Survival strategies and pathogenicity of Ralstonia solanacearum phylotype II subjected to prolonged starvation in environmental water microcosms

Abstract

Survival strategies exhibited over 4 years by Ralstonia solanacearum phylotype (ph) II biovar (bv) 2 in environmental water microcosms were examined. The bacterium is a devastating phytopathogen whose ph II bv 2 causes bacterial wilt in solanaceous crops and ornamental plants. Outbreaks of the disease may originate from dissemination of the pathogen in watercourses, where it has to cope with prolonged nutrient limitation. To ascertain the effect of long-term starvation on survival and pathogenicity of R. solanacearum in natural water microcosms, survival experiments were conducted. Microcosms were prepared from different sterile river water samples, inoculated separately with two European strains of ph II at 10(6) c.f.u. ml(-1) and maintained at 24 degrees C for 4 years. In all assayed waters, starved R. solanacearum remained in a non-growing but culturable state during the first year, maintaining approximately the initial numbers. Thereafter, part of the population of R. solanacearum progressively lost the ability to form colonies, and non-culturable but metabolically active cells appeared. During the whole period, the bacterium remained pathogenic on host plants and underwent a transition from typical bacilli to small cocci which tended to aggregate. Some starved R. solanacearum cells filamented and formed buds. Starvation response, viable but non-culturable state, morphological changes and aggregation have not previously been reported for this pathogen as survival mechanisms induced in oligotrophic conditions. The potential existence of long-starved pathogenic cells in environmental waters may raise new concerns about the epidemiology of bacterial wilt disease.