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- Title
PCR-based detection ofXanthomonas campestrispathovars inBrassicaseed.
- Authors
Berg, T.; Tesoriero, L.; Hailstones, D. L.
- Abstract
Xanthomonas campestrisis a seedborne bacterium that causes black rot of crucifers. Substantial crop losses may result from the rapid spread of the bacteria under favourable conditions, especially those occurring during seedling production. A PCR-based method has been developed for the rapid and sensitive detection of the pathovars ofX. campestristhat affect crucifers. Primers were designed to specifically amplify a 619 bp fragment of thehrpFgene fromX. campestris. Amplification products were not detected from otherXanthomonasspecies, or from other pathogenic or epiphytic bacteria occurring on these plants. To avoid false-negative results arising from the presence of amplification inhibitors in plant extracts, primers targeting a 360 bp section of the internal transcribed spacer (ITS) region fromBrassicaspp. were included in a multiplex PCR. The assay readily detectedX. campestrispv.campestrisinfections in diseased plants and from bacterial colonies isolated on growth media, and was more sensitive and specific than traditional plating methods and a commercially available ELISA. A seed-washing protocol was optimized to allow the detection of a single artificially infected seed among 10 000 healthy seeds using the multiplex PCR.
- Subjects
XANTHOMONAS campestris; DISEASED plants; PLANT physiology; PLANT growth; AGRICULTURAL pests; PLANT diseases
- Publication
Plant Pathology, 2005, Vol 54, Issue 3, p416
- ISSN
0032-0862
- Publication type
Article
- DOI
10.1111/j.1365-3059.2005.01186.x