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United States Department of Agriculture

Agricultural Research Service

Research Project: RESEARCH TO DEVELOP STRATEGIES AND TECHNOLOGIES FOR PRESERVING PLANT GENETIC DIVERSITY IN EX SITU GENEBANKS

Location: Plant Germplasm Preservation Research Unit

Title: Shoot regeneration and embryogenesis in lily shoot tips cryopreserved by droplet vitrification

Authors
item Yin, Zhen-Fang -
item Chen, Long -
item Bi, Wen-Lu -
item VOLK, GAYLE
item Wang, Qiao-Chun -

Submitted to: International Society for Horticultural Science Meeting
Publication Type: Abstract Only
Publication Acceptance Date: August 11, 2013
Publication Date: August 11, 2013
Citation: Yin, Z.F., Chen, L., Bi, W., Volk, G.M., Wang, Q. 2013. Shoot regeneration and embryogenesis in lily shoot tips cryopreserved by droplet vitrification. International Society for Horticultural Science Meeting. p.55.

Technical Abstract: Shoot regeneration and embryogenesis were, for the first time, achieved directly in shoot tips of Lilium Oriental hybrid ‘Siberia’ following cryopreservation by droplet-vitrification. Shoot tips (2 mm in length) including 2-3 leaf primordia were excised from 4-week-old adventitious shoots directly regenerated from basal leaf segments and precultured on MS containing 0.5 M sucrose for 1 day. The precultured shoot tips were treated with a loading solution composed of solid MS containing 0.4 M sucrose and 2 M glycerol for 20 min at room temperature and dehydrated for 4 h by PVS2 at 0 °C. Dehydrated shoot tips were transferred onto droplets made on sterile aluminum foil (7 x 20 mm), each droplet containing 3.5 µl PVS2 and single shoot tip, prior to direct immersion into liquid nitrogen for 1 h. Following cryoexposure, foil strips with shoot tips were incubated for 20 min in an unloading solution comprised of MS containing 1.2 M sucrose at room temperature. Cryoexposed shoot tips were post-cultured on recovery media in the dark for 3 days and then transferred to light conditions for shoot recovery. Dark conditions were also used to stimulate embryogenic callus formation. Shoots were directly regenerated from cryoexposed shoot tips that were post-cultured on recovery medium containing TDZ and NAA and the highest shoot regrowth rate was about 90%. Embryogenic callus was obtained on recovery medium containing KT and NAA, and the highest frequency of embryogenic callus formation was about 70%. Embryogenic callus formation in cryopreserved shoot tips is significant for studies on plant development, regeneration and genetic transformation. Applicability of the droplet-vitrification cryopreservation method developed in the present study is being tested in five other commercially important Lilium species or hybrids.

Last Modified: 9/29/2014
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