Direct detection of bacterial phytopathogens in plant extracts
1994
Maes, M. | Garbeva, P. | Virgula, A. | Crepel, C. (Rijksstation voor Plantenziekten, Merelbeke (Belgium))
For the detection of the quarantine bacterial phytopathogens Xanthomonas campestris translucens and Erwinia amylovora, target specific rDNA oligonucleotide sequences were used as primers in PCR. The Xanthomonas campestris translucens primers recognized the phytobacteria belonging to the Xanthomonas translucens complex, producing a single 156 bp PCR fragment. The Erwinia amylovora primers were only complementary to the bacteria from the Erwinia amylovora species, initiating the amplification of a single 565 bp fragment in PCR. In both systems, taxonomically related or commonly present phytobacteria did not produce a PCR fragment and less than 4000 cfu target bacteria per n-d pure culture dilution were detected. Xanthomonas translucens infection was diagnosed in extracts of imported (Idaho, USA) wheat and barley seed. PCR mediated pathogen diagnosis was hindered when seeds were treated with fungicides. Erwinia amylovora bacteria could be detected when added to the epiphytical bacterial population present on the leafs, flowers and early fruit formations of pear. For Erwinia amylovora detection inside the shoots and branches of host plants, a PVPP treatment of the extract was required to eliminate PCR blocking agents.
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