Genus- and isolate-specific real-time PCR quantification of Erwinia on leaf surfaces of English oaks (Quercus robur L.).
In order to quantify pathogenic epiphytic bacteria on leaf surfaces of the important European forest tree Quercus robur without time-intensive cultivation and separation of microorganisms, methods were developed to selectively quantify DNA copy numbers of the genus Erwinia in DNA isolated from the leaf surface. By using the combination of the two different real-time PCR techniques SYBR-Green and TaqMan, methods were developed not only to allow quantification of the total DNA copy number of Erwinia on the oak leaf surface, but also to distinguish between two significantly different groups of Erwinia strains. In the present work, these techniques were successfully applied to quantify the copy number of the genus Erwinia and its subgroups compared with the total bacteria number in DNA samples extracted from the upper leaf surface of English oaks collected on the 4th of June 2001 (Julian day 155).
Institute for Meteorology and Climatology, Environmental Research (IMK-IFU), Research Centre Karlsruhe, Kreuzeckbahnstr. 19, D-82467 Garmisch-Partenkirchen, Germany. thomasheuser@gmx.net
Thomas
T
Heuser
Wolfgang
W
Zimmer
D001431:Bacteriological Techniques; D001483:Base Sequence; D003001:Cloning, Molecular; D015169:Colony Count, Microbial / Q000379:methods; D004269:DNA, Bacterial / Q000302:isolation & purification; D021903:DNA, Ribosomal Spacer / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D004885:Erwinia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016680:Genome, Bacterial; D018745:Genome, Plant; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D029963:Quercus / Q000382:microbiology*; D012680:Sensitivity and Specificity; D017385:Sequence Homology
D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer