Fusarium head blight evaluation in wheat transgenic plants expressing the maize b-32 antifungal gene

The maize gene b-32, normally expressed in the maize (Zea mays) endosperm, encodes for a RIP (Ribosome Inactivating Protein) characterised by antifungal activity. Transgenic wheat plants were obtained via biolistic transformation, in which the b-32 gene is driven by the 35SCaMV promoter in association with the bar gene as a selectable marker. Plants were brought to homozygosity through genetic analysis of progeny and pathogenicity tests were performed on the fourth generation. Six homozygous b-32 wheat lines were characterised. All plants had a normal phenotype, not distinguishable from the control cv. Veery except for slightly smaller size, flowered and set seeds. Western blot analyses confirmed b-32 expression during the plant life cycle in the various tissues. Each line differed in the b-32 content in leaf protein extracts and the transgenic protein expression level was maintained at least up to 10 days after anthesis. Pathogenicity tests for Fusarium head blight (FHB) were performed on the b-32 transgenic wheat lines in comparison to the parental cv. Veery. Resistance to FHB was evaluated by the single floret injection inoculation method on immature spikes with spores of Fusarium culmorum. In all the transgenic lines, a similar reduction in FHB symptoms, not dependent on the level of b-32 protein, has been observed (20% and 30% relative to the control, respectively 7 and 14 days after inoculation). Percentage of tombstone kernels at maturity was also recorded; in all transgenic lines disease control for this parameter was around 25%. The data obtained indicate that maize b-32 was effective as in vivo antifungal protein reducing FHB symptoms in wheat lines expressing the maize RIP protein.