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1 Biotechnologia

1 2001

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Cryopreservation of plant tissue culture

100%

Makowska Z. , Kotlinska T.

Biotechnologia

2001

nr 2

126-134

Three methods of cryopreservation: vitrification, encapsulation and slow freezing were discussed. The factors having influence on long-term storage in liquid nitrogen as selection of plant explants, media, optimal condition for regeneration and growth of plants were described. Practical utilisation of vitrification method and obtained results on garlic (Allium sativum L.) are given as an example. Vitrification involves treatment of samples with cryoprotective substances, dehydration with highly concentrated vitrification solution, aimed at water removal from the cells in order to protect them against destruction because of liquid nitrogen influence. This method offers a possibility to store plant material for a long time without any modification and contamination. The aim of the study is to develop a useful method for cryopreservation of apical meristems of garlic (Allium sativum L.) bulbils. The apical meristems of garlic bulbils were treated with PVS 3 solution containing 50% sucrose and 50% glicerol. The time of treatment of explants with PVS 3 was from 0 min to 240 min, then samples were plunged into liquid nitrogen (LN2) where they were kept for 1 hour or 30 days. Rapid heat up was achieved by plunging the samples in a 45oC water-bath until cryoprotectant solution became liquid. After cryopreservation, survival and regrowth on MS (Murashige and Skoog) medium were observed. Results showed that the best kind of explants giving high percentage of regeneration were meristems with two leaves primordia about 1 mm of diameter and about 3 mm in length.