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Plant and Cell Physiology Advance Access originally published online on August 24, 2005
Plant and Cell Physiology 2005 46(11):1779-1786; doi:10.1093/pcp/pci190
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Differential Distribution of Proteins Expressed in Companion Cells in the Sieve Element-Companion Cell Complex of Rice Plants

Akari Fukuda1,4,*, Syu Fujimaki1,5, Tomoko Mori1,6, Nobuo Suzui1,5, Keiki Ishiyama2,7, Toshihiko Hayakawa2, Tomoyuki Yamaya2, Toru Fujiwara1,3,8, Tadakatsu Yoneyama1 and Hiroaki Hayashi1,9

1 Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo, 113-8657 Japan
2 Department of Applied Plant Science, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai, 981-8555 Japan
3 PRESTO, JST, Honcho, Kawaguchi, Saitama, 332-0012 Japan

* Corresponding author: E-mail, akfukuda{at}affrc.go.jp; Fax, +81-187-66-2639.

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 unclear whether or not all proteins produced in companion cells had the same distribution pattern in the sieve element-companion cell complex. In this study, the distribution pattern of four proteins, ß-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 TRXh 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 hand, TRXh and GST appeared 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.

4 Present address: Department of Paddy Farming, National Agricultural Research Center for Tohoku Region, Yotsuya, Daisen, Akita, 014-0102 Japan.

5 Present address: Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, Gunma, 370-1207 Japan.

6 Present address: Fuji Photo Film Co. Senzui, Asaka, Saitama, 351-8585 Japan.

7 Present address: RIKEN Yokohama Institute, Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa, 230-0045 Japan.

8 Present address: Biotechnology Research Center, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo, 113-8657 Japan.

9 Present address: 694 Futago, Aki, Higashikunisaki, Oita, 873-0356 Japan.

(Received May 13, 2005; Accepted August 15, 2005)
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