Chitosanase and chitinase activities in tomato roots during interactions with arbuscular mycorrhizal fungi or Phytophthora parasitica

Abstract

New chitosanase acidic isoforms have been shown in Glomus mosseae-colonized tomato roots and their induction, together with the previously described mycorrhiza-related chitinase isoform, has been further corroborated in plants colonized with another Glomus species (G. intraradices), as well as in tomato roots colonized in vitro by Gigaspora rosea. The induction of these chitosanase isoforms appears as a specific response to the arbuscular mycorrhizal (AM) symbi- osis, and does not correspond to unspecific defence mechanisms, since these isoforms were not induced by the pathogen Phytophthora parasitica. Analysis by isoelectrofocusing showed two closely migrating chitinase isoforms, specific to mycorrhizal plants colonized either with G. mosseae or G. intraradices, and their isoelectric points were estimated to be 4.5 and 4.7. The estimated molecular mass of chitosan- ases was 20 kDa, and after isoelectrofocusing, the chitosanase activities were detected along the acidic pH range (6.5–3.5). Constitutive and induced isoforms were also investigated during a time-course study. In some experiments, chitin and chitosan were embed- ded together as substrates in polyacrylamide gels with the aim of studying the capacity of some isoforms to display both chitinase and chitosanase activities. In extracts from plants colonized with either G. mosseae or G. intraradices, some constitutive chitinases and the previously described mycorrhiza-related chitinase isoform, appeared to display chitosanase activity, while this bifunctional character was not found for the chitinases from non-mycorrhizal tissue, nor in Phytophthora-infected plants. These results suggest some diversity in the chitinase activities concerning substrate specificity in mycorrhizal plants. The possible implications of these observations in the functioning of the symbiosis is discussed. Key