Biocontrol of potato wilt by selective rhizospheric and endophytic bacteria associated with potato plant

Ralstonia solanacearum is the causative agent of wilt disease in plants, which constitutes a severe problem to agricultural crops, particularly for potato production in Madagascar. The present study focuses on the isolation, in vitro and in vivo assays of potential rhizospheric and endophytic bacteria associated with healthy potato plant, capable to inhibit the growth of Ralstonia solanacearum for controlling potato bacterial wilt. A total of 77 bacteria strains were isolated from six soil rhizospheric samples and six vegetal material samples of healthy potatoes in the district of Antsirabe II. Forty of them were telluric actinomycetes, 25 were endophytic actinomycetes and 12 were fluorescent Pseudomonas spp. An additional 30 phytopathogenic isolates were obtained from six rhizopsheric soil samples of diseased potatoes. Morphological, cultural, biochemical characterization and molecular identification with the Ralstonia solanacearum specific primers 759/760 revealed that 24 of the pathogenic isolates belong to the Ralstonia solanacearum species, biovar two; the causal agent of potato bacterial wilt. Isolates from healthy plants were, then, examined in vitro and in vivo for their antagonistic activity against Ralstonia solanacearum strain for their potential to improve potato plant growth. In vitro antagonism of actinomycete and Pseudomonas isolates against Ralstonia solanacearum development was performed using agar diffusion technique, while in vivo tests were conducted under greenhouse conditions. Ten antagonistic strains including two Pseudomonas, four telluric actinomycetes, and four endophytic actinomycetes inhibited the tested Ralstonia strain. Four strains, E7, E13 (endophytic actinomycete from root potatoes), S25 (telluric actinomycetes) and P7 (fluorescent Pseudomonas), showed high antagonistic activity against the pathogen with zones of inhibition from 23 to 40 mm. Of the fours strains tested in greenhouse, E7 significantly reduced (p < 0.05) the percentage of Ralstonia solanacearum that infected plants by 72.04%. The isolates E13 and S25 have also been demonstrated to improve plant growth by increase of plant height to 44.63% and 44.84%, fresh weight to 68.75% and 75.85% and dry weight to 86.17% and 115.42%, respectively compared with non-treated control. Morphological and cultural characterization of these three active isolates showed that they belong to the genus Streptomyces. The antagonism of these isolates against Ralstonia solanacearum according to in vitro and in vivo tests results, along with their high efficiency as regards the improvement of plant development, suggests that these three actinomycete strains E7, E13 and S25 could be useful for biocontrol of potato bacterial wilt.Key words: Potato, Ralstonia, Actinomycetes, Pseudomonas, Biocontrol