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Plant and Cell Physiology Advance Access published online on February 2, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci041
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received November 29, 2004
Revised December 22, 2004
Accepted January 7, 2005

Regular Paper

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

Mikihisa Umehara 1*, Shinjiro Ogita 2, Hamako Sasamoto 3, Hiroyuki Koshino 4, Tadao Asami 5, Shozo Fujioka 5, Shigeo Yoshida 5, and Hiroshi Kamada 6

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

* To whom correspondence should be addressed.
Mikihisa Umehara, E-mail: umehara{at}farc.pref.fukuoka.jp


  Abstract

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 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: Larix leptolepis; Somatic embryogenesis; Embryo proper; Suspensor Inhibitory factor; Vanillyl benzyl ether.
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