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Background:    Ralstonia  spp. is a major pathogenic microbe for tomato, which invades the roots of diverse plant hosts and colonizes xylem vessels causing wilt, especially in tropical, subtropical and warm-temperate regions.  Ralstonia  spp.   produces several virulence factors helping it to invade the plant’s natural defense mechanism. Native isolates of  Trichoderma spp., Pseudomonas fluorescens  and  Bacillus subtilis  can be used as biocontrol agents to control the bacterial wilt and combined application of these beneficial microbes can give better results.    Methods:  Bacterial wilt infection in the field was identified by field experts and the infected plant part was used to isolate  Ralstonia  spp.   in CPG media and was positively identified. Subsequently, the efficacy of the biocontrol agents was tested and documented using agar well diffusion technique and digital microscopy. 2ml of the microbial concentrate (10  9  cells/ml) was mixed in one liter of water and was applied in the plant root at the rate of 100 ml per plant as a treatment method.    Results:  It was observed that the isolated  Trichoderma  spp. AA2 and  Pseudomonas fluorescens  PFS were most potent in inhibiting the growth of  Ralstonia  spp.  ,  showing ZOI 20.67 mm and 22.33 mm, respectively. Digital microscopy showed distinct inhibitory effect on the growth and survival of  Ralstonia  spp  .  The results from the field data indicated that  Trichoderma  spp. and  Pseudomonas fluorescens  alone were able to prevent 92% and 96% of the infection and combination of both were more effective, preventing 97% of infection. Chemical control methods prevented 94% of infection.  Bacillus subtilis  could only prevent 84 % of the infection.      Conclusions:  Antagonistic effect against  Ralstonia spp.  shown by native isolates of   Trichoderma  spp. and  P. fluorescens  manifested the promising potential as biocontrol agents. Combined application gave better results. Results shown by  Bacillus subtilis  were not significant.

Evaluation of Trichoderma spp., Pseudomonas fluorescens and Bacillus subtilis for biological control of Ralstonia wilt of tomato