Development of a Bio-PCR Protocol for the Detection of Xanthomonas arboricola pv. pruni

Ballard, E. L., Dietzgen, R. G., Sly, L. I., Gouk, C., Horlock, C. and Fegan, M. (2011) Development of a Bio-PCR Protocol for the Detection of Xanthomonas arboricola pv. pruni. Plant Disease, 95 9: 1109-1115. doi:10.1094/PDIS-09-10-0650


Author Ballard, E. L.
Dietzgen, R. G.
Sly, L. I.
Gouk, C.
Horlock, C.
Fegan, M.
Title Development of a Bio-PCR Protocol for the Detection of Xanthomonas arboricola pv. pruni
Formatted title
Development of a Bio-PCR protocol for the detection of Xanthomonas arboricola pv. pruni
Journal name Plant Disease   Check publisher's open access policy
ISSN 0191-2917
1943-7692
Publication date 2011-09-01
Sub-type Article (original research)
DOI 10.1094/PDIS-09-10-0650
Open Access Status DOI
Volume 95
Issue 9
Start page 1109
End page 1115
Total pages 7
Place of publication St. Paul, MN, United States
Publisher American Phytopathological Society
Collection year 2012
Language eng
Formatted abstract
A real-time SYBR Green I assay was developed and evaluated as a
biological and enzymatic polymerase chain reaction (Bio-PCR) protocol
for the detection of Xanthomonas arboricola pv. pruni. Suppression
subtractive hybridization was used to generate a X. arboricola pv.
pruni-specific subtracted DNA library, using X. arboricola pv. corylina
as the driver strain. Primer pair 29F/R, designed from cloned sequence,
showed no homology to GenBank sequences and amplified a 344-bp
product in all X. arboricola pv. pruni isolates. Compared with other
published X. arboricola pv. pruni primers, this primer pair was shown
to be the only one capable of differentiating X. arboricola pv. pruni
from all other X. arboricola pathovars. A real-time assay was
developed and shown to be capable of detecting less than 10 CFU and
0.1 pg of DNA. Epiphytic bacteria isolated from plum tissue was used
to further evaluate the specificity of the assay. A Bio-PCR protocol,
developed for field evaluation, confirmed X. arboricola pv. pruni isolation
from asymptomatic and symptomatic plum tissue over a 9-week
period between host flowering and the first appearance of leaf and fruit
symptoms in an orchard. Dilution plating enabled X. arboricola pv.
pruni numbers to be quantified, providing supportive evidence for the
usefulness of the Bio-PCR protocol in plant pathology and quarantine
surveillance.
Keyword Suppression Subtractive Hybridization
Polymerase-Chain-Reaction
Helicobacter-Pylori
Genetic-Differences
Bacterial Spot
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
School of Chemistry and Molecular Biosciences
 
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