Functional Expression of Cf9 and Avr9 Genes in Brassica napus Induces Enhanced Resistance to Leptosphaeria maculans

Abstract

The tomato Cf9 resistance gene induces an Avr9-dependent hypersensitive response (HR) in tomato and transgenic Solanaceae spp. We studied whether the Cf9 gene product responded functionally to the corresponding Avr9 gene product when introduced in a heterologous plant species. We successfully expressed the Cf9 gene under control of its own promoter and the Avr9 or Avr9R8K genes under control of the p35S₁ promoter in transgenic oilseed rape. We demonstrated that the transgenic oilseed rape plants produced the Avr9 elicitor with the same specific necrosis-inducing activity as reported for Cladosporium fulvum. An Avr9-dependent HR was induced in Cf9 oilseed rape upon injection of intercellular fluid containing Avr9. We showed Avr9-specific induction of PR1, PR2, and Cxc750 defense genes in oilseed rape expressing Cf9. Cf9 × Avr9 oilseed rape did not result in seedling death of the F₁ progeny, independent of the promoters used to express the genes. The F₁ (Cf9 × Avr9) plants, however, were quantitatively more resistant to Leptosphaeria maculans. Phytopathological analyses revealed that disease development of L. maculans was delayed when the pathogen was applied on an Avr9-mediated HR site. We demonstrate that the Cf9 and Avr9 gene can be functionally expressed in a heterologous plant species and that the two components confer an increase in disease resistance.