Summary
The pAN7.1 plasmid containing the E. coli hygromycin B phosphotransferase gene was used to transform protoplasts of the ectomycorrhizal fungus Hebeloma cylindrosporum. Hygromycin-resistant transformants were selected at a frequency of one to five per μg of transforming DNA. Southern blot analyses revealed multiple copy integration of the transforming plasmid into the genome. The selection system was used to introduce other genes of interest by co-transformation. Two plasmids, one containing tryptophan biosynthesis genes and the other the NADP-glutamate dehydrogenase gene from the saprophytic basidiomycete Coprinus cinereus, were successfully introduced into the H. cylindrosporum genome with up to 70% efficiency of co-transformation. The hygromycin resistance phenotype was stably maintained during growth of transformants on hygromycinfree medium. All tranformants retained their ability to form mycorrhizae with the habitual host plant Pinus pinaster, making them suitable for future physiological studies.
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References
Barrett V, Lemke PA, Dixon RK (1989) Appl Microbiol Biotechnol 30: 381–387
Barrett V, Dixon RK, Lemke PA (1990) Appl Microbiol Biotechnol 33: 313–316
Bruchet G (1980) Rev Forest Fr 32: 294–295
Casselton LA, de la Fuente Herce A (1989) Curr Genet 16: 35–40
Chilvers GA, Douglass PA, Lapeyrie F (1986) New Phytol 103: 397–402
Cross SH, Little PFR (1986) Gene 49: 9–22
Debaud J-C, Gay G (1987) New Phytol 105: 429–435
Debaud J-C, Gay G, Bruchet G (1986) In: Gianinazzi-Pearson V, Gianinazzi S (eds) Physiological and genetical aspects of mycorrhizae. INRA publications, Paris, pp 581–588
Fincham JRS (1989) Microbiol Rev 53: 148–170
Harley JL, Smith SE (1983) Mycorrhizal symbiosis. Academic Press, London
Hébraud M, Fèvre M (1987) Can J Microbiol 34: 157–161
Hilbert J-L, Martin F (1988) New Phytol 110: 339–346
Le Tacon F, Valdenaire J-M (1980) Rev Fores Fr 32: 281–293
Malajczuk N, Lapeyrie F, Garbaye J (1989) New Phytol 114: 627–631
Martin F, Botton B (1991) Adv Plant Pathol (in press)
Mellon FM, Casselton LA (1988) Curr Genet 14:451–456
Mellon FM, Little PFR, Casselton LA (1987) Mol Gen Genet 210: 352–357
Mooibroek H, Kuipers AGJ, Sietsma JH, Punt PJ, Wessels JGH (1990) Mol Gen Genet 222: 41–48
Mutasa ES, Tymon AM, Göttgens B, Mellon FM, Little PFR, Casselton LA (1990) Curr Genet 16: 35–40
Punt PJ, Oliver RP, dingemanse MA, Pouwels PH, van den Hondel CAMJJ (1987) Gene 56: 117–124
Rao PS, Niederpruem DJ (1969) J Bacteriol 100: 1222–1228
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Slankis V (1973) In: Marks GC, Koslowksi TT (eds) Ectomycorrhizae. Academic Press, London, pp 231–298
Valjalo J (1979) In Delmas J (ed) Mushroom science, part 1. Center for Agricultural Publication and Documentation, Wageningen, pp 903–918
Veal D, Casselton LA (1985) Arch Microbiol 142: 157–163
Wagner F (1988) PhD thesis, University of Lyon, France
Zolan ME, Pukkila PJ (1986) Mol Cell Biol 6: 195–200
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Communicated by C. P. Hollenberg
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Marmeisse, r., Gay, G., Debaud, JC. et al. Genetic transformation of the symbiotic basidiomycete fungus Hebeloma cylindrosporum . Curr Genet 22, 41–45 (1992). https://doi.org/10.1007/BF00351740
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DOI: https://doi.org/10.1007/BF00351740