Germination of Bacillus spores by LiCl

Spores of Bacillus subtilis have been found to germinate when incubated with LiCl, but not with other monovalent or divalent metal cations. Bacillus megaterium spores also germinated with LiCl, but B. cereus spores did not. In B. subtilis, the LiCl germination was via the activation of spores' GerA germinant receptor (GR), and in B. megaterium, it was the GerU GR. Notably, LiCl germination was much slower than normal physiological germinant triggered GR germination. In B. subtilis spores, rates of LiCl germination were increased in spores with a more fluid IM and decreased in spores with a less fluid IM. Analyses of the GerA germinant binding site suggested that Li⁺ could bind in a specific site in the B. subtilis GerAB subunit where normally a Na⁺ likely binds. Importantly, NaCl strongly inhibited LiCl germination of B. subtilis spores, much more so than the larger cation in KCl, although neither salt inhibited L-alanine germination via the GerA GR. These findings increase the understanding of features of mechanisms of germination of Bacillus spores.IMPORTANCEThe ability of some bacteria to form spores upon nutrient starvation confers properties of metabolic dormancy and enhanced resistance to environmental stressors that would otherwise kill vegetative cells. Since spore-forming bacteria include several notable pathogens and economically significant spoilage organisms, insight into how spores are stimulated to germinate and form new vegetative cells is important. Here, we reveal that relatively high concentrations of the inorganic salt lithium chloride trigger the germination of Bacillus subtilis and Bacillus megaterium spores by stimulating one of the spores of each species cohort of nutrient germinant receptors. This is significant since novel germinants and increased knowledge of the germination process should provide opportunities for improved control of spores in healthcare, food, and environmental sectors.