Root-specific expression of defensin in transgenic tobacco results in enhanced resistance against Phytophthora parasitica var. nicotianae

Abstract

Phytophthora species are soil-borne pathogens that damage plants in both agro- and natural ecosystems. To suppress the devastating pathogen, we generated a root-specific expression system using a specific promoter (pPRP3) conferring elevated expression of the target gene in roots that are very susceptible to soil-borne pathogens. To verify root-specific expression, we compared β-glucuronidase (GUS) expression driven by a constitutive or root-specific promoters in shoots and roots. In histochemical and fluorometric assays, GUS activity was detected in whole tobacco plants when GUS expression was driven by p35S, but was detected only in the roots by pPRP3. We then expressed a pepper defensin (J1–1) gene in tobacco to elucidate its effect on plant resistance. The accumulation of J1–1 was also tissue-specific in transgenic tobacco plants. Finally, transgenic plants carrying GUS or J1–1 genes in combination with p35S or pPRP3 were inoculated with Phytophthora parasitica var. nicotianae and Pythium aphanidermatum. Disease symptoms were significantly suppressed in transgenic plants that accumulated J1–1, regardless of the promoter used. Furthermore, the expression of PR genes was induced in J1–1 transgenic plants, exhibiting much higher levels in p35S-driven J1–1 plants than in pPRP3::J1–1 plants. These results demonstrated that J1–1 transgenic plants were primed for enhanced expression of PR genes, which provided synergistic effects with the defensin for disease resistance.