Rodent tuber (Typhonium flagelliforme Lodd.) is an herbal medicinal plant with anticancer activity. The genetic diversity of rodent tuber is low due to vegetative propagation. Somatic cell population of rodent tuber from Bogor had been irradiated with gamma ray to increase genetic diversity. There were 37 clones of first generation putative mutant (MV1) which had been analyzed based on morphological and RAPD markers. Out of those 37 MV1 clones, there were 17 clones which had undergone genetic mutation and had a diversified genetic profile. MV1 had been regenerated to fourth generation putative mutant clones (MV4). This research was aimed to analyze the mutation stability of MV4 based on morphological and RAPD markers. Clone 6-1-2 had the highest increase of the number of shoots and leaves than control and the other MV4 clones, with 4.7 and 19.7 shoots and leaves, respectively. Clone 6-1-1-6 obtained the highest increase in plant height than control and the other MV4 clones, i.e. 25.2 cm. Clone 6-9-5 had the weightiest fresh and dry weight, i.e. 41.67 gram and 12.01 gram respectively. RAPD molecular marker analysis of MV4 by using 15 primers had produced 64 polymorphic DNA bands out of 146 total bands. OPD-10 primer produced the highest number of polymorphic bands, i.e. 15 polymorphic bands out of 17 total bands with sizes 200-2000 bp. RAPD profile of MV4 had showed 5 main clusters at similiarity coefficient cut-off 0.91. Morphological characterization and RAPD analysis had proved the stability of genetic mutation of MV4.
Objective: The objective of this study was to determine the new bioactive compounds through gas chromatography–mass spectrometry analysis and the cytotoxic activity of two rodent tuber mutant plants against breast cancer cells (MCF-7).
Methods: The bioactive compounds in rodent tuber mutant plants were successfully increased by somaclonal variation using gamma rays irradiation technique. Further, the cytotoxicity activity of rodent tuber mutant plants was tested on breast cancer cell line (MCF-7) performed by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide assay method.
Results: This results study confirmed that the presence of phytochemical composition in the tuber of rodent tuber mutant plants KB 6–1–3–4 and KB 6–9–5 was found six bioactive compounds from fatty acid groups which have the potential as an anticancer compound, such as octadecanoic acid, hexadecanoic acid, hexadecanoic acid methyl ester, 9-octadecanoic acid, linolelaidic acid methyl ester, and butanoic acid. The results showed that extracts from rodent tuber mutant plants had a cytotoxicity effect on MCF-7 cancer cells with half maximal inhibitory concentration (IC50) values that were lower than the control (mother plant). In vitro tests of KB 6–1–3–4 and KB 6–9–5 against MCF-7 cancer cell lines have IC50 values of 12.482 μg/mL and 7.043 μg/mL, respectively, while it had a lower cytotoxicity effect with the IC50 value of control plant was 19.113 μg/mL. The mutant plants of KB 6-9-5 have 3 times more effective than control.
Conclusion: The results of this study clearly indicated that rodent tuber mutant plants have shown promising as an anticancer drug on breast cancer.
Morphological Characterization on Abnormalities of Oilpalm (Elaeis guineensis Jacq) Embryo Somatic Generated from Leaf Explant. Nesti F. Sianipar, Gustav A. Wattimena, Hajrial Aswidinnoor, Maggy Thenawidjaya S., Nurita Toruan-Mathius, and Gale Ginting. Somatic embryogenesis is the development of somatic cells to form a structure alike zygotic embryo direct or indirectly. Somatic embryos from young leaf explants could be induced from primary callus formed surrounding the palm-leaf rib. Embryogenic callus will develop to be somatic embryos which grew nonuniformly. Embryo somatic growth pattern of globular, asymmetric heart shape, and cotyledon ary stage produced different morphological variation. Morphological variability of in vitro somatic embryos could be due to high application of growth regulator 2,4-D at the beginning of initiation, subculture frequency, loaded cells, and polysomic cells from certain tissues. From the three clones used, which were clone 638, 636, and 558, there were different variation at each step of development stages, grouping morphologically into normal and abnormal based on the development of somatic embryos. The percentage of abnormality from the three clone used was clone 27% (638), 30% (636), and 46% (558). The normal somatic embryos at globular stage were round and bipolar shaped; while the abnormal embryos were oval and no bipolar. At heart-shape stage, the normal somatic embryos had symmetrical polarized surface; while the abnormal embryos had asymmetrical polarized surface. At the cotyledon stage, the normal embryos had monocot-tyledon; the abnormal ones were more than one cotyledon.
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