Intrasegmental recombination does not contribute to the long-term evolution of group A rotavirus

•0.7% of rotavirus sequences in GenBank have evidence of intragenic recombination.•There was no evidence for the sustained transmission of recombinant sequences.•Spurious causes of intrasegmental recombination should be considered.•The lack of intrasegmental recombination is evidence of an evolutionary constraint.

Rotavirus is a genetically diverse pathogen with an eleven-segmented, double-stranded RNA genome. Intrasegmental recombination has been proposed as a potential mechanism to generate antigenic diversity and a possible route of escape from vaccine-imposed selective pressure. Here intrasegmental recombination was studied by performing a genome-wide scan across the eleven genome segments of 797 publically available rotavirus strains. Sixty-two sequences, or 0.7% of sequences analyzed, have evidence of intrasegmental homologous recombination. None of the specific recombination events is seen in more than one sequence. This uniqueness is consistent with either a spurious finding of recombination or the possibility that recombinant sequences arise naturally but are rapidly purged from the rotavirus population through selection. Arguments for the former explanation are presented. This analysis finds no evidence that intrasegmental recombination leads to ongoing transmission or plays a constructive role in rotavirus evolution. These results have practical implications for phylogenetic analyses and suggest a fundamental constraint that may have shaped rotavirus genome structure and evolution.