Taxus callus cultures: Initiation, growth optimization, characterization and taxol production

Enaksha R.M. Wickremesinhe, Richard N. Arteca

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

Callus was induced from Taxus baccata cv. Repandens Parsons ex Rehd., T. brevifolia Nutt., T. cuspidata Sieb. & Zucc., and T. x media cvs. Hicksii and Densiformis Rehd. using different concentrations of 2,4-d-(2,4-dichlorophenoxyacetic acid), IBA (indole-3-butyric acid), or NAA α-naphthalene acetic acid in combination with kinetin. All cultures grew slowly following the first subculture, and a majority turned brown and ceased growth within the next six to twelve months. The callus cultures which lived, continued to grow very slowly for one to two years before the growth rate improved. Initiation of roots and shoot primordia-like structures occurred on some cultures maintained in the dark, and 16 h light/8 h dark, respectively. A fast-growing, habituated callus line (CR-1) derived from T. x media Rehd. cv. Hicksii was established from callus initiated in 1986. Supplementing the medium with casein hydrolysate and both fructose and glucose enhanced the growth rate. A great deal of heterogeneity was found among and within the callus, with respect to the amount of taxol produced. The callus exhibited levels of taxol ranging from 0.1 to 13.1 mg kg-1 (0.0001 to 0.0131%) on a dry weight basis. Overall, the older brown-colored callus produced more taxol than the younger pale yellow-colored callus. The presence of taxol in callus samples was established by high performance liquid chromatography, its biological activity confirmed by a microtubule-stabilizing bioassay and its structure confirmed using one-and two-dimensional 1H and 13C nuclear magnetic resonance spectroscopy.

Original languageEnglish (US)
Pages (from-to)181-193
Number of pages13
JournalPlant Cell, Tissue and Organ Culture
Volume35
Issue number2
DOIs
StatePublished - Nov 1 1993

Fingerprint

Taxus
paclitaxel
callus culture
callus
naphthaleneacetic acid
Taxus brevifolia
Taxus cuspidata
shoot primordia
Taxus baccata
root primordia
casein hydrolysates
indole butyric acid
kinetin
microtubules
2,4-D
nuclear magnetic resonance spectroscopy
bioactive properties
fructose
high performance liquid chromatography
bioassays

All Science Journal Classification (ASJC) codes

  • Horticulture

Cite this

Wickremesinhe, Enaksha R.M. ; Arteca, Richard N. / Taxus callus cultures : Initiation, growth optimization, characterization and taxol production. In: Plant Cell, Tissue and Organ Culture. 1993 ; Vol. 35, No. 2. pp. 181-193.
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Taxus callus cultures : Initiation, growth optimization, characterization and taxol production. / Wickremesinhe, Enaksha R.M.; Arteca, Richard N.

In: Plant Cell, Tissue and Organ Culture, Vol. 35, No. 2, 01.11.1993, p. 181-193.

Research output: Contribution to journalArticle

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