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Plant and Cell Physiology Advance Access published online on November 27, 2006
Plant and Cell Physiology, doi:10.1093/pcp/pcl039
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Co-regulation of brassinosteroid biosynthesis-related genes during xylem cell differentiation

Ryo Yamamoto1,4, Shozo Fujioka2, Kuninori Iwamoto1, Taku Demura1,5, Suguru Takatsuto3, Shigeo Yoshida2 and Hiroo Fukuda1

1 Department of Biological Sciences, Graduate School of Science, the University of Tokyo, Tokyo, 113-0033 Japan
2 Discovery Research Institute, RIKEN, Wako, 351-0198 Japan
3 Department of Chemistry, Joetsu University of Education, Joetsu, 943-8512 Japan

Corresponding author: Ryo Yamamoto Soybean Physiology Research Team, National Institute of Crop Science, 2-1-18, Kannondai, Tsukuba, Ibaraki 305-8518, Japan Tel: +81-029-838-8392 Fax: +81-029-838-8392 E-mail: brassin{at}affrc.go.jp, Hiroo Fukuda Department of Biological Sciences, Graduate School of Science, the University of Tokyo, Tokyo, 113-0033 Japan Tel: +81-03-5841-4461 Fax: +81-03-3812-4929 E-mail: fukuda{at}biol.s.u-tokyo.ac.jp


  Abstract

To understand the regulatory mechanisms of brassinosteroid (BR) biosynthesis in specific plant developmental processes, we first investigated the accumulation profiles of BRs and sterols in xylem differentiation in a Zinnia culture. The amounts of many substances in the late C28 sterol biosynthetic pathway to campesterol (CR), such as episterol and 24-methylenecholesterol, as well as those in the BR-specific biosynthetic pathway from CR to brassinolide (BL), were elevated in close association with tracheary element differentiation. Among them, 6-deoxotyphasterol (6-deoxoTY) was unusually highly accumulated within cells cultured in tracheary element-inductive medium, while castasterone (CS) were not elevated either within or out of cells. To identify the molecular basis of this co-upregulation of BRs and C28 sterols, we isolated Zinnia genes for the key enzymes of BR biosynthesis, ZeSTE1, ZeDIM, ZeDWF4, ZeCPD1 and ZeCPD2. RNA gel blot analysis of these genes indicated a coordinated increase in transcripts for ZeSTE1, ZeDIM, ZeDWF4 and ZeCPD1 and a tracheary element differentiation-specific increase in transcripts for ZeDWF4 and ZeCPD1. In situ hybridization experiments of ZeDWF4 and ZeCPD1 mRNAs revealed their preferential accumulation in procambium cells, immature xylem cells and xylem parenchyma cells. These results suggest that BR biosynthesis during tracheary element differentiation may be regulated by the a coordinated regulation of broad sterol biosynthesis and specific regulation of BR biosynthesis, which occurs in part by elevated transcript levels of genes encoding BR biosynthetic enzymes, specifically ZeDWF4 and ZeCPD1. These data provide new insights into the regulation of BR biosynthesis and BR signaling during plant development.

Keywords: Brassinosteroid biosynthesis - Auxin - Cytokinin - Gene expression - Tracheary element differentiation - Zinnia elegans

The nucleotide sequences reported in this paper have been submitted to the DDBJ database under accession numbers ZeCPD1 (AB231153 [GenBank] ), ZeCPD2 (AB231154 [GenBank] ), ZeDWF4 (AB231155 [GenBank] ), ZeDIM (AB231156 [GenBank] ), and ZeSTE1 (AB231157 [GenBank] )

4 Current address: National Institute of Crop Science (NICS), Tsukuba, 305-8518 Japan

5 Current address: Plant Science Center, RIKEN, Yokohama, 230-0045 Japan


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