title: The pea gene NA encodes ent-kaurenoic acid oxidase.
abstract: The gibberellin (GA)-deficient dwarf na mutant in pea (Pisum sativum) has severely reduced internode elongation, reduced root growth, and decreased leaflet size. However, the seeds develop normally. Two genes, PsKAO1 and PsKAO2, encoding cytochrome P450 monooxygenases of the subfamily CYP88A were isolated. Both PsKAO1 and PsKAO2 had ent-kaurenoic acid oxidase (KAO) activity, catalyzing the three steps of the GA biosynthetic pathway from ent-kaurenoic acid to GA(12) when expressed in yeast (Saccharomyces cerevisiae). In addition to the intermediates ent-7alpha-hydroxykaurenoic acid and GA(12)-aldehyde, some additional products of the pea KAO activity were detected, including ent-6alpha,7alpha-dihydroxykaurenoic acid and 7beta-hydroxykaurenolide. The NA gene encodes PsKAO1, because in two independent mutant alleles, na-1 and na-2, PsKAO1 had altered sequences and the five-base deletion in PsKAO1 associated with the na-1 allele cosegregated with the dwarf na phenotype. PsKAO1 was expressed in the stem, apical bud, leaf, pod, and root, organs in which GA levels have previously been shown to be reduced in na plants. PsKAO2 was expressed only in seeds and this may explain the normal seed development and normal GA biosynthesis in seeds of na plants.
authors.affiliation: School of Plant Science, University of Tasmania, GPO Box 252-55, Hobart, Tasmania, 7001, Australia.
authors.forename: Sandra E
authors.initials: SE
authors.lastname: Davidson
authors.forename: Robert C
authors.initials: RC
authors.lastname: Elliott
authors.forename: Chris A
authors.initials: CA
authors.lastname: Helliwell
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Poole
authors.forename: James B
authors.initials: JB
authors.lastname: Reid
mesh_terms: D015152:Blotting, Northern; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D020224:Expressed Sequence Tags; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D005875:Gibberellins / Q000096:biosynthesis*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D018532:Pisum sativum / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development; D018547:Plant Stems / Q000235:genetics / Q000254:growth & development; D018514:Plant Structures / Q000235:genetics* / Q000254:growth & development; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012639:Seeds / Q000235:genetics / Q000254:growth & development
chemical_list: D005875:Gibberellins; D010940:Plant Proteins; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C423619:ent-kaurenoic acid oxidase