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P L A N T C E L L , O R G A N , A N D T IS S U E C O L L E C T IO N S , T H E M E T H O D S O F

G E R M P L A S M P R E S E R V A T IO N

CRYOPRESERVATIONOFRASPBERRY (

RUBUS IDAEUSL

.)APICAL

SHOOTTIPS

O .N .Vysotskaya, A .I.M ochammed, R .G .Butenko

Timiriasev Institute o f PlantPhysiology Russian Academy ofSciences, Russia

In vitro

plants storage based on periodically change o f cultural medium

is w ide ly used fo r maintenance o f patogennegative clones o f vegetative

propagated horticultural crops. Long term cryopreservation o f plant material

excluding periodically change o f nutrient media helps to reduce partly labour

expediture.

The experimental results have showen that cold hardening (4-8°C,

darkness) o f raspberry mother plants on agar medium (5 -6% sugars and

cytokinins) provided the succesful cryopreservation o f excised apices (0,5-

2,0 mm ). The explants isolated from hardened

in vitro

plantlets were

precultivated on liguid medium with 5-7% dimethylsulfoxide at 18-22°C fo r

18 h.. Then they were transferred to cold cryoprotectant mixture (5% sugars

and 7-9% dimethylsulfoxide) in cryotubes (Nunc) and frosen slowly or rapidly

by fina lly immersed to liguid nitrogen at -196°C . In a hour later o r more

samples were taken out liquid nitrogen and thawed in 40°C water. A fte r

cryoconservation viable meristems o f raspberry (cvs. Skromnitza, Lastochka,

Poland) regenerated w ithout callus formation stage. A fte r such treatment

they produced multiply viab ily progeny on cytokinin containing nutrient

medium.

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