Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 136, Issue 3, pp 499–510 | Cite as

Soluble sugar, starch and phenolic status during rooting of easy- and difficult-to-root magnolia cultivars

  • Agnieszka WojtaniaEmail author
  • Edyta Skrzypek
  • Agnieszka Marasek-Ciolakowska
Original Article


The aim of the study was to determine the effect of indole-3-butyric acid (IBA) and exogenous sucrose concentrations on in vitro rooting, soluble sugar, starch and phenolic production, and ex vitro survival of four magnolia cultivars. There was a significant linear increase in rooting of most magnolia genotypes with an increase in IBA concentration in the medium from 1 to 6 mg L−1. A further increase of IBA concentration to 10 mg L−1 decreased (‘Elizabeth’, ‘Burgundy’) or had no effect on rooting frequency (‘Spectrum’). The effect of IBA on rooting of magnolia shoots was modified by sucrose supply. The three out of four magnolia cultivars showed the highest rooting efficiency in the presence of 6 mg L−1 IBA and 30 g L−1 of sucrose. Generally, decreasing and increasing the sucrose supply from 30 g L−1 significantly lowered the rooting frequency. In ‘Yellow Bird’, sucrose at 40 g L−1 totally blocked root formation. It has been found that the poor rooting ability of ‘Yellow Bird’ coincided with a low soluble sugar content, and high production of starch and phenolics in the shoot bases during the whole rooting period as compared to easy-to-root cultivars. After 5 weeks of the growth on IBA medium, rooted and unrooted shoots were transferred to ex vitro conditions. Both types of shoots showed a high survival and rooting rate (85.4–100%), but they differed in their growth activity and quality. Sucrose concentration in the rooting medium had a post-effect on ex vitro root formation and survival of magnolia plantlets. Ex vitro establishment (13.3%) of recalcitrant ‘Yellow Bird’ was obtained only when the shoots were taken from rooting medium containing the lowest level of sucrose (20 g L−1).


Acclimatization Auxin concentrations In vitro Root formation Sucrose concentrations 



This research was supported by the Polish Ministry of Science and Higher Education as part of the statutory activities (Grant No. 10.1.3) of the Department of Applied Biology, Research Institute of Horticulture in Skierniewice, Poland.

Author contributions

In this experiment Wojtania A. designed and carried out the experiment, analyzed data and wrote the paper. Skrzypek E. was responsible for biochemical analysis. Marasek-Ciołakowska A. was responsible for histological analysis of tissue, including material fixation, slides preparation and photo documentation.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.


  1. Aslmoshtaghi E, Shahsavar AR (2010) Endogenous soluble sugars, starch contents and phenolic compounds in easy- and difficult-to-root olive cuttings. J Biol Environ Sci 49:83–86Google Scholar
  2. Bandurski RS, Cohen JD, Slovin JP, Reneike DM (1995) Auxin biosynthesis and metabolism. In: Davies PJ (ed) Plant hormones. Kluwer Academic Publishers, Dordrecht, pp 39–65CrossRefGoogle Scholar
  3. Biedermann IEG (1987) Factors affecting establishment and development of magnolia hybrids in vitro. Acta Hortic 212:625–629CrossRefGoogle Scholar
  4. Boudet AM (2007) Evolution and current status of research in phenolic compounds. Phytochemistry 68:2722–2735CrossRefGoogle Scholar
  5. Calamar A, De Klerk GJ (2002) Effect of sucrose on adventitious root regeneration in apple. Plant Cell Tiss Org Cult 70:207–212CrossRefGoogle Scholar
  6. Chaidaroon S, Ungvichian I, Ratanathavornkiti K (2004) In vitro root initiation of ‘Champi Sirindhorn’ (Magnolia sirindhorniae Noot. & Chalermglin). Assumpt Univ J Tech 129–132Google Scholar
  7. Chenevard D, Jay-Allemand C, Gendraud M, Frossard JS (1995) The effect of sucrose on the development of hybrid walnut microcutting (Juglans nigra × Juglans regia). Consequences on their survival during acclimatization. Ann Sci For 147:147–156CrossRefGoogle Scholar
  8. Chu ED, Tavares AR, Kanashiro S, Giampaoli P, Yokota ES (2010) Effects of auxins on soluble carbohydrates, starch and soluble protein content in Aechmea blanchetiana (Bromeliacea) cultured in vitro. Sci Hortic 125:451–455CrossRefGoogle Scholar
  9. De Klerk GJ, Guan PH, Marinova S (2011) Effects of phenolic compounds on adventitious root formation and oxidative decarboxylation of applied indoleacetic acid in Malus ‘Jork’. Plant Growth Regul 63:175–185CrossRefGoogle Scholar
  10. Denaxa N, Vemmos SN, Roussos PA (2012) The role of endogenous carbohydrates and seasonal variation in rooting ability of cutting of an easy and a hard to root olive cultivars (Olea europaea L.). Sci Hortic 143:19–28CrossRefGoogle Scholar
  11. Dirr MA (2011) Magnolia. In: Dirr’s encyclopedia of trees and shrubs. Timber Press, Portland, pp 472–494Google Scholar
  12. Dubois M, Gilles KA, Hamilton JK, Roberts PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356CrossRefGoogle Scholar
  13. Eveland AL, Jackson DP (2011) Sugars, signalling, and plant development. J Exp Bot 63:3367–3377CrossRefGoogle Scholar
  14. Faivre-Rampant H, Charpentier JP, Kevers JP, Dommes J, Van Onckelen H, Jay-Allemand C, Gaspar T (2002) Cuttings of the non-rooting rac tobacco mutant overaccumulate phenolic compounds. Funct Plant Biol 29:63–71CrossRefGoogle Scholar
  15. Ferreira WM, Suzuki RM, Pescador R, Figueiredo-Ribeiro RCL, Kerbauy GB (2011) Propagation, growth, and carbohydrates of Dendrobium second love (Orchidaceace) in vitro as affected by sucrose, light, and dark. In Vitro Cell Dev Biol Plant 47:420–427CrossRefGoogle Scholar
  16. Fuentes G, Talavera C, Oropeza C, Desjardins Y, Santamaria JM (2005) Exogenous sucrose can decrease in vitro photosynthesis but improve field survival and growth of coconut (Cocos nucifera L.) in vitro plantlets. Vitro Cell Dev Biol-Plant 41:69–76CrossRefGoogle Scholar
  17. Husen A (2012) Changes in soluble sugars and enzymatic activities during adventitious rooting in cuttings of Grewia optiva as affected by age of donor plants and auxin treatments. Am J Plant Physiol 7:1–16CrossRefGoogle Scholar
  18. Ibrahim O, Gercheva P, Nacheva L, Ivanova V (2011) Rooting of in vitro-raised microcuttings of Magnolia grandiflora, L., and Magnolia × soulangiana Soul.-Bod. J Mt Agric Balk 4:854–868Google Scholar
  19. Ju D, Sun Y, Xia CL, Shi K, Zhou YH, Yu JQ (2007) Physiological basis of different allelopathic reactions of cucumber and figleaf gourd plants to cinnamic acid. J Exp Bot 58:3765–3773CrossRefGoogle Scholar
  20. Kadleček P, Tichá I, Haisel D, Čapková V, Schäfer C (2001) Importance of in vitro pretreatment for ex vitro acclimatization and growth. Plant Sci 161:695–701CrossRefGoogle Scholar
  21. Kamenicka A (1998) Influence of selected carbohydrates in rhizogenesis of shoots saucer magnolia in vitro. Acta Physiol Plant 20:425–429CrossRefGoogle Scholar
  22. Kamenicka A, Lanakova M (2000) Effect of medium composition and type of vessel closure on axillary shoot production of magnolia in vitro. Acta Physiol Plant 22:129–134CrossRefGoogle Scholar
  23. Kozai T (1991) Micropropagation under photoauthotrophic conditions. In: Zimmerman DP, Richard H (eds) Micropropagation: technology and application. Kluwer Akademic Publishers, Dordrecht, 447–469CrossRefGoogle Scholar
  24. Kozlowski TT (1992) Carbohydrate sources and sinks in woody plants. Bot Rev 58:107–222CrossRefGoogle Scholar
  25. Kracke H, Marangoni B, Cristoferi G (1982) Carbohydrate, protein, and phenol changes during rooting of grapevine rootstock. In: Dinsmoor WA (ed) Grape and Wine Centennial Symposium Proceedings. University of California, Davis, CA, pp 56–60Google Scholar
  26. Kromer K, Gamian A (2000) Analysis of soluble carbohydrates, protein and lipids in shoots of M7 apple rootstock cultured in vitro during regeneration of adventitious roots. J Plant Physiol 156:775–782CrossRefGoogle Scholar
  27. Lattier JD, Touchell DH, Ranney TG (2014) Micropropagation of an interspecific hybrid dogwood (Cornus ‘NCCH1’). Propag Ornam Plants 14:184–190Google Scholar
  28. Li M, Leung DWM (2000) Starch accumulation is associated with adventitious root formation in hypocotyl cutting of Pinus radiata. J Plant Growth Regul 19:423–428CrossRefGoogle Scholar
  29. Licea-Moreno RJ, Contreras A, Morales AV, Urban I, Daquinta M, Gomez L (2015) Improved walnut mass micropropagation through the combined use of phloroglucinol and FeEDDHA. Plant Cell Tiss Organ Cult 123:143–154CrossRefGoogle Scholar
  30. Ling WX, Zhong Z (2012) Seasonal variation in rooting of the cutting from Tetraploid Locust in relation to nutrients and endogenous plant hormones of the shoot. Turk J Agric For 36:257–266Google Scholar
  31. Matysiak N, Gabryszewska E (2016) The effect of in vitro culture conditions on the pattern of maximum photochemical efficiency of photosystem II during acclimatisation of Helleborus niger plantlets to ex vitro conditions. Plant Cell Tiss Organ Cult 125:585–593CrossRefGoogle Scholar
  32. Mishra BS, Singh M, Aggrawal P, Laxmi A (2009) Glucose and auxin signaling interaction in controlling Arabidopsis thaliana seedlings root growth and development. PLoS ONE 4:e4502CrossRefGoogle Scholar
  33. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  34. Osterc G, Trobec M, Usenik V, Solar A, Štampar F (2004) Changes in polyphenols in leafy cutting during the root initiation phase regarding various cutting types in Castanea. Phyton (Horn Austria) 44:109–119Google Scholar
  35. Otiende M, Nyabundi JO, Ngamau K, Opala P (2017) Effects of cutting position of rose rootstock cultivars on rooting and its relationship with mineral content and endogenous carbohydrates. Sci Hortic 225:204–212CrossRefGoogle Scholar
  36. Palanisamy K, Ansari AA, Kumar P, Gupta BN (1998) Adventitious rooting in shoot cuttings of Azadirachta indica and Pongamia pinnata. New For 16:81–88CrossRefGoogle Scholar
  37. Parris JK, Touchel DH, Ranney TG, Adelberg J (2012) Basal salt composition, cytokinins, and phenolic binding agents influence in vitro growth and ex vitro establishment of Magnolia ‘Ann’. HortScience 47:1625–1629CrossRefGoogle Scholar
  38. Pierik RLM, Oosterkamp J, Ebbing MAC (1997) Factors controlling adventitious root formation of explants from juvenile and adult Quercus robur ‘Fastigiata’. Sci Hortic 71:87–92CrossRefGoogle Scholar
  39. Podwyszyńska M, Cieślińska M (2018) Rooting shoots of apple varieties and their tetraploids obtained by the in vitro technique. Acta Sci Pol-Hortoru 17:49–62CrossRefGoogle Scholar
  40. Premkumar A, Barcelo-Muñoz A, Quesada MA, Mercado JA, Pliego-Alfaro F (2003) Influence of sucrose concentration on in vitro rooting, growth, endogenous sugars and ex vitro survival of juvenile avocado. J Hortic Sci Biotechnol 78:45–50CrossRefGoogle Scholar
  41. Radomir AM (2012) Comparative study on the in vitro multiplication potential of Magnolia stellata and Magnolia × soulangiana species. J Hortic Forest Biotechnol 16:39–44Google Scholar
  42. Ruzin SE (1999) Plant microtechnique and microscopy. Oxford University Press, New York, 322 pGoogle Scholar
  43. Sharma J, Knox GW, Ishida ML (2006) Adventitious rooting of stem cuttings of yellow-flowered magnolia cultivars is influenced by time after budbreak and indole-3-butyric acid. HortScience 41:202–206CrossRefGoogle Scholar
  44. Singh HP, Kaur S, Batish DR, Kohli RK (2009) Caffeic acid inhibits in vitro rooting in mung bean [Vigna radiata (L.)] hypocotyls by inducing oxidative stress. Plant Growth Regul 57:21–30CrossRefGoogle Scholar
  45. Singh HP, Kaur S, Batish DR, Kohli RK (2014) Ferulic acid impairs rhizogenesis and root growth, and alters associated biochemical changes in mung bean (Vigna radiata) hypocotyls. J Plant Interact 9:267–274CrossRefGoogle Scholar
  46. Singleton VS, Rossi JJA (1965) Colorimetry of total phenolics with phosphomolybdic phosphotungstic acid reagent. Am J Enol Vitic 16:144–158Google Scholar
  47. Vahdati K, Leslie C, Zamani Z, McGranahan G (2004) Rooting and acclimatization of in vitro-grown shoots from mature trees of three Persian walnut cultivars. HortScience 39:324–327CrossRefGoogle Scholar
  48. Veierskov (B) (1988) Relations between carbohydrates and adventitious root formation. In: Davis TD, Haissing BE, Sankhla N (eds) Adventitious root formation in cuttings. Dioscorides Press, Portland, pp 70–78Google Scholar
  49. Wojtania A, Matysiak B (2018) In vitro propagation of Rosa ‘Konstancin’ (R. rugosa × R. beggeriana), a plant with high nutritional and pro-health value. Folia Hortic 30:259–267CrossRefGoogle Scholar
  50. Wojtania A, Gabryszewska E, Podwyszyńska M (2011) The effect of growth regulators and sucrose concentration on in vitro propagation of Camellia japonica L. Propag Ornam Plants 11:177–183Google Scholar
  51. Wojtania A, Skrzypek E, Gabryszewska E (2015) Effect of cytokinin, sucrose and nitrogen salts concentrations on the growth and development and phenolics content in Magnolia × soulangiana ‘Coates’ shoots in vitro. Acta Sci Pol-Hortoru 14:51–62Google Scholar
  52. Wu HC, du Toit ES, Reinhardt CF, Rimando AM, van der Koody F, Meyer JJM (2007) The phenolic, 3,4-dihydroxybenzoic acid, is an endogenous regulator of rooting in Protea cynaroides. Plant Growth Regul 52:207–215CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Agnieszka Wojtania
    • 1
    Email author
  • Edyta Skrzypek
    • 2
  • Agnieszka Marasek-Ciolakowska
    • 1
  1. 1.Research Institute of HorticultureSkierniewicePoland
  2. 2.The F. Górski Institute of Plant PhysiologyPolish Academy of SciencesKrakówPoland

Personalised recommendations