The genetic basis of vascular differentiation and function is relatively poorly understood, partly due to the difficulty of screening for mutants defective in internal vascular tissues. Here we present an approach based on a predicted increase in vascular-related gene expression in response to an auxin transport inhibitor-induced vascular overgrowth. We used microarray analyses to identify 336 genes that were up-regulated > or =2-fold in shoot tissues of Arabidopsis thaliana showing vascular overgrowth. Promoter-marker gene fusions revealed that 38 out of 40 genes with > or =4-fold up-regulation in vascular overgrowth tissues had vascular-related expression in transgenic Arabidopsis plants. Obtained expression patterns included cambial tissues and differentiating xylem, phloem and fibers. A total of 15 genes were found to have vascular-specific expression patterns in the leaves and/or inflorescence stems. This study provides empirical evidence of the efficiency of the approach and describes for the first time the in situ expression patterns of the majority of the assessed genes.