Abstract

SALAS, Carolina  and  ZAMORA, Humberto M. ß (1,3) GLUCANASE FROM INTERCELLUAR SPACE OF TOMATO LEAVES (LYCOPERSICON ESCULENTUM CERACIFORME) AFTER INFECCIÓN WITH PHYTOPHTHORA INFESTANS. Rev.Colomb.Quim. [online]. 2009, vol.38, n.1, pp.43-59. ISSN 0120-2804.

In this paper was analysed the intercellular washing fluid of Lycopersicon esculentum ceraciforme L1568 belonging to the national germplasm with resistance to P. infestans in field which showed the presence of four ß (1,3) glucanases after infection with the pathogen. For this purpose the plants were inoculated with the pathogen to induce a concentration increase of PR proteins and other defence-related substances, 15 days after inoculation with the parasite was extracted the intercellular washing fluid of tomato leaves using water. The proteins with ß(1,3) glucanase activity were extracted, freeze-dried and re-suspended in buffer solution and were purified using exclusion chromatography, ion exchange chromatography and preparative electrophoresis. The results of partial characterization showed the presence of a basic ß (1,3) glucanase with a molecular weight of 36.8 kDa and pi 9.2 and three ß (1,3) glu-canase acid 35.4, 30.1 and 7.2 kDa with isoelectric points of 3.8, 3.6 and 5.0 respectively. Additionally it was determined the kinetic properties of each enzyme, KM,Vmax, and Eo,K3, according to the results, the basic protein has not been reported in the literature related to tomato.

Keywords : intercellular space; ß (1,3) glucanases; PR (pathogenesis related) proteins; leaves; tomato.

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