Shoot tips of two in vitro grown interspecific Prunus rootstocks, Fere
ley-Jaspi (R) and Ferlenain-Plumina (R), were successfully cryopreserv
ed using a two step freezing method. Excised shoot tips from hardened
plantlets were precultured for 24 h on solid medium (modified half str
ength Murashige and Skoog medium (MS)) supplemented with 5% dimethylsu
lfoxide and 2% proline. They were transferred to 1.2-ml cryotubes and
treated with 0.5 ml of modified PVS2 cryoprotective solution for 20-40
min. Cryotubes were then cooled at 1 degrees C/min to -40 degrees C b
efore being immersed in liquid nitrogen (LN(2)), After 1 min warming a
t 40 degrees C, shoot tips were rinsed with MS medium containing 1.2 M
sucrose and then plated. First regrowth occurred after 5 days and was
evaluated 21 days after plating. The average rate of shoot formation
was 69% and 74% for Fereley and Ferlenain respectively, compared with
86% for controls of both hybrids, The roles of several factors, especi
ally hardening procedures and meristem size, in the improvement of sur
vival rates, were discussed.