Skip to main content
Log in

Dormancy development during cold hardening of in vitro cultured Malus domestica Borkh. plants in relation to their frost resistance and cryotolerance

  • Original Paper
  • Published:
Trees Aims and scope Submit manuscript

Abstract

The development of dormancy, frost resistance and cryotolerance of in vitro apple plants (Malus domestica Borkh.), cv. Greensleeves during their exposure to cold hardening was studied. In vitro cultures were cold hardened at 4°C under a short photoperiod up to 25 weeks. The dormancy status, non-structural saccharides, proline, water content and frost resistance were evaluated for optimization of cryopreservation. According to regrowth tests, in vitro cultures exhibited endogenous dormancy after the maximal frost resistance was reached. The highest regeneration ability of shoot tips after cryopreservation by encapsulation–dehydration method coincided with the period of the plant’s dormant state and maximum of frost resistance. All studied saccharides and proline exhibited the maximal values at the beginning of cold hardening and/or the dormancy phase. Contrary to the accumulation of saccharides and proline, water content showed the inverse time behaviour. According to these results, the cold hardening-induced endodormancy, high frost resistance and accumulation of saccharides and proline are the important prerequisites for the successful cryopreservation of shoot tips of in vitro grown apple plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Aguettaz P, Paffen A, Delvallée I, Piet Linde P, Geert-Jan Klerk GJ (1990) The development of dormancy in bulblets of Lilium speciosum generated in vitro. Plant Cell Tiss Organ Cult 22(3):167–172

    Article  Google Scholar 

  • Allona A, Ramos I, Ibáñez A, Contreras A, Casado A, Aragoncillo C (2008) Molecular control of winter dormancy establishment in trees. Span J Agric Res 6(Sp.Iss.SI):201–210

    Google Scholar 

  • Arora R, Rowland LJ, Panta GR (1997) Chill-responsive dehydrins in blueberry: are they associated with cold hardiness or dormancy transitions? Physiol Plant 101(1):8–16

    Article  CAS  Google Scholar 

  • Arora R, Rowland LJ, Tanino K (2003) Induction and release of bud dormancy in woody perennials: a science comes of age. HortScience 38:911–921

    Google Scholar 

  • Baldwin BD, Bandara MS, Tanino KK (1998) Is tissue culture a viable system with which to examine environmental and hormonal regulation of cold acclimation in woody plants? Physiol Plant 102:201–209

    Article  CAS  Google Scholar 

  • Barthe P, Bulard C (1982) Influence of agar and sucrose on the behaviour of dormant apple embryos cultured in vitro. New Phytol 91:517–529

    Article  CAS  Google Scholar 

  • Barthe P, Bulard C (1983) Anaerobiosis and release from dormancy in apple embryos: leaching of (±) [14C] abscisic acid and its metabolites under aerobic and anaerobic conditions. Plant Physiol 72(4):1005–1010

    Article  PubMed  CAS  Google Scholar 

  • Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207

    Article  CAS  Google Scholar 

  • Bewley JD, Black M (1994) Seeds: physiology of development and germination, 2nd edn. Plenum Press, New York

    Google Scholar 

  • Borkowska B (1981) Dormancy and development of apple axillary buds investigated in vitro. Acta Hort (ISHS) 120:161–166

    Google Scholar 

  • Borkowska B (1986) Dormancy of sour cherry plantlets propagated by in vitro culture technique. Acta Hort (ISHS) 179:175–176

    Google Scholar 

  • Borkowska B, Michalczuk L (2000) Charakterystyka spoczynku truskawek rozmnażanych in vitro. Zeszyty Problemowe Postępów Nauk Rolniczych 473:39–45

    Google Scholar 

  • Bubán T, Faust M (1995) New aspects of bud dormancy in apple trees. Acta Hort (ISHS) 395:105–112

    Google Scholar 

  • Caswell KL, Tyler NJ, Stushnoff C (1986) Cold hardening of in vitro apple and Saskatoon shoot cultures. HortScience 21:1207–1209

    CAS  Google Scholar 

  • Chang Y, Reed BM (2000) Extended alternating-temperature cold acclimation and culture duration improve pear shoot cryopreservation. Cryobiology 40:311–322

    Article  PubMed  CAS  Google Scholar 

  • Cox SE, Stushnoff C (2001) Temperature-related shifts in soluble carbohydrate content during dormancy and cold acclimation in Populus tremuloides. Can J For Res 31:730–737

    CAS  Google Scholar 

  • Debeaujon I, Koornneef M (2000) Gibberellin requirement for Arabidopsis seed germination is determined both by testa characteristics and embryonic abscisic acid. Plant Physiol 122:415–424

    Article  PubMed  CAS  Google Scholar 

  • Delvallée I, Paffen A, De Klerk GJ (1990) The development of dormancy in bulblets of Lilium speciosum generated in vitro. II. The effect of temperature. Physiol Plant 80(3):431–436

    Article  Google Scholar 

  • Dereuddre J, Hassen N, Blandin S, Kaminski M (1991) Resistance of alginate-coated somatic embryos of carrot (Daucus carota L.), to desiccation and freezing in liquid nitrogen. 2. Thermal analysis. Cryo Lett 12:135–148

    Google Scholar 

  • Du YC, Nose A (2002) Effects of cold-hardening temperature on the activity of enzymes of sucrose synthesis and the accumulation of saccharides in leaves of three sugarcane cultivars differing in cold sensitivity. Photosynthetica 40(3):389–395

    Article  CAS  Google Scholar 

  • Duncan DR, Widholm JM (1987) Proline accumulation and its implication in cold tolerance of regenerable maize callus. Plant Physiol 83:703–708

    Article  PubMed  CAS  Google Scholar 

  • Faust M, Liu D, Line MJ, Stutte GW (1995a) Conversion of bound to free water in endodormant buds of apple is an incremental process. Acta Hort (ISHS) 395:113–118

    Google Scholar 

  • Faust M, Liu D, Wang SY, Stutte GW (1995b) Involvement of apical dominance in winter dormancy of apple buds. Acta Hort (ISHS) 395:47–56

    Google Scholar 

  • Halmagyi A, Valimareanu S, Coste A, Deliu C, Isac V (2010) Cryopreservation of Malus shoot tips and subsequent plant regeneration. Romanian Biotechnol Lett 15(1) Suppl. S:79–85

    Google Scholar 

  • Hauagge R, Cummins JN (1991) Genetics of length of dormancy period in Malus vegetative buds. J Am Soc Hortic Sci 116:121–126

    Google Scholar 

  • Hausman JF, Evers D, Thiellement H, Jouve L (2000) Compared responses of poplar cuttings and in vitro raised shoots to short-term cold-hardening treatments. Plant Cell Rep 19:954–960

    Article  CAS  Google Scholar 

  • Heide OM, Prestrud AK (2005) Low temperature, but not photoperiod, controls growth cessation, dormancy induction and release in apple and pear. Tree Physiol 25:109–114

    Article  PubMed  CAS  Google Scholar 

  • Imanishi HT, Suzuki T, Masuda K, Harada T (1998) Accumulation of raffinose and stachyose in shoot apices of Lonicera caerulea L. during cold acclimation. Sci Hortic 72(3–4):255–263

    Article  CAS  Google Scholar 

  • Jones KS, Paroschy J, McKersie BD, Bowley SR (1999) Carbohydrate composition and freezing tolerance of canes and buds in Vitis vinifera. J Plant Physiol 155:101–106

    Article  CAS  Google Scholar 

  • Kalcsits LA, Silim S, Tanino K (2009) Warm temperature accelerates short photoperiod-induced growth cessation and dormancy induction in hybrid poplar (Populus × ssp.). Trees 23:971–979

    Article  Google Scholar 

  • Keller JD, Loescher WH (1989) Nonstructural carbohydrate partitioning in perennial parts of sweet cherry. J Amer Soc Hortic Sci 114:969–975

    CAS  Google Scholar 

  • Kushad MM, Yelenosky G (1987) Evaluation of polyamine and proline levels during low temperature acclimation of citrus. Plant Physiol 84:692–695

    Article  PubMed  CAS  Google Scholar 

  • Kushnarenko SV, Romadanova NV, Reed BM (2009) Cold acclimation improves regrowth of cryopreserved apple shoot tips. CryoLetters 30(1):47–54

    PubMed  CAS  Google Scholar 

  • Kwang-Soo Kim K-S, Davelaar E, De Klerk G-J (1994) Abscisic acid controls dormancy development and bulb formation in lily plantlets regenerated in vitro. Physiol Plant 90:59–64

    Article  Google Scholar 

  • Lang GA, Early JD, Martin GC, Darnell RL (1987) Endo-, para-, and ecodormancy: physiological terminology and classification for dormancy research. HortScience 22:371–377

    Google Scholar 

  • Lavee S, May P (1997) Dormancy of grapevine buds—facts and speculation. Aust J Grape Wine Res 3:31–46

    Article  CAS  Google Scholar 

  • Li C, Viherä-Aarnio A, Puhakainen T, Junttila O, Heino P, Palva ET (2003) Ecotype-dependent control of growth, dormancy and freezing tolerance under seasonal changes in Betula pendula Roth. Trees Struct Funct 17(2):127–132

    CAS  Google Scholar 

  • Maksimović V, Jevremović S, Trifunović M, Subotić A (2008) Effect of low temperature on rooting rate and carbohydrate content of Frittilaria Meleagris bulbs formed in culture in vitro. Arch Biol Sci 60(1):5–6

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497

    Article  CAS  Google Scholar 

  • Nougarède A, Silveira CE, Rondet P (1996) In nature dormant buds and in vitro dormant-like buds of Fraxinus excelsior L. Protoplasma 190(1–2):16–24

    Article  Google Scholar 

  • Palonen P, Lindén L (1999) Dormancy, cold hardiness, dehardening and rehardening in selected red raspberry cultivars. J Amer Soc Hortic Sci 124:341–346

    Google Scholar 

  • Palonen P, Buszard D, Donnelly D (2000) Changes in carbohydrates and freezing tolerance during cold acclimation of red raspberry cultivars grown in vitro and in vivo. Physiol Plant 100:393–401

    Article  Google Scholar 

  • Piola F, Label P, Vergne P, von Aderkas P, Rohr R (1998) Effects of endogenous ABA levels and temperature on cedar (Cedrus libani Loudon) bud dormancy in vitro. Plant Cell Rep 18:279–283

    Article  CAS  Google Scholar 

  • Rohde A, Bhalerao RP (2007) Plant dormancy in the perennial context. Trends Plant Sci 12:217–223

    Article  PubMed  CAS  Google Scholar 

  • Sedlák J, Paprštein F, Bilavčík A, Zámečník J (2001) Adaptation of apple and pear plants to in vitro conditions and to low temperature. Acta Horticulturae (ISHS) 560:457–460

    Google Scholar 

  • Simpson GM (1990) Seed dormancy in grasses. Cambridge University Press, Cambridge

    Book  Google Scholar 

  • Tantau H, Balko C, Brettschneider B, Melz G, Dörfling K (2004) Improved frost tolerance and winter survival in winter barley (Hordeum vulgare L.) by in vitro selection of proline overaccumulating lines. Euphytica 139:19–32

    Article  CAS  Google Scholar 

  • Travert S, Valerio L, Fouraste I, Boudet AM, Teulieres C (1997) Enrichment in specific soluble sugars of two cell-suspension cultures by various treatments enhances their frost tolerance via a noncolligative mechanism. Plant Physiol 114:1433–1442

    PubMed  CAS  Google Scholar 

  • Vágújfalvi A, Kerepesi I, Galiba G, Tischner T, Sutka J (1999) Frost hardiness depending on carbohydrate changes during cold acclimation in wheat. Plant Sci 144:85–92

    Article  Google Scholar 

  • Vertucci CW, Stushnoff C (1992) The state of water in acclimating vegetative buds from Malus and Amelanchier and its relationship to the winter hardiness. Physiol Plant 86(4):503–511

    Article  Google Scholar 

  • Welling A, Palva ET (2006) Molecular control of cold acclimation in trees. Physiol Plant 127:167–181

    Article  CAS  Google Scholar 

  • Xu RY, Niimi Y, Han DS (2006) Changes in endogenous abscisic acid and soluble sugars levels during dormancy-release in bulbs of Lilium rubellum. Sci Hortic 111(1):28–72

    Article  Google Scholar 

  • Yelenosky G (1979) Accumulation of free proline in citrus leaves during cold-hardening of young trees in controlled temperature regimes. Plant Physiol 64:425–427

    Article  PubMed  CAS  Google Scholar 

  • Yepes LM, Aldwinckle HS (1994) Micropropagation of thirteen Malus cultivars and rootstocks, and effect of antibiotics on proliferation. Plant Growth Regul 15:55–67

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by 0002700604 project of the Ministry of Agriculture of the Czech Republic and the OC08062 project financed by the Ministry of Education, Youth and Sport of the Czech Republic. Authors are thankful to Naďa Kantnerová, student of Czech Agricultural University, Prague who assisted in experimental part of in vitro cultures, to Dr. Helena Lipavská and Dr. Hana Konrádová, Charles University, Prague for estimations of saccharides content in plantlets and to Prof. Paul T. Lynch, University Derby, UK providing us by Greensleeves apple cultivar.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jiří Zámečník.

Additional information

Communicated by D. Treutter.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bilavčík, A., Zámečník, J., Grospietsch, M. et al. Dormancy development during cold hardening of in vitro cultured Malus domestica Borkh. plants in relation to their frost resistance and cryotolerance. Trees 26, 1181–1192 (2012). https://doi.org/10.1007/s00468-012-0694-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00468-012-0694-7

Keywords

Navigation