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Plant and Cell Physiology Advance Access published online on January 19, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci017
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights researved.
Received March 3, 2004
Accepted November 5, 2004

Regular Paper

Involvement of Extracellular Dilignols in Lignification During Tracheary Element Differentiation of Isolated Zinnia Mesophyll Cells

Naohito Tokunaga 1, Norikazu Sakakibara 2, Toshiaki Umezawa 2, Yasuko Ito 3, Hiroo Fukuda 4, and Yasushi Sato 1*

1 Department of Biology and Earth Sciences, Faculty of Science, Ehime University, Matsuyama 790-8577, Japan
2 Laboratory of Biochemical Control, Wood Research Institute, Kyoto University, Uji, Kyoto 611-0011, Japan
3 Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan
4 Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan; Plant Science Center, RIKEN, 1-7-22 Suehiro, Tsurumi-ku, Yokohama-shi, Kanagawa, 230-0045, Japan

* To whom correspondence should be addressed.
Yasushi Sato, E-mail: ysato{at}sci.ehime-u.ac.jp


  Abstract

During differentiation of isolated Zinnia mesophyll cells into tracheary elements (TEs), lignification on TEs progresses by supply of monolignols from not only TEs themselves but also from surrounding xylem parenchyma-like cells through culture medium [Hosokawa et al. (2001) Plant Cell Physiol. 42: 959-968]. However, how lignin polymerizes from the secreted monolignols has not been resolved. In this study, we analyzed phenol compounds in cultured medium with reversed-phase HPLC, gas chromatography-mass spectrometry and nuclear magnetic resonance spectrometry, and found 12 phenolic compounds including coniferyl alcohol and four dilignols, i.e., erythro-guaiacylglycerol-ß-coniferyl ether, threo-guaiacylglycerol-ß-coniferyl ether, dehydrodiconiferyl alcohol and pinoresinol in the medium in which TEs were developing. Coniferyl alcohol applied to TE-inductive cultures during TE formation rapidly disappeared from the medium, and caused a sudden increase in dilignols. Addition of the dilignols promoted lignification of TEs in which monolignol biosynthesis was blocked by an inhibitor of phenylalanine anmmonia-lyase (PAL), L-{alpha}-aminooxy-ß-phenylpropionic acid (AOPP). These results suggested that dilignols can act as intermediates of lignin polymerization.

Keywords: Coniferyl alcohol; Dilignol; Lignification; Tracheary element; Zinnia elegans.
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