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Simultaneous PCR Detection of the Two Major Bacterial Pathogens of Geranium
Author(s) -
GLICK D. L.,
COFFEY C. M.,
SULZINSKI M. A.
Publication year - 2002
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1046/j.1439-0434.2002.00716.x
Subject(s) - biology , ralstonia solanacearum , geranium , polymerase chain reaction , multiplex polymerase chain reaction , primer (cosmetics) , bacterial wilt , multiplex , microbiology and biotechnology , pathogen , genetics , virology , gene , botany , chemistry , organic chemistry , cultivar
Xanthomonas campestris pv. pelargonii ( Xcp ) and Ralstonia solanacearum ( Rs ) are the two most important bacterial pathogens of commercially cultivated geraniums ( Pelargonium spp.), both causing bacterial wilt and leaf spot. Asymptomatic infections are important reservoirs of infections in commercial growing facilities. Our objective was to design a multiplex PCR (Polymerase Chain Reaction) assay to detect infection by either or both of these pathogens. We used a previously characterized PCR primer pair for Xcp that amplifies a region of 200 bp. In addition, we designed a new primer pair specific for Rs that amplifies a region of 822 bp. With these two primer pairs, we could detect either or both pathogens. As geranium tissue extracts frequently contain inhibitors of the PCR process, a negative PCR could result from either an accurate indication that the plant was pathogen‐free or from a false negative assay. We therefore designed `amplification competence' primers, targeting a portion of the geranium 18 s rRNA gene, and generating a 494‐bp amplification product that confirms amplification competence and validates a negative assay result. Thus, the triple primer pair multiplex PCR screens for the two most important bacterial pathogens of geraniums simultaneously confirms amplification competence for each geranium sample.