Embryogenic callus culture of pereskia grand/folia and pereskia bleo and determination of their cytotoxicity effect

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Date
2007
Authors
Eng Leng, Chuah
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Abstract
Aseptic leaf explants of Pereskia grandifolia and Pereskia bleo could be established via surface-sterilization process using 150 mg/L sodium dichloroisocyanurate (NaOCC) for 15 minutes. Only embryogenic callus could be induced from P. grandifolia mid-vein leaf explants after culturing on Gamborg (85) medium supplemented with 6.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-0) for four weeks. The embryogenic callus of P. grandifolia could be multiplied by subculturing every four weeks 0.5 g callus biomass on 20 ml of solid media with the same constituents. Embryogenic cells of P. grandifolia could be propagated by culturing the friable embryogenic callus in liquid 85 medium supplemented with 6.5 mg/L 2,4-0. Subculturing of embryogenic cells was carried out every 12 days. Leaf explants of P. bleo did not produced any embryogenic callus when they were culture on culture medium with the same formulation. 85 medium containing 8A and NAA (2 - 10 mg/L) induced the formation of green filamentous structure (GFS) from the leaf explants of both Pereskia species, P. grandifo/ia and P. bleo. Microscopy study showed that GFS was made up of chains of long cells and contained chlorophyll granules in each cell. Hexane extracts of P. grandifolia and P. bleo mother plants showed cytotoxicity effect with EC50 values of 5.92 llg/ml (76.0 %) and 15.95 11g/ml (80.7 %) respectively towards lung cancer cells (NCI-H23). Hexane extract of in vitro embryogenic cells also showed anti-cancer activities towards lung cancer cells but level of effectiveness was less than 50 % and the ECso value could not be determined. The hexane extracts of the mother plants of both Pereskia species also showed ability to inhibit the growth of ovarian cancer cells (Caov-3) but its cytotoxicity effect was lower as compared to the effect towards NCI-H23 cells. The chloroform and butanol extracts for both the mother plants of P. grandifolia and P. bleo and the embryogenic calluses possessed low cytotoxicity effects towards both the lung and ovarian cancer cells. However, the GFS did not showed any cytotoxicity effect towards both of the cancer cells.
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Keywords
Callus culture , Pereskia bleo , Cytotoxicity effect
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