Evaluation Of Factors Affecting The Cell Suspension Cultures And The Production Of Bioactive Compounds Of Spilanthes acmella L.
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Date
2010-12
Authors
Lim, Fung Hui
Journal Title
Journal ISSN
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Publisher
Universiti Sains Malaysia
Abstract
In this study, five lines of the callus culture were established from the petioles of Spilanthes acmella L. The culture was initiated on callus induction medium, Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2, 4-D. The petiole explants were also cultured on MS medium with 0.5 mg/L 2, 4-D and different concentrations of picloram (0.0 – 2.0 mg/L). Callus initiated on the medium with the presence of picloram induced lesser callus when compared with the callus initiated with only 2, 4-D. Compact and granular calluses were continuously produced with each subculture cycle. The soft and friable callus line L4 was used as the initial material for the establishment of cell suspension culture in the liquid medium with the same composition. Observation on growth kinetic of the cells showed a sigmoid curve and both fresh weight and dried weight reached the highest cell mass after 15 days of culture. The yellow-coloured cells produced nearly ten fold cell mass from the initial weight of 0.5 g at the 8th subculture cycle. The addition of sucrose (30 g/L) in the medium was found to support the highest cell biomass compared to other concentrations of sucrose. Cells treated with casein hydrolysate (3.0 g/L) proved to enhance fresh and dried cell mass, when the elicitor was added on the first and the 8th day of culture. The growth of cells was improved when the yeast extract treatment was applied on the 8th day of culture. But, the addition of chitosan into the cell culture showed negative effect as it did not encourage any cell growth. Ethanol extracts from the in vitro plantlets, control cells
and elicited cells were analyzed by GC/MS. Compounds such as N-isobutyl-(2E, 4Z, 8Z, 10E)-dodecatetraenamide, stigmasterol, butylated hydroxytoluene and 9, 12-octadecadienoic acid were found in all the tested samples.
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Keywords
Evaluation of factors affecting the cell suspension cultures , and the production of bioactive compounds of Spilanthes acmella l