Plant Cell Advance Online Publication
Published on June 18, 2004; 10.1105/tpc.022178
Received February 27, 2004
Accepted April 8, 2004
A Dominant Negative Mutant of Cyclin-Dependent Kinase A Reduces Endoreduplication but Not Cell Size or Gene Expression in Maize Endosperm
João T. Leiva-Neto 1, Gideon Grafi 2, Paolo A. Sabelli 1, Ricardo A. Dante 1, Young-min Woo 3, Sheila Maddock 4, William J. Gordon-Kamm 4, and Brian A. Larkins 1*
1 Department of Plant Sciences, University of Arizona, Tucson, Arizona 85721
2 Department of Plant Sciences, Weizmann Institute of Science, Rehovot 76100, Israel
3 Kumho Life and Environmental Science Laboratory, Gwangju, Korea
4 Pioneer Hi-Bred International, Johnston, Iowa 50131-1004
* To whom correspondence should be addressed. E-mail: larkins{at}ag.arizona.edu.
Cells in maize (Zea mays) endosperm undergo multiple cycles of endoreduplication, with some attaining DNA contents as high as 96C and 192C. Genome amplification begins around 10 d after pollination, coincident with cell enlargement and the onset of starch and storage protein accumulation. Although the role of endoreduplication is unclear, it is thought to provide a mechanism that increases cell size and enhances gene expression. To investigate this process, we reduced endoreduplication in transgenic maize endosperm by ectopically expressing a gene encoding a dominant negative mutant form of cyclin-dependent kinase A. This gene was regulated by the 27-kD  -zein promoter, which restricted synthesis of the defective enzyme to the endoreduplication rather than the mitotic phase of endosperm development. Overexpression of a wild-type cyclin-dependent kinase A increased enzyme activity but had no effect on endoreduplication. By contrast, ectopic expression of the defective enzyme lowered kinase activity and reduced by half the mean C-value and total DNA content of endosperm nuclei. The lower level of endoreduplication did not affect cell size and only slightly reduced starch and storage protein accumulation. There was little difference in the level of endosperm gene expression with high and low levels of endoreduplication, suggesting that this process may not enhance transcription of genes associated with starch and storage protein synthesis.
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