Expression of the Arabidopsis AtAux2-11 auxin-responsive gene in transgenic plants.
Five constructions containing deletions of the promoter from an auxin-inducible gene of Arabidopsis thaliana, AtAux2-11, were fused to the coding region of the reporter gene LacZ, which encodes beta-galactosidase, and a polyadenylation 3'-untranslated nopaline synthase sequence from Agrobacterium. These chimeric genes were introduced into Arabidopsis by Agrobacterium tumefaciens-mediated transformation, and expression of the gene was examined by spectrophotometric and histochemical analyses. A 600 bp fragment from the AtAux2-11 promoter conferred histochemical patterns of staining similar to the longest 5' promoter tested, a 3.0 kb fragment. Localization of AtAux2-11/LacZ activity in the transgenic plants revealed spatial and temporal expression patterns that correlated with tissues and cells undergoing physiological processes modulated by auxin. LacZ activity was expressed in the elongating region of roots, etiolated hypocotyls, and anther filaments. Expression was detected in the vascular cylinder of the root and the vascular tissue, epidermis, and cortex of the hypocotyl, and filament. The AtAux2-11/LacZ gene was preferentially expressed in cells on the elongating side of hypocotyls undergoing gravitropic curvature. Expression of the chimeric gene in the hypocotyls of light-grown seedlings was less than that in etiolated seedling hypocotyls. The AtAux2-11/LacZ gene was active in the root cap, and expression in the root stele increased at sites of lateral root initiation. Staining was evident in cell types that develop lignified cell walls, e.g. trichomes, anther endothecial cells, and especially developing xylem. The chimeric gene was not expressed in primary meristems. While the magnitude of expression increased after application of exogenous auxin (2,4-D), the histochemical localization of AtAux2-11/LacZ remained unchanged. Transgenic plants with a 600 bp promoter construct (-0.6 kb AtAux2-11/LacZ) had higher levels of basal and auxin-inducible expression than plants with a 3.0 kb promoter construct. Transgenic plants with a -500 bp promoter had levels of expression similar to the -3.0 kb construct. The -0.6 kb AtAux2-11/LacZ gene responded maximally to a concentration of 5 x 10(-6) to 5 x 10(-5) M 2,4-D and was responsive to as little as 5 x 10(-8) M. The evidence presented here suggests that this gene may play a role in several auxin-mediated developmental and physiological processes.
Wyatt
R E
RE
Department of Botany, University of Georgia, Athens 30602.
Ainley
W M
WM
Nagao
R T
RT
Conner
T W
TW
Key
J L
JL
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D017360:Arabidopsis / Q000033:anatomy & histology / Q000235:genetics* / Q000254:growth & development; D001483:Base Sequence; D004247:DNA; D005786:Gene Expression Regulation; D017343:Genes, Plant; D007210:Indoleacetic Acids / Q000502:physiology*; D007763:Lac Operon; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012231:Rhizobium / Q000201:enzymology / Q000235:genetics; D012639:Seeds / Q000378:metabolism; D014170:Transformation, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D007210:Indoleacetic Acids; D004247:DNA; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase; D001616:beta-Galactosidase