In vitro sucrose concentration affects growth and acclimatization of Alocasia amazonica plantlets

  • Eun-A Jo
  • Rajesh Kumar Tewari
  • Eun-Joo Hahn
  • Kee-Yoeup Paek
Original Paper


Plantlets of Alocasia amazonica were regenerated on the MS medium supplemented with different concentrations (0–9%) of sucrose. An absence of sucrose in the growth medium induced generation of leaves, however, it decreased multiplication. On contrary, sucrose supply of 6% or 9% enhanced multiplication but hampered photoautotrophic growth (generation of leaves). Increasing sucrose supply also increased sugars and starch content and number of stomata and decreased water potential and size of stomata during in vitro growth period. During ex vitro acclimatization, shoot length, root length, leaf number and root number of Alocasia plantlets grown with 3% sucrose, were found to be better among the other studied sucrose concentrations. Under ex vitro acclimatization, number of stomata, contents of various carbohydrates in the leaves were increased but size of stomata decreased with increasing sucrose supply during in vitro growth period. Moreover, water potential of leaves of plantlets, which have been grown with a sucrose concentration other than 3%, was decreased. During in vitro growth, net CO2 assimilation rate (PN), transpiration (E), stomatal conductance (gs) and variable fluorescence to maximum fluorescence ratio (Fv/Fm) were unaffected, however, during acclimatization these were changed and maximum PN, E, and gs were observed in the plantlets micropropagated with 3% sucrose. Fv/Fm was decreased severely in the plantlets micropropagated with 6% sucrose during acclimatization. Thus a sucrose concentration of 3% in the medium is appeared to be better among studied concentrations for both in vitro growth and ex vitro acclimatization of A. amazonica plantlets.


In vitro culture Carbohydrates concentration Stomatal development Osmotic stress Photosynthesis Transpiration 



Benzyl adenine


Days after acclimatization




Variable fluorescence to maximum fluorescence ratio


Stomatal conductance


Photon flux density


Net CO2 assimilation rate


Relative humidity


  1. Adelberg J, Toler J (2004) Comparison of agar and thin-film liquid system for micropropagation of ornamental Alocasia and Colocasia. HortScience 39:1088–1092Google Scholar
  2. Amiard V, Mueh KE, Demmig-Adams B, Ebbert V, Turgeon R, Adams WW III (2005) Anatomical and photosynthetic acclimation to the light environment in species with differing mechanisms of phloem loading. Proc Natl Acad Sci USA 102:12968–12973. doi:10.1073/pnas.0503784102 PubMedCrossRefGoogle Scholar
  3. Capellades M, Lemeus R, Debergh P (1991) Effect of sucrose on starch accumulation and rate of photosynthesis of Rosa cultivated in vitro. Plant Cell Tissue Organ Cult 25:21–26. doi:10.1007/BF00033908 CrossRefGoogle Scholar
  4. Desjardins Y, Hdider C, De Riek J (1995) Carbon nutrition in vitro—regulation and manipulation of carbon assimilation in micropropagated systems. In: Aitken-Christie J, Kozai T, Smith MLA (eds) Automation and environmental control in plant tissue culture. Kluwer Academic Publishers, Netherlands, pp 441–471Google Scholar
  5. Gray JD, Kolesik P, Hoj PB, Coombe BG (1999) Confocal measurement of the three-dimensional size and shape of plant parenchyma cells in a developing fruit tissue. Plant J 19:229–236. doi:10.1046/j.1365-313X.1999.00512.x PubMedCrossRefGoogle Scholar
  6. Hazarika BN (2003) Acclimatization of tissue-cultured plants. Curr Sci 85:1704–1712Google Scholar
  7. Hazarika BN, Parthasarathy VA, Nagaraju V, Bhowmik G (2000) Sucrose induced biochemical changes in in vitro microshoots of Citrus species. Indian J Hortic 57:27–31Google Scholar
  8. Hdider C, Desjardins Y (1994) Effects of sucrose on photosynthesis and phosphoenolpyruvate carboxylase activity of in vitro cultured strawberry plantlets. Plant Cell Tissue Organ Cult 36:27–33. doi:10.1007/BF00048312 CrossRefGoogle Scholar
  9. Heo JW, Shin KS, Kim SK, Paek KY (2006a) Light quality affects in vitro growth of grape ‘Teleki 5BB’. J Plant Biol 49:276–280Google Scholar
  10. Heo JW, Lee CW, Paek KY (2006b) Influence of mixed LED radiation on the growth of annual Plants. J Plant Biol 49:286–290CrossRefGoogle Scholar
  11. Jo UA, Murthy HN, Hahn EJ, Paek KY (2008a) Micropropagation of Alocasia amazonica using semisolid and liquid cultures. In Vitro Cell Dev Biol Plant 44:26–32. doi:10.1007/s11627-007-9081-2 Google Scholar
  12. Jo EA, Tewari RK, Hahn EJ, Paek KY (2008b) Effect of photoperiod and light intensity on in vitro propagation of Alocasia amazonica. Plant Biotechnol Rep 2:207–212. doi:10.1007/s11816-008-0063-6 CrossRefGoogle Scholar
  13. Kilb B, Wietoska H, Godde D (1996) Changes in the expression of photosynthetic genes precede loss of photosynthetic activities and chlorophyll when glucose is supplied to mature spinach leaves. Plant Sci 115:225–235. doi:10.1016/0168-9452(96)04362-2 CrossRefGoogle Scholar
  14. Kooten OV, Snel JFH (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth Res 25:147–150. doi:10.1007/BF00033156 CrossRefGoogle Scholar
  15. Kwa SH, Wee YC, Lim TM, Kumar PP (1995) Establishment and physiological analyses of photoautotrophic callus cultures of the fern Platycerium coronarium (Koenig) Desv under CO2 enrichment. J Exp Bot 46:1535–1542. doi:10.1093/jxb/46.10.1535 CrossRefGoogle Scholar
  16. Lane WD (1978) Regeneration of apple plants from shoot meristem tips. Plant Sci Lett 13:281–285. doi:10.1016/0304-4211(78)90107-4 CrossRefGoogle Scholar
  17. Langford PJ, Wainwright H (1987) Effect of sucrose concentration on the photosynthetic ability of rose shoots in vitro. Ann Bot (Lond) 60:633–640Google Scholar
  18. Lee SH, Tewari RK, Hahn EJ, Paek KY (2007) Photon flux density and light quality induce changes in growth, stomatal development, photosynthesis and transpiration of Withania somnifera (L.) Dunal. plantlets. Plant Cell Tissue Organ Cult 90:141–151. doi:10.1007/s11240-006-9191-2 CrossRefGoogle Scholar
  19. Marino G, Bertazza G, Magnanini E, Altan AD (1993) Comparative effects of sorbitol and sucrose as main carbon energy sources in micropropagation of apricot. Plant Cell Tissue Organ Cult 34:235–244. doi:10.1007/BF00029712 CrossRefGoogle Scholar
  20. Mehta UJ, Krishnamurthy KV, Hazra S (2000) Regeneration of plants via adventitious bud formation from zygotic embryo axis of tamarind (Tamarindus indica L.). Curr Sci 78:1231–1234Google Scholar
  21. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497. doi:10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
  22. Pospíšilová J, Tichá I, Kadleček P, Haisel D, Plzáková Š (1999) Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42:481–497. doi:10.1023/A:1002688208758 CrossRefGoogle Scholar
  23. Puthur JT, Thomas TD (2004) High frequency in vitro regeneration of Kigelia pinnata L. via organogenesis. J Plant Biol 47:48–51Google Scholar
  24. Rolland F, Moore B, Sheen J (2002) Sugar sensing and signaling in plants. Plant Cell 14:S185–S205PubMedGoogle Scholar
  25. Serret MD, Trillas MI, Matas J, Araus JL (1997) Development of photoautotrophy and photoinhibition of Gardenia jasmoides plantlets during micropropagation. Plant Cell Tissue Organ Cult 45:1–16. doi:10.1007/BF00043422 CrossRefGoogle Scholar
  26. Thao NTP, Miyajima I, Ureshino K, Ozaki Y, Okubo H (2003a) Micropropagation of ornamental Alocasia. J Fac Agric Kyushu Univ 47:277–282Google Scholar
  27. Thao NTP, Ureshino K, Miyajima I, Ozaki Y, Okubo H (2003b) Induction of tetraploids in ornamental Alocasia through colchicine and oryzalin treatments. Plant Cell Tissue Organ Cult 72:19–25. doi:10.1023/A:1021292928295 CrossRefGoogle Scholar
  28. Van Huylenbroeck JM, Debergh PC (1996) Impact of sugar concentration in vitro on photosynthesis and carbon metabolism during ex vitro acclimatization of Spathiphyllum plantlets. Physiol Plant 96:298–304. doi:10.1034/j.1399-3054.1996.960220.x CrossRefGoogle Scholar
  29. Van Huylenbroeck JM, Piqueras A, Debergh PC (2000) The evolution of photosynthetic capacity and the antioxidant enzymatic system during acclimatization of micropropagated Calathea plants. Plant Sci 155:59–66. doi:10.1016/S0168-9452(00)00201-6 PubMedCrossRefGoogle Scholar
  30. Wainwright H, Scrace J (1989) Influence of in vitro preconditioning with carbohydrates during the rooting of microcuttings on in vivo establishment. Sci Hortic (Amsterdam) 38:261–267. doi:10.1016/0304-4238(89)90073-3 CrossRefGoogle Scholar
  31. Yeo UD, Pandey DM, Kim KH (2004) Long term effects of growth regulators on growth and turnover of symplastic and apoplastic sugars in the suspension subculture of kidney bean. J Plant Biol 47:21–26. doi:10.1023/A:1027316511865 CrossRefGoogle Scholar
  32. Zapata EV, Morales GS, Lauzardo ANH, Bonfil BM, Tapia GT, Sánchez ADJ, Valle MVD, Aparicio AJ (2003) In vitro regeneration and acclimatization of plants of Turmeric (Curcuma longa L.) in a hydroponic system. Biotecnol Apl 20:25–31Google Scholar
  33. Zimmerman RH (1983) Factors affecting in vitro propagation of apple cuttings. Acta Hortic 131:171–178Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Eun-A Jo
    • 1
  • Rajesh Kumar Tewari
    • 1
    • 2
  • Eun-Joo Hahn
    • 1
  • Kee-Yoeup Paek
    • 1
  1. 1.Research Center for the Development of Advanced Horticultural TechnologyChungbuk National UniversityCheongjuRepublic of Korea
  2. 2.Laboratory of Plant Nutrition, Graduate School of HorticultureChiba UniversityChibaJapan

Personalised recommendations