Temporal and spatial activity of a promoter from a pea enzyme inhibitor gene and its exploitation for seed quality improvement

The promoter from one of the two seed-expressed genes encoding trypsin/chymotrypsin inhibitors (TI) has been isolated and characterised in transgenic pea lines, following its re-introduction by Agrobacterium-mediated transformation, as a TI promoter-β-glucuronidase (GUS) gene fusion. The promoter from this gene ( TI1) directed expression of GUS enzyme at late stages of embryogenesis, comparable to those determined for activity of the homologous native TI genes. GUS expression was detected in roots of plants subjected to drought stress conditions, indicating that the TI1 gene, normally seed-specific in its expression, can be induced under these conditions. A second gene construct utilised the TI1 gene promoter to direct expression of an antisense TI gene. Seed TI activities in some lines transformed with this construct were reduced significantly. A limitation of the pea transformation methodology for antisense manipulations, in particular, is the observed frequency of non-transmission of transgenes from primary transformants (up to 80%).