Analysis of SCARECROW expression using a rapid system for assessing transgene expression in Arabidopsis roots.
Although the introduction of foreign genes into Arabidopsis has become routine, the production of transgenic Arabidopsis plants still requires several months. A transgene expression system (TES) has been developed that allows characterization of gene expression patterns and the effects of foreign genes in the Arabidopsis root in 2-4 weeks. The method is based on regeneration of stably transformed roots directly from callus tissue. TES has been used to study the expression of the SCARECROW gene, which is involved in establishing radial patterning in the root. The 2.5 kb region directly upstream of the SCARECROW coding region was found to be sufficient to confer cell-type specific expression. Furthermore, this promoter is active in the scr mutant background, indicating that factors essential for cell-type specific expression are present even in the absence of correct radial patterning. Finally, TES was used to demonstrate that the SCARECROW gene under control of this promoter complements the root organization defect of the scr mutant. These experiments demonstrate the utility of the TES system for studying gene expression in roots in wild-type and mutant backgrounds and for molecular complementation of root mutant phenotypes. It is possible that the method will also be applicable to other organs.
Malamy
J E
JE
New York University, Biology Department, NY 10003, USA.
Benfey
P N
PN
D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D016350:Leucine Zippers / Q000235:genetics*; D016296:Mutagenesis; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D029681:Arabidopsis Proteins; D010940:Plant Proteins; C100978:SCR protein, Arabidopsis