Physiology and Molecular Biology of Plants

, Volume 25, Issue 6, pp 1457–1467 | Cite as

Biochemical analyses of Dendrobium Sabin Blue PLBs during cryopreservation by vitrification

  • Jessica Jeyanthi James AntonyEmail author
  • Suhana Zakaria
  • Rahmad Zakaria
  • Jorim Anak Ujang
  • Nurulhasanah Othman
  • Sreeramanan SubramaniamEmail author
Research Article


Dendrobium Sabin Blue is an important orchid hybrid that has been grown extensively as cut flower, potted plant and is also popular for its deep purplish blue flowers.  The most efficient long term conservation method of this hybrid is through cryopreservation. Cryopreservation involving the vitrification method consists of explants exposure to highly concentrated cryoprotective solution followed by freezing rapidly in liquid nitrogen. However, these treatments involved highly concentrated cryoprotectant that could incur toxicity to the explants. Hence, cryopreservation protocol requires biochemical analyses in understanding the damages or injuries occurred during cryopreservation treatments. In this study, biochemical analyses revealed a general reduction in chlorophyll, carotenoid and porphyrin content to 0.40 µg/g F W (thawing stage), 31.50 µg/g F W unloading stage and 2230.41 µg/g F W (thawing stage), respectively in comparison to the control treatments. In addition, increased level in proline content were obtained at different cryopreservation stages with highest level (5.42 µmole/g F W) recorded at the PVS2 dehydration stage. Fluctuated outcomes were obtained in catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POX) enzyme activities in PLBs exposed to different cryopreservation stages. Lowest values recorded for CAT enzyme activity were obtained at the dehydration stage (3.94 U/g). Lowest POX enzyme activities were obtained at the dehydration (122.36 U/g) and growth recovery (106.40 U/g) stages. Additionally, lowest APX enzyme activities values were recorded at the thawing (7.47 U/g) and unloading (7.28 U/g) stages. These have contributed to low regeneration of Dendrobium Sabin Blue protocorm like bodies (PLBs) following cryopreservation. Hence, in the future experimental design, exogenous antioxidant could be included in the cryopreservation procedures to improve the existing protocol.


Cryopreservation Catalase (CAT) Ascorbate peroxidase (APX) Peroxidase (POX) Chlorophyll Carotenoid Porphyrin Proline 



Abscisic acid


Ascorbate peroxidase




Liquid nitrogen


Protocorm like bodies




Plant vitrification solution 2


Reactive oxygen species


Superoxide dismutase



The authors would like to thank financial support from Malaysian Ministry of Higher Education for FRGS 2014 Grant (203/PBIOLOGI/6711456) and My Ph.D. Scholarship. Authors also present gratitude Universiti Sains Malaysia for supporting this study.

Authors contribution

Sreeramanan, Rahmad and Nurulhasanah provided the resources and supervision for the study and writing. Jessica, Suhana, Jorim and Sreeramanan designed the methodology for the study. Jessica carried out the experiments and wrote the article.


This research did not receive any specific Grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this article.


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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2019

Authors and Affiliations

  1. 1.School of Biological SciencesUniversiti Sains Malaysia (USM)GeorgetownMalaysia
  2. 2.Department of Crop Science, Faculty of Agricultural and Food SciencesUniversiti Putra MalaysiaGeorgetownMalaysia
  3. 3.Institute for Research in Molecular Medicine (INFORMM)Universiti Sains MalaysiaGeorgetownMalaysia

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