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Plant and Cell Physiology Advance Access originally published online on February 2, 2005
Plant and Cell Physiology 2005 46(3):445-453; doi:10.1093/pcp/pci041
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JSPP © 2005

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

Mikihisa Umehara1,6,7, Shinjiro Ogita2, Hamako Sasamoto3, Hiroyuki Koshino4, Tadao Asami5, Shozo Fujioka5, Shigeo Yoshida5 and Hiroshi Kamada6

1 Department of Biotechnology, Fukuoka Agricultural Research Center, Yoshiki 587, Chikushino, Fukuoka, 818-8549 Japan
2 Enzyme Chemistry Department, Biotechnology Research Center, Toyama Prefectural University, Kurokawa 5180, Kosugi, Toyama, 939-0398 Japan
3 Graduate School of Environmental and Information Sciences, Yokohama National University, Tokiwadai 79-7, Hodogaya, Yokohama, Kanagawa, 240-8501 Japan
4 Molecular Characterization Team, Advanced Development and Supporting Center, RIKEN, Hirosawa 2-1, Wako, Saitama, 351-0198 Japan
5 Plant Function Laboratory, RIKEN, Hirosawa 2-1, Wako, Saitama, 351-0198 Japan
6 Gene Research Center, Graduate School of Life and Environmental Science, University of Tsukuba, Ten-noudai 1-1-1, Tsukuba, Ibaraki, 305-8572 Japan

7 Corresponding author: E-mail, umehara{at}farc.pref.fukuoka.jp; Fax, +81-92-924-2981.

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.

Received October 2, 2004; Accepted December 22, 2004
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