Auxin-independent growth of maize tissue culture cells

Wall-regenerating protoplasts and suspension culture cells of Zea mays L. 'Black Mexican Sweet' were co-cultivated] with Ti plasmid-containing Agrobacterium tumefaciens strain ACH5. After elimination of the bacteria, putative transformants were selected for their ability to grow in the absence of added auxin. Althoug some of the treated cells grew, untreated controls and cells treated with the Ti plsmidless strain ACH5C3 also proliferated. The frequency of auxin-independent growth was similar in all treatments. Because none of 75 candidate transforms contained detectable T-DNA, the hormone-independent phenotype appears to be a consequence of habituation. Although some of the habituated colonies grew as undifferentiated callus, others produced rootlike structures. Miaze cells subcultured in liquid media containing progressively reduced concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) also became habituated.