Induction and characterization of pathogenesis-related proteins in roots of cocoyam (Xanthosoma sagittifolium [L] Schott) infected with Pythium myriotylum

Abstract

Although Pythium myriotylum is a very destructive root pathogen of cocoyam, the host defense response in this plant-pathogen interaction has not been fully studied. Four cocoyam germplasm accessions were inoculated with P. myriotylum, and their induced defense responses were characterized. The induction and spatio-temporal accumulation of chitinase and b-1,3-glucanase were determined by enzymatic activity assays of crude root extracts from inoculated and non-inoculated (control) plants, sampled at 0, 2, 4, 6 and 8 days post inoculation (dpi). Furthermore, induced proteins were extracted from roots of inoculated and control tolerant (RO1054 and RO3015) and susceptible (RO2063) accessions at 8 dpi, and characterized by isoelectric focusing (IEF), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analyses. Chitinase and b-1,3-glucanase were consistently produced in high amounts in the roots of the tolerant accession RO1054, 8 days after inoculation. SDS-PAGE and immunoblotting showed that induced chitinases (37, 35 and 33 kDa) in the tolerant cocoyams were immunologically related to PR-3a purified from barley leaves inoculated with Erysiphe graminis f. sp. hordei, and induced osmotins (42-45 kDa) were immunologically related to osmotins purified from cultured NaCl-adapted tobacco protoplasts. These results suggest that tolerance in cocoyam infected with P. myriotylum may be associated in part with the production of pathogenesis-related (PR) proteins including one hydrolytic enzyme of known antifungal activity (PR-3).

Key words: b-1,3-glucanase, chitinase, cocoyam, PR protein, Pythium myriotylum, osmotin.