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THE PLANT CELL, Vol 8, Issue 8 1291-1304, Copyright © 1996 by American Society of Plant Biologists
Cloning and Characterization of CER2, an Arabidopsis Gene That Affects Cuticular Wax Accumulation
Y. Xia, B. J. Nikolau and P. S. Schnable
Department of Zoology and Genetics, Iowa State University, Ames, Iowa 50011
Cuticular waxes are complex mixtures of very long chain fatty acids and
their derivatives that cover plant surfaces. Mutants of the ECERIFERUM2
(cer2) gene of Arabidopsis condition bright green stems and siliques,
indicative of the relatively low abundance of the cuticular wax crystals
that comprise the wax bloom on wild-type plants. We cloned the CER2 gene
via chromosome walking. Three lines of evidence establish that the cloned
sequence represents the CER2 gene: (1) this sequence is capable of
complementing the cer2 mutant phenotype in transgenic plants; (2) the
corresponding DNA sequence isolated from plants homozygous for the cer2-2
mutant allele contains a sequence polymorphism that generates a premature
stop codon; and (3) the deduced CER2 protein sequence exhibits sequence
similarity to that of a maize gene (glossy2) that also is involved in
cuticular wax accumulation. The CER2 gene encodes a novel protein with a
predicted mass of 47 kD. We studied the expression pattern of the CER2 gene
by in situ hybridization and analysis of transgenic Arabidopsis plants
carrying a CER2-[beta]-glucuronidase gene fusion that includes 1.0 kb
immediately upstream of CER2 and 0.2 kb of CER2 coding sequences. These
studies demonstrate that the CER2 gene is expressed in an organ- and
tissue-specific manner; CER2 is expressed at high levels only in the
epidermis of young siliques and stems. This finding, is consistent with the
visible phenotype associated with mutants of the CER2 gene. Hence, the
1.2-kb fragment of the CER2 gene used to construct the
CER2-[beta]-glucuronidase gene fusion includes all of the genetic
information required for the epidermis-specific accumulation of CER2 mRNA.
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