Transformation of the mycorrhizal fungus laccaria bicolor by using agrobacterium tumefaciens

Abstract

Most boreal and temperate forest trees form a mutualistic symbiosis with soil borne fungi called ectomycorrhiza (EC M). In this association both partners benefit due to nutrient exchange at the symbiotic interface. Laccaria bicolor is the first mycorrhizal fungus with its genome sequenced thus making possible for the first time to analyze genome scale gene expression profiles of a mutualistic fungus. However, in order to be able to take full advantage of the genome sequence, reverse genetic tools are needed. Among them a high throughput transformation system is crucial. Herein we present a detailed protocol for genetic transformation of Laccaria bicolor by means of Agrobacterium tumefaciens with emphasis on critical steps affecting the success and efficiency of the approach.