Abstract
Two T-DNA tagging vectors, pMHA2 and pΔgusBin19, were employed for isolation ofin vivo gene fusions of a promoterlessgus (uidA) reporter gene to low-temperature-(LT)-and abscisic-acid- (ABA)-responsive genes or regulatory sequences (RS) ofArabidopsis thaliana. Characterization of T2 progeny of 1200 T-DNA-tagged transgenic lines byin-situ staining of the leaf tissues for GUS activity indicated the presence ofin-vivo gene fusions to constitutively expressed genes ofA. thaliana in 16.1 percent of the lines (193/1200). A total of 1200 T2 lines was tested for LT-and 841 lines for ABA-induced GUS activity, and two of these exhibited expression of theuidA reporter gene in response to these stimuli. In one of the lines (274) GUS activity was predominantly induced by LT, whereas in line 317 it was mainly responsive to ABA. The induction properties of these fusions were verified by fluorometric assay of the GUS activity and by northern analysis of the corresponding transcript. These results demonstrate that promoter trapping provides a viable alternative for identification of particular classes of plant genes or RS that respond to specific environmental stresses;e.g., low temperature, drought, insect injuries and pathogen attack, or that are regulated hormonally or developmentally.
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