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Journal of Virology, December 2007, p. 12979-12984, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.00524-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Distribution of Fitness and Virulence Effects Caused by Single-Nucleotide Substitutions in Tobacco Etch Virus{triangledown} ,{dagger}

Purificación Carrasco, Francisca de la Iglesia, and Santiago F. Elena*

Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-UPV, 46022 València, Spain

Received 13 March 2007/ Accepted 16 September 2007

Little is known about the fitness and virulence consequences of single-nucleotide substitutions in RNA viral genomes, and most information comes from the analysis of nonrandom sets of mutations with strong phenotypic effect or which have been assessed in vitro, with their relevance in vivo being unclear. Here we used site-directed mutagenesis to create a collection of 66 clones of Tobacco etch potyvirus, each carrying a different, randomly chosen, single-nucleotide substitution. Competition experiments between each mutant and the ancestral nonmutated clone were performed in planta to quantitatively assess the relative fitness of each mutant genotype. Among all mutations, 40.9% were lethal, and among the viable ones, 36.4% were significantly deleterious and 22.7% neutral. Not a single case of beneficial effects was observed within the level of resolution of our measures. On average, the fitness of a genotype carrying a deleterious but viable mutation was 49% smaller than that for its unmutated progenitor. Deleterious mutational effects conformed to a beta probability distribution. The virulence of a subset of viable mutants was assessed as the reduction in the number of viable seeds produced by infected plants. Mutational effects on virulence ranged between 17% reductions and 24.4% increases. Interestingly, the only mutations showing a significant effect on virulence were hypervirulent. Competitive fitness and virulence were uncorrelated traits.

* Corresponding author. Mailing address: Instituto de Biología Molecular y Celular de Plantas (CSIC-UPV), Avenida de los naranjos s/n, 46022 València, Spain. Phone: 34 963 877 895. Fax: 34 963 877 859. E-mail: sfelena{at}ibmcp.upv.es

{triangledown} Published ahead of print on 26 September 2007.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, December 2007, p. 12979-12984, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.00524-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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