Expression of theShpx2peroxidase gene ofStylosanthes humilisin transgenic tobacco leads to enhanced resistance toPhytophthora parasiticapv.nicotianaeandCercospora nicotianae

Publisher Website
Google Scholar
Abstract
Previous research indicated that the constitutive expression of a pathogen-inducible peroxidase gene (Shpx6a) from Stylosanthes humilis in transgenic plants resulted in enhanced resistance to fungal pathogens ( Kazan, K., Goulter, K.C., Way, H.M. and Manners, J.M. (1998) Expression of a pathogenesis-related peroxidase of Stylosanthes humilis in transgenic tobacco and canola and its effect on disease development. Plant Sci. 136, 207–217). We have now investigated another pathogen-inducible peroxidase gene of S. humilis, termed Shpx2, which is highly divergent from Shpx6a. Constitutive expression of the Shpx2 cDNA was obtained in tobacco using the 35S CaMV promoter, and up to a 12-fold increase in total peroxidase activity was observed in the leaves of transgenic plants compared to nontransgenic controls. Disease development was evaluated after inoculations in T1 and T2 transgenic lines expressing Shpx2. Lesion expansion was significantly (P < 0.05) reduced by up to 25% and 50% on leaves and stems, respectively, of transgenic plants expressing high levels of peroxidase compared to nontransgenic controls, following inoculation with Phytophthora parasitica pv. nicotianae, the cause of black shank disease. In addition, plant survival and recovery were greatly enhanced in transgenic plants following stem inoculations of plants with this plant pathogen. A significant (55%, P < 0.05) reduction in lesion number was observed in transgenic plants with high levels of peroxidase activity following inoculation with the fungus Cercospora nicotianae, the cause of frog-eye disease. No significant differences in disease development were observed between the lines expressing Shpx2 and controls following inoculation with the bacterium Pseudomonas syringae pv. tabaci, the cause of wildfire disease. These results provide evidence for a role for this peroxidase gene in plant defence, and suggest that diverse peroxidase genes may be employed as components of strategies aimed at the engineering of disease resistance. Map of the T-DNA region of the binary vector construct for pHW10. The peroxidase gene, Shpx2 (striped box), was flanked by the 5′ region of the cauliflower mosaic virus promoter 35S RNA gene (P35S) and the transcription termination region of genes 5 and 7 of pTiA6 (5′ and 7′, respectively). The nptII gene was flanked by the 5′ (Pnos) and the 3′ ends of the nopaline synthase promoter gene (nos polyA). The HindIII site used in cloning is shown. LB, T-DNA left border; RB, T-DNA right border.