Establishment Of Cell Cultures For Narciclasine And Tazettine Production And Pharmacological Evaluation Of Hymenocallis Littoralis (Jacq.) Salisb
| dc.contributor.author | Noormi, Rosli | |
| dc.date.accessioned | 2018-07-18T02:22:17Z | |
| dc.date.available | 2018-07-18T02:22:17Z | |
| dc.date.issued | 2015-06 | |
| dc.description.abstract | A study was carried out to determine narciclasine and tazettine distribution in vivo intact plants from juvenile and flowering stages; and in vitro cultures namely callus, cell suspension, root and treated root cultures of Hymenocallis littoralis (Jacq.) Salisb plant. Phytohormone 2,4-D at 2.0 mg/L was the best auxin for the callus induction and growth proliferation in all different types of explants. Half continuous light with full strength of MS medium, pH 5.5 and sucrose at 3 % (w/v) was the best condition for callus growth and proliferation. The initiation of Hymenocallis littoralis cell suspension cultures in bulb explants was carried out by using 2.0 mg/L 2,4-D. Cell suspension cultures at four weeks cultivation period with half continuous light, full strength of MS medium, pH 5.5, sucrose at 3 % (w/v) and 2.0 mg/L 2,4-D produced the highest amount of fresh weight biomass. Roots could be induced from in vitro root culture using MS medium supplemented with 1.4 mg/mL IAA. Highest biomass production for root proliferation was obtained at fourth week under half continuous light with full strength of MS medium, 4% (w/v) sucrose and pH 6. The histology of control and treated root demonstrated the present of clear vascular bundle system in treated root which indicated it can be easily multiply within short period of time. Twelve (12) samples crude extracts of different explants from juvenile and flowering stages of Hymenocallis littoralis (Jacq.) Salisb wild plants were used to screen the antimicrobial and antioxidant activities. In antimicrobial assay, flowering stage explants showed pronounced inhibition of microbe’s growth of the cultures compared to the juvenile stage with different range on inhibition zone. Roots from the flowering stage produced highest activity against Bacillus subtilis (zone of inhibition was 29.0 + 0.04 mm). Best MIC was obtained at 6.25 mg/mL from the juvenile roots and flowering old leaves against Micrococcus spp.; flowering old leaves and stems against Escherichia coli hospital strain; and juvenile old leaves, flowering old leaves and stems against Candida albicans. For time kill study results, growth profile curve shows the fungicidal activity exhibited by H. littoralis flowering old leaf extract against C. albicans. H. littoralis flowering old leaf extract inhibited both growth of C. albicans and E. coli at different incubation durations with extract was more fungicidal against C. albicans compared to bactericidal against E. coli. SEM analysis for H. littoralis treated leaf extract against C. albicans and E. coli after 16 h of treatment resulted distorted cells with the presence of invaginations and convolutions incidence. Higher degree of collapsed cells was obtained at 20h. The antioxidant activities presented in all 12 samples of Hymenocallis littoralis juvenile and flowering stages based on the results obtained from DPPH, ABTS and FRAP methods. Young leaf at juvenile stage produced highest antioxidant activities (DPPH (44.2%), ABTS (99.64%) and FRAP (630 mgFeSO4/g DW extract)) and the total phenolic content (1119 mg Garllic acid/g DW extract). Results for analgesic activity based on hot plate and tail flick assays demonstrated significant analgesic properties of Hymenocallis littoralis (Jacq.) Salisb plant’s extracts. Mice treated with 400 mg/kg flowering root extracts produced 65.37 % of analgesic activity and was the highest analgesic activity within in all the tested samples. For toxicity studies, the results obtained showed that toxicity of the plant extracts increases accordingly with the extract concentrations. Chloroform methanolic juvenile root extract [bioactivity by LC50 :-1.118 (6 h), -3.583 (12 h), -4.245 (18 h), 0 (24 h)] exhibited the highest toxicity among all extracts. Results on wound healing activity indicated that juvenile bulb extract [56.40 + 2.640 d (1 μg/mL), 150.21 + 2.44c,d (10 μg/mL)] was the most effective for the wound healing activity based on scratch assay with Hs-27 cell lines within 12 hours compared to control treatment, negative group and other types of explant’s extracts. Qualitative analysis using TLC revealed that narciclasine and tazettine production was highest using in vivo root of Hymenocallis littoralis (Jacq.) Salisb plant from juvenile and flowering stages. Callus culture produced the highest narciclasine and tazettine under in vitro condition. The results of HPLC analysis based on chloroform methanolic extracts confirmed that flowering stage produced the highest concentration of narciclasine (74.07+ 0.017 mg/g DW tissue). Meanwhile, callus cultures produced highest narciclasine (11.87 + 0.021 mg/g DW tissue) under in vitro condition. The highest concentration of tazettine was detected in flowering root (7.264 + 0.003 mg/g DW tissue). Similarly, callus culture contained highest tazettine (5.456 + 0.003 mg/g DW tissue) under in vitro condition. As a comparison, the content of narciclasine was significant tested in all samples higher at ten times compare to the content of tazettine. The production of narciclasine and tazettine in vitro will be enhanced with large scale through bioreactor technology system. | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/5949 | |
| dc.subject | Narciclasine | en_US |
| dc.subject | Pharmacological Evaluation | en_US |
| dc.title | Establishment Of Cell Cultures For Narciclasine And Tazettine Production And Pharmacological Evaluation Of Hymenocallis Littoralis (Jacq.) Salisb | en_US |
| dc.type | Thesis | en_US |
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