Frost-related dieback of willows. Comparison of epiphytically and endophytically isolated bacteria from different Salix clones, with emphasis on ice nucleation activity, pathogenic properties and seasonal variation

Swedish Salix plantations for biomass production have been suffering severe dieback during the past 10 years, possibly due to the combination of frost and bacterial disease. As opposed to summer and winter, spring and autumn are periods when epiphytic populations of ice nucleation active (INA) bacteria are generally high. The culturable bacterial floras from stems of diseased plants of four Salix viminalis clones were compared in spring and autumn. Both epiphytic and endophytic bacteria were isolated (i.e. from plant surface and from tissues beneath the bark, respectively), characterised and tested for ice nucleating activity and pathogenicity. Some strains were also identified with BIOLOG and 16S rRNA. Endophytically isolated communities were generally more stable than epiphytes, both in number of isolates and type of bacteria. More types were found in autumn than in spring the same year, although the total number of strains isolated was rather constant. In contrast, more strains (and a higher percentage of the total community) expressed ice nucleating activity in spring than in autumn. The overall number of pathogenic strains remained stable but their proportion among the community tested on plants increased. A close relationship was observed between the dieback rates in the field and the percentage of pathogenic strains found in the different clones. The dominating bacterial type isolated, Sphingomonas spp., also contained the highest percentage of ice nucleation active pathogenic strains. (C) 2004 Elsevier Ltd. All rights reserved.