RESUMEN

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).

Asunto(s)

Mitragyna/efectos de los fármacos , Reguladores del Crecimiento de las Plantas/farmacología , Alcaloides de Triptamina Secologanina/metabolismo , Ácido 2,4-Diclorofenoxiacético/farmacología , Compuestos de Bencilo , Iridoides/farmacología , Cinetina/farmacología , Mitragyna/crecimiento & desarrollo , Mitragyna/metabolismo , Ácidos Naftalenoacéticos/farmacología , Purinas , Técnicas de Cultivo de Tejidos , Triptófano/farmacología

RESUMEN

Mitragynine is a pharmacologically-active terpenoid indole alkaloid found in Mitragyna speciosa leaves. Treatment with methyl jasmonate (10 µM) for 24 h and yeast extract (0.1 mg/ml) for 12 h were the optimum conditions of elicitation of mitragynine accumulation in a M. speciosa shoot culture. The former elicitor gave 0.11 mg mitragynine/g dry wt. Tryptophan decarboxylase and strictosidine synthase mRNA levels were enhanced in accordance with mitragynine accumulation.

Asunto(s)

Acetatos/farmacología , Ciclopentanos/farmacología , Mitragyna/efectos de los fármacos , Mitragyna/metabolismo , Oxilipinas/farmacología , Alcaloides de Triptamina Secologanina/metabolismo , Levaduras/química , Descarboxilasas de Aminoácido-L-Aromático/genética , Descarboxilasas de Aminoácido-L-Aromático/metabolismo , Liasas de Carbono-Nitrógeno/genética , Liasas de Carbono-Nitrógeno/metabolismo , Medios de Cultivo , Técnicas de Cultivo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Mitragyna/crecimiento & desarrollo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo