A comprehensive strategy for identifying long-distance mobile peptides in xylem sap
A comprehensive strategy for identifying long-distance mobile peptides in xylem sap
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DOI：
10.1111/tpj.13015
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发表时间：
2015-11-01
期刊：
影响因子：
7.2
通讯作者：
Matsubayashi, Yoshikatsu
中科院分区：
文献类型：
--
作者：
Okamoto, Satoru;Suzuki, Takamasa;Matsubayashi, Yoshikatsu
越来越多的人认识到分泌肽介导高等植物器官间的通讯。木质部汁液肽组学是一种有效但具有挑战性的鉴定长距离移动的肽的方法。在这项研究中，我们开发了一个简单的，无凝胶的纯化系统，结合邻氯苯酚萃取与HPLC分离。利用该系统，我们成功地从大豆木质部汁液分泌物中鉴定出7个具有一个或多个转录后修饰的寡肽：糖基化、硫酸化和/或羟基化。RNA测序和定量PCR分析表明，编码肽的基因在多种组织中表达。我们进一步分析了长距离易位的7个肽的基因编码肽与单个氨基酸取代，并确定这四个肽作为潜在的根到芽移动的寡肽。启动子-GUS分析表明，所有四个肽编码基因表达的根内皮层的内部组织。此外，我们发现这些肽编码基因中的一些对生物和/或非生物因素有反应。这些结果表明，我们的纯化系统提供了一个全面的方法，有效地识别内源性小肽，并加强了概念，高等植物采用各种肽的根到地上部的信号。
There is a growing awareness that secreted peptides mediate 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.