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Research Article

Efficient Cryopreservation of Lilium spp. Shoot Tips using Droplet-vitrification

Jung Yoon Yi1,*, Gi An Lee1, Jong Wook Chung1, Sok Young Lee1, Ki Byung Lim2
Plant Breeding and Biotechnology 2013;1(2):131-136.
Published online: June 30, 2013

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*Corresponding author: JungYoon Yi, naaeskr@korea.kr, Tel: +82-31-299-1886, Fax: +82-31-299-1839
• Received: June 14, 2013   • Revised: June 19, 2013   • Accepted: June 19, 2013

Copyright © 2013 The Korean Society of Breeding Science

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Efficient Cryopreservation of Lilium spp. Shoot Tips using Droplet-vitrification
Plant Breed. Biotech.. 2013;1(2):131-136.   Published online June 30, 2013
DOI: https://doi.org/10.9787/PBB.2013.1.2.131
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Efficient Cryopreservation of Lilium spp. Shoot Tips using Droplet-vitrification
Plant Breed. Biotech.. 2013;1(2):131-136.   Published online June 30, 2013
DOI: https://doi.org/10.9787/PBB.2013.1.2.131
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Efficient Cryopreservation of Lilium spp. Shoot Tips using Droplet-vitrification
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Fig. 1 Cryopreservation of lily shoot tips using droplet-vitrification. A) bulb-scale culture for producing shoot tips for cryopreservation; B) dehydration of shoot tips in vitrification solution droplets on aluminum foil strips; and C) plant regeneration from cryopreserved shoot tips 2 weeks after cryopreservation.

Fig. 1

Efficient Cryopreservation of Lilium spp. Shoot Tips using Droplet-vitrification

Compositions of the vitrification and loading solutions used in this study.

Solution Composition Reference
PVS2 30%Gz) + 15%DMSOy) + 15%EGx) + 13.7%Sucw) in MS Sakai et al. 1990
Modified PVS2 37.5%G + 15%DMSO + 15%EG + 22.5%Suc in MS -
PVS3 50%G + 50%Suc in MS Nishizawa et al. 1993
LD1 35% of PVS3
LD2 40% of PVS3

z)G, glycerol;

y)DMSO, dimethyl sulfoxide;

x)EG, ethylene glycol;

w)Suc, sucrose

Effect of cold-hardening on the survival and regeneration percentage of shoot tips of the five accessions of Lilium spp.

Accession No.
GBL0089 GBL0099 GBL0202 GBL0474 GBL0518
Survival rate (%) Cold-hardened 89.5 ± 5.83z) 67.9 ± 5.31 83.3 ± 4.33 57.7 ± 3.67 67.9 ± 4.87
Control 43.3 ± 2.21 31.3 ± 2.87 43.3 ± 1.22 24.3 ± 1.13 40.2 ± 3.25
Regeneration rate (%) Cold-hardned 87.5 ± 3.32 64.3 ± 4.34 77.8 ± 2.41 52.7 ± 2.83 60.7 ± 4.63
Control 33.3 ± 1.11 28.5 ± 2.17 29.5 ± 3.33 19.9 ± 1.82 28.2 ± 2.17

z)Mean ± standard deviation

The effect of different loading solutions and treatment times on the survival and regeneration rates of the control (−LN) and cryopreserved (+LN) lily shoot tips of the Lilium hybrid Carmina.

Loading solution −LN +LN
Survival (%) Regeneration (%) Survival (%) Regeneration (%)
Non-loading 98.3 97.3 45.3dz) 38.3d
LD1 40 min 99.3 97.8 80.3a 78.5ab
LD1 60 min 98.2 97.7 89.5a 87.5a
LD2 40 min 97.9 96.9 66.3b 63.5b
LD2 60 min 99.1 98.3 53.2c 50.3bc

z)Mean separation within columns by Duncan’s multiple range test at 5% level

The effect of different vitrification solutions and treatment times on the survival and regeneration rates of the control (−LN) and the cryopreserved (+LN) shoot tips of the Lilium hybrid Carmina.

Vitrification solution & time −LN +LN
Survival (%) Regeneration (%) Survival (%) Regeneration (%)
PVS2 90 min 99.9 95.3 25.5c 18.3d
Modified PVS2 90 min 97.1 96.9 49.8bc 32.3c
Modified PVS2 150 min 95.3 93.7 51.3bc 33.3c
PVS3 180 min 99.3 98.1 63.3b 56.8bc
PVS3 240 min 98.2 97.7 89.5a 87.5a
PVS3 360 min 95.7 92.5 73.1ab 67.8b

z; Mean separation within columns by Duncan’s multiple range test at 5% level

Survival and regeneration rates of the shoot tips of the five lily accessions cryopreserved using droplet-vitrification.

Accession no. Scientific name Accession name Survival rate (%) Regeneration rate (%)
GBL0089 Lilium hybrid Carmina 89.5 ± 4.43z) 87.5 ± 5.89
GBL0099 Lilium hybrid Crystal Light 67.9 ± 5.05 64.3 ± 0.01
GBL0202 Lilium callosum Sinomartagon section 83.3 ± 7.86 77.8 ± 5.46
GBL0474 Lilium hybrid Santander 57.7 ± 7.36 52.7 ± 5.33
GBL0518 Lilium hybrid Marrero 67.9 ± 5.05 60.7 ± 3.15

z)Mean ± standard deviation

Table 1 Compositions of the vitrification and loading solutions used in this study.

G, glycerol;

DMSO, dimethyl sulfoxide;

EG, ethylene glycol;

Suc, sucrose

Table 2 Effect of cold-hardening on the survival and regeneration percentage of shoot tips of the five accessions of Lilium spp.

Mean ± standard deviation

Table 3 The effect of different loading solutions and treatment times on the survival and regeneration rates of the control (−LN) and cryopreserved (+LN) lily shoot tips of the Lilium hybrid Carmina.

Mean separation within columns by Duncan’s multiple range test at 5% level

Table 4 The effect of different vitrification solutions and treatment times on the survival and regeneration rates of the control (−LN) and the cryopreserved (+LN) shoot tips of the Lilium hybrid Carmina.

z; Mean separation within columns by Duncan’s multiple range test at 5% level

Table 5 Survival and regeneration rates of the shoot tips of the five lily accessions cryopreserved using droplet-vitrification.

Mean ± standard deviation

Table 1

Table 2

Table 3

Table 4

Table 5

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