Anti-Fungal Analysis of Bacillus subtilis DL76 on Conidiation, Appressorium Formation, Growth, Multiple Stress Response, and Pathogenicity in Magnaporthe oryzae.

Abstract

In recent years, biological control has gained more attention as a promising method to combat plant disease. Such severe diseases cited include rice blasts caused by Magnaporthe oryzae. However, more effective microbial strains with strong adaptability still need to be identified. Therefore, we sought to assess the conidia germination, and formation of appressorium of DL76 in Magnaporthe oryzae. Besides, we also aimed at understanding the growth, multiple stress response and pathogenicity in Magnaporthe oryzae. We isolated Bacillus subtilis DL76 from a rice farm, which observed a strong antimicrobial effect on M. oryzae. The sterilized culture filtrate of DL76 inhibited the growth of M. oryzae, which motivated us to deduce the influence of DL76 on the pathogenicity of M. oryzae. We screened the effect of Bacillus subtilis DL76 on M. oryzae guy11. It demonstrated that sterilized culture filtrate (1 × 107 CFU/mL) of DL76 can delay and even suppress the germination of conidia and (1 × 107 and 1 × 106 CFU/mL) prevent the formation of appressorium in vitro and in vivo. DL76 became hypersensitive to osmotic, oxidative, and cell wall degrading agents. In addition, the relative transcript levels of stress-responsive genes oxidative and osmotic were down-regulated by DL76 except for sod1, cat1, and cat2. In vivo assessment of the antifungal activity of Bacillus subtilis using conidia suspension of DL76 reduced the incidence and severity of rice blast. Conclusively, our results show that DL76 is essential for controlling rice blast by inhibiting conidiation, growth, multiple stress tolerance, and pathogenicity in M. oryzae.