The effects of plant growth regulators on accumulation of pinocembrin and cardamonin of adventitious root suspension cultures of boesenbergia rotunda (L.) mansf. / Siti Zulaiha Ahamad Azhar

• Main Author: Ahamad Azhar, Siti Zulaiha
• Format: Thesis
• Language: English
• Published: 2020
• Subjects: Medicinal plants (Culture only)
• Online Access: https://ir.uitm.edu.my/id/eprint/60039/1/60039.pdf; https://ir.uitm.edu.my/id/eprint/60039/

Boesenbergia rotunda is an important medicinal plant. The rhizome requires a long period of time to produce in a large scale for this plant. In vitro culture is the best method as a continuous source of supply of disease free planting material for commercial utilization because it offers an opportunity to exploit the cell, tissue, organ or entire organism by growing them in vitro. Secondary metabolites of B. rotunda possess many biological and pharmacological activities such as antioxidant, antibacterial, antitumor, anti-inflammatory and anti-cancer activities. Pinocembrin has shown cytotoxicity against certain cancer cell lines whereas cardamonin was reported to have strong inhibition of HIV-1 protease activity. There were a few studies on secondary metabolites of B. rotunda in cell culture and callus culture. However, this plant has low content of secondary metabolites in cell suspension cultures and callus culture. This study was conducted to establish the protocol for adventitious root induction from shoot meristem of B. rotunda and to determine the effects of PGRs on the biomass and accumulation of pinocembrin and cardamonin in adventitious root suspension cultures of B. rotunda. Surface sterilization with 60% clorox for 30 minutes, followed by 20% clorox for 15 minutes was the best treatment to establish the aseptic explants with mean value of 4.667 ± 0.724. For the adventitious root induction, half strength of Murashige and Skoog (MS) medium was supplemented with various concentrations of auxins; 1-naphthaleneacetic acid (NAA), indole-3- butyric acid (IBA), and 2,4-dichlorophenoxyacetic acid (2,4-D) at a range of 0.5 mg/L-3.0 mg/L either alone or in combination of cytokinins; kinetin (Kn) and thidiazuron (TDZ) at a range of 0.1 to 0.5 mg/L. The most effective treatment for adventitious root induction was supplemented with 0.5 mg/L NAA which produced the highest number and length of roots with a mean value of 7.667 ± 2.082 and 1.733 ± 0.416 respectively. The most effective combination of auxin and cytokinin for adventitious root induction was 0.5 mg/L NAA+ 0.1 mg/L Kn that produced the highest number and length of roots with a mean value of 3.400 ± 2.408 and 1.330 ± 1.021 respectively. The best treatment for the biomass production of adventitious root of B. rotunda was 0.5 mg/L NAA + 0.1 mg/L TDZ with a mean value of 3.684 ± 1.233 g fresh weight and also produced the highest number of shoots with a mean value of 3.167 ± 1.169. Histological examinations of adventitious root of B. rotunda revealed the presence of important root anatomic structures including epidermis, endodermis, pericycle, phloem, cortex, late metaxylem and protoxylem pole. HPLC analysis showed that both pinocembrin and cardamonin were present in the extract of adventitious root suspension cultures of B. rotunda. The highest compound contents were 13.09 ± 3.92 mg/g for pinocembrin whereas 16.95 ± 4.01 mg/g for cardamonin when the adventitious root were respectively cultured into 0.5 mg/L NAA + 0.3 mg/L TDZ and 0.5 mg/L NAA + 0.1 mg/L TDZ. The result shows that the PGRs (NAA and TDZ) is a good approach for enhancing the secondary metabolite content in adventitious root cultures of B. rotunda. Further research using bioreactor, elicitors, application of hairy root culture, histological analysis at cellular level and pharmacological properties should be done as B. rotunda is an important aromatic and medicinal plant.