T-DNA activation tagging.

Abstract

T-DNA activation tagging is a method to generate dominant mutations in plants or plant cells by random insertion of a T-DNA carrying constitutive enhancer elements, which can cause transcriptional activation of flanking plant genes. The method consists of generating a large number of transformed plants or plant cells using a specialized T-DNA construct, followed by selection for the desired phenotype. Subsequently, the activated plant gene is rescued from selected mutant transformants for further functional analysis. Since the exact procedure depends on the plant material and the selected phenotype, this chapter describes one specific example of T-DNA activation tagging of suspension-cultured cells, including, where possible, cross-references to more general applications of the technique.