type virus. Experiments with wild-type and mutated infectious clones showed that reverse mutation events from mutant to wild-type occur and that the wild-type virus does not have a replicative advantage over the mutant. These results suggest that reverse mutation events occur, with subsequent selection pressure acting in favour of the wild-type virus in the growing plant parts, possibly related to a lower long-distance movement efficiency of the mutant

Genetic diversity of the coat protein of olive latent virus 1 isolates

The CP gene variability among 21 olive latent virus 1 (OLV-1) isolates obtained from different hosts and locations and at different times was assessed. Amplicons obtained by RT-PCR were cloned, and at least 10 sequences from each isolate were analyzed and compared. OLV-1 sequences available in GenBank were included. The encoded CPs consisted of 270 amino acids, except those of isolates G1S and C7 (269 aa) and G6 (271 aa). Comparison of CP genomic sequences of the isolates under study showed very low values of nucleotide diversity, 0.02, and maximum nucleotide distances between (0.087) or within isolates (0.001). Although very few nucleotide sequence differences were observed among the isolates, olive isolates exhibited lower diversity (0.012). In addition, at position 158 (157 in C7 and G1S and 159 in G6) of the deduced aa sequences, an alanine residue was found to be conserved among the olive isolates. In citrus and tulip isolates, a threonine residue was present at position 158, whereas a valine was present at this same position in tomato isolates. Phylogenetic analysis indicated that OLV-1 isolates clustered in five groups according to original host. However, G6, originally recovered from olive but repeatedly inoculated and maintained in N. benthamiana plants for 8 years in our laboratory, was separated from other isolates. This may be attributable to adaptation to the experimental host over time. There was no correlation of phylogenetic grouping of isolates based on geographical location or year of collection. Strong negative selection may have contributed to the low diversity among the OLV-1 CP isolates