Skip to Main content Skip to Navigation
Journal articles

The dominant Hc. Sdh R carboxin-resistance gene of the ectomycorrhizal fungus Hebeloma cylindrosporum as a selectable marker for transformation

Abstract : In an attempt to get a marker gene suitable for genetical transformation of the ectomycorrhizal fungus Hebeloma cylindrosporum, the gene Hc.Sdh (R) that confers carboxin-resistance was isolated from a UV mutant of this fungus. It encodes a mutant allele of the Fe-S subunit of the succinate dehydrogenase gene that carries a single amino acid substitution known to confer carboxin-resistance. This gene was successfully used as the selective marker to transform, via Agrobacterium tumefaciens, monokaryotic and dikaryotic strains of H. cylindrosporum. We also successfully transformed hygromycin-resistant insertional mutants. Transformation yielded mitotically stable carboxin-resistant mycelia. This procedure produced transformants, the growth of which was not affected by 2 mu g l(-1) carboxin, whereas wild-type strains were unable to grow in the presence of 0.1 mu g l(-1) of this fungicide. This makes the carboxin-resistance cassette much more discriminating than the hygromycin-resistance one. PCR amplification and Southern blot hybridisation indicated that more than 90% of the tested carboxin-resistant mycelia contained the Hc.Sdh (R) cassette, usually as a single copy. The AGL-1 strain of A. tumefaciens was a much less efficient donor than LBA 1126; the former yielded ca. 0-30% transformation frequency, depending on fungal strain and resistance cassette used, whereas the latter yielded ca. 60-95%.
Document type :
Journal articles
Complete list of metadatas

https://hal-univ-lyon1.archives-ouvertes.fr/hal-02555064
Contributor : Depot 1 Lyon 1 <>
Submitted on : Monday, April 27, 2020 - 9:36:21 AM
Last modification on : Friday, May 29, 2020 - 11:18:02 PM

Links full text

Identifiers

Collections

Citation

C. Ngari, J. P. Combier, J. Doré, Roland Marmeisse, G. Gay, et al.. The dominant Hc. Sdh R carboxin-resistance gene of the ectomycorrhizal fungus Hebeloma cylindrosporum as a selectable marker for transformation. Current Genetics, Springer Verlag, 2009, 55 (2), pp.223-231. ⟨10.1007/s00294-009-0231-4⟩. ⟨hal-02555064⟩

Share

Metrics

Record views

17