Selection Of Elite Cell Lines, Optimization And Elicitation Of Cell Suspension Culture Of Misai Kucing (Orthosiphon Stamineus Benth.) For The Production Of Rosmarinic Acid

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dc.contributor.authorBordbar, Laleh
dc.date.accessioned2017-11-01T02:03:22Z
dc.date.available2017-11-01T02:03:22Z
dc.date.issued2014-06
dc.description.abstractPlants from tropical rain forests have incredible abilities to produce biological and chemical defense resources, which make them suitable leads for the synthesis of new drugs and pharmaceutical products. Orthosiphon stamineus Benth., commonly known as Misai Kucing is a very important medicinal plant that has been used for the treatment of many ailments, especially as a remedy for removal of kidney stones. Many chemical compounds were successfully isolated from O. stamineus and rosmarinic acid (RA) is one of the major secondary metabolites. However, the production of most secondary metabolites from plants varies due to the location and environmental factors. Hence, in vitro culture techniques could be used as the alternative for consistent production of these useful medicinal compounds. In the present study, attempts were made to produce rosmarinic acid from cell suspension culture of O. stamineu. Two callus lines of O. stamineus were induced from the in vitro plants. The induced callus lines with another eight callus lines maintained from the previous studies were selected and categorized based on the growth index into three groups of fast, intermediate and slow growing lines. The factors influencing cell biomass and secondary metabolite production which include, agitation, sucrose concentration, medium pH, photoperiod and temperature, were studied on the categorized cell lines. Effect of the MS medium nutrients (macronutrients and micronutrients) was also studied in which five macronutrients of MS were individually studied and the micronutrients were tested in different concentrations of MS strengths (0, ½ MS, MS, 1.5MS and 2MS strengths). The impact of different elicitors including silver nitrate, salicylic acid and methyl jasmonate and also the effects of flask size (100, 250, 500 and 1000 mL) on cell biomass and RA production were evaluated. The results indicated that the slow and fast growing cell lines produced high fresh cell biomass when agitated at 120 rpm. However, the intermediate growing cell line produced no significantly different cell biomass when grown with agitation at 100 and 120 rpm. The fast and intermediate growing cell lines produced higher fresh cell biomass when cultured in cell proliferation medium with 30 g/L sucrose compared to 45 g/L sucrose except it was not significantly different in the slow growing cell line. The most suitable medium pH was 5.65 - 5.8 for cell biomass and RA production of the three cell lines. O. stamineus cell lines showed differences in terms of temperature and photoperiod requirment. Based on the medium modification results two media were formulated, Bio-Mod and RA-mod media. When the O. stamineus cells were cultured in Bio-Mod and RA-Mod media for the production of cell biomass and RA respectively, it was found that the cells cultured in the cell proliferation medium (MS + 1 mg/L NAA and 1 mg/L 2,4-D) produced higher cell biomass and RA content compared to the two modified media (Bio-mod and RA-mod). In the elicitation study,en_US
dc.identifier.urihttp://hdl.handle.net/123456789/5198
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectCulture of Misai kucingen_US
dc.subjectproduction of rosmarinic aciden_US
dc.titleSelection Of Elite Cell Lines, Optimization And Elicitation Of Cell Suspension Culture Of Misai Kucing (Orthosiphon Stamineus Benth.) For The Production Of Rosmarinic Aciden_US
dc.typeThesisen_US
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