Use este identificador para citar ou linkar para este item: http://repositorio.ufla.br/jspui/handle/1/12487
Título: Otimização da técnica de PCR para a detecção de Xanthomonas axonopodis pv. phaseoli em sementes de feijão
Título(s) alternativo(s): Optimization of PCR technique for detection of Xanthomonas axonopodis pv. phaseoli in bean seeds
Palavras-chave: Crestamento bacteriano comum
Curtobacterium flaccumfaciens pv. flaccumfaciens
Lotes comerciais
Feijoeiro - Bactérias
Common bacterial blight
Commercial batches
Common bean - Bacteria
Data do documento: Jan-2013
Editor: Grupo Paulista de Fitopatologia
Citação: SILVA, F. C. et al. Otimização da técnica de PCR para a detecção de Xanthomonas axonopodis pv. phaseoli em sementes de feijão. Summa Phytopathologica, Botucatu, v. 39, n. 1, p. 45-50, jan./mar. 2013.
Resumo: Common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli (Xap), is the major bacteriosis affecting bean plants in Brazil and is mainly transmitted by seeds. This study aimed to improve a technique, using different methods of extract preparation, for the detection of Xap alone and simultaneously with Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) in bean seed extracts, via PCR. Samples of bean seeds artificially inoculated with Xap and commercial batches were used to evaluate: crude extract obtained directly from the seeds, extract concentrated by filtration in 0.22ìM-diameter membrane, extract concentrated by centrifugation and extract plated on semiselective XCP1 medium with and without antibiotics (BIO-PCR). The simultaneous presence of Xap and Cff in 10 commercial batches of bean seeds was assessed through multiplex reaction, using the primers X4c, X4e, and CffFOR2 CffREV4. By using crude extract, extract concentrated by centrifugation and by filtration in millipore® membrane, detection of Xap in bean seeds artificially contaminated or in 47 commercial batches of bean seeds/grains was not possible. BIO-PCR technique allowed the detection of Xap in extracts from artificially contaminated bean seeds and in 18 of the 47 commercials batches. The technique of simultaneous detection of Xap and Cff on the same gel is feasible, for amplifying DNA fragments typical of each phytobacterium. The use of XCP1 culture medium without adding antibiotics allowed the detection of Xap with shorter incubation period in one day, compared to detection using the culture medium with antibiotics.
URI: http://repositorio.ufla.br/jspui/handle/1/12487
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