Comparisons have been made between a sycamore callus (S4) isolated in 1970 and one first isolated in 1958 (S2). Incubation of S4 on media containing different concentrations of auxin and kinetin showed that the ratio of the hormones was the factor which controlled the growth pattern within the tissue. High ratios of auxin: cytokinin favoured a white friable tissue, whereas a compact callus with a much harder texture was formed at low ratios. Roots were differentiated at intermediate values.
Callus (S2) has not been induced to undergo cellular differentiation. No soluble factors which were capable of stimulating differentiation in S2 were produced during the differentiation of S4, and S2 did not affect the ability of S4 to form roots.
Divisions seen in suspension cultures of S4 callus were mainly diploid; the cells of S2 were predominantly tetraploid. When S2 suspensions were grown in the cytokinin-free medium used for S4 a considerable number of diploid cells were seen. A number of S2 clones have been isolated and all these developed chloroplasts on exposure to continuous light and, when grown on the medium used to induce differentiation in S4, five of them showed a growth pattern in which the cells were more closely packed than is usually the case in S2 callus.
A greater degree of cell contact has also been induced by including polyethylene glycol in the medium and so restricting the availability of water to the callus. Under these conditions the cells must have a greater capacity for interaction with one another. The effect of tissue texture on the extent of cell contact and adhesion between the cells is discussed in relation to the composition of the plant cell wall and ideas on the totipotency of plant tissue cultures.