Phenylalanine Ammonia-Lyase Genes Involved in Anthocyanin Synthesis and the Regulation of its Expression in Suspension Cultured Carrot Cells

Byline: Yoshihiro Ozeki (1), Yoshio Ito (1), Nobuhiro Sasaki (1), Mikiko Oyanagi (1), Hirofumi Akimoto (1), Yukie Chikagawa (1), Junko Takeda (2) Keywords: Keywords: Anthocyanin, Carrot, Differentiation, myb, Phenylalanine ammonia-lyase (PAL) Abstract: and is also induced rapidly and transiently by transfer of cells to fresh medium and lowering the cell density. From the carrot genomic library, four clones of PAL genes, gDcPAL1,2,3 and 4, were obtained. Analyses of nucleotide sequences revealed that only the gDcPAL3 gene is responsible for the induction of anthocyanin synthesis by 2,4-D. Several cis-elements, boxes M, P, A, L, and G, exist in the proximal promoter region of gDcPAL3. Transient expression experiments in carrot protoplasts using deletion mutants of the proximal promoter region of gDcPAL3 gene showed that boxes M and L, both of which contain core sequences of the Myb binding sites, might play an important role in gDcPAL3 promoter activity. Four myb cDNAs, Dcmyb8,10,12 and 14 were obtained from a carrot subtracted cDNA library and their structure and expression patterns were analyzed. In addition to the analysis of the proximal region of gDcPAL3 promoter, the possibility of the regulation of gene expression by genomic DNA structure and chromatin modification in metabolic differentiation is discussed. Author Affiliation: (1) Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo, 184--8588 Japan, JP (2) Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606--8502 Japan, JP Article note: Received 10 June 2000/ Accepted in revised form 1 July 2000