Molecular analysis of a new member of the opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylase gene family

• Published in: Plant physiology (Bethesda) Vol. 110; no. 1; pp. 43 - 49
• Main Authors: Maldonado-Mendoza, I. E., Lopez-Meyer, M., Galef, J. R., Burnett, R. J., Nessler, C. L.
• Format: Journal Article
• Language: English
• Published: Rockville, MD American Society of Plant Physiologists 1996
• Subjects: actividad enzimatica; activite enzymatique; Alkaloids; Amino Acid Sequence; Amino acids; analisis cuantitativo; analyse quantitative; anatomia de la planta; anatomie vegetale; Base Sequence; Biochemistry and Enzymology; Biological and medical sciences; cell culture; chemical composition; composicion quimica; composition chimique; cultivo de celulas; culture de cellule; culture media; dopa; Dopa Decarboxylase - genetics; Enzymes; enzymic activity; Escherichia coli - genetics; etapas de desarrollo de la planta; expresion genica; expression des genes; feuille; fleur; flores; flowers; Fundamental and applied biological sciences. Psychology; Gene expression; gene transfer; Genes; Genes, Plant; genetica; genetics; genetique; Genomic Library; hojas; Latex; leaves; liasas; lyase; lyases; medio de cultivo; Metabolism; milieu de culture; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Multigene Family; Nicotiana - genetics; nicotiana tabacum; Nitrogen metabolism; Opium; Papaver - enzymology; Papaver - genetics; papaver somniferum; plant anatomy; plant developmental stages; Plant physiology and development; Plant Proteins - genetics; Plant roots; plantas transgenicas; plante transgenique; Plants; Plants, Genetically Modified; Plants, Medicinal; Plants, Toxic; plantulas; plantule; quantitative analysis; racine; raices; Recombinant Proteins - metabolism; roots; Seedlings; Sequence Analysis, DNA; Sequence Homology, Amino Acid; stade de developpement vegetal; Stems; Substrate Specificity; tallo; tige; tirosina; Tissue Distribution; transferencia de genes; transfert de gene; transgenic plants; tyrosine; Tyrosine Decarboxylase - genetics; Root; Enzyme; Germination; Tissue specificity; Papaver somniferum; Lyases; Gene expression; Papaveraceae; Medicinal plant; Carbon-carbon lyases; Enzymatic activity; Complementary DNA; Carboxy-lyases; Dicotyledones; Substrate specificity; Angiospermae; Aromatic-L-amino-acid decarboxylase; Spermatophyta; Transgenic plant; Aminoacid sequence
• DOI: 10.1104/pp.110.1.43
• ISSN: 0032-0889

An aromatic amino acid decarboxylase DNA fragment was generated from opium poppy (Papaver somniferum L.) genomic DNA by the PCR using primers designed from conserved amino acid sequences of other aromatic amino acid decarboxylase genes. Using this fragment as a probe, a genomic clone was isolated that encodes a new member of the opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylase gene family (TyDC5). The predicted TyDC5 amino acid sequence shares extensive identity with other opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylases (84%), and when expressed in Escherichia coli, it is active against tyrosine and to a lesser extent against 3,4-dihydroxyphenylalanine. Ribonuclease protection assays indicate that TyDC5 is expressed primarily in the roots of mature poppy plants. A peak of TyDC5 expression was also observed during germination, coincident with the emergence of the radicle from the seed coat. Parallel results were obtained in transgenic tobacco using a TyDC5 promoter fragment (-2060) translationally fused to the beta-glucuronidase reporter gene (GUS). In TyDC5::GUS tobacco, GUS activity transiently appeared in all parts of the seedling during germination, but was limited to the roots in older plants. These results indicate that TyDC5 expression is transcriptionally regulated and suggest that the TyDC5 enzyme may play an important role in providing precursors for alkaloid synthesis in the roots and germinating seedlings of opium poppy.