RAPD analyses and rDNA intergenic-spacer sequences discriminate Brazilian populations of Triatoma rubrovaria (Reduviidae: Triatominae)

Abstract

Triatoma rubrovaria, a member of the 'infestans' subgroup, is a potential vector of Trypanosoma cruzi in southern Brazil. Surveillance data indicate a growing domiciliary and peridomiciliary invasion by Tri. rubrovaria in the rural areas of Rio Grande do Sul (RS). In fact, following effective control of Tri. infestans, Tri. rubrovaria, which seems to have pre-adaptative characteristics for anthropic ecotopes, has become the most frequent species of triatomine bug to be collected in these areas. To explore the intraspecific variability and domiciliation of Tri. rubrovaria, the ribosomal DNA (rDNA) of two RS populations of Tri. rubrovaria that were geographically separated by only 220 km was investigated. The RAPD profiles and nucleotide sequences of the intergenic region of the rDNA, including the internal transcribed spacers 1 and 2 (ITS-1 and ITS-2) and the 5.8S gene, were analysed. In the RAPD study, the use of three decameric primers revealed polymorphisms reflecting both genetic differences between the two populations and heterogeneity within each. A phenetic dendrogram of the Tri. rubrovaria specimens, based on the three-primer consensus and a simple-matching coefficient of similarity, showed two clusters, clearly differentiating the bugs from the two localities studied. The rDNA sequencing revealed four different nucleotide sequences, with two different genotypes in each locality. The level of intraspecific variability detected within ITS-1 and ITS-2 of the Tri. rubrovaria, which was remarkably high considering the physical closeness of the two populations sampled, may indicate that the two collection sites are separated by geographical barriers that ensure the reproductive isolation of each population. The ITS sequences, like the RAPD results, clearly distinguished the two populations while showing that there is heterogeneity within each of them. The present study appears to be the first to reveal ITS length differences between populations of the same triatomine species without any associated difference in the number of microsatellite repeats. These results are in agreement with those of earlier studies on iso-enzymes, chromatic patterns, the ecological effects of environmental modification by humans, and bloodmeal sources.