Abstract
In vitro regeneration of black nightshade (Solanum nigrum L.) plants was achieved through callus-mediated shoot organogenesis followed by 30 d indoor ex vitro adaptation to nutritional stress under environmental ambience and thereafter 6-d outdoor acclimatization in pots prior to field establishment. Relevant physiological parameters including pigment content, chlorophyll a fluorescence, net photosynthetic rate (PN), transpiration rate (E), and stomatal conductance (gs) of in vitro-regenerated plants were investigated during the course of ex vitro adaptation. During the first 4 d of indoor transplantation to potting substrate, there was a marginal reduction in the leaf chlorophyll and carotenoid contents but PN and E were strongly reduced. The stomatal conductance and E/PN ratio were significantly higher in plants up to 20 d of indoor adaptation than those of comparable age grown naturally from seeds. The shape of the OJIP fluorescence transient varied significantly with acclimatization, and the maximum change was observed at 2.0 ms. The 2.0 ms variable fluorescence (Vj), 30 ms relative fluorescence (M0), photon trapping probability (TR0/Abs), and photosystem II (PSII) trapping rate (TR0/RC) showed initial disturbance and subsequent stabilization during 30 d of indoor acclimatization. Energy dissipation (DI0/RC) and electron transport probability (ET0/TR0) showed an initial phase of increase during the 4 d after plants were transplanted outdoors. During the 6-d outdoor acclimatization after transfer of plants to soil, no significant change in total chlorophylls and carotenoids, E, and gs were observed, but PN improved after reduction on the first d. The OJIP-derived parameters experienced change on the first d but were stabilized quickly thereafter. There was no significant difference between outdoor acclimatized plants and those of the seed-grown plants of comparable age with respect to photosynthetic and fluorescence parameters. Direct transfer of plants without indoor acclimatization, however, showed a completely different trend with respect to PN, E, and OJIP fluorescence transients. The bearing of this study on optimizing micropropagation is discussed.
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References
Antal T.; Rubin A. In vivo analysis of chlorophyll a fluorescence induction. Photosynth Res 96: 217–226; 2008.
Blanke M. M.; Belcher A. R. Stomata of apple leaves cultured in vitro. Plant Cell Tissue Organ Cult 19: 85–89; 1989.
Buddendorf-Joosten J. M. C.; Woltering E. J. Components of the gaseous environment and their effects on plant growth and development in vitro. Plant Growth Regul 15: 1–16; 1994.
Desjardins Y. Photosynthesis in vitro—on the factors regulating CO2 assimilation in micropropagation systems. Acta Hort 393: 45–61; 1995.
Fila G.; Ghashghaie J.; Hoarau J.; Cornic G. Photosynthesis, leaf conductance and water relations of in vitro cultured grapevine rootstock in relation to acclimatization. Physiol Plant 102: 411–418; 1998.
Force L.; Critchley C.; Van Rensen J. J. S. New fluorescence parameters for monitoring photosynthesis in plants; 1. The effect of illumination on the fluorescence parameters of the JIP-test. Photosynth Res 90: 1–19; 2003.
Gomez K. A.; Gomez A. A. Statistical procedures for agricultural researches. John Willey & Sons, New York, p 680; 1984.
Hazarika B. N. Acclimatization of tissue cultured plants. Curr Sci 85(12): 1704–1712; 2003.
Kozai T. Micropropagation under photoautotrophic conditions. In: Debergh P. C.; Zimmerman R. H. (eds) Micropropagation technology and applicaton. Kluwer Academic Publishers, Dodrecht, pp 447–469; 1991.
Kozai T.; Smith M. A. L. Environmental control in plant tissue culture—general introduction and overview. In: Aitken-Christie J.; Kozai T.; Smith M. L. (eds) Automation and environmental control in plant tissue culture. Kluwer Academic Publishers, Dodrecht, pp 301–318; 1991.
Krüger G. H. J.; Tsimilli-Michael M.; Strasser R. J. Light stress provokes plastic and elastic modifications in structure and function of photosystem II in Camellia leaves. Physiol Plant 101: 265–277; 1997.
Lamhamedi M. S.; Chamberland H.; Tremblay F. M. Epidermal transpiration, ultrastructural characteristics and net photosynthesis of white spruce somatic seedlings in response to in vitro acclimatization. Physiol Plant 118: 554–561; 2003.
Mohapatra P. K.; Patra S.; Samantray P. K.; Mohanty R. C. Effect of pyrethoroid insecticide cypermethrin on photosynthetic pigments of the cyanobacterium Anabaena doliolum Bhar. Pol J Environ Stud 12(2): 207–212; 2003.
Murashige T.; Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15: 473–497; 1962.
Porra R. J.; Thomson W. A.; Kriederman P. E. Determination of accurate extinction coefficients and simultaneous equation of assaying Chl a and Chl b extracted with four different solvents. Verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975: 384–385; 1989.
Pospisilova J.; Solarava J.; Catsky J. Photosynthetic responses to stresses during in vitro cultivation. Photosynthetica 26: 3–18; 1992.
Pospisilova J.; Synkova H.; Haisel D.; Batkova P. Effect of abscisic acid on photosynthetic parameters during ex vitro transfer of micropropagated tobacco plantlets. Biol Plant 53(1): 11–20; 2009.
Pospisilova J.; Synkova H.; Haisel D.; Catsky J.; Wilhelmova N.; Sramek F. Effect of elevated CO2 concentration on acclimatization of tobacco plantlets to ex vitro culture. Photosynthetica 33: 403–412; 1997.
Pospisilova J.; Ticha I.; Kadlecek P.; Haisel D.; Plzakova S. Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42(4): 481–497; 1999.
Rodriguez R.; Aragon C. E.; Escalona M.; Gonzalez-Olmedo J. L.; Desjardins V. Carbon metabolism in the micropropagated sugarcane during acclimatization phase. In Vitro Cell Dev Biol Plant 44: 533–539; 2008.
Srivastava A.; Guisse B.; Greppin H.; Strasser R. J. Regulation of antennae structures and electron transport in photosystem II of Pisum sativum under elevated temperature probed by the fast polyphasic chlorophyll a fluorescence transients OJIP. Biochim Biophys Acta 1320: 95–106; 1997.
Strasser B. J. Donor side capacity of photosystem II probed by chlorophyll a fluorescence transients. Photosynth Res 52: 147–155; 1997.
Strasser R. J.; Srivastava A.; Tsimilli-Michael M. The fluorescence transients as a tool to characterize and screen photosynthetic samples. In: Yunus M.; Pathre U.; Mohanty P. (eds) Probing photosynthesis (Mechanism, Regulation and Adaptation) Taylor and Francis. London, pp 445–483; 2000.
Strasser B. J.; Strasser R. J. Measuring fast fluorescence transients to address environmental questions: the JIP–test. In: Mathis P. (ed) Photosynthesis (From Light to Biosphere). Vol. 5. Kluwer Academic Publishers, Dodrecht, pp 977–980; 1995.
Strasser R. J.; Tsimilli-Michael M. Stress in plants, from daily rhythm to global changes, detected and quantified by the JIP-test. Chim Nouv (SRC) 75: 3321–3326; 2001.
Strasser R. J.; Tsimilli-Michale M.; Srivastava A. Analysis of the chlorophyll a fluorescence transient. In: Papageorgiou G. C.; Govindjee (eds) Chlorophyll fluorescence: a signature of photosynthesis. Kluwer Academic Publishers, London, pp 321–362; 2004.
Synkova H. Sucrose affects the photosynthetic apparatus and the acclimation of transgenic tobacco to ex vitro culture. Photosynthetica 33: 403–412; 1997.
Tsimilli-Michael M.; Strasser R. J. Fingerprints of climate changes on the photosynthetic apparatus behavior, monitored by the JIP-test. In: Walther G.-R.; Burga C. A.; Edwards P. J. (eds) Fingerprints of climate changes (adapted behavior and shifting species ranges). Kluwer Academic Publishers, New York, pp 229–247; 2001.
Van Heerden P. D. R.; Tsimilli-Michael M.; Krüger G. H.; Strasser R. J. Dark chilling effects on soybean genotypes during vegetative development; parallel studies of CO2 assimilation, chlorophyll a fluorescence kinetics, OJIP and nitrogen fixation. Physiol Plant 117: 476–491; 2003.
Van Huylenbroeck J. M.; Debergh P. C. Impact of sugar concentration in vitro on photosynthesis and carbon metabolism during ex vitro acclimatization of Spathiphyllum plantlets. Physiol Plant 96: 298–304; 1996.
Wu F. Z.; Bao W. K.; Li F. L.; Wu N. Effect of water stress and nitrogen supply on leaf gas exchange and fluorescence parameters of Sophora dardii seedings. Photosynthetica 46(1): 40–48; 2008.
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Funding support by the University Grants Commission, India through SAP-DRS program is gratefully acknowledged.
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Editor: D. T. Tomes
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Swain, S.S., Tripathy, T., Mohapatra, P.K. et al. Photosynthetic and transpiration responses of in vitro-regenerated Solanum nigrum L. plants to ex vitro adaptation. In Vitro Cell.Dev.Biol.-Plant 46, 134–141 (2010). https://doi.org/10.1007/s11627-009-9269-8
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DOI: https://doi.org/10.1007/s11627-009-9269-8