Journal
SOUTH AFRICAN JOURNAL OF BOTANY
Volume 77, Issue 2, Pages 397-403Publisher
ELSEVIER
DOI: 10.1016/j.sajb.2010.10.005
Keywords
Apical meristem; Cryopreservation; Droplet-vitrification method; Lily (Lilium L); Orthogonal experimental design
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Funding
- National Key Technology R&D Program in accordance with the 11th five year plan of P. Rep. of China [2006BA013B10]
- BasiC scientific Research Operation for Central Commonwealth Scientific Research Academy
- Ministry of Finance, P. Rep. of China [092060302-7]
- Ministry of Agriculture, People's Republic of China [NB08-2130135]
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In this study, apical meristems from adventitious buds of three lily (Lillium L.) cultivars were successfully cryopreserved by droplet-vitrification. The most effective techniques were as follows. Excised meristems from in vitro plantlets which had been sub-cultured for about 2 months were cold-hardened at 4 degrees C for I week, precultured on MS medium supplemented with 0.3 mol L-1 sucrose for 2 days, osmoprotected in loading solution for 20-40 min at room temperature and then soaked in PVS2 solution for 90-120 min at 0 degrees C, frozen in microdroplets of vitrification solution placed on aluminium foils, which were immersed rapidly in liquid nitrogen. The meristems were then rapidly rewarmed by dilution solution, transferred to regeneration medium and stored in the dark for two weeks at 20 degrees C, and then cultured under white fluorescent light at an intensity of 2000 lux, with a 16 h photoperiod at 20 degrees C. The highest post-thaw survival percentages of three cultivars 'Siberia' (Lilium x siberia), Lilium lancifolium Thunb. and 'Snow Queen' Lilium x longiflorum were 65.0%, 83.8% and 43.3%, and regeneration percentages were 62.0%, 67.6% and 35.0%, respectively. The study demonstrated that cryopreservation by droplet-vitrification increased survival and regeneration percentages of certain lily cultivars compared with vitrification. Thus to cryopreserve lily meristems, droplet-vitrification method is preferable to the vitrification method. Crown Copyright (C) 2010 Published by Elsevier B.V. on behalf of SAAB. All rights reserved.
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