Dieback, caused by Erwinia psidii is currently one of the most severe emerging diseases of Eucalyptus spp. in Brazil. Because of its recent report, little is known about the mechanisms underlying bacterial infection. We studied the colonization and movement of E. psidii in host tissue using a strain labeled with green fluorescent protein. We were able to transform E. psidii with pGreen-TIR and to demonstrate plasmid stability in the absence of antibiotic selection both in vitro and in vivo. We demonstrate that tissue colonization by E. psidii is not restricted to the inoculation point (leaf axil). E. psidii colonizes the xylem vessels, sclerenchyma and parenchyma of leaves and stems of eucalypt. At 35 days after inoculation, the bacterium was found at 5 cm above the inoculation point, indicating that it colonizes the plant acropetally. Confocal microscopy analysis revealed that when rootinoculated the bacterium penetrates the primary and secondary roots and reaches the xylem, but it was never found in the root crown or stem, irrespective of the evaluation time. Our results provide insights into the biology of the E. psidii-eucalypt interaction, which requires a better understanding in order to design efficient strategies for pathogen control and disease management.

Dieback and wilt, caused by Erwinia psidii (Ep), is one of the most important emergent diseases of Eucalyptus spp. in Brazil. Currently, pathogen detection relies on isolation of bacteria from infected plant tissue and either identification based on morphological, physiological and biochemical tests or DNA amplification using the polymerase chain reaction (PCR), which in many cases is laborious and cumbersome. Considering the need for a simpler and more rapid, yet reliable, method for detecting the pathogen, we obtained a polyclonal antibody (anti‐Ep) and developed an agglutination test for specific detection of E. psidii. The antiserum was produced against the E. psidii strain LPF534 and tested against 101 E. psidii isolates from Eucalyptus spp.; three E. psidii isolates from Psidium guajava; 23 Ralstonia solanacearum and 18 Xanthomonas axonopodis isolates pathogenic to Eucalyptus spp.; and seven endophytic isolates from Eucalyptus spp., three of which are phylogenetically related to the genus Erwinia. Results of direct ELISA indicated that a concentration as low as 3.5 µg/ml of the anti‐Ep antibody was able to detect the E. psidii antigen and that the antibody did not cross‐reacted with other bacteria pathogenic and non‐pathogenic to Eucalyptus spp. In the agglutination test, the anti‐Ep antibody showed positive reaction with all strains of E. psidii tested whereas cross‐reaction with none of the strains that belong to other taxonomic groups was observed. The agglutination test showed a detection limit of 105 colony‐forming units (CFU)/ml, and its specificity was the same as that obtained by PCR amplification using E. psidii‐specific primers. These results demonstrate that the agglutination test developed here is a useful tool for specific, fast and inexpensive detection of E. psidii although only operational on pure bacterial suspensions and not yet directly from infected tissues.