Development of novel in vitro systems for natural anthocyanin production
Juthangkoon, Suthin
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https://hdl.handle.net/2142/21306
Description
Title
Development of novel in vitro systems for natural anthocyanin production
Author(s)
Juthangkoon, Suthin
Issue Date
1995
Doctoral Committee Chair(s)
Smith, Mary Ann Lila
Department of Study
Crop Sciences
Discipline
Crop Sciences
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Agriculture, Food Science and Technology
Agriculture, Plant Culture
Biology, Plant Physiology
Language
eng
Abstract
A reliable in vitro system for natural anthocyanin production in Ajuga pyramidalis 'Metallica Crispa' was developed. We found that 2,4-D was necessary to induce callus culture. WPM salts supplemented with sucrose 30%, IAA 2.26 $\mu$M, zeatin 3.49 $\mu$M, and pH 5.7 (adjusted prior autoclaving) was optimal for growth and anthocyanin production in callus cultures. In cell culture suspensions, the medium composition was similar to callus cultures, but sucrose concentration was changed to 50%. Suspension cultures were capable of producing anthocyanins at 41-42 mg $\cdot$ 100 g FW as compared to 7-8 mg in callus cultures and 10-12 mg in pigmented leaf tissue in vivo. The physical microenvironment including temperature at $25\sp\circ$C, and irradiance at a PPF of 150 $\mu$mol m$\sp{-2}$ s$\sp{-1}$ provided by cool-white fluorescent lamps was optimal for the system. Callus cultures of Begonia, Ocimum, and Tradescantia were established for the system. Anthocyanin expression was observed in callus cultures of Begonia and Ocimum in the dark. However, fast growing and friable cultures were not obtained for these two crops. We did obtain friable callus from Tradescantia, but the cultures grew very slowly and never expressed anthocyanin. Anthocyanin from Ajuga suspension cultures was more stable than in vivo leaf extracts. Radiation stability of in vitro anthocyanin from Begonia was not different from in vivo. Stability of anthocyanin from both sources was low compared to Ajuga and Tradescantia. In vivo anthocyanin from Tradescantia was more stable than in vitro. PAL activity was observed in colorless callus cultures of Tradescantia. This result suggested that the lack of anthocyanin expression in the cultures might relate to other enzymes in the anthocyanin biosynthetic pathway.
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