CRYOPRESERVATION OF DORMANT VEGETATIVE BUDS AND GENETIC STABILITY ANALYSIS OF REGENERATED PLANTLETS IN PERSIMMON

A procedure was established for cryopreservation of dormant vegetative buds of persimmon (Diospyros kaki Thunb.). Intact buds, attached to shoot segments and subjected to preliminary stepwise freezing down to -20℃ (12 h at each of 0, -5, -10 and -20℃)，were immersed in liquid nitrogen for storage.
After thawing in agitated water at 40℃, shoot-tips of 1.5-2.0 mm in size were excised from shoot segments surface-sterilized with 0.1% HgCl₂. The shoot-tips were then plated on the modified MS medium with a half-strength nitrate supplemented with 0.2 mg L^-1 CPPU, 1.0 mg L^-1BA, 0.05 mg L^-1 IAA and 500 mg L^-1 soluble PVP. They were grown for one week in darkness before exposure to light.
The survival of shoot-tips was higher than 60%. The genetic stability of regenerated plantlets from cryopreserved shoot-tips of dormant buds was assessed by cytological and AFLP analysis.
The results showed that there were no variations in chromosome number and AFLP patterns.
This simple and reliable protocol provided a new strategy for long-term preservation of persimmon germplasm.

III International Symposium on Persimmon

P. Ai, Z. Luo

Diospyros kaki Thunb., AFLP analysis, in vitro conservation, vitrification

https://doi.org/10.17660/ActaHortic.2005.685.8