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Plant and Cell Physiology Advance Access published online on August 24, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci190
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received May 13, 2005
Accepted August 15, 2005

Regular Paper

Differential Distribution of Proteins Expressed in Companion Cells in the Sieve Element-Companion Cell Complex of Rice Plants

Akari Fukuda 1*, Syu Fujimaki 2, Tomoko Mori 3, Nobuo Suzui 2, Keiki Ishiyama 4, Toshihiko Hayakawa 5, Tomoyuki Yamaya 5, Toru Fujiwara 6, Tadakatsu Yoneyama 7, and Hiroaki Hayashi 8

1 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Present address: Department of Paddy Farming, National Agricultural Research Center for Tohoku Region, Yotsuya, Daisen, Akita, 014-0102 Japan
2 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Present address: Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, Gunma 370-1207, Japan
3 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Present address: Fuji Photo Film Co. Senzui, Asaka, Saitama, 351-8585, Japan
4 Department of Applied Plant Science, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555, Japan; Present address: RIKEN Yokohama Institute, Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa, 230-0045, Japan
5 Department of Applied Plant Science, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555, Japan
6 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; PRESTO, JST, Japan; Present address: Biotechnology Research Center, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657 Japan
7 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
8 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Present address: 694 Futago, Aki, Higashikunisaki, Oita 873-0356 Japan

* To whom correspondence should be addressed.
Akari Fukuda, E-mail: akfukuda{at}affrc.go.jp


  Abstract

Sieve tubes are comprised of sieve elements, enucleated cells that are incapable of RNA and protein synthesis. The proteins in sieve elements are supplied from the neighboring companion cells through plasmodesmata. In rice plants, it was not clear whether all proteins produced in companion cells had same distribution pattern in sieve element-companion cell complex or not. In this study, the distribution pattern of four proteins, {beta}-glucuronidase (GUS), green fluorescent protein (GFP), thioredoxin h (TRXh) and glutathione S-transferase (GST) were analyzed. The foreign proteins GUS and GFP were expressed in transgenic rice plants under the control of the thioredoxin h gene promoter (PTRXh), a companion cell-specific promoter. Analysis of leaf cross-sections of PTRXh-GUS and PTRXh-GFP plants indicated high accumulation of GUS and GFP, respectively, in companion cells rather than in sieve elements. GUS and GFP were also detected in phloem sap collected from leaf sheaths of the transgenic rice plants, suggesting these proteins could enter sieve elements. Relative amounts of GFP and endogenous phloem proteins, TRXh and GST, in phloem sap and total leaf extracts were compared. Compared to TRXh and GST, GFP content was higher in total leaf extracts, but lower in phloem sap, suggesting that GFP accumulated mainly in companion cells rather than in sieve elements. On the other hands, TRXh and GST were suggested to accumulate in sieve elements rather than in companion cells. These results indicate the evidence for differential distribution of proteins between sieve elements and companion cells in rice plants.

Keywords: companion cell; Oryza sativa; phloem; rice; sieve element.
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