Detection of RAPD polymorphisms inGladiolus cultivars with differing sensitivities toFusarium oxysporum f. sp.gladioli

Abstract

Fusarium oxysporum Schlecht. Fr. F. sp.gladioli (FOG) is the most important gladiolus pathogen. One of the most environmentally friendly methods to control its spread is to use cultivars that are minimally sensitive to the pathogen. Infected corm tissues in more resistant varieties have been shown to produce suberin layers that inhibit fungal iphae growth. RAPD analysis of genomes from 9 selected gladiolus cvs, chosen to be the most resistant and sensitive to FOG, were performed to verify DNA polymorphism levels. Total nucleic acid extraction was carried out with a chloroform-phenol method from tissues of plants in 3 growth stages. RAPD experiments were performed using 14 primers with varyingTaq polymerases and primer concentrations. Five of the primers tested gave no polymorphic profiles. Five primers produced polymorphic bands, allowing us to obtain RAPD profiles typical for one or more of the more resistant cvs. All the tested growth stages provided repeatable results, indicating the reliability of detected polymorphisms. Cloning the more interesting polymorphic DNA fragments in the future will verify the presence of specific genes related to FOG resistance mechanisms in gladiolus.