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