In vitro photoautotrophic acclimatization, direct transplantation and ex vitro adaptation of rubber tree (Hevea brasiliensis)

  • Rujira Tisarum
  • Thapanee Samphumphung
  • Cattarin Theerawitaya
  • Wittaya Prommee
  • Suriyan Cha-um
Original Article


We investigated the effect of carbon dioxide (CO2)-ambient (350 µmol CO2 mol−1) and CO2-enriched (1500 µmol CO2 mol−1) conditions of in vitro photoautotrophic system on two cultivars, ‘RRIM600’ and ‘RRIT413’ of rubber tree (Hevea brasiliensis) in an acclimatization process of 45 days. Survival percentage of in vitro rubber tree plantlets derived from somatic embryos under ambient CO2 was better than those under CO2-enriched conditions, especially in cv. ‘RRIT413’. Subsequently, the survival rate of ex vitro transplanted plantlets was similar to the in vitro plantlets and abnormal morphological characters such as light-green leaves (SPAD), small leaves in cv. ‘RRIT413’ acclimatized under CO2-enriched conditions were demonstrated 30 days after the plantlets were transferred into the soil. Maximum quantum yield of PSII, photon yield of PSII, stomatal conductance and transpiration rate in cv. ‘RRIT413’ acclimatized under CO2-enriched conditions were sharply declined by 39.0, 50.6, 47.1 and 45.8%, respectively as compared to those acclimatized under ambient CO2 conditions. In contrast, the in vitro acclimatized plantlets of cv. ‘RRIM600’ were un-responsive under both ambient- and enriched-CO2 conditions. In conclusion, genotypic dependent in response to CO2 enriched conditions in in-vitro acclimatization of rubber tree plantlets was evidently demonstrated as a key result to regulate plant growth and development in ex vitro environments. Interestingly, soluble sugar contents (sucrose, glucose and fructose) were increased after transplanting the plantlets of cv. ‘RRIM600’ acclimatized under CO2-enriched condition into the soil and thus, can be considered as an adaptive indicator of ex vitro adaptation.


Chlorophyll fluorescence CO2 enrichment Net photosynthetic rate SPAD Survival percentage 



The authors wish to thank Rubber Research Institute of Thailand, Department of Agricultural, Ministry of Agricultural and Cooperative, as funding source and partially support by National Science and Technology Development Agency.

Author Contributions

The experiment design, statistical analysis and manuscript preparation were prepared by SC, somatic embryogenesis of rubber tree was provided by WP, in vitro acclimatization and transplantation was processed by TS, soluble sugar assay was done by CT and physiological and morphological data was recorded by RT. All authors were involved with editors of all versions, and agreed to the final version for publication, assuming public responsibility for the results.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  1. 1.National Center for Genetic Engineering and Biotechnology (BIOTEC)National Science and Technology Development Agency (NSTDA)Pathum ThaniThailand
  2. 2.Chachoengsao Rubber Research CenterRubber Research InstituteChachoengsaoThailand
  3. 3.National Center for Genetic Engineering and Biotechnology (BIOTEC)National Science and Technology Development Agency (NSTDA)Pathum ThaniThailand

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