Genetic diversity of Ralstonia solanacearum infecting solanaceous vegetables from India reveals the existence of unknown or newer sequevars of Phylotype I strains

Bacterial wilt is one of the important constraints in the cultivation of solanaceous vegetables in India. The disease is caused by Ralstonia solanacearum, a soil bacterium. We have collected 232 isolates of R. solanacearum infecting solanaceous vegetables (eggplant, tomato and chilli) and other crops from different parts of India. Pathogenicity of the isolates was tested on eggplant, tomato and chilli and the pathogen was confirmed by PCR. Multiplex PCR and biochemical tests indicated that all the isolates were phylotype I and biovar 3. Ninety-five representative isolates selected based on geographical region, host range and pathogenicity were subjected to further phylogenetic and diversity analysis. Sequence analysis of egl, pga and hrpB genes of 95 isolates and genetic diversity of 50 representative isolates was reported and discussed. Indian isolates within the Phylotype I did not group based on the host or geographical location, except clustering of isolates from the Andaman Islands. Indian isolates clustered into two sub groups based on egl and pga trees indicating the presence of two major population groups. Sub group 1 is the dominant group in the data set and consists of unknown/newer sequevars, and sub group 2 consist of mainly the isolates which are designated with sequevar numbers based on egl sequences. In the hrpB based tree, the sub group 2 is the dominant group in the data set and it is the same for the sub group 1 of the egl tree. Indian phylotpe I R. solanacearum strains are phenotypically diverse including the previously described sequevars 14, 17, 44, 47 and 48. Our studies indicated the existence of R. solanacearum isolates with unknown/newer sequevars; the diversity existing among the phylotype I isolates might be due to a continuous evolutionary process. To our knowledge this is the first detailed report on the diversity of phylotype I R. solanacearum strains infecting solanaceous vegetables and the existence of unknown/newer sequevars in India.