Abstract
Boesenbergia rotunda is an important medicinal herb that contains useful bioactive compounds. Although micropropagation of ginger has undergone spectacular development in recent years, gradual loss of plant regeneration capacity in prolonged cell suspension culture continues to be a significant limitation. In this study, biochemical changes and proteins associated with plant totipotency over 9 months of culture for B. rotunda cell suspension culture were investigated. The established cell line B showed 42.2 and 53.8% decrease in growth after 6 and 9 months of culture respectively compared to 0 month-old cells. About 67, 57 and 47% of cell-derived shoot-like-structures for 0, 6 and 9 month-old cells, respectively, regenerated into shoots. The activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) for 6 month-old cells were 14.40 Unit SOD/mg protein, 74.38 µmol H2O2/mg protein, and 2.23 µmol AsA/mg protein, respectively, higher than 0-month-old cells. The difference for hydrogen peroxide (H2O2) content between 0 and 6 month-old cells was 5.27 µmol H2O2/mg protein. While the activities of SOD, H2O2 and APX decreased from 6 to 9 month-old cells, CAT activity was increased. We applied a gel-based proteomic technique to analyze protein changes for 0, 6 and 9 month-old cells. A total of 13 protein spots showed significant differential expression and 8 protein spots were successfully identified. They were classified as protein synthesis, energy metabolism, defense and stress responses, and catabolic proteins.
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Acknowledgements
This project was supported by University Teknologi MARA (UiTM) under Grants 600RMI/RAGS5/3(193/2012) and 600RMI/FRGS5/3(84/2015), the Ministry of Science, Technology and Innovation (MOSTI), Malaysia, under Escience Grant (02-01-03-SF0999) and University of Malaya under CEBAR RU Grant (RU015-2015, PV022-2016 and PV005-2017).
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NZK, BCT and NAY conceived the idea, designed the experiments and edited the manuscript. AFZ conducted the experiments and wrote the manuscript. AFZ and BCT analyzed the data. All authors read and approved the final manuscript.
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Supplementary figure 1—Proteins selected for mass spectrometry analysis and changes of protein abundance over 0–9 month-old cell suspension cultures (TIF 45 KB)
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Aziz, A.F., Yusuf, N.A., Tan, B.C. et al. Prolonged culture of Boesenbergia rotunda cells reveals decreased growth and shoot regeneration capacity. Plant Cell Tiss Organ Cult 130, 25–36 (2017). https://doi.org/10.1007/s11240-017-1201-z
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DOI: https://doi.org/10.1007/s11240-017-1201-z