Detection of Xanthomonas fragariae and presumptive detection of Xanthomonas arboricola pv. fragariae, from strawberry leaves, by real-time PCR.
Real-time (TaqMan) PCR assays were developed to detect the strawberry angular leaf spot pathogen Xanthomonas fragariae (Xf) and the strawberry bacterial blight pathogen Xanthomonas arboricola pv. fragariae (Xaf). The Xf PCR (Xf gyrB) was designed within regions of the gyraseB gene, unique to Xf, after generating gyraseB DNA sequence data from Xf and other closely related strains. The Xaf PCR (Xaf pep) was designed within regions of the pep prolyl endopeptidase gene that were unique to Xaf, after generating pep DNA sequence data from Xf and Xaf strains. The Xf gyrB PCR detected only Xf strains amongst a panel of 20 Xanthomonas-related spp. and pathovars. The Xaf pep PCR assay detected all Xaf strains tested plus two other (of three tested) X. arboricola pathovars. An existing genomic DNA extraction protocol was modified to facilitate detection of both pathogens to 10(3) cells per strawberry leaf disc.
Weller
S A
SA
Central Science Laboratory, DEFRA, Sand Hutton, York, YO41 1LZ, UK. s.weller@csl.gov.uk
Beresford-Jones
N J
NJ
Hall
J
J
Thwaites
R
R
Parkinson
N
N
Elphinstone
J G
JG
D027081:DNA Gyrase / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D031985:Fragaria / Q000382:microbiology*; D018515:Plant Leaves / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D000085822:Prolyl Oligopeptidases; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases / Q000235:genetics; D014974:Xanthomonas / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases; D027081:DNA Gyrase