Rapid identification of pseudomonas avellanae field isolates, causing hazelnut decline in central italy, by repetitive PCR genomic fingerprinting

Pseudomonas avellanae is the main cause of hazelnut "Corylus avellana L[# decline\ the so called {moria|\ in cen! tral Italy where it has already killed more than 29 999 trees[ Its current identi_cation is very long requiring biochemi! cal\ physiological and nutritional tests as well as patho! genicity tests and takes not less than 5 months for its completion[ In the present study the reliability of the repetitive polymerase chain reaction "rep!PCR# technique for a rapid and accurate identi_cation of such a pathogen was compared with the traditional identi_cation method[ In order to assess the variability of the pathogen\ REPBOX and ERIC primer sets were used in preliminary work to generate genomic _ngerprints of 59 P[ avellanae reference strains previously isolated from di}erent areas of hazelnut cultivation[ ERIC primers yielded the most discriminative clustering of strains that were grouped according to their geographic origin[ Sixty _eld isolates collected from hazelnut orchards of central Italy\ planted with di}erent cultivars\ during the years 0885 87 were submitted to either the traditional identi_cation methods or to rep!PCR by using ERIC primers[ The latter tech! nique accurately identi_ed all the isolates that were also identi_ed by the traditional methods[ Whole!cell protein analysis by means of sodium dodecyl sulphate!poly! acrylamide gel electrophoresis con_rmed this achievement[ Rep!PCR can be successfully adopted for the rapid and accurate identi_cation of P[ avellanae in central Italy and it constitutes a very useful tool for the sanitation of the area