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Molecular transformation of Fusarium solani with an antibiotic resistance marker having no fungal DNA homology

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Summary

A vector was constructed for transformation of the plant pathogenic fungus Fusarium solani. The promoter 35Sp, from cauliflower mosaic virus, was fused to the bacterial gene APH(3′)II, which confers resistance to the aminoglycoside antibiotic G418. Two transformation procedures were developed: one using isolated fungal protoplasts, the other using germinated fungal spores. A transformation frequency of 3.3 G418-resistant colonies were obtained per microgram DNA. Of 14 colonies analyzed, 12 had vector sequences integrated into their high molecular weight DNA, and 2 were untransformed. Integration was sometimes accompanied by rearrangements of both the vector and flanking fungal DNAs. Primer-extension analysis of the mRNA from one transformant revealed two putative transcription initiation sites in the chimeric APH(3′)II gene. Both sites differed from the normal initiation site in plants. This vector will be useful in transformation systems in which integration by non-homologous recombination is desired.[/P ]

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References

  • Aviv H, Leder P (1972) Proc Natl Acad Sci USA 69:1408–1412

    Google Scholar 

  • Beck E, Ludwig G, Awerswald EA, Reiss B, Schaller H (1982) Gene 19:327–336

    Google Scholar 

  • Boel E, Hansen MT, Hjort I, Hoegh I, Fill NP (19844) EMBO J 3:1581–1585

  • Calzone FJ, Britten RJ, Davidson EH (1987) In: Berger SL, Kimmel AR (eds) Guide to molecular cloning techniques. Academic Press, pp 611–633

  • Case ME, Schweizer M, Kushner SR, Giles NH (1979) Proc Natl Acad Sci USA 76:5259–5263

    Google Scholar 

  • Chen EY, Seeburg PH (1985) DNA 4:165–170

    Google Scholar 

  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Biochemistry 18:5294–5299

    Google Scholar 

  • Choi GH, Richey MG, Turbek CSM, Smith DA (1987) Phys Mol Plant Pathol 30:215–224

    Google Scholar 

  • Dhwale SS, Paietta JV, Marzluf GA (1984) Curr Genet 8:77–79

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) Anal Biochem 132:6–13

    Google Scholar 

  • Garber RC, Yoder OC (1983) Anal Biochem 135:416–422

    Google Scholar 

  • Gorman C, Padmanabhan R, Howard BH (1983) Science 221:551–553

    Google Scholar 

  • Guilley H, Dudley RK, Jonard G, Balazs E, Richards KE (1982) Cell 30:763–773

    Google Scholar 

  • Gritz L, Davies J (1983) Gene 25:179–188

    Google Scholar 

  • Hanahan D (1983) J Mol Biol 1670:557–580

    Google Scholar 

  • Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933

    Google Scholar 

  • Hynes MJ (1986) Exp Mycol 10:1–8

    Google Scholar 

  • Ito HY, Kukuda K, Murata K, Kimura A (1978) J Bacteriol 153:163–168

    Google Scholar 

  • Jimenez A, Davies J (1980) Nature 287:869–871

    Google Scholar 

  • Johnson DA, Gautsch JW, Sportsman JR, Elder JH (1984) Gene Aust Techn 1:3–8

    Google Scholar 

  • Kinnaird JH, Fincham JRS (1983) Gene 26:253–260

    Google Scholar 

  • Kistler HC, Benny UK (1988) Curr Genet 13:145–149

    Google Scholar 

  • Kuhn PJ, Smith DA (1978) Ann Appl Biol 89:362–370

    Google Scholar 

  • Kurz MB, Cortelyou MW, Miller SM, Lai M, Kirsch DR (1987) Mol Cell Biol 7:209–217

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 88–95

    Google Scholar 

  • Mirocha CJ, Pathre SV, Christensen CM (1977) In: Wyllie TD, Morehouse LG (eds) Marcel Dekker, New York, Basel

  • Mullaney EJ, Hamer JE, Roberti KA, Yelton MM, Timberlake WE (1985) Mol Gen Genet 199:37–45

    Google Scholar 

  • Nelson PE, Toussoun TA, Cook RJ (1981) Fusarium: diseases, biology and taxonomy. The Pensylvania State University Press, University Park London

    Google Scholar 

  • Odell JT, Knowlton S, Lin W, Mauvais CJ (1988) Plant Mol Biol 10:263–272

    Google Scholar 

  • Odell JT, Nagy F, Chua N (1985) Nature 313:810–812

    Google Scholar 

  • Parsons KA, Chumley FG, Valent B (1987) Proc Natl Acad Sci USA 84:4161–4165

    Google Scholar 

  • Punt PJ, Oliver RP, Dingemanse MA, Pouwels PH, Van den Hondel CAMJJ (1987) Gene 56:117–124

    Google Scholar 

  • Rogers SG, Horsch RB, Fraley RT (1986) Methods Enzymol 118:627–640

    Google Scholar 

  • Santerre RF, Allen NE, Hobbs Jr, JN, Rao RN, Schmidt J (1984) Gene 30:147–156

    Google Scholar 

  • Schardl CL, Byrd AD, Benzion G, Altschuler MA, Hildebrand DF, Hunt AG (1987) Gene 61:1–11

    Google Scholar 

  • Smith DA, Banks SW (1986) Phytochemistry 25:979–995

    Google Scholar 

  • Smith DA, Harrer JM, Cleveland TE (1982) Phytopathology 72:1319–1323

    Google Scholar 

  • Southern EM (1975) J Mol Biol 98:503–517

    Google Scholar 

  • Sreekrishna K, Webster TD, Dickson RC (1984) Gene 28:73–81

    Google Scholar 

  • Turgeon BG, Garber RC, Yoder OC (1987) Mol Cell Biol 7:3297–3305

    Google Scholar 

  • Turpen TH, Griffith OM (1986) Biotechniques 4:11–15

    Google Scholar 

  • Wang J, Holden DW, Leong SA (1988) Proc Natl Acad Sci USA 85:865–869

    Google Scholar 

  • Webster TD, Dickson RC (1983) Gene 26:243–252

    Google Scholar 

  • Yelton MM, Hamer JE, Timberlake WE (1984) Proc Natl Acad Sci USA 81:1470–1474

    Google Scholar 

Download references

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The investigation reported in this paper (No. 89-11-42) is in connection with a project of the Kentucky Agricultural Experiment Station and is published with approval of the director

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Marek, E.T., Schardl, C.L. & Smith, D.A. Molecular transformation of Fusarium solani with an antibiotic resistance marker having no fungal DNA homology. Curr Genet 15, 421–428 (1989). https://doi.org/10.1007/BF00376799

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  • DOI: https://doi.org/10.1007/BF00376799

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