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Plant and Cell Physiology, 1993, Vol. 34, No. 7 1029-1037
© 1993

Structure and Expression of Chalcone Synthase Gene in Carrot Suspension Cultured Cells Regulated by 2,4-D

Yoshihiro Ozeki1, Eric Davies2 and Junko Takeda3

1Department of Biology, College of Arts and Sciences, The University of Tokyo Komaba, Tokyo, 153 Japan
2School of Biological Sciences, University of Nebraska-Lincoln Lincoln, NE 68588-0118, U.S.A.
3Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University Kyoto, 606 Japan

Two nearly full-length chalcone synthase (EC 2.3.1.74 [EC] ) cDNAs, designated CHS2 and CHS9, were isolated from a cDNA library of mRNA prepared from anthocyanin-synthesizing cells of carrot suspension cultures and showed 98% nucleotide sequence identity to each other. The genomic sequence (gCHS2), which corresponds to CHS2, was isolated from a carrot genomic library. Primer extension identified the transcription start site of gCHS2 at 72 bp upstream from the initiation codon ATG, and showed that gCHS2 was activated during anthocyanin synthesis. A chimeric gene carrying the gCHS2 putative promoter sequence from –2,357 bp to +92 bp connected to the firefly luciferase gene and Nos terminator was introduced by electroporation into protoplasts prepared from anthocyanin-synthesizing carrot cells and the effects of light and 2,4-D on the promoter activity were examined by a transient assay for luciferase activity. Luciferase activity was induced by light exposure in the medium lacking 2,4-D but suppressed by the addition of 2,4-D and/or dark treatment.

(Received May 24, 1993; Accepted July 21, 1993)
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