Abstract

FRIAS-PIZANO, Jesús et al. Detection of Clavibacter michiganensis ssp. michiganensis by PCR in in tomato plants (Lycopersicon esculentum Mill.). Rev. Mex. Cienc. Agríc [online]. 2016, vol.7, n.6, pp.1347-1357. ISSN 2007-0934.

Bacterial canker caused by Clavibacter michiganensis ssp. michiganensis (Cmm) is a devastating disease of tomato around the world. The aim of this study was to describe a rapid procedure based on PCR for the detection of pathogenic Cmm in tomato plants. The DNA was isolated by boiling method and PCR was performed to amplify pat-1 gene (serine protease) using the oligonucleotides CMM5F and CMM6R whose amplicon size was 608 bp. The fragment was cloned, sequenced and compared with the database from NCBI (National Center for Biotechnology Information), showing 100% identity with sequences pat-1 gene of Cmm. Additionally, from the sequence obtained two pairs of nested oligonucleotides were designed to produce amplicons of 250 bp to an annealing temperature of 62 ° C. The results described in this paper will allow rapid and accurate identification of Cmm by PCR in seed and tomato plant tissue by reducing the analysis time from 24 to 4 h approximately.

Keywords : Clavibacter michiganensis ssp.; michiganensis; Lycopersicon esculentum Mill.; bacterial canker; boiled; nested oligonucleotides.

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