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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Establishment of a Protein Reference Map for Soybean Root Hair Cells^1,[W],[OA]

National Center for Soybean Biotechnology, Division of Plant Sciences, University of Missouri, Columbia, Missouri 65211 (L.B., S.S., T.H.N.N, M.L., G.S.); United States Department of Agriculture-Agricultural Research Service, Beltsville, Maryland 20705 (J.L., B.C.); Computer Science Department and Christopher S. Bond Life Sciences Center, University of Missouri, Columbia, Missouri 65211 (Z.S., T.J., D.X.); United States Department of Agriculture-Agricultural Research Service, University of Missouri, Columbia, Missouri 65211 (N.O.); Charles W. Gehrke Proteomic Center, University of Missouri, Columbia, Missouri 65211 (B.M.); and Center for Sustainable Energy, Division of Biochemistry and Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, Missouri 65211 (G.S.)

Root hairs are single tubular cells formed from the differentiation of epidermal cells on roots. They are involved in water and nutrient uptake and represent the infection site on leguminous roots by rhizobia, soil bacteria that establish a nitrogen-fixing symbiosis. Root hairs develop by polar cell expansion or tip growth, a unique mode of plant growth shared only with pollen tubes. A more complete characterization of root hair cell biology will lead to a better understanding of tip growth, the rhizobial infection process, and also lead to improvements in plant water and nutrient uptake. We analyzed the proteome of isolated soybean (Glycine max) root hair cells using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and shotgun proteomics (1D-PAGE-liquid chromatography and multidimensional protein identification technology) approaches. Soybean was selected for this study due to its agronomic importance and its root size. The resulting soybean root hair proteome reference map identified 1,492 different proteins. 2D-PAGE followed by mass spectrometry identified 527 proteins from total cell contents. A complementary shotgun analysis identified 1,134 total proteins, including 443 proteins that were specific to the microsomal fraction. Only 169 proteins were identified by the 2D-PAGE and shotgun methods, which highlights the advantage of using both methods. The proteins identified are involved not only in basic cell metabolism but also in functions more specific to the single root hair cell, including water and nutrient uptake, vesicle trafficking, and hormone and secondary metabolism. The data presented provide useful insight into the metabolic activities of a single, differentiated plant cell type.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Gary Stacey (staceyg{at}missouri.edu).

^[W] The online version of this article contains Web-only data.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.108.131649

^* Corresponding author; e-mail staceyg{at}missouri.edu.

Received October 24, 2008; accepted November 24, 2008; published November 26, 2008.