Nonmycorrhizal (myc-) mutants of Hebeloma cylindrosporum obtained through insertional mutagenesis

Molecular Plant-microbe Interactions : MPMI
Jean-Philippe CombierGilles Gay

Abstract

Polyethylene glycol-mediated transformation of protoplasts was used as a method for insertional mutagenesis to obtain mutants of the ectomycorrhizal fungus Hebeloma cylindrosporum impaired in symbiotic ability. Following restriction enzyme-mediated integration or conventional plasmid insertion, a library of 1,725 hygromycin-resistant monokaryotic transformants was generated and screened for the symbiotic defect, using Pinus pinaster seedlings as host plants. A total of 51 transformants displaying a dramatically reduced mycorrhizal ability were identified. Among them, 29 were nonmycorrhizal (myc-), but only 10 of them had integrated one or several copies of the transforming plasmid in their genome. Light and scanning electron microscopy observations of pine roots inoculated with myc- mutants suggested that we selected mutants blocked at early stages of interaction between partners or at the stage of Hartig net formation. Myc- mutants with plasmid insertions were crossed with a compatible wild-type monokaryon and allowed to fruit. Monokaryotic progenies were obtained in three independent crosses and were analyzed for symbiotic activity and plasmid insertion. In all three progenies, a 1:1 myc-:myc+ segregation ratio was observed, ...Continue Reading

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Citations

Mar 6, 2007·The New Phytologist·Luisa Lanfranco
Feb 1, 2007·FEMS Microbiology Letters·David RekangaltLaurence Fraissinet-Tachet
Feb 10, 2006·Cell Research·Richard J WeldHayley J Ridgway
Jun 27, 2018·The New Phytologist·Adeline BecquerClaude Plassard
Jul 11, 2006·Environmental Biosafety Research·Chi ZhangUwe Nehls

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