Regrowth of embryogenic tissues of <Emphasis Type="Italic">Pinus nigra</Emphasis> following cryopreservation

Embryogenic tissues of a conifer species Pinus nigra Arn. have been cryopreserved by a slow-freezing method. The effect of cryoprotective compounds such as maltose, sucrose or sorbitol (each 0.5 M) combined with 7.5% (v/v) DMSO has been tested. After thawing, the following parameters have been investigated: tissue regrowth 6 weeks after thawing, and post-thaw growth evaluated as fresh mass accumulation as well as genetic stability in post-thaw period. The parameters mentioned have been compared in both cryopreserved and non-cryopreserved tissues. Out of eight cell lines used in experiments, seven survived cryopreservation (87.5% regrowth), although cell line and treatment effects were observed. In most cell lines, sucrose or maltose pretreatments were superior over sorbitol. In the regrown cell lines, the post-thaw growth as fresh mass accumulation was not negatively influenced by cryopreservation. No genetic variation was observed in cryopreserved tissues using a RAPD approach.