In-vitro callus formation in leaf and petiole sections of ashitaba (Angelica keiskei Koidz.) and enhancement of total phenols in callus cultures in response to UV-B radiation

Date

12-2015

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Evalour T. Aspuria

Abstract

The high phenolic content of Ashitaba (Angelica keiskei KOIDZ.) extracts and their derivatives are of pharmacological interest because of their cancer fighting properties. This study was conducted to study how UV-B may enhance in callus cultures of Ashitaba. Callus cultures were initiated from leaf and petiole explants in Murashige and Skoog (MS) medium supplemented with 1-2 mg/L 2, 4-D and 0.5-2.0 mg/L BA. White friable callus induced in leaf explants between 34 and 45 days of inoculation in all treatments. On the other hand, either green compact or white friable callus was induced in petiole explants between 21 and 36 days. Green compact calli gave rise to white friable calli. The highest percentage of callus formation (75.05%) was noted for treatments with 2.0 mg/L 2, 4-D + 1.0 mg/L BA and a remarkable percentage callus formation of 100% was obtained from petiole explants on all MS media formulations supplemented with 2, 4-D and BA. White friable calli that formed from leaf and petiole explants eventually turned brown after sometime in culture medium. The best medium for callus induction of leaf and petiole explants based from statistical analysis of data gathered was MS medium supplemented with 1.0 mg/L 2, 4-D and 0.5 mg/L BA. Shoot-like and or bud-like structures from leaf-derived calli were also observed in selected MS media formulations. It is possible to have micropropagation studies of Ashitaba in these media formulations through indirect organogenesis. The induced calli were then subjected to UV-B treatments for 3- or 6-hour durations for 7 days to enhance phenolic compounds. Quantitative determination of phenols in leaf and callus extract were carried out using spectrophotometric analysis with Gallic acid as the standard. The calli irradiated in 3-hour and 6-hour UV-B exhibited a 41.20% (380.65 ug GAE/g) and a 54.86% (417.46 ug GAE/ g) increase in total phenolic content, respectively, based on the levels found in the control cultures (269.56 ug GAE/g). However, enhancement of phenolic compounds of petiole-derived calli through UV-B light irradiation was only effective in MS medium supplemented with 1 mg/L 2, 4-D and 0.5 mg/L BA and MS medium supplemented with 2 mg/L 2,4-D and 0.5 mg/L BA irradiated for 3 hours. These results showed that in-vitro production of phenolic compounds in Ashitaba using callus is possible and may be further explored as an alternative system for plant secondary metabolite production.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2015 A127 /A24

Document Type

Thesis

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