Induction and analysis of the alkaloid mitragynine content of a Mitragyna speciosa suspension culture system upon elicitation and precursor feeding.
This study aimed to determine the effects of different concentrations and combinations of the phytohormones 2,4-dichlorophenoxy acetic acid (2,4-D), kinetin, 6-benzylaminopurine (BAP), and 1-naphthaleneacetic acid (NAA) on callus induction and to demonstrate the role of elicitors and exogenous precursors on the production of mitragynine in a Mitragyna speciosa suspension culture. The best callus induction was achieved from petiole explants cultured on WPM that was supplemented with 4 mg L⁻¹ 2,4-D (70.83%). Calli were transferred to liquid media and agitated on rotary shakers to establish Mitragyna speciosa cell suspension cultures. The optimum settled cell volume was achieved in the presence of WPM that contained 3 mg L⁻¹ 2,4-D and 3% sucrose (9.47 ± 0.4667 mL). The treatment of cultures with different concentrations of yeast extract and salicylic acid for different inoculation periods revealed that the highest mitragynine content as determined by HPLC was achieved from the culture treated with 250 mg L⁻¹ yeast extract (9.275 ± 0.082 mg L⁻¹) that was harvested on day 6 of culturing; salicylic acid showed low mitragynine content in all concentrations used. Tryptophan and loganin were used as exogenous precursors; the highest level of mitragynine production was achieved in cultures treated with 3  μM tryptophan and harvested at 6 days (13.226 ± 1.98 mg L⁻¹).
School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia.
Nor Nahazima
NN
Mohamad Zuldin
Ikram Md
IM
Said
Normah
N
Mohd Noor
Zamri
Z
Zainal
Chew
C
Jin Kiat
Ismanizan
I
Ismail
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D001593:Benzyl Compounds; D039823:Iridoids / Q000494:pharmacology; D007701:Kinetin / Q000494:pharmacology; D032065:Mitragyna / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D009280:Naphthaleneacetic Acids / Q000494:pharmacology; D010937:Plant Growth Regulators / Q000494:pharmacology*; D011687:Purines; D046948:Secologanin Tryptamine Alkaloids / Q000378:metabolism*; D046509:Tissue Culture Techniques; D014364:Tryptophan / Q000494:pharmacology
D001593:Benzyl Compounds; D039823:Iridoids; D009280:Naphthaleneacetic Acids; D010937:Plant Growth Regulators; D011687:Purines; D046948:Secologanin Tryptamine Alkaloids; D015084:2,4-Dichlorophenoxyacetic Acid; C034182:1-naphthaleneacetic acid; D014364:Tryptophan; C001801:mitragynine; C059516:loganin; C480551:benzylaminopurine; D007701:Kinetin