BIOCTA: Novel approach to biocontrol of recently described plant tumorogenic Rhizobium spp. using autochthonous microbial solutions

Abstract

Introduction: A novel group of Rhizobiumspp. strains belonging to the “tumorigenes” clade has recently been described on blackberry in Serbia and Germany and on rhododendron in Germany. The BIOCTA project aimed to characterize efficient plant-associated bacterial strains for biocontrol of crown gall, thus providing an environmentally friendly alternative to pesticides that would contribute to the development of sustainable agriculture. Methods: Antagonistic potential of 37 biocontrol strains against two R. tumorigenes strains 932 and 1078 and Rhizobium sp. strain rho-6.2 was evaluated in vitro using the “well diffusion” method, as well as in vivo on tomato plants, using two inoculation strategies (co-inoculation and preventive). DNA metabarcoding approach was used to analyze the phytobiome of treated and non-treated tomato plants. Results: Based on the determined in vitro antagonistic potential, seven strains – Bacillus spp. (B. amyloliquefaciens ID084 and GT28.3, B. velezensis X5-2, and B. subtilis GD1), Pseudomonas sp. (R-6.10 and R11-20) and Agrobacterium rosae rho-6.1 were selected for further in vivo experiments. Of all tested strains/treatments, two Pseudomonas strains were the most efficient, showing up to 92.86%efficacy in suppressing tumors caused by Rhizobiumsp. strain rho-6.2 when applied in a co-inoculation strategy. Based on the DNA metabarcoding analysis, genera Pseudolabrys and Asanoa prevailed in the co-inoculation strategy, while Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium prevailed in positive control. Conclusion: Crown gall tumors have shown to be a valuable source of antagonistic isolates. Pseudomonas strains R-6.10 and R11-20 could be proposed for the efferent control of crown gall caused by newly described Rhizobium spp. strains in nurseries.