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- Title
Quantification of viable cells of Clavibacter michiganensis subsp. michiganensis using a DNA binding dye and a real-time PCR assay.
- Authors
Luo, L. X.; Walters, C.; Bolkan, H.; Liu, X. L.; Li, J. Q.
- Abstract
Viable cells of Clavibacter michiganensis subsp. michiganensis (CMM), the causal agent of bacterial canker of tomato, were discriminated from the dead cells by quantitative real-time polymerase chain reaction (PCR), after the bacterial solution was treated with the DNA binding dye ethidium monoazide (EMA). The primers and TaqMan probe, based on the 16S-23S rDNA spacer sequences, were highly specific for CMM at the subspecies level. The detection limit of the direct real-time PCR was 103 colony forming units per mL (cfu mL−1) in samples and with an apparent sensitivity of 2 cfu of target cells in PCR reaction solution. Application of this method allows for selective quantification of viable cells of CMM and facilitates monitoring the pathogen in tomato seeds.
- Subjects
PHYTOPATHOGENIC bacteria in host plants; TOMATO disease &; pest resistance; SEED viability; PLANT-pathogen relationships; POLYMERASE chain reaction; DNA polymerases; GENETICS
- Publication
Plant Pathology, 2008, Vol 57, Issue 2, p332
- ISSN
0032-0862
- Publication type
Article
- DOI
10.1111/j.1365-3059.2007.01736.x