title: Evolution of the indole alkaloid biosynthesis in the genus Hordeum: distribution of gramine and DIBOA and isolation of the benzoxazinoid biosynthesis genes from Hordeum lechleri.
abstract: Two indole alkaloids with defense related functions are synthesized in the genus Hordeum of the Triticeae. Gramine (3(dimethyl-amino-methyl)-indole) is found in H. spontaneum and in some varieties of H. vulgare, the benzoxazinoid 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one (DIBOA) is detected in H. roshevitzii, H. brachyantherum, H. flexuosum, H. lechleri. Biosynthesis of DIBOA and of gramine was found to be mutually exclusive in wild Hordeum species, indicating that there was selection against simultaneous expression of both pathways during evolution. The full set of genes required for DIBOA biosynthesis in H.lechleri was isolated and the respective enzyme functions were analyzed by heterologous expression. The cytochrome P450 genes Bx2-Bx5 demonstrate a monophyletic origin for H. lechleri, Triticum aestivum and Zea mays. HlBx2-HlBx5 share highest homology to the orthologous genes of T. aestivum. In contrast, the branch point enzyme of the DIBOA pathway, the indole-3-glycerol phosphate lyase BX1, might have evolved independently in H. lechleri. In all Hordeum species that synthesize DIBOA, DNA sequences homologous to Bx genes are found. In contrast, these sequences are not detectable in the genomes of H. vulgare and H. spontaneum that do not synthesize benzoxazinoids.
authors.affiliation: Institute for Biochemical Plant Pathology, GSF-National Research Center for Environment and Health, Ingolstädter Landstrasse 1, D-85746 Neuherberg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Grün
authors.forename: Monika
authors.initials: M
authors.lastname: Frey
authors.forename: Alfons
authors.initials: A
authors.lastname: Gierl
mesh_terms: D000470:Alkaloids / Q000096:biosynthesis* / Q000737:chemistry; D000595:Amino Acid Sequence; D048588:Benzoxazines; D019143:Evolution, Molecular; D001467:Hordeum / Q000201:enzymology* / Q000235:genetics*; D026121:Indole Alkaloids / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010078:Oxazines / Q000737:chemistry / Q000378:metabolism*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D000470:Alkaloids; D048588:Benzoxazines; D026121:Indole Alkaloids; D010078:Oxazines; D010940:Plant Proteins; C104846:2,4-dihydroxy-1,4-benzoxazin-3-one; C007884:gramine