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Plant Physiol, February 2000, Vol. 122, pp. 295-318

Patterns of Protein Synthesis and Tolerance of Anoxia in Root Tips of Maize Seedlings Acclimated to a Low-Oxygen Environment, and Identification of Proteins by Mass Spectrometry1

William W.P. Chang, Lan Huang, Min Shen, Cecelia Webster, Alma L. Burlingame, and Justin K.M. Roberts*

Department of Biochemistry, University of California, Riverside, California 92521 (W.W.P.C., C.W., J.K.M.R.); and Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143 (L.H., M.S., A.L.B.).

Tolerance of anoxia in maize root tips is greatly improved when seedlings are pretreated with 2 to 4 h of hypoxia. We describe the patterns of protein synthesis during hypoxic acclimation and anoxia. We quantified the incorporation of [35S]methionine into total protein and 262 individual proteins under different oxygen tensions. Proteins synthesized most rapidly under normoxic conditions continued to account for most of the proteins synthesized during hypoxic acclimation, while the production of a very few proteins was selectively enhanced. When acclimated root tips were placed under anoxia, protein synthesis was depressed and no "new" proteins were detected. We present evidence that protein synthesis during acclimation, but not during subsequent anoxia, is crucial for acclimation. The complex and quantitative changes in protein synthesis during acclimation necessitate identification of large numbers of individual proteins. We show that mass spectrometry can be effectively used to identify plant proteins arrayed by two-dimensional gel electrophoresis. Of the 48 protein spots analyzed, 46 were identified by matching to the protein database. We describe the expression of proteins involved in a wide range of cellular functions, including previously reported anaerobic proteins, and discuss their possible roles in adaptation of plants to low-oxygen stress.

1 This work was supported by the U.S. Department of Agriculture National Research Initiative-Competitive Grants Program (grant no. 98351006146 to J.K.M.R.) and by the National Institutes of Health NCRR (grant no. RR 01614 to A.L.B.).

* Corresponding author; e-mail jkmr{at}ucrac1.ucr.edu; fax 909-787-3590.

© 2000 American Society of Plant Physiologists


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