Epsilon Open Archive

Abstract

This thesis summarises the author's work on the effectiveness of two control methods against the necrotrophic fungal pathogen Heterobasidion annosum s.l. (Fr.) Bref. The studied control methods were stump treatments through the application of a fungal biocontrol agent (Phlebiopsis gigantea) and application of urea solution. An additional component to this thesis was to develop a useful Agrobacterium tumefaciens-mediated transformation system for the H. annosum s.l. The potential risk for a build-up in the capacity of H. annosum s.l. to resist overgrowth by P. gigantea was investigated by utilising a P. gigantea strain used in the commercial preparation Rotstop and different strains of H. annosum s.l. The impact on intraspecific genetic diversity from the P. gigantea used in Rotstop four years after stump treatment was estimated using both microsatellite markers and AP-PCR. The long-term effect of urea treatment during first rotation thinning of Picea abies stands was evaluated 15 years post-treatment. An A. tumefaciens-mediated transformation system was developed using two different transformation plasmids and a North American H. annosum strain. The results revealed differences between H. annosum s.l. strains in their ability to resist overgrowth by P. gigantea. This trait was found to be heritable which suggests that H. annosum s.l. has the capacity to develop more tolerant strains against the biocontrol agent if the conditions are met. Also, one QTL effect associated with this trait was identified. Genetic diversity was lower within Rotstop treated plots and the resident population was more genetically similar to the applied P. gigantea strain than other populations sampled in Sweden. This effect on genetic structure was not detected 100-200 m from the treated plot which suggests that stump treatments with P. gigantea only has a local effect on genetic diversity. Urea treated plots exhibited less rot than untreated plots and the urea treatments also significantly reduced the occurrence of windthrown trees. This shows that urea treatment during thinning operations is an efficient method to reduce root rot in P. abies stands. A total of 18 H. annosum s.l. transformants were obtained from the A. tumefaciens-mediated transformation system. The transformants expressed GFP throughout the mycelia and had a normal growth rate. This system will be very useful for genomic work and interaction studies.