Plants react to wounding by transcriptionally activating specific defence-related genes.
In this project we will develop an approach based on gene activation by T-DNA tagging in order to isolate genes which are regulatory steps for defence-gene expression. This technique is based on the ability of the T-DNA, derivating from the Agrobacterium Ti plasmid, to randomly integrates into plant genome. Activation of tagged genes is achieved by using a T-DNA tagging vector which contains four CaMV 35S transcriptional enhancers. In order to isolate wound-regulatory genes, tomato transgenic lines carrying wound-inducible promoters fused to a marker gene, luciferase or bar (selection on the herbicide basta) genes, will be obtained. Activation tagging will be carried out on leaf protoplasts isolated from these marker lines. Screening of mutants will be done by recording luciferase activity or selection for basta resistance. Tagged genes will be characterised.