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2014 | 36 | 07 |
Tytuł artykułu

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

Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
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.
Słowa kluczowe
Wydawca
-
Rocznik
Tom
36
Numer
07
Opis fizyczny
p.1683-1692,fig.,ref.
Twórcy
autor
  • State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Genetic Improvement of al Crops of Northwest China, College of Horticulture, Northwest A and F University, 712100 Yangling, Shaanxi, People's Republic of China
autor
  • State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Genetic Improvement of al Crops of Northwest China, College of Horticulture, Northwest A and F University, 712100 Yangling, Shaanxi, People's Republic of China
autor
  • State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Genetic Improvement of al Crops of Northwest China, College of Horticulture, Northwest A and F University, 712100 Yangling, Shaanxi, People's Republic of China
autor
  • State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Genetic Improvement of al Crops of Northwest China, College of Forestry, Northwest A and F University, 712100 Yangling, Shaanxi, People's Republic of China
autor
  • USDA-ARS National Center for Genetic Resources Preservation, 111 S. Mason St., Fort Collins, 80521 CO, USA
autor
  • State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Genetic Improvement of al Crops of Northwest China, College of Horticulture, Northwest A and F University, 712100 Yangling, Shaanxi, People's Republic of China
Bibliografia
  • Bacchetta L, Remotti PC, Bernardini C, Saccardo F (2003) Adventitious shoot regeneration from leaf explants and stem nodes of Lilium. Plant Cell Tissue Organ Cult 74:37–44
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  • Chen XL, Li JH, Xin X, Zhang ZE, Xin PP, Lu XX (2011) Cryopreservation of in vitro-grown apical meristems of Lilium by droplet-vitrification. S Afr J Bot 77:397–403
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  • Kim HH, Lee YG, Park SU, Lee SC, Baek HJ, Cho EG, Engelmann F (2009) Development of alternative loading solutions in droplet-vitrification procedures. Cryo Lett 30:291–299
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  • Scowcroft WR (1984) Genetic variability in tissue culture: impact on germplasm conservation and utilization. IBPGR, Rome, Italy
  • Wang QC, Valkonen JPT (2009) Cryotherapy of shoot tips: novel pathogen eradication method. Trends Plant Sci 14:119–122
  • Wang QC, Panis B, Engelmann F, Lambardi M, Valkonen JPT (2009) Cryotherapy of shoot tips: a technique for pathogen eradication to produce healthy planting materials and prepare healthy plant genetic resources for cryopreservation. Ann Appl Biol 154:351–363
  • Wang B, Zhang ZB, Yin ZF, Feng CH, Wang QC (2012) Novel and potential application of cryopreservation to plant genetic transformation. Biotech Adv 30:604–612
  • Yin ZF, Zhao B, Bi WL, Chen L, Wang QC (2013) Direct shoot regeneration from basal leaf segments of Lilium and assessment of genetic stability in regenerants by ISSR and AFLP markers. In Vitro Cell Dev Biol Plant 49:333–342
Typ dokumentu
Bibliografia
Identyfikatory
Identyfikator YADDA
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