Abstract:
【Objective】This study aimed to identify
Bacillus strains that showed effective antagonistic activity against the ginger bacterial wilt causative pathogen
Ralstonia solanacearum, and to optimize the fermentation conditions of these strains, thereby providing theoretical foundation for enriching the resources of biocontrol
Bacillus strains and further developing and applying biocontrol agents for ginger wilt disease. 【Method】Ginger rhizosphere soil samples were research object,ginger
R. solanacearum strains were targets, dilution coating method and antibacterial circle method were employed to screen strains exhibiting potent antagonistic effects against
R. solanacearum. Antagonistic strains were then classified based on morphological observations, physiological and biochemical testing, as well as molecular biological identification. The antibacterial activity of the selected strains against
R. solanacearum was quantified using the Oxford cup method. Subsequently, both single-factor and orthogonal experiments were conducted to refine their fermentation conditions. 【Result】Strain HC-5, isolated from the ginger rhizosphere soil and exhibiting strong antagonistic properties against
R. solanacearum formed an inhibition zone of 0.71 cm in diameter. Integrating morphological features, physiological and biochemical characteristics and molecular identification results, HC-5 was classified as
Bacillus velezensis. The results of single factor and orthogonal experiments showed that, optimal growth medium for strain HC-5 was established to contain glutinous rice flour(5.0 g/L), yeast extract(7.0 g/L), and zinc sulfate(11.0 g/L). The best fermentation conditions were determined as temperature of 35 ℃, fermentation duration of 36 h, inoculation amount of 1.0%, rotation speed of 180r/min, initial pH of 5.5 and liquid volume of 100 mL. The diameter of the antibacterial zone of the sterile supernatant of the optimized strain HC-5 against
R. solanacearum increased to 3.03 cm, which was 76.2% and 62.9% larger than under LB and fermentation medium conditions, respectively. 【Conclusion】Strain HC-5 demonstrates a substantial antagonistic effect against
R. solanacearum. The enhanced antibacterial activity of its sterile supernatant following fermentation condition optimization suggests its potential as a biocontrol agent in combating ginger bacterial wilt.