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Plant Physiol, February 2001, Vol. 125, pp. 585-594

Altered Patterns of Sucrose Synthase Phosphorylation and Localization Precede Callose Induction and Root Tip Death in Anoxic Maize Seedlings1,2

Chalivendra C. Subbaiah3* and Martin M. Sachs

Department of Crop Sciences, University of Illinois, Urbana, Illinois 61801 (C.C.S., M.M.S.); and United States Department of Agriculture-Agricultural Research Service, Urbana, Illinois 61801 (M.M.S.)

Root extracts made from maize (Zea mays) seedlings submerged for 2 h showed an increased 32P-labeling of a 90-kD polypeptide in a Ca2+-dependent manner. This protein was identified as sucrose synthase (SS) by immunoprecipitation and mutant analysis. Metabolic labeling with 32Pi indicated that the aerobic levels of SS phosphorylation were maintained up to 2 h of anoxia. In contrast, during prolonged anoxia the protein was under-phosphorylated, and by 48 h most of the protein existed in the unphosphorylated form. In seedlings submerged for 2 h or longer, a part of SS became associated with the microsomal fraction and this membrane localization of SS was confined only to the root tip. This redistribution of SS in the root tip preceded callose induction, an indicator of cell death. The sh1 mutants showed sustained SS phosphorylation and lacked the anoxia-induced relocation of SS, indicating that it was the SH1 form of the enzyme that was redistributed during anoxia. The sh1 mutants also showed less callose deposition and greater tolerance to prolonged anoxia than their non-mutant siblings. EGTA accentuated anoxic effects on membrane localization of SS and callose accumulation, whereas Ca2+ addition reversed the EGTA effects. These results indicate that the membrane localization of SS is an important early event in the anoxic root tip, probably associated with the differential anoxic tolerance of the two SS mutants. We propose that beside the transcriptional control of genes encoding SS, the reversible phosphorylation of SS provides a potent regulatory mechanism of sugar metabolism in response to developmental and environmental signals.


1 This work is supported by the National Research Initiative Competitive Grants Program (grant no. 96-35100-3143 to M.M.S. and C.C.S.) from the U.S. Department of Agriculture and by the Illinois Council of Food and Agricultural Research (grant no. 00I-062-3 to C.C.S.).

2 Product names are necessary to report factually on available data; however, neither the U.S. Department of Agriculture nor the University of Illinois guarantees or warrants the standard of the product, and the use of the names implies no approval of the product to the exclusion of others that may be suitable.

3 Present address: DNA Plant Technology Corporation, 6701 San Pablo Avenue, Oakland, CA 94608-1239.

* Corresponding author; e-mail Chalivendra{at}dnap.com; fax 510-547-2817.

© 2001 American Society of Plant Physiologists



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