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Summa Phytopathologica

versão impressa ISSN 0100-5405

Resumo

ARAUJO JUNIOR, Artur Teixeira de et al. Use of RFLP-PCR method to detect kresoxim methyl-resistant Venturia inaequalis isolates from the South of Brazil. Summa phytopathol. [online]. 2015, vol.41, n.3, pp.224-226. ISSN 0100-5405.  https://doi.org/10.1590/0100-5405/2085.

The principal mechanism in the fungus Venturiainaequalis which confers resistance to oxidation inhibitors (quinol QoIs) is the occurrence of mutations in the cytochrome b gene CYTB (G143A). The aim of this study was to implement the PCR-RFLP methodology to characterize the reaction of V. inaequalis isolates to the presence of QoIs. Seven monosporic isolates of V. inaequaliswere collected from commercial orchards in Santa Catarina and supplied by EPAGRI Experimental Station at São Joaquim, Santa Catarina. The isolates were classified as sensitive or resistant after their growth on PDA medium containing 10 μg.mL-1 kresoxim methyl. To determine the presence of G143A mutation, the specific markers ViCytB-5F and ViCytB-6071R were used. The DNA of isolates was amplified by PCR and separated on 1.5% agarose gel. The fragments were then digested with the restriction enzyme Fnu4HI, mutant identifier, and the digestion products were analyzed by means of electrophoresis on 1.0% agarose gel. The revealed genotypes were associated with the phenotype established by the growth of isolates in the presence of the fungicide, indicating that 6 isolates already had the resistance allele. The use of PCR-RFLP techniques allowed rapid and reliable assessment in the detection of V. inaequalis resistance to kresoxim methyl.

Palavras-chave : G143A; Qols; Fungo; Marcador molecular; G143A; QoIs; Fungus; Molecular marker.

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