Identification of an agar constituent responsible for hydric control in micropropagation of radiata pine

Tissue cultured Pinus radiata grown on media containing agar as the gelling agent display toxic symptoms and poor long-term shoot survival, however it does have the attribute of hydric control, through a mechanism which, until now, has not been elucidated. Gelrite as an alternative gelling agent is clearly non-toxic but results in hyperhydric (vitrified) tissues. In an effort to overcome these problems, the controlling mechanism found in agar was examined. Hydric control was shown to be effected by a non-gelling, cold-water soluble constituent of a commercial agar, rather than by physical properties of the gel. It could be separated from low molecular weight components of the agar responsible for the toxic symptoms by dialysis. It was identified as being an agaroid-type xylogalactan bearing pyruvate and sulphate substituents. Improved management of gelling agents in culture medium has contributed substantially to a thirty fold increase in propagation rates.