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Photosynthetic parameters of Ulmus minor plantlets affected by irradiance during acclimatization


In order to set up large-scale acclimatization protocols of micropropagated plants, an in-depth knowledge of their physiological responses during in vitro to ex vitro transfer is required. This work describes the photosynthetic performance of Ulmus minor micropropagated plants during acclimatization at high irradiance (HI; 200 ± 20 μmol m−2 s−1 or low irradiance (LI; 100 ± 20 μmol m−2 s−1). During this experiment, leaf pigment content, chlorophyll a fluorescence, gas exchange, stomata morphology, the activity of the Calvin cycle enzymes and saccharides were measured in persistent and new leaves. The results indicated that HI induces a higher photosynthetic performance compared to LI. Therefore, plants acclimatized under HI are likely to survive better after field transfer.

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dry mass

ci/ca :

ratio of intercellular to atmospheric CO2 concentration


transpiration rate

Fv/Fm :

variable to maximum chlorophyll fluorescence (maximal efficiency of PSII)

gs :

stomatal conductance


high irradiance


low irradiance

PN :

net photosynthetic rate


photosystem 2


non-photochemical quenching


photochemical quenching


relative humidity


ribulose-1,5-bisphosphate carboxylase/oxygenase


stromal fructose-1,6-bisphosphatase


effective quantum efficiency of PSII


  • Amâncio, S., Rebordão, J.P., Chaves, M.M.: Improvement of acclimatization of micropropagated grapevine: photosynthetic competence and carbon allocation. — Plant Cell Tissue Organ Cult. 58: 31–37, 1999.

    Article  Google Scholar 

  • Brito, G., Costa, A., Coelho, C., Santos, C.: Large-scale field acclimatization of Olea maderensis micropropagated plants: morphological and physiological survey. — Trees 23: 1019–1031, 2009.

    Article  Google Scholar 

  • Brüggemann, W., Klaucke, S., Maas-Kantel, K.: Long-term chilling of young tomato plants under low light. V. Kinetic and molecular properties of two key enzymes of the Calvin cycle in Lycopersicon esculentum Mill. and L. peruvianum Mill. — Planta 194: 160–168, 1994.

    Article  Google Scholar 

  • Carvalho, L.C., Esquível, M.G., Martins, I., Pinto Ricardo, C., Amâncio, S.: Monitoring the stability of Rubisco in micropropagated grapevine (Vitis vinifera L.) by twodimensional electrophoresis. — J. Plant Physiol. 162: 365–374, 2005.

    Article  PubMed  CAS  Google Scholar 

  • Carvalho, L.C., Osório, M.L., Chaves, M.M., Amâncio, S.: Chlorophyll fluorescence as an indicator of photosynthetic functioning of in vitro grapevine and chestnut plants under ex vitro acclimatization. — Plant Cell Tissue Organ Cult. 67: 271–280, 2001.

    Article  Google Scholar 

  • Choudhury, N.K., Behera, R.K.: Photoinhibition of photosynthesis: role of carotenoids in photoprotection of chloroplasts. — Photosynthetica 39: 481–488, 2001.

    Article  CAS  Google Scholar 

  • Conde, P., Loureiro, J., Santos, C.: Somatic embryogenesis and plant regeneration from leaves of Ulmus minor Mill. — Plant Cell Rep. 22: 632–639, 2004.

    Article  PubMed  CAS  Google Scholar 

  • Conde, P., Sousa, A., Costa, A., Santos, C.: A protocol for Ulmus minor Mill. micropropagation and acclimatization. — Plant Cell Tissue Organ Cult. 92: 113–119, 2008.

    Article  Google Scholar 

  • Correia, M.J., Fonseca, F., Azedo-Silva, J., Dias, C., David, M.M., Barrote, I., Osório, M.L., Osório, J.: Effects of water deficit on the activity of nitrate reductase and contents of sugars, nitrate and free amino acids in the leaves and roots of sunflower and with lupin plants growing under two nutrient supply regimes. — Physiol. Plant. 124: 61–70, 2005.

    Article  CAS  Google Scholar 

  • Dias, M.C., Brüggemann, W.: Photosynthesis under drought stress in Flaveria species with different degrees of development of the C4 syndrome. — Photosynthetica 45: 75–84, 2007.

    Article  Google Scholar 

  • Dias, M.C., Brüggemann, W.: Limitations of photosynthesis in Phaseolus vulgaris under drought stress: gas exchange, chlorophyll fluorescence and Calvin cycle enzymes. — Photosynthetica 48: 96–102, 2010.

    Article  CAS  Google Scholar 

  • Dias M.C., Pinto G., Santos C.: Acclimatization of micropropagated plantlets induces an antioxidative burst: a case study with Ulmus minor Mill. — Photosynthetica 49: 259–266, 2011.

    Article  Google Scholar 

  • Dunn, C.P.: The Elms — Breeding, Conservation and Disease Management. — Kluwer Academic Publishers, Dordrecht 2000.

    Google Scholar 

  • Estrada-Luna, A.A., Davies, F.T., Egilla, J.N.: Physiological changes and growth of micropropagated Chile ancho pepper plantlets during acclimatization and post-acclimatization. — Plant Cell Tissue Organ Cult. 66: 17–24, 2001.

    Article  CAS  Google Scholar 

  • Faisal, M., Anis, M.: Changes in photosynthetic activity, pigment composition, electrolyte leakage, lipid peroxidation, and antioxidant enzymes during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets. — Plant Cell Tissue Organ Cult. 99: 125–132, 2009.

    Article  CAS  Google Scholar 

  • Fila, G., Badeck, F.W., Meyer, S., Cerovic, Z., Ghashghai, J.: Relationships between leaf conductance to CO2 diffusion and photosynthesis in micropropagated grapevine plants, before and after ex vitro acclimatization. — J. exp. Bot. 57: 2687–2695, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Fuentes, G., Talavera, C., Opereza, C., Desjardins, Y., Santamaria, J.: Exogenous sucrose can decrease in vitro photosynthesis but improve field survival and growth of coconut (Cocos nucifera L.) in vitro plantlets. — In vitro cell. dev. Biol. Plant. 41: 69–76, 2005.

    Article  CAS  Google Scholar 

  • Grout, B.W.: Photosynthesis of regenerated plantlets in vitro and the stresses of transplanting. — Acta Hort. 230: 129–135, 1988.

    Google Scholar 

  • Harvengt, L., Meier-Dinkel, A., Dumas, E., Collin, E.: Establishment of a cryopreserved gene bank of European elms. — Can. J. Forest. Res. 34: 43–55, 2004.

    Article  Google Scholar 

  • Hazarika, B.N.: Morpho-physiological disorders in in vitro culture of plants. — Sci. Hort. 108: 105–120, 2006.

    Article  CAS  Google Scholar 

  • Hdider, C.Y., Desjardins, L.: Reduction of ribulose 1-5 bisphosphate carboxylase/oxygenase efficiency by the presence of sucrose during the tissue culture of strawberry plantlets. — In Vitro cell. dev. Biol. Plant 31: 165–170, 1995.

    Article  CAS  Google Scholar 

  • Jones, M.G.K., Outlaw, W.H., Lowry, O.H.: Enzymatic assay of 10-7 to 10-14 moles of sucrose in plant tissue. — Plant Physiol. 60: 379–383, 1977.

    Article  PubMed  CAS  Google Scholar 

  • Lilley, R.M., Walker, D.A.: An improved spectrophotometric assay for ribulose-bisphosphate carboxylase. — Biochem. biophys. Acta 358: 226–229, 1974.

    Article  PubMed  CAS  Google Scholar 

  • Merkel, S.A., Nairn, C.J.: Hardwood tree biotechnology. — In Vitro cell. dev. Biol. 41: 602–619, 2005.

    Google Scholar 

  • Osório, M.L., Gonçalves, S., Osório, J., Romano, A.: Effects of CO2 concentration on acclimatization and physiological responses of two cultivars of carob tree. — Biol. Plant. 49: 161–167, 2005.

    Article  Google Scholar 

  • Osório, M.L., Osório, J., Romano, A.: Chlorophyll fluorescence in micropropagated Rhododendron ponticum subsp. baeticum plants in response to different irradiances. — Biol. Plant. 54: 415–422, 2010.

    Article  Google Scholar 

  • Park, Y-S.: Implementation of conifer somatic embryogenesis in clonal forestry: technical requirements and deployment considerations. — Ann. Forest. Sci. 59: 651–656, 2002.

    Article  Google Scholar 

  • Pinto, G., Park, Y-S., Silva, S., Araújo, C., Neves, L., Santos, C.: Factors affecting maintenance, proliferation, and germination of secondary somatic embryos of Eucalyptus globulus Labill.. — Plant Cell Tissue Organ Cult. 95: 69–78, 2008.

    Article  CAS  Google Scholar 

  • Pinto, G., Silva, S., Loureiro, J., Costa, A., Dias, M.C., Araújo, C., Neves, L., Santos, C.: Acclimatization of secondary somatic embryos derived plants of Eucalyptus globulus Labill.: an ultrastructural approach. — Trees 25: 383–392, 2011.

    Article  Google Scholar 

  • Pospíšilová, J., Synková, H., Haisel, D., Baťková, P.: Effect of abscisic acid on photosynthetic parameters during ex vitro transfer of micropropagated tobacco plantlets. — Biol. Plant. 53: 11–20, 2009.

    Article  Google Scholar 

  • Pospíšilová, J., Synková, H., Haisel, D., Čatský, J., Wilhelmová, N., Šrámek, F.: Effect of elevated CO2 concentrations on acclimation of tobacco plantlets to ex vitro conditions. — J. exp. Bot. 50: 119–126, 1999.

    Google Scholar 

  • Rybczyński, J.J., Borkowska, B., Fiuk, A., Gawńroska, H., Śliwińska, E., MikuŁa, A.: Effect of sucrose concentration on photosynthetic activity of in vitro cultures Gentiana kurroo (Royle) germlings. — Acta Physiol. Plant. 29: 445–453, 2007.

    Article  Google Scholar 

  • Schreiber, U., Bilger, W., Neubauer: Chlorophyll flourescence as a non-destructive indicator for rapid assessment of in vivo photosynthesis. — In: Schulze, E.D., Caldwell, M.M. (ed.): Ecophysiology of Photosynthesis. Pp. 49–70, Springer-Verlag, Berlin 1995.

    Chapter  Google Scholar 

  • Sims, D.A., Gamon, J.A.: Relationships between leaf pigment content and spectral reflectance across a wide range of species, leaf structures and developmental stages. — Remote Sensing Environ. 81: 337–354, 2002.

    Article  Google Scholar 

  • Stitt, M., Bulpin, P.V., Rees, T.A.: Pathway of starch breakdown in photosynthetic tissues of Pisum sativum. — Biochem. Acta 544: 200–214, 1978.

    Article  CAS  Google Scholar 

  • Van Huylenbroeck, J.M., Piqueras, A., Debergh, P.C.: The evolution of photosynthesis capacity and the antioxidant enzymatic system during acclimatization of micropropagated Calathea plants. — Plant Sci. 155: 59–66, 2000.

    Article  PubMed  Google Scholar 

  • Van Kooten, O., Snel, J.F.H.: The use of chlorophyll fluorescence nomenclature in plant stress physiology. — Photosynth. Res. 25: 47–150, 1990.

    Article  Google Scholar 

  • Von Caemmerer, S., Farquhar, G.D.: Some relationships between the biochemistry of photosynthesis and the gas exchange of leaves. — Planta 153: 376–387, 1981.

    Article  Google Scholar 

  • Walters, R.G., Horton, P.: Resolution of components of nonphotochemical chlorophyll fluorescence quenching in barley leaves. — Photosynth. Res. 27:121–133, 1991.

    Article  CAS  Google Scholar 

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Correspondence to M. C. Dias.

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Acknowledgement: This work was supported by the Portuguese Foundation for Science and Technology through a post-doctoral fellowship of M.C. Dias (SFRH/BPD/41700/2007) and S. Silva (SFRH/BPD/74299/2010). The authors acknowledge Dr. K. Bennett for her assistance in proofreading the manuscript.

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Dias, M.C., Pinto, G., Correia, C.M. et al. Photosynthetic parameters of Ulmus minor plantlets affected by irradiance during acclimatization. Biol Plant 57, 33–40 (2013).

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Additional key words

  • Calvin cycle enzymes
  • chlorophyll
  • elm
  • fluorescence
  • micropropagation
  • net photosynthetic rate
  • stomata
  • transpiration rate