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Research Article

Cryopreservation of Winter-dormant Apple Buds Using Two-step Freezing

Jung Yoon Yi1,*, Gi An Lee1, Sok Young Lee1, Jong Wook Chung1, Sup Shin2
Plant Breeding and Biotechnology 2013;1(3):283-289.
Published online: September 30, 2013

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*Corresponding author: JungYoon Yi, naaeskr@korea.kr, Tel: +82-31-299-1886, Fax: +82-31-299-1839
• Received: September 4, 2013   • Revised: September 24, 2013   • Accepted: September 24, 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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Cryopreservation of Winter-dormant Apple Buds Using Two-step Freezing
Plant Breed. Biotech.. 2013;1(3):283-289.   Published online September 30, 2013
DOI: https://doi.org/10.9787/PBB.2013.1.3.283
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Cryopreservation of Winter-dormant Apple Buds Using Two-step Freezing
Plant Breed. Biotech.. 2013;1(3):283-289.   Published online September 30, 2013
DOI: https://doi.org/10.9787/PBB.2013.1.3.283
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Cryopreservation of Winter-dormant Apple Buds Using Two-step Freezing
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Fig. 1 Plant regenerated from cryopreserved winter bud of ‘Hongro’ apple stem segments: A) shoot formation from cryopreserved apple winter bud by in vitro cutting; B) budding of cryopreserved apple winter bud onto young seedling; C) comparison of the growth two months after budding of winter apple buds cryopreserved without LN (left) and with LN treatment (right).

Fig. 1

Cryopreservation of Winter-dormant Apple Buds Using Two-step Freezing

Effect of desiccation on the regeneration rate of winter buds of ‘Hongro’ and ‘Chukwang’ cryopreserved in liquid nitrogen.

Treatment Regeneration Rate (%)
‘Hongro’ ‘Chukwang’
−LN +LN −LN +LN
Non-Dry, Non-freezing 100.0 - 100.0 -
Non-Dry, freezing 97.7 44.6 d z 97.8 47.3 d
30%, freezing 95.9 70.3 b 98.2 67.3 b
35%, freezing 98.3 85.4 a 96.2 81.5 a
40% freezing 96.9 67.7 c 99.9 60.3 c

zMean separation within columns by Duncan’s multiple range test at 5% level by R project (R version 3.0.1) for statistical computing.

Effect of the pre-freezing temperature on the regeneration rate of winter buds of ‘Hongro’ and ‘Chukwang’ cryopreserved in liquid nitrogen.

Pre-freezing Temperature Regeneration rates (%)
‘Hongro’ ‘Chukwang’
−LN +LN −LN +LN
−25°C 97.5 55.3 c z 98.3 59.3 c
−30°C 100 73.3 b 98.1 71.2 b
−35°C 97.7 85.4 a 100 81.5 a
−40°C 96.5 58.3 c 97.2 44.3 d

zMean separation within columns by Duncan’s multiple range test at 5% level by R project (R version 3.0.1) for statistical computing

Effect of the pre-freezing rate to −35°C on the regeneration rate of winter buds of ‘Hongro’ and ‘Chukwang’ cryopreserved in liquid nitrogen.

Pre-freezing Rates Recovery rates (%)
‘Hongro’ ‘Chukwang’
−LN +LN −LN +LN
0.5°C/h 95.9 61.2 b z 98.8 58.3 b
1°C/h 97.4 85.4 a 100 81.5 a
2°C/h 97.1 55.2 b 98.2 59.3 b

zMean separation within columns by Duncan’s multiple range test at 5% level by R project (R version 3.0.1) for statistical computing

List of regenerated winter buds of 14 apple cultivars cryopreserved in liquid nitrogen vapor.

Accession number Scientific name Accession name Recovery rates (%) No. of accessions conserved
249799 Malus domestica Hongso 41.1±3.09 150
225513 Malus domestica Seohong 43.3±5.77 226
249804 Malus domestica Picnic 43.5±6.27 90
226029 Malus domestica Red Gold 43.5±8.84 65
255854 Malus domestica Hwangok 43.9±6.26 250
254759 Malus domestica Hwarang 46.7±0.01 320
249800 Malus domestica Hongan 48.6±9.90 100
249803 Malus domestica Greenball 52.2±8.01 30
249255 Malus domestica Gamhong 55.0±0.01 400
225511 Malus domestica Seokwang 56.9±2.74 560
255536 Malus domestica Yeohong 63.1±3.37 280
225602 Malus domestica Hwahong 65.8±5.98 100
225578 Malus domestica Chukwang 81.5±4.41 100
225596 Malus domestica Hongro 85.4±6.01 100
Average 55.0
Table 1 Effect of desiccation on the regeneration rate of winter buds of ‘Hongro’ and ‘Chukwang’ cryopreserved in liquid nitrogen.

Mean separation within columns by Duncan’s multiple range test at 5% level by R project (R version 3.0.1) for statistical computing.

Table 2 Effect of the pre-freezing temperature on the regeneration rate of winter buds of ‘Hongro’ and ‘Chukwang’ cryopreserved in liquid nitrogen.

Mean separation within columns by Duncan’s multiple range test at 5% level by R project (R version 3.0.1) for statistical computing

Table 3 Effect of the pre-freezing rate to −35°C on the regeneration rate of winter buds of ‘Hongro’ and ‘Chukwang’ cryopreserved in liquid nitrogen.

Mean separation within columns by Duncan’s multiple range test at 5% level by R project (R version 3.0.1) for statistical computing

Table 4 List of regenerated winter buds of 14 apple cultivars cryopreserved in liquid nitrogen vapor.

Table 1

Table 2

Table 3

Table 4

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