Agrobacterium-mediated insertional mutagenesis in the mycorrhizal fungus Laccaria bicolor

Agrobacterium -mediated gene transfer (AMT) is extensively employed as a tool in fungal functional genomics and accordingly, in previous studies we used AMT on a dikaryotic strain of the ectomycorrhizal basidiomycete Laccaria bicolor . The interest in this fungus derives from its capacity to establi...

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Bibliographic Details
Published in:Current genetics 2017-05, Vol.63 (2), p.215-227
Main Authors: Stephan, B. I., Alvarez Crespo, M. C., Kemppainen, M. J., Pardo, A. G.
Format: Article
Language:English
Subjects:
Agrobacterium - genetics
Base Sequence
Basidiomycetes
Binding Sites - genetics
Biochemistry
Biomedical and Life Sciences
Blotting, Southern
Cell Biology
Deoxyribonucleic acid
DNA
DNA, Bacterial - genetics
DNA, Fungal - genetics
Forest ecosystems
Fungal Proteins - genetics
Genome, Fungal - genetics
Laccaria - genetics
Laccaria bicolor
Life Sciences
Microbial Genetics and Genomics
Microbiology
Mutagenesis, Insertional
Mycorrhizae - genetics
Nutrient cycles
Original Article
Plant Sciences
Proteomics
Sequence Analysis, DNA
Symbiosis
Transformation, Genetic
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Summary:Agrobacterium -mediated gene transfer (AMT) is extensively employed as a tool in fungal functional genomics and accordingly, in previous studies we used AMT on a dikaryotic strain of the ectomycorrhizal basidiomycete Laccaria bicolor . The interest in this fungus derives from its capacity to establish a symbiosis with tree roots, thereby playing a major role in nutrient cycling of forest ecosystems. The ectomycorrhizal symbiosis is a highly complex interaction involving many genes from both partners. To advance in the functional characterization of fungal genes, AMT was used on a monokaryotic L. bicolor . A collection of over 1200 transgenic strains was produced, of which 200 randomly selected strains were analyzed for their genomic T-DNA insertion patterns. By means of insertional mutagenesis, a number of transgenic strains were obtained displaying differential growth features. Moreover, mating with a compatible strain resulted in dikaryons that retained altered phenotypic features of the transgenic monokaryon. The analysis of the T-DNA integration pattern revealed mostly similar results to those reported in earlier studies, confirming the usefulness of AMT on different genetic backgrounds of L. bicolor . Taken together, our studies display the great versatility and potentiality of AMT as a tool for the genetic characterization of L. bicolor .
ISSN:0172-8083
1432-0983
DOI:10.1007/s00294-016-0627-x