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1

Cryopreservation of plant tissue culture

100%

Makowska Z., Kotlinska T.

Biotechnologia

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2001

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issue 2

126-134

EN

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.

2

Methods of preservation of in vitro plant cell culture

88%

Marecik R., Kroliczak P.

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issue 1

105-116

EN

Tissue culture has been utilized for producing virus free plants for vegetative propagation and genetic engineering. In addition, altered cells lines showing higher productivity of secondary metabolities can be obtained using tissue culture. The wide use of tissue culture requires the development of new preservation techniques for in vitro culture material. One of this method is growth reduction achieved by modifying various parameters such as: temperature, culture medium, gasous environment. However, crypreservation (i. e. storage in liquid nitrogen. -196?C) is the only method available nowadays for long-term conservation. New cryopreservation techniques such as encapsulation-dehydratation, vitrification and desiccation helped expend the list of plant species that can tolerate low temperatures and are characterized by a normal rate of growth. Each step of the cryopreservation procces requires specific conditions.

3

Vitrification of mammalian oocytes and embryos

63%

Smorag Z., Gajda B.

Biotechnologia

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1995

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issue 3

68-83

EN

Vitrification is a new approach to oocyte and embryo cryoconservation.It consists in the solidification of a solution caused by draastic increase in viscosity during cooling and not by crystalization.The application of this approach to cryoconservation of oocytes and embryos of different species depends upon the development of proper procedures and non-toxic media.From the technical point of view, the vitrification method is simple and relatively easily applicable under field conditions.The authors review the current procedures applied to oocytes and embryos of laboratory and farm animals.

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3 Biotechnologia

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1 Gajda B.

1 Kotlinska T.

1 Kroliczak P.

1 Makowska Z.

1 Marecik R.

1 Smorag Z.

Cryopreservation of plant tissue culture

Methods of preservation of in vitro plant cell culture

Vitrification of mammalian oocytes and embryos