Acta Biológica Colombiana (Jul 2005)
Polyphenoloxidase and β-1,3-Glucanase Differential Induction in Carnation (Dianthus caryophyllus) Infected by Fusarium oxysporum f. sp. dianthi race 2
Abstract
We evaluated the changes on the dynamics of polyphenoloxidase (PFO) and β-1,3- glucanase (Glu) enzymes in carnation steems, which had been previously inoculatedwith the pathogen Fusarium oxysporum f. sp. dianthi race 2 (Fod). We established the experimental conditions in order to obtain the extract of the enzymes from the carnation steems with the aim to evaluate their enzymatic activity. The best results for PFO were obtained when acetone powder formation, before the extraction with phosphate buffer pH 6,5 aditionated with 3% PVPP, was used, and for Glu, phosphate buffer pH 6.5. The PFO activity quantification was done using catecol at pH 7.0, 37 ºC and measuring the products on 420 nm and, for Glu activity using "laminarina digitata" at 37 ºC and pH 5.5. Then, carnation's cutting from a highly tolerant variety (Carolina) and a susceptible (Uconn) to Fod race 2, were inoculated with the pathogen, then submitted to the enzymes analysis at different post-inoculation time-lapses. For the susceptible variety, the PFO activity was not affected, whereas in the highly illness resistant variety, there was an important inducement 12 h and 24 h post-inoculation, meaning that this enzyme could be playing a significant rol in the defense response, in metabolisms related with the lignification and synthesis of phenolic precursors. Alternatively, the Glu enzyme showed activity inducement in both varieties which seems to comprise a fraction of a non-specific response, uncorrelated with the active defense mechanisms of the carnation against this pathogen.
