Sterol binding by methyl-β-cyclodextrin and nystatin--comparative analysis of biochemical and physiological consequences for plants

The dependence of membrane function on its sterol component has been intensively studied with model lipids and isolated animal membranes, but to a much lesser extent with plant membranes. Depleting membrane sterols could be predicted to have a strong effect on membrane activity and have harmful physiological consequences. In this study, we characterized membrane lipid composition, membrane permeability for ions, some physiological parameters, such as H2O2 accumulation, formation of autophagosomal vacuoles, and expression of peroxidase and autophagic genes, and cell viability in the roots of wheat (Triticum aestivum L.) seedlings in the presence of two agents that specifically bind to endogenous sterols. The polyene antibiotic nystatin binds to endogenous sterols, forming so-called 'nystatin pores' or 'channels' in the membrane, and methyl-β-cyclodextrin has the capacity to sequester sterols in its hydrophobic core. Unexpectedly, although application of both methyl-β-cyclodextrin and nystatin reduced the sterol content, their effects on membrane permeability, oxidative status and autophagosome formation in roots differed dramatically. For comparison, we also tested the effects of the antibiotic gramicidin S, which does not bind to sterols but forms nonspecific channels in the membrane. Gramicidin S considerably increased membrane permeability, caused oxidative stress, and reduced cell viability. Our results suggest that a decrease in the sterol content is, in itself, not sufficient to have deleterious effects on a cell. The disturbance of membrane integrity, rather than the decrease in the sterol content, is responsible for the toxicity of sterol-binding compounds.