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Aims  This study examined the contribution of GltB on biofilm formation and biocontrol efficiency of  B .  subtilis  Bs916.    Methods and Results  The  gltB  gene was identified through a biofilm phenotype screen and a bioinformatics analysis of serious biofilm formation defects, and then a  gltB  single knockout mutant was constructed using homologous recombination. This mutant demonstrated severe deficits in biofilm formation and colonisation along with significantly altered production ofγ-polyglutamate (γ-PGA) and three lipopeptide antibiotics (LPs) as measured by a transcriptional analysis of both the wild type  B .  subtilis  Bs916 and the  gltB  mutant. Consequently, the mutant strain retained almost no antifungal activity against  Rhizoctonia solani  and exhibited decreased biocontrol efficiency against rice sheath blight. Very few  gltB  mutant cells colonised the rice stem, and they exhibited no significant nutrient chemotaxis compared to the wild type  B .  subtilis  Bs916. The mechanism underlying these deficits in the  gltB  mutant appears to be decreased significantly in production of γ-PGA and a reduction in the production of both bacillomycin L and fengycin. Biofilm restoration of  gltB  mutant by additionγ-PGA in the EM medium demonstrated that biofilm formation was able to restore significantly at 20 g/L.    Conclusions  GltB regulates biofilm formation by altering the production ofγ-PGA, the LPs bacillomycin L and fengcin and influences bacterial colonisation on the rice stem, which consequently leads to poor biocontrol efficiency against rice sheath blight.    Significance and Impact of Study  This is the first report of a key regulatory protein (GltB) that is involved in biofilm regulation and its regulation mechanism and biocontrol efficiency by  B .  subtilis .

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Loss of GltB Inhibits Biofilm Formation and Biocontrol Efficiency of Bacillus subtilis Bs916 by Altering the Production of γ-Polyglutamate and Three Lipopeptides