IMPROVING SHOOT TIP CRYOPRESERVATION PROTOCOL TO SET UP A CRYOGENIC BANK FOR POTATO CULTIVARS AND HYBRIDS
Main Article Content
Authors
S.V. Kushnarenko
Institute of Plant Biology and Biotechnology, 45, Timiryazev str., Almaty, 050040, Kazakhstan
N.V. Romadanova
Institute of Plant Biology and Biotechnology, 45, Timiryazev str., Almaty, 050040, Kazakhstan
M.O. Bekebaeva
Institute of Plant Biology and Biotechnology, 45, Timiryazev str., Almaty, 050040, Kazakhstan
G.N. Matakova
Institute of Plant Biology and Biotechnology, 45, Timiryazev str., Almaty, 050040, Kazakhstan
Abstract
Cryopreservation of cells, tissues, and organs in liquid nitrogen at -196°C is widely practiced around the world for long-term conservation of plant genetic resources. Cryopreservation of shoot tips is particularly relevant for the conservation of vegetatively propagated crops including potatoes because liquid nitrogen exposure not only allows to obtain a genetic copy of the donor material, but also improves plants from pathogens like bacteria, fungi, and viruses. An effective cryopreservation protocol for potato shoot tips based on PVS2-vitrification is presented in this paper. About 1.5-2.0 mm long shoot tips dissected from plantlets, grown in vitro, were precultured for a day, on Murashige-Skoog medium supplemented with 0.3 M sucrose, in a plant growth room at 24°C and subjected to 16 h light/8 h dark photoperiod (light intensity of 25 µmol/m2/s). Subsequently, the shoot tips were loaded in 2 M glycerol with 0.4 M sucrose for 20 min. After treatment in PVS2 cryoprotectant for 30 min at room temperature, shoot tips were immersed in liquid nitrogen. Recovery of shoot tips following cryopreservation in these conditions varied between 43.1 to 59.8% depending on the genotype. This elaborated protocol of shoot tips cryopreservation is recommended to set up a cryogenic bank in Kazakhstan for foreign potato cultivars and hybrids.
Keywords
potato, cryopreservation, shoot tips, PVS2-vitrification method, cryogenic bank
Article Details
References
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