Pseudomonas syringae pv. actinidiae detection in kiwifruit plant tissue and bleeding sap

摘要: The rapid spreading of the disease during last few years highlighted need a quick, sensitive and reliable method for Pseudomonas syringae pv. actinidiae (Psa) detection, to find possible inoculum sources limit pathogen spreading. A PCR method, using new primers designed on gene encoding putative outer membrane protein P1, was developed detect Psa in symptomatic asymptomatic tissue; nested-PCR also applied. Bleeding sap samples, collected early spring from orchards with trees, were used both assays isolation identification. nested methods able presence at very low concentration plant pollen extracts; RFLP analyses BclI amplicons confirmed assay specificity, while digestion BfmI AluI allowed discriminate strains isolated before 2008 those after 2008. Furthermore, crude bleeding samples detected 3 5 15 assayed respectively. Direct same bacterial identification results molecular analysis.