Abstract
Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC) and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS), Woody Plant Medium (WPM) and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D). Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume) of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC) method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.
Highlights
Barringtonia racemosa, (Lecythidaceae family) with natural habitat of the West Coast of India, Sundarbans, Assam, Andaman Islands and Malaysia (Thomas et al, 2002) is a traditional medicine in Malaysia
During the third week some calli developed in WPM medium supplemented with 2 mg L–1 2,4-D, (20 days ± 1sd) (Table 1)
WPM resulted in a higher percentage of callus induction (Table 1); WPM medium was supplemented with 2 mg L–1 2,4-D treatment which was observed to yield more callus (90%, dry weight: 12.4 ± 0.4) compared to other treatments in dark condition (p < 0.05)
Summary
Barringtonia racemosa, (Lecythidaceae family) with natural habitat of the West Coast of India, Sundarbans, Assam, Andaman Islands and Malaysia (Thomas et al, 2002) is a traditional medicine in Malaysia. The active compound responsible for the anti-oxidant and anti-inflammatory properties of the leaves was found to be lycopene and B. racemosa is one of the most important sources of this compound compared to any other reported plants (Behbahani et al., 2007b). This most predominant carotenoid is a potent in vitro scavenger of singlet oxygen as an antioxidant
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