Application of Pb(II) to probe the physiological responses of fungal intracellular vesicles.

Vesicles (Ves) within fungal cells are the critical linkage between intracellular and extracellular systems. This study explored the application of Pb2+ to probe the physiology of intracellular Ves in Rhodotorula mucilaginosa (Rho). At low Pb2+ levels (0-500 mg/L), there was no evident change in the content of extracellular polymeric substances (EPS) or microbial activity. At medium-high levels (1000-2000 mg/L), the sizes of Ves within the Rho cells were significantly enlarged, with abundant lead nano-particles (Pb NPs) formed either on the cell surface or interior, whereas the EPS content and bioactivity were still stable. At a high level (2500 mg/L), the Rho cells were severely deformed, with cell counts reduced by more than 99%. However, the EPS contents and the respiration rate of the surviving cells dramatically increased to the maximum values (i.e., 1785 mg/1010 cells and 37 mg C 10-10 cells h-1, respectively). The Ves surface adsorbed Pb cations with higher density, compared with the cell membrane. Moreover, fusion of some Ves to the membrane (functioning in transport) was observed under transmission electron microscope (TEM). Three pathways of detoxification via intracellular Ves were finally proposed, i.e., Ve-mediated transport (from intracellular to extracellular) of EPS components, absorption of Pb NPs on the Ve surface, and accumulation of Pb NPs within Ves. This study sheds light on the possibility of exploring microbial physiology via Pb2+ cations.