Genetic transformation of filamentous fungi by Agrobacterium tumefaciens

Abstract

Agrobacterium tumefaciens-mediated transformation of filamentous fungi consists of (i) induction of A. tumefaciens culture harbouring a binary vector, (ii) co-incubation of bacteria with fungal spores on a solid support, and (iii) selection of transformants. During the induction, vir genes on the helper component of the binary vector are activated, conditioning A. tumefaciens for transformation. During co-cultivation, T-DNA part of the binary vector system is transferred into fungal nucleus and inserted into the genome. Transformants are selected on a medium with appropriate antibiotic. In order to maximize the number of transformants, the ratio of A. tumefaciens cells to fungal spores and the duration of the co-cultivation need be optimized. The procedure takes two to three weeks for fast-growing fungi.