Abstract
Phylogenetic analysis through morphological and cultural traits is considered inconsistent and hard to be scientifically accepted once there is no way to establish the direction of evolution on morphological traits. Molecular markers are suitable for phylogenetic analysis. The sequencing of ITS1 and ITS2 (internal transcribed spacers) regions and of 5.8S gene from ribosomal DNA were used to estimate the genetic variation and distance of 10 Phytophthora capsici isolates. The amount of genetic variation amongst isolated P. capsici from distinct regions of Sao Paulo State was 0.1 to 1.6 and 0.1 to 1.1% at ITS1 and ITS2, respectively, and a phylogenetic distance of about 78.5% between P. capsici and Phytophthora spp. was observed
Highlights
O gênero Phytophthora é estudado desde o início da fitopatologia (Erwin e Ribeiro, 1996) e abrange patógenos destrutivos a inúmeras espécies de plantas no mundo (Lee e Taylor, 1992)
As seqüências de nucleotídeos das regiões internas transcritas (ITS1 e ITS2) do gene ribossomal 5.8S rDNA, bem como o próprio gene 5.8S rDNA têm sido utilizadas na taxonomia de Phytophthora (Berbee e Taylor, 1992; Lee e Taylor, 1992), visando a um rápido diagnóstico de P
LEE, S.B.; TAYLOR, J.W. Phylogeny of five fungus-like protoctistan Phytophthora species, inferred from the internal transcribed spacers of ribosomal DNA
Summary
DNA, assim como as seqüências obtidas junto ao GenBank, estão listados na tabela 1. Procedeu-se à reação de PCR dos isolados de Phytophthora capsici utilizando-se os oligonucleotídeo ITS4 e ITS5 (White et al, 1990) para ITS (Internal Transcribed Spacers) e gene 5.8S rDNA As reações de seqüenciamento foram realizadas para um volume de 10μL, utilizando-se 75ng do produto de PCR purificado ou do produto de clonagem, 2,0μL de “Big Dye Terminator” da Amersham, 3,5μL de água ultrapura Milliq e 1mM de cada um dos “oligonucleotídeo” ITS2, ITS3, ITS4 e ITS5, de acordo com o protocolo para “Amersham Premix Terminator”. As quatro seqüências obtidas pelos quatro oligonucleotídeos, para cada isolado, foram analisadas e alinhadas utilizando-se “Phred/Phrap/Consed” (Ewing et al, 1998; Gordon et al., 1998). A seqüênciaconsenso de todos os isolados de Phytophthora capsici mais as seqüências da região ITS-5.8S rDNA dos isolados do GenBank (Tabela 1) foram alinhadas usando o software.
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