Ribosomal DNA ITS-1 Intergenic Spacer Polymorphism in Triatomines (Triatominae, Heteroptera)

Abstract

The length polymorphism of ribosomal DNA ITS-1 intergenic spacer was analyzed in eight species of triatomines belonging to Triatoma, Rhodnius, and Panstrongylus genera. The analyzed species were Rhodnius domesticus, R. neivai, R. robustus, Triatoma brasiliensis, T. infestans, T. vitticeps, Panstrongylus megistus, and P. herreri. These insects are vectors of Chagas' disease, one of the most prominent public health problems among South American countries. This work allowed the differentiation between species of the Triatomini and Rhodniini tribes through the analysis of ITS-1 length polymorphism by PCR and RFLP techniques. The species of the Triatoma and Panstrongylus genera presented an amplified ITS-1 fragment between 600 and 1000 bp, whereas Rhodnius presented a less variable ITS-1 length fragment, around 300 bp, which could reflect the monophyletic origin of the Rhodniini tribe. Species belonging to this genus were further differentiated by RFLP with HaeIII and AluI endonucleases. Our results corroborate the hypothesis of polyphyletic origin in this group of insects and contribute to knowledge about evolutionary relationships in triatomines.

Vetores da doença de Chagas no Brasil

“…A técnica de PCR-RFLP foi empregada em apenas três trabalhos (Tartarotti & Ceron 2005, Naegele et al 2006, López et al 2007), e com o barateamento dos custos do sequenciamento de DNA, a tendência é a de que ela caia em desuso em muito pouco tempo.…”

Phylogenetic approach to the study of Triatomines (Triatominae, Heteroptera)

“…The hypothesis of polyphyletic evolution is corroborated by several studies on the Rhodiniini and Triatomini tribes. Analyses of sequences of mitochondrial (Stothard et al, 1998;Lyman, 1999) and ribosomal DNA (Bargues et al, 2000;Marcilla et al, 2001) and analysis of polymorphism length of intergenic transcribed rDNA (Tartarotti & Ceron 2005), enzymatic studies, morphological analyses and taxonomic (Carcavallo, 1999), ecological studies (Schofield, 1988), all support the theory of polyphyletic evolution. Thus, the Triatominae subfamily may be defined as a grouping based on hematophagous feeding with predatorial Reduviidae ancestry dispersed to several areas of the world.…”

“…Due to the high variation between the sequence alignments the tribes were considered separately (Marcilla et al, 2001). AFLP analysis of ITS-1 ribosomal transcribed DNA intergenic spacer on Rhodnius, Triatoma and Panstrongylus species reinforced the idea of polyphyletic origin between Rhodniini and Triatomini tribes (Tartarotti & Ceron, 2005).…”

Análise das relações taxonômicas e sistemáticas entre espécies de triatomíneos (Hemiptera: Reduviidae) de colônias mantidas pelo Serviço Especial de Saúde de Araraquara, inferida de seqüências de genes mitocondriais

“…Diversas técnicas baseadas na observação direta do DNA foram aplicadas aos triatomíneos, tanto nos níveis supra-específicos, de gêneros ou tribos, quanto nos intraespecíficos e forneceram novas interpretações sistemáticas e inferências para hipóteses evolucionárias do grupo, dando início a era cladística na análise filogenética desses insetos (GARCÍA e POWELL, 1998, STOTHARD et al, 1998, LYMAN et al, 1999HYPŠA et al, 2002, GALVÃO 2003ABAD-FRANCH e MONTEIRO, 2005;PAULA et al, 2005 (CARLIER et al, 1996, DUJARDIN et al, 1998a, GARCÍA et al, 1998, NOIREAU et al, 2000a, BORGES et al, 2000, JARAMILLO et al, 2001, BRENIÈRE et al 2002, PACHECO et al, 2003RAMÍREZ et al, 2005), SSPC, do inglês: Single Strand Conformational Polymorphism, ou análise única, ou simples, de polimorfismo conformacional (STOTHARD et al, 1998), caracterização de marcadores polimórficos de microssatélites (HARRY et al, 1998, ANDERSON et al, 2002MARCET et al, 2006;PFEILER et al, 2006), PCR-RFLP (Restriction Fragement Length Polymorphism) (JUSTINO 2004;TARTAROTTI et al, 2005;NAEGELE et al 2006) e mais recentemente, Multiplex-PCR (PAVAN e MONTEIRO, 2007).…”