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Plant and Cell Physiology Advance Access originally published online on March 27, 2008
Plant and Cell Physiology 2008 49(5):767-790; doi:10.1093/pcp/pcn049
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Nano Scale Proteomics Revealed the Presence of Regulatory Proteins Including Three FT-Like proteins in Phloem and Xylem Saps from Rice

Toshihiko Aki1,2, Mikao Shigyo1, Ryouhei Nakano1,2, Tadakatsu Yoneyama1 and Shuichi Yanagisawa1,2,*

1Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657 Japan
2Japan Science and Technology Agency (JST), Core Research for Evolutional Science and Technology (CREST), Kawaguchi, 332-0012 Japan

*Corresponding author: E-mail, asyanagi{at}mail.ecc.u-tokyo.ac.jp; Fax, +81-5841-8032.


  Abstract

The main physiological roles of phloem and xylem in higher plants involve the transport of water, nutrients and metabolites. They are also involved, however, in whole plant events including stress responses and long-distance signaling. Phloem and xylem saps therefore include a variety of proteins. In this study, we have performed a shotgun analysis of the proteome of phloem and xylem saps from rice, taking advantage of the complete and available genomic information for this plant. Xylem sap was prepared using the root pressure method, whereas phloem sap was prepared with a unique method with the assistance of planthoppers to ensure the robustness of the detected proteins. The technical difficulties caused by the very limited availability of rice samples were overcome by the use of nano-flow liquid chromatography linked to a mass spectrometer. We identified 118 different proteins and eight different peptides in xylem sap, and 107 different proteins and five different peptides in phloem sap. Signal transduction proteins, putative transcription factors and stress response factors as well as metabolic enzymes were identified in these saps. Interestingly, we found the presence of three TERMINAL FLOWER 1/FLOWERING LOCUS T (FT)-like proteins in phloem sap. The detected FT-like proteins were not rice Hd3a (OsFTL2) itself that acted as a non-cell-autonomous signal for flowering control, but they were members of distinct subfamilies of the FT family with differential expression patterns. These results imply that proteomics on a nano scale is a potent tool for investigation of biological processes in plants.

Keywords: FLOWERING LOCUS T - Phloem sap - Proteomics - Rice - Xylem sap

Abbreviations: 1D, one-dimensional; 2D, two-dimensional; DAS, days after sowing; DTT, dithiothreitol; E, electrophoresis; FT, flowering locus T; LC, liquid chromatography; MS, mass spectrometry; RT–PCR, reverse transcription–PCR; SAM, shoot apical meristem; TFE, 2,2,2-trifluoroethanol; TFL, terminal flower.

(Received January 29, 2008; Accepted March 23, 2008)
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