Journal Article

Identification of a Novel factor, Vanillyl Benzyl Ether, Which Inhibits Somatic Embryogenesis of Japanese Larch (<i>Larix leptolepis</i> Gordon)

Mikihisa Umehara, Shinjiro Ogita, Hamako Sasamoto, Hiroyuki Koshino, Tadao Asami, Shozo Fujioka, Shigeo Yoshida and Hiroshi Kamada

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 3, pages 445-453
Published in print March 2005 | ISSN: 0032-0781
Published online March 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci041
Identification of a Novel factor, Vanillyl Benzyl Ether, Which Inhibits Somatic Embryogenesis of Japanese Larch (Larix leptolepis Gordon)

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In contrast to angiosperms, some gymnosperms form well-developed suspensors in somatic embryogenesis. This characteristic makes it easy to study suspensor biology. In cultures with high cell densities, somatic embryogenesis of Japanese larch, especially the suspensor development, is strongly inhibited due to factor(s) that are released by the cells into the culture medium. In this study, we purified and identified one of the inhibitory factors present in high-cell-density conditioned medium (HCM) of larch cells. The factor with the strongest inhibitory activity was purified by dialysis, extraction by ethyl acetate, octadecylsilyl (ODS) column chromatography and high-performance liquid chromatography (HPLC). The inhibitory factor was identified as vanillyl benzyl ether (VBE) by physicochemical analysis. This compound was first isolated from natural resources. Authentic VBE inhibited somatic embryo formation in Japanese larch, and the inhibitory effect in the suspensor was stronger than in the embryo proper. Furthermore, quantification of VBE by HPLC demonstrated that VBE accumulates at high concentrations in HCM. These results suggest that VBE is a novel negative regulator of somatic embryogenesis.

Keywords: Embryo proper; Inhibitory factor; Larix leptolepis; Somatic embryogenesis; Suspensor; Vanillyl benzyl ether; 6BAP, 6-benzyl amino purine; CM, conditioned medium; DI, dialyzed inner solution; DMF, dimethylformamide; DMSO, dimethyl sulfoxide; DO, dialyzed outer solution; EC, embryogenic cell; HCM, high-cell-density conditioned medium; HR-EI-MS, high resolution electron impact mass spectrometry; mCD, modified Campbell and Durzan’s medium; MS, mass spectrometry; NMR, nuclear magnetic resonance; NOE, nuclear Overhauser effect; ODS, octadecylsilyl; PCV, packed cell volume centrifuged at 100×g; pHBA, p-hydroxybenzyl alcohol; VBE, vanillyl benzyl ether

Journal Article.  6014 words.  Illustrated.

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

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