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Regulation of NADPH-cytochrome P450 reductase expressed during Douglas-fir germination and seedling development

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Abstract

NADH-cytochrome P450 is a key enzyme that transfers electrons from NADPH to the cytochrome P450 family of enzymes. To begin to determine the regulation of CPR gene expression and enzyme activity in Douglas-fir a full-length cDNA was isolated from a seedling λZAP cDNA library and the ORF was used to develop a synthetic CPR-peptide-based antiserum. Northern blot analysis indicated CPR expression was regulated both developmentally prior to seed maturation and during germination, and differentially in the cotyledons, radicle and megagametophyte of seed and seedling tissues. The CPR-peptide antiserum detected a single CPR in seed and seedling microsomes analyzed by western blot of two-dimensional SDS-polyacrylamide gels. In microsomal extracts from whole seeds and seedlings, the amount of CPR protein remained constant while NADPH:cytochrome c reductase activity increased during stratification, germination and early seedling development. In contrast to cotyledons and megagametophyte, the level of CPR protein detected in radicles was higher than expected when compared to the amount of CPR transcript.

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Author notes

  1. Timothy J. Tranbarger

    Present address: Biochimie et Physiologie Moléculaire des Plantes, INRA/ENSA-M/CNRS URA 2133, Place Viala, 34060, Montpellier Cedex 1, France

Authors and Affiliations

  1. Department of Biochemistry and Microbiology, Centre for Forest Biology, University of Victoria, P.O. Box 3055 STN CSC, Victoria, British Columbia, V8W 3P6, Canada

    Benjamin S. Forward & Santosh Misra

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  1. Timothy J. Tranbarger
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  2. Benjamin S. Forward
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Tranbarger, T.J., Forward, B.S. & Misra, S. Regulation of NADPH-cytochrome P450 reductase expressed during Douglas-fir germination and seedling development. Plant Mol Biol 44, 141–153 (2000). https://doi.org/10.1023/A:1006425025702

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