A comprehensive strategy for identifying long-distance mobile peptides in xylem sap.
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.
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.
Satoru
S
Okamoto
Division of Biological Science, Graduate School of Science, Nagoya University Chikusa, Nagoya, 464-8602, 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.
Takamasa
T
Suzuki
Division of Symbiotic Systems, National Institute for Basic Biology, Myodaiji, Okazaki, Aichi, 444-8585, Japan.
Masayoshi
M
Kawaguchi
Division of Biological Science, Graduate School of Science, Nagoya University Chikusa, Nagoya, 464-8602, 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.|Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.
Tetsuya
T
Higashiyama
Division of Biological Science, Graduate School of Science, Nagoya University Chikusa, Nagoya, 464-8602, Japan.
Yoshikatsu
Y
Matsubayashi
D000595:Amino Acid Sequence; D005591:Chemical Fractionation / Q000379:methods; D002733:Chlorophenols / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000378:metabolism; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods; D021381:Protein Transport; D012323:RNA Processing, Post-Transcriptional; D017423:Sequence Analysis, RNA; D013025:Glycine max / Q000378:metabolism*; D052584:Xylem / Q000378:metabolism*
D002733:Chlorophenols; D010455:Peptides; D010940:Plant Proteins; C030683:2-chlorophenol
C-terminally encoded peptide; CLV3/ESR-related; long-distance; peptidomics; sulfated peptide; xylem sap