Enhancement of callose production by a combination of aluminum and iron in suspension-cultured tobacco (Nicotiana tabacum) cells

In nutrient medium, aluminum (Al) enhances ferrous ion [Fe(II)]-mediated peroxidation of lipids, which results in the loss of the plasma membrane integrity and the accumulation of Al in tobacco cells. Under these conditions, the mechanism of callose production and possible involvement of callose in the accumulation of Al were investigated. Callose production was enhanced by both Al and Fe(II), but not by Al or Fe(II) alone, and the enhancement was inhibited by a lipophilic antioxidant, suggesting that the enhancement of callose production is caused by the Al-enhanced, Fe(II)-mediated peroxidation of lipids. The enhancement of callose production depended on the presence of external Ca²⁺ in the treatment medium. The activity of β-1,3-glucan synthase in the microsomes was increased several times by the addition of Ca²⁺ in the assay medium, although the activity in the microsomes was reduced by the treatment of cells with Al and Fe(II) together. Therefore, it is likely that callose production is enhanced by exogenous Ca²⁺ via the Al-enhanced, Fe(II)-mediated peroxidation of lipids. During the exposure of the cells to Al and Fe(II), callose production started and increased simultaneously with Al accumulation. However, the digestion of callose in the cell wall materials prepared from the Al-treated cells by laminarinase did not release Al, suggesting that callose is not involved in the binding or trapping of Al.