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Establishment of Regenerative Callus, Cell Suspension System, and Molecular Characterization of Taxus wallichiana Zucc. for the in vitro production of Taxol

Gauchan, Dhurva Prasad and Bhuju, Sudina and Lamichhane, Janardan and Shakya, Rajani and Garcia Gil, Rosario (2021). Establishment of Regenerative Callus, Cell Suspension System, and Molecular Characterization of Taxus wallichiana Zucc. for the in vitro production of Taxol. Journal of applied pharmaceutical science. 11 :6 , 22-34
[Research article]

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Abstract

Taxus wallichiana, an endangered Taxus species native to Nepal, is a source of Taxol (paclitaxel), an anticancer drug, which if produced via plant cell culture in vitro not only reduces the pressure on natural habitat but also allows continuous production throughout the seasons. Gamborg’s B5 medium was the culture medium to develop callus and suspension culture. Quantification of Taxol was carried out via high performance liquid chromatography (HPLC) using C18 column, water-acetonitrile mobile phase, and 1 ml/minute flowrate at 227 nm wavelength. Genetic variation among population was determined using the POPGEN32, version 1.31. Best callus response was in medium with 1 mg/l 2,4-D for both stem and needle explants. For cell suspension culture, callus from needle explants showed better response compared to stem. Fructose and sucrose proved to be the best carbon sources. Cell count increased as carbon concentration increased. 2,4-D at low concentration (< 2 mg/l) and naphthaleneacetic acid at high concentration (>5 mg/l) showed higher cell growth rate. Inoculum volume was directly proportional to cell count. Extra addition of macronutrients had inhibitory effect on cell count, but the effect was reversed for MgSO4. Using HPLC, for bark, the highest Taxol yield was 0.057 mg/g distilled water (DW), while for needle was 0.056 mg/g DW. For suspension cultured HPLC samples, Taxol yield ranged at 0.004%. Size range of amplified Deoxyribonucleic acids was between 200 and 2,000 bp. Thus, this investigation generated a concrete data for Taxus population of Nepal providing efficient protocols for establishment of callus and cell suspension culture with the aim of producing Taxol in in vitro conditions.

Authors/Creators:Gauchan, Dhurva Prasad and Bhuju, Sudina and Lamichhane, Janardan and Shakya, Rajani and Garcia Gil, Rosario
Title:Establishment of Regenerative Callus, Cell Suspension System, and Molecular Characterization of Taxus wallichiana Zucc. for the in vitro production of Taxol
Series Name/Journal:Journal of applied pharmaceutical science
Year of publishing :2021
Volume:11
Number:6
Page range:22-34
Number of Pages:13
Language:English
Publication Type:Research article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 3 Medical and Health Sciences > 301 Basic Medicine > Pharmacology and Toxicology
Keywords:Taxus wallichiana, Taxol, cell suspension culture, HPLC, RAPD-PCR.
URN:NBN:urn:nbn:se:slu:epsilon-p-114601
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-114601
Additional ID:
Type of IDID
DOI10.7324/JAPS.2021.110603
ID Code:26937
Faculty:S - Faculty of Forest Sciences
Department:(S) > Dept. of Forest Genetics and Plant Physiology
Deposited By: SLUpub Connector
Deposited On:08 Feb 2022 14:26
Metadata Last Modified:08 Feb 2022 14:31

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