Adh1-0: A selectable marker in Zea mays cell culture

Summary

Rapidly-dividing, well-dispersed cell cultures were established from a Zea mays seedling homozygous for Adh1-0. The seedling was among the progeny of selfed first backcross (BC₁) plants produced during a programme to transfer Adh1-0 into the Black Mexican Sweet background by backcrossing. Electrophoretic analysis showed the complete absence in the cultured cells of the major ADH activities (Set I and Set II bands) and the presence of a faint band due to the second ADH gene, Adh2. The extractable ADH activity of mutant cells was typically 7–8% of control wildtype cells. Adh1-0 cells were shown to be more sensitive to anaerobic conditions than wildtype cells, the exposure time giving 50% kill being 14 h for mutant and 52 h for wildtype cells. Adh1-0 cells were also more sensitive to the respiratory inhibitor, antimycin A, than wildtype cells but less sensitive to allyl alcohol. Thus conditions were established to select for or against Adh1-0 cells in culture. ADH⁺ cells introduced into Adh1-0 cell cultures could be rescued by plating in presence of antimycin A.