Biofilm-overproducing Bacillus amyloliquefaciens strain B9ΔywcC reduces cadmium uptake in lettuce by upregulating the expression of proteins associated with root surface biofilm formation and cell membrane homeostasis.

In this study, Bacillus amyloliquefaciens strain B9 and its biofilm-overproducing B9ΔywcC mutant (ΔywcCM) were characterized for their effects on Cd accumulation in Italian lettuce and the underlying molecular mechanisms under hydroponic conditions. Both B9 and ΔywcCM significantly reduced Cd contents in lettuce tissues compared with the controls. Furthermore, ΔywcCM exhibited significantly decreased Cd accumulation and increased root surface biofilm biomass and biofilm-mediated Cd immobilization compared with B9. Proteomic analysis of lettuce root surface biofilms revealed that ΔywcCM significantly upregulated the expression of several proteins compared to B9. These included IolC and IolD (associated with inositol metabolism), GlpK (associated with glycerol metabolism), TyrS (associated with tRNA synthesis and transport), and LepA and PepT (associated with protein translation and modification), which are associated with biofilm formation; ArgS and AspS (associated with tRNA synthesis and transport), LepA (associated with protein translation and modification), and GcvT (associated with aminomethyltransferase), which are involved in cell membrane homeostasis. Furthermore, ΔywcCM significantly upregulated the expression of iolD, iolI, and pepT in the root surface biofilms relative to B9. These findings demonstrated that the biofilm-overproducing ΔywcCM enhances root surface biofilm formation and stability, thereby promoting Cd immobilization and reducing Cd accumulation in lettuce leaves grown in Cd-contaminated solutions.