Journal Article

Purification from Conditioned Medium and Chemical Identification of a Factor That Inhibits Somatic Embryogenesis in Carrot

Toshihiro Kobayashi, Katsumi Higashi, Kazuo Sasaki, Tadao Asami, Shigeo Yoshida and Hiroshi Kamada

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 41, issue 3, pages 268-273
Published in print March 2000 | ISSN: 0032-0781
Published online March 2000 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/41.3.268
Purification from Conditioned Medium and Chemical Identification of a Factor That Inhibits Somatic Embryogenesis in Carrot

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Somatic embryogenesis is strongly inhibited in cultures of carrot (Daucus carota L.) cells when the cell density is high. The inhibition is caused by factors that are released by cells into the medium of such cultures. In this study, we purified and identified one of the inhibitory factors found in the medium of high-cell-density cultures of carrot cells. The inhibitory factor with the strongest apparent activity was purified by fractionation with ethylacetate, chromatography on an octadecylsilyl (ODS) silica gel-column and HPLC. The inhibitory factor had a single peak of absorbance at 280 nm and was identified as 4-hydroxybenzyl alcohol by mass spectrometry and 1H- and 13C-NMR spectroscopy. Authentic 4-hydroxybenzyl alcohol strongly inhibited the formation of somatic embryos at a concentration equal to that in high-cell-density cultures. These results suggest that 4-hydroxybenzyl alcohol is a major factor that accumulates in high-cell-density cultures of carrot cells and inhibits somatic embryogenesis.

Keywords: Daucus carota; Inhibitory conditioning factor; 4-hydroxybenzyl alcohol; Somatic embryogenesis

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Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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