Molecular detection of a bacterial contaminant Bacillus pumilus in symptomless potato plant tissue cultures

Abstract

An aberrant random amplified polymorphic DNA (RAPD) marker in genomic DNA of tissue culture plantlets was frequently observed during a comparison of DNA fingerprints derived from potato germplasm grown in tissue culture and the field. The RAPD marker was cloned, sequenced and determined to be of bacterial origin. A bacterial contaminant was isolated from the tissue culture plants and identified as a Bacillus pumilus. A set of sequence characterised amplified region (SCAR) primers were designed from the sequence of the cloned fragment and tested for the specific detection of B. pumilus. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) were also used to generate B. pumilus profiles specific to our isolate in order to test and confirm the sequence homology of amplified markers generated from a range of DNA samples isolated from tissue culture plants and pure isolates of B. pumilus-like bacteria.

DOI: 10.1007/s00299-003-0583-z

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Cite this paper

@article{Isenegger2003MolecularDO, title={Molecular detection of a bacterial contaminant Bacillus pumilus in symptomless potato plant tissue cultures}, author={Daniel Isenegger and Paul William James Taylor and Kathleen B Mullins and Glenn R. McGregor and Martin Barlass and James Hutchinson}, journal={Plant Cell Reports}, year={2003}, volume={21}, pages={814-820} }