Induction of different chitinase andΒ-1,3-glucanase isoenzymes in sunflower (Helianthus annuus L.) seedlings in response to infection byPlasmopara halstedii

Abstract

Chitinase andΒ-1,3-glucanase activities were assayed in roots, hypocotyls and cotyledons of downy mildewsusceptible and -resistant sunflower (Helianthus annuus L.) cultivars. While the highestΒ-1,3-glucanase activity was in roots, that of chitinase activity was in hypocotyls. Inoculation of both sunflower cultivars withPlasmopara halstedii resulted in a marked increase of chitinase andΒ-1,3-glucanase activities. The increase was observed earlier in incompatible than in compatible reactions. Both enzymes occurred in root tissue as a complex mixture of isoenzymes. At least three different peaks with chitinase activity and three with glucanase activity could be resolved by gel filtration chromatography on Sephacryl S-100 and chromatofocusing on PBE 94 (pH 7-4). Following ammonium sulfate precipitation and ion-exchange on CM- and DEAE-Trisacryl, three glucanase and chitinase fractions, referred to as basic, neutral and acidic, were separated on the basis of their Chromatographic behaviour. A different pattern of distribution of chitinase andΒ-1,3-glucanase fractions was observed between inoculated and non-inoculated plants in both resistant (cv. RS-105) and susceptible (cv. Peredovik) cultivars. In healthy plantsΒ-1,3-glucanase was mainly found in the basic (cv. Peredovik) and neutral (cv. RS-105) fractions, whereas chitinase was in the basic fraction for both cultivars. The neutral and acidic fractions of chitinases were induced in the compatible and incompatible reactions. Inoculation of the plants induced the neutralΒ-1,3-glucanase fraction in resistant and susceptible cultivars and the acidic only in the susceptible one. Induction of the basic fraction of both activities was not observed in any case.