Abstract
New methods to establish various types of plant genetic collection are developed in Ukraine in order to preserve the existing species and varieties. This research deals with the effects of short-term hypothermic (4°C) and long-term low temperature (–196°C) storage on viability of the collection samples of garlic ( Allium sativum L. ). In the first case the culture of garlic sprout of Duchess variety was stored in vitro at the temperature of 4°C in the Murashige-Skoog (MS) medium with different mineral composition (1/2 MS and MS) in the dark for 3, 6, 9 and 12 months as well as in MS medium with various concentrations of sucrose (4.5, 6, 9 and 12%) at 22°C. The material for long-term low temperature storage of the garlic samples in liquid nitrogen were the meristems of different development type, Duchess (winter) and Manuilivskyy (spring) varieties. The apices were dehydrated in plant vitrification solution (1 M sucrose + 2 M glycerol + 2.5 M ethylene glycol) for 120 min, placed into cryovials and immersed into liquid nitrogen. The samples were stored at the temperature of –196°C for 5 years. The storage at 4°C in 1/2 MS medium provided the maximum preservation of the regenerated plants during a year (94.4%). During culturing at 22°C the highest viability (85%) was observed in the medium, containig 12% sucrose. Long-term storage of garlic meristem at –196°C did not affect the viability and growth rate if compared to the apices, thawed an hour after plunging into liquid nitrogen. The viability of vitrified-warmed garlic meristems of Duchess and Manuilivskyy varieties was about 60%. Probl Cryobiol Cryomed 2017; 27(2): 110–120
Highlights
Results and discussionOne of the modern approaches to preserve the gene pool is the in vitro storage of plant at low positive temperatures
New methods to establish various types of plant genetic collection are developed in Ukraine in order to preserve the existing species and varieties
Since there are no reports about the possible terms of in vitro storage of garlic plants allowing to preserve considerable amount viable samples sufficient for breeding, the first stage of our work was to clear this issue
Summary
One of the modern approaches to preserve the gene pool is the in vitro storage of plant at low positive temperatures. The IVAG bank of garlic collection could be created using the cultivation of plants in nutritive media 1/2 MS and MS at 4°C for 12 months without transfer to fresh nutritive medium Using this storage temperature during the whole observation period allowed to reduce the number of formed leaves to 2–3 that was considerably lower comparing to the control (9–10 pieces). 3 місяці показало, що висока концентрація сахарози rated plants we transferred the in vitro garlic plants у середовищі для депонування позитивно впливала into fresh nutritive medium MS with standard sucrose на підтримання життєздатності пробіркових рос- content (4.5%). Regenerated garlic plants which were trasferred у живильному середовищі свідчить про його при- after storage at 4°C to fresh nutritive MS medium for датність для тривалого депонування колекційних further propagation in culture, after 15 days of cultivaзразків часнику in vitro. Розвиток рослин-регенерантів часнику після 6-місячного депонування у культурі in vitro на етапі рекультивування протягом 15 діб
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