DEVELOPMENTAL AND TISSUE-SPECIFIC EXPRESSION OF A TOMATO ANIONIC PEROXIDASE (TAP-1) GENE BY A MINIMAL PROMOTER, WITH WOUND AND PATHOGEN INDUCTION BY AN ADDITIONAL 5'-FLANKING REGION

The tomato anionic peroxidase genes (tap1 and tap2) are induced by wounding and pathogen attack. The 5'-flanking region of tap1 confers wound- and pathogen-inducible beta-glucuronidase (GUS) expression in tobacco plants transformed with a tap1/GUS chimeric fusion gene construct. A series of nested 5' promoter deletions in the tap1/GUS fusion gene construct was created, and introduced into tobacco protoplasts via polyethylene glycol-mediated DNA transfer. A -202 construct (where the transcriptional start site is denoted + 1) and larger tap1 promoter constructs showed constitutive GUS expression. A 2-fold increase in GUS expression over the high constitutive levels was observed with -358 bp and larger tap1 constructs when protoplasts were incubated with elicitor preparations from Verticillium albo-atrum. In tobacco plants transformed with the tap1 promoter deletion/GUS fusion gene constructs, wounding caused induction of GUS expression by 20 h that increased 6- to 18-fold by 72 h. The region between -202 and -358 of the tap1 promoter conferred wound responsiveness. GUS was also found to be expressed in the epidermis and trichomes in the aerial parts of transgenic plants. High-level GUS expression was observed in the nodal region of stems that was associated with the leaf traces. GUS that was absent in very young flower buds was found in the subsequent developmental stages in the pistils, ovaries and anthers. The developmentally regulated tissue-specific expression of GUS was found with all constructs containing the -202 and larger promoters whereas wound and pathogen induction required -358 or larger promoter. These results suggest that the tap1 gene, which was heretofore thought to be expressed only upon wounding or pathogen attack, plays a role in normal developmental processes of the plant and this gene acquired additional 5'-flanking promoter for the purpose of responding to wounding and fungal attack.