NDLI: An efficient, widely applicable cryopreservation of Lilium shoot tips by droplet vitrification

Content Provider	Springer Nature Link
Author	Yin, Zhen Fang Bi, Wen Lu Chen, Long Zhao, Bing Volk, Gayle M. Wang, Qiao Chun
Copyright Year	2014
Abstract	We report a straightforward and widely applicable cryopreservation method for Lilium shoot tips. This method uses adventitious shoots that were induced from leaf segments cultured for 4 weeks on a shoot regeneration medium containing 1 mg/l α-naphthaleneacetic acid and 0.5 mg/l thidiazuron. Shoot tips (1.5–2 mm in length) including 2–3 leaf primordia were precultured on Murashige and Skoog (MS; 1962) medium with 0.5 M sucrose for 1 day and then treated with a loading solution containing 0.4 M sucrose and 2 M glycerol for 20 min, followed by a Plant Vitrification Solution 2 (PVS2) treatment for 4 h at 0 °C. Dehydrated shoot tips were transferred onto 2.5 µl PVS2 droplets on aluminum foil strips, prior to a direct immersion into liquid nitrogen for 1 h. Frozen shoot tips were re-warmed in MS medium containing 1.2 M sucrose for 20 min at room temperature, followed by post-thaw culture for shoot regrowth. Shoot regrowth levels ranged from 42.5 % for L. longiflorum × Oriental ‘Triumphator’ to 87.5 % for L. Oriental hybrid ‘Siberia’, with a mean shoot regrowth level of 67.1 % across the six diverse Lilium genotypes tested. Histological observations found that the survival patterns were similar in cryopreserved shoot tips of ‘Triumphator’ and ‘Siberia’. Assessments using inter-simple sequence repeat markers found no differences in regenerants recovered from the control stock cultures and from cryopreserved shoot tips in ‘Triumphator’ and ‘Siberia’. This Lilium droplet-vitrification cryopreservation method is efficient, simple and widely applicable for the long-term conservation of lily genetic resources.
Starting Page	1683
Ending Page	1692
Page Count	10
File Format	PDF
ISSN	01375881
Journal	Acta Physiologiae Plantarum
Volume Number	36
Issue Number	7
e-ISSN	18611664
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2014-04-30
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Adventitious shoot Genetic stability Lilies Histological study Plant Physiology Plant Genetics & Genomics Plant Biochemistry Plant Pathology Plant Anatomy/Development Agriculture
Content Type	Text
Resource Type	Article
Subject	Physiology Plant Science Agronomy and Crop Science

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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