Abstract
The growth ofGardenia jasminoides Ellis plantlets and the development of photoautotrophy during two successive culture stages (shoot multiplication and root induction)in vitro was analyzed. We examined the effects of changes in growth conditions (type of tube closure, light, and sugar levels) on the development of photoautotrophy and growth during micropropagation and sought to establish whether they affected later acclimation to conditionsex vitro. During the two stagesin vitro, plantlets were grown in tubes under two different PPFD (50 and 110 µmol m−2 s−1), in media with three different sucrose concentrations (0, 1.5, and 3.0%, w/v) and with two different CO2 levels inside the tubes (controlled by either tightly closed caps or loosely sealed caps, and with an external CO2 concentration of 750 µmol mol−1). The development of photoautotrophy was assessed by determining the difference between the stable carbon isotope composition (δ13C) of sugar cane sucrose used as a heterotrophic carbon source and that of leaflets grownin vitro. Plantlets from the root-induction stage showed a more highly developed photoautotrophy than those from the shoot- multiplication stage. At both stages, utilization of closed caps was the treatment which most stimulated development of photoautotrophy in plantlets. Also, lowering PPFD or sucrose concentration induced a greater degree of photoautotrophic development, the strongest effect being observed in plantlets cultured inside loosely sealed tubes. During acclimationex vitro, plantlets taken from loosely sealed tubesin vitro performed better than those cultured inside tightly sealed tubes. The former, as well as recording a larger increase in fresh weight during this stage, also showed more negative δ13C in the newly developed leaves, which would seem to indicate a better water status during acclimation. Present results validate the usefulness of δ13C analysis of leaflets as a simple technique in assessing the development of photoautotrophy during culturein vitro. In addition, δ13C analysis can be extended to evaluate growth conditions during acclimation toex vitro conditions.
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Abbreviations
- LS-T:
-
culture in loosely-sealed tubes under an external CO2 concentration of 750 µmol mol−1
- TS-T:
-
culture in tightly-sealed tubes at external CO2 concentration of 750 µmol mol−1
- Δ:
-
stable carbon isotope discrimination
- δ13C:
-
ratio of13C/12C relative to PeeDee belemnite standard
- MS medium:
-
Murashige and Skoog medium
- p i /p a :
-
ratio of intercellular to atmospheric partial pressure of CO2
- PPFD:
-
photosynthetic photon flux density
- L-PPFD:
-
culture with 50±5 µmol m−2 s−1 PPFD
- H-PPFD:
-
culture with 110±10 µmol m−2 s-1 PPFD
- N-Suc:
-
medium with no sucrose
- L-Suc:
-
medium with 0.5% sucrose
- H-Suc:
-
medium with 3% sucrose
- IAA:
-
indole-3-acetic acid
- BA:
-
N6-benzyladenine
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Serret, M.D., Trillas, M.I., Matas, J. et al. The effect of different closure types, light, and sucrose concentrations on carbon isotope composition and growth ofGardenia jasminoides plantlets during micropropagation and subsequent acclimationex vitro . Plant Cell Tiss Organ Cult 47, 217–230 (1997). https://doi.org/10.1007/BF02318976
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DOI: https://doi.org/10.1007/BF02318976