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The gene for the major cuticular wax‐associated protein and three homologous genes from broccoli ( Brassica oleracea ) and their expression patterns
Author(s) -
Pyee Jaeho,
Kolattukudy P.E.
Publication year - 1995
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1995.07010049.x
Subject(s) - biology , gene , complementary dna , genetics , intron , gene expression , brassica oleracea , microbiology and biotechnology , petal , cdna library , gene family , botany
The gene for the major protein (WAX9) found in surface wax of broccoli, designated wax9D , and three homologous genes ( wax9A, B and C ) were isolated from a genomic library using the previously isolated cDNA encoding the WAX9 protein as the probe; all four genes were sequenced. Genomic Southern blot analysis using the WAX9 cDNA as a probe showed the presence of at least four homologous genes in broccoli genome. The sequence of the originally isolated WAX9 cDNA matched with that of gene D . All four genes have an intron two codons before the stop codon. The putative promoter regions of the four genes, beyond the first 200 bp immediately 5′ to the translation start sites, are quite different. Essential elements such as TATA and CAAT boxes and several regions homologous to the promoter regions of other plant ltp genes were identified. The expression patterns of the genes were determined by RT‐PCR with gene‐specific primers and sequencing of the PCR products. All the genes were expressed in leaves and flower buds. While genes A, B and D also were expressed in stems and open flowers, expression of gene C was not detected in these organs. None of them were expressed in roots. The 972 bp 5′‐flanking region of wax9D when fused to β‐glucuronidase (GUS) gene, directed tissue‐specific GUS expression in transgenic tobacco plants; GUS expression was found in the epidermis of leaves, stems and flower petals, sepals, trichomes, and ovules but not in roots.