The Open Conference Proceedings Journal

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ISSN: 2210-2892 ― Volume 10, 2020

Cell Suspension and In Vitro Production of Colchicine in Wild Colchicum Hierosolymitanum Feib

The Open Conference Proceedings Journal, 2012, 3: 52-59

Nidal Q. Daradkeh, Rida A. Shibli, Ibrahim M. Makhadmeh, Feras Alali, Tamara S. Al-Qudah

Deaprtment of Horticulture and Agronomy, Faculty of Agriculture, The University of Jordan, Amman, Jordan.

Electronic publication date 28/12/2012
[DOI: 10.2174/1876326X01203020052]


Callus was induced from seeds of Colchicum hierosolymitanum Feib inoculated on the surface of MS media supplemented with 0.45 μM 2, 4-dichlorophenoxyacetic acid under dark conditions. Then callus was cultured on MS media supplemented with 4.52 μM 2, 4-dichlorophenoxyacetic for growth and maintenance. Friable callus from the fourth generation was transferred to liquid MS media supplemented with 0.54 μM 1-naphthaleneacetic acid to form cell suspension. Cells were successfully subcultured every 27 days on the same liquid media supplemented with 0.54 μM 1- naphthaleneacetic acid. Higher concentration (9 μM) of 6-benzyladenine with 0.45 μM 2, 4-dichlorophenoxyacetic acid resulted in higher cells fresh weight, while 1-naphthaleneacetic acid combinations with 6-benzyladenine had no effect on cell growth. On the other hand, the time for subculturing the cells into new fresh liquid media was determined to be after 27 days of incubation. (-) -Colchicine was identified in callus and cell suspension of C. hierosolymitanum by performing HPLC analysis against standard. Different ratios of NH+4 : NO-3 were used to study their effect on (-)-colchicine content, the highest colchicine content of 0.070 mg g-1DW was obtained at 30 mM NH4 + of total nitrogen. Colchicine alkaloid was highest, 0.090 mg g-1 DW, at 0.1 M of sucrose after 4 weeks incubation. (-) - Colchicine alkaloid was not detected in callus grown on sucrose free media. Maximum production of colchicine, 0.235 mg g-1 DW, was obtained in callus extracts of 60 days old callus grown under dark conditions. Cell suspension had 0.012 mg g-1 DW (-) -colchicine from suspended cells grown under dark. (-) -Colchicine content of callus incubated under dark (0.095 mg g-1 DW) was higher than light (0.070 mg g-1 DW) condition.

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