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Metabolism and Enzymology

Heat Inducible Expression of a Chimeric Maize hsp70CAT Gene in Maize Protoplasts ¹

Department of Biological Sciences, Stanford University, Stanford, California 94305-5020

The response of maize (Zea mays L.) protoplasts to high temperature stress was investigated. After isolation and electroporation, protoplasts were preincubated for 12 hours at 26°C then incubated for 6 hours at elevated temperatures. The pattern of polypeptides synthesized by these protoplasts during the last hour was monitored by in vivo labeling with ³⁵S-methionine. Incubation at 40° and 42°C resulted in the synthesis of polypeptides not detectable at 26°C. Introduction of a chimeric maize heat shock protein 70 promoter-chloramphenicol acetyltransferase coding region gene into protoplasts via electroporation resulted in the temperature-dependent induction of chloramphenicol acetyltransferase activity with maximal activity at 40°C. In the same protoplasts, a second chimeric gene, in which the firefly luciferase coding region was under the control of the 35S promoter from cauliflower mosaic virus, did not show an increase in expression after incubation at higher temperatures. Maize protoplasts provide a system to study molecular responses to high temperature stress.

³ Present address: USDA-PGEC, 800 Buchanan St., Albany, CA 94710.

¹ Supported by National Institutes of Health grant GM32422 to V. W. and by a gift from Pioneer Hi-Bred International.

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