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Plant J. 2015 Nov;84(3):611-20. doi: 10.1111/tpj.13015. Epub 2015 Oct 7.

A comprehensive strategy for identifying long-distance mobile peptides in xylem sap.

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Division of Biological Science, Graduate School of Science, Nagoya University Chikusa, Nagoya, 464-8602, Japan.
Research Fellow of the Japan Society for the Promotion of Science, 5-3-1 Kojimachi, Chiyoda-ku, Tokyo, 102-0083, Japan.
RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro, Tsurumi, Yokohama, Kanagawa, 230-0045, Japan.
Japan Science and Technology Agency (JST) Exploratory Research for Advanced Technology (ERATO) Higashiyama Live-Holonics Project, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.
Division of Symbiotic Systems, National Institute for Basic Biology, Myodaiji, Okazaki, Aichi, 444-8585, Japan.
Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.


There is a growing awareness that secreted pemediate organ-to-organ communication in higher plants. Xylem sap peptidomics is an effective but challenging approach for identifying long-distance mobile peptides. In this study we developed a simple, gel-free purification system that combines o-chlorophenol extraction with HPLC separation. Using this system, we successfully identified seven oligopeptides from soybean xylem sap exudate that had one or more post-transcriptional modifications: glycosylation, sulfation and/or hydroxylation. RNA sequencing and quantitative PCR analyses showed that the peptide-encoding genes are expressed in multiple tissues. We further analyzed the long-distance translocation of four of the seven peptides using gene-encoding peptides with single amino acid substitutions, and identified these four peptides as potential root-to-shoot mobile oligopeptides. Promoter-GUS analysis showed that all four peptide-encoding genes were expressed in the inner tissues of the root endodermis. Moreover, we found that some of these peptide-encoding genes responded to biotic and/or abiotic factors. These results indicate that our purification system provides a comprehensive approach for effectively identifying endogenous small peptides and reinforce the concept that higher plants employ various peptides in root-to-shoot signaling.


C-terminally encoded peptide; CLV3/ESR-related; long-distance; peptidomics; sulfated peptide; xylem sap

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