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Title: Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum

Abstract

Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e., roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag 8-plex ITRAQ, enriched using Ni-NTA magnetic beads and subjected to nRPLC-MS/MS analysis using HCD and decision tree guided CID/ETD strategy. A total of 1,625 unique phosphopeptides, spanning 1,659 non-redundant phosphorylationmore » sites, were detected from 1,126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5 fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
1064583
Report Number(s):
PNNL-SA-86415
44703; 47733; KP1704020
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Molecular & Cellular Proteomics. MCP, 11(11):1140-1155
Additional Journal Information:
Journal Name: Molecular & Cellular Proteomics. MCP, 11(11):1140-1155
Country of Publication:
United States
Language:
English
Subject:
Environmental Molecular Sciences Laboratory

Citation Formats

Nguyen, Tran H., Brechenmacher, Laurent, Aldrich, Joshua T., Clauss, Therese RW, Gritsenko, Marina A., Hixson, Kim K., Libault, Marc, Tanaka, Kiwamu, Yang, Feng, Yao, Qiuming, Pasa-Tolic, Ljiljana, Xu, Dong, Nguyen, Henry T., and Stacey, Gary. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum. United States: N. p., 2012. Web. doi:10.1074/mcp.M112.018028.
Nguyen, Tran H., Brechenmacher, Laurent, Aldrich, Joshua T., Clauss, Therese RW, Gritsenko, Marina A., Hixson, Kim K., Libault, Marc, Tanaka, Kiwamu, Yang, Feng, Yao, Qiuming, Pasa-Tolic, Ljiljana, Xu, Dong, Nguyen, Henry T., & Stacey, Gary. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum. United States. doi:10.1074/mcp.M112.018028.
Nguyen, Tran H., Brechenmacher, Laurent, Aldrich, Joshua T., Clauss, Therese RW, Gritsenko, Marina A., Hixson, Kim K., Libault, Marc, Tanaka, Kiwamu, Yang, Feng, Yao, Qiuming, Pasa-Tolic, Ljiljana, Xu, Dong, Nguyen, Henry T., and Stacey, Gary. Sun . "Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum". United States. doi:10.1074/mcp.M112.018028.
@article{osti_1064583,
title = {Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum},
author = {Nguyen, Tran H. and Brechenmacher, Laurent and Aldrich, Joshua T. and Clauss, Therese RW and Gritsenko, Marina A. and Hixson, Kim K. and Libault, Marc and Tanaka, Kiwamu and Yang, Feng and Yao, Qiuming and Pasa-Tolic, Ljiljana and Xu, Dong and Nguyen, Henry T. and Stacey, Gary},
abstractNote = {Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e., roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag 8-plex ITRAQ, enriched using Ni-NTA magnetic beads and subjected to nRPLC-MS/MS analysis using HCD and decision tree guided CID/ETD strategy. A total of 1,625 unique phosphopeptides, spanning 1,659 non-redundant phosphorylation sites, were detected from 1,126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5 fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.},
doi = {10.1074/mcp.M112.018028},
journal = {Molecular & Cellular Proteomics. MCP, 11(11):1140-1155},
number = ,
volume = ,
place = {United States},
year = {2012},
month = {11}
}