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Agrobacterium tumefaciens-mediated transformation of severalRubus genotypes and recovery of transformed plants

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Abstract

Experiments in shoot regeneration and virulentAgrobacterium tumefaciens-mediated transformation were used to develop a protocol forRubus transformation. This protocol was then used to produce transformedRubus plants fromin vitro internodes inoculated with anAgrobacterium tumefaciens encoding neomycin phosphotransferase on its disarmed T-DNA. Two transformed plants were selected from 800 inoculations on a medium containing 10 µg ml−1 kanamycin. Results indicated that this level of kanamycin successfully selected against non-transformed cells but did not reduce the number of transformed, kanamycin-resistant, shoots formed. Enzyme assays and Southern blot analysis verified the presence of the β-glucuronidase gene in the plant genome.

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Abbreviations

BA:

benzyladenine

CAT:

chloramphenicol acetyl transferase

CSR:

chlorsulfuron resistance

2,4-D:

2,4-dichlorophenoxyacetic acid

β-GUS:

β-glucuronidase

IAA:

indoleacetic acid

IBA:

indolebutyric acid

NAA:

naphthaleneacetic acid

NPT:

neomycin phosphotransferase

TDZ:

thidiazuron

References

  • Alt-Moerbe J, Neddermann P, von Lintig J, Weiler EW & Schroder J (1988) Temperature-sensitive step in Ti plasmidvir-region induction and correlation with cytokinin secretion byAgrobacteria. Mol. Gen. Genet. 213: 1–8

    Article  Google Scholar 

  • Bolton GW, Nester EW & Gordon MP (1986) Plant phenolic compounds induce expression of theAgrobacterium tumefaciens loci needed for virulence. Science 232: 983–985

    PubMed  Google Scholar 

  • Burr TJ, Katz BH & Bishop AL (1987) Populations ofAgrobacterium in vineyard and nonvineyard soils and grape roots in vineyards and nurseries. Plant Dis. 71: 617–620

    Google Scholar 

  • Colby SM & Meredith CP (1990) Kanamycin sensitivity of cultured tissues ofVitis. Plant Cell Rep. 9: 237–240

    Article  Google Scholar 

  • Colby SM, Juncosa AM & Meredith CP (1991) Cellular differences inAgrobacterium susceptibility and regenerative capacity restrict the development of transgenic grapevines. J. Amer. Soc. Hort. Sci. 116: 356–361

    Google Scholar 

  • Davies JA, Addison CF, Delaney SF, Sunkel C & Glover DM (1986) Expression of a prokaryotic gene for chloramphenicol acetyl transferase inDrosophila under the control of larval serum protein 1 gene promoters. J. Mol. Biol. 189: 13–24

    Article  PubMed  Google Scholar 

  • Ditta G, Stanfield S, Corbin D & Helinski D (1980) Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank ofRhizobium meliloti. Proc. Natl. Acad. Sci. USA. 77: 7347–7351

    PubMed  Google Scholar 

  • Fillatti JJ, Sellmer J, McCown B, Haissig B & Comai L (1987)Agrobacterium mediated transformation and regeneration ofPopulus. Mol. Gen. Genet. 206: 192–199

    Article  Google Scholar 

  • Fiola JA, Hassan MA, Swartz HJ, Bors RH & McNicol RJ (1990) The effect of thidiazuron, light fluence rate and kanamycin on shoot regeneration from excisedRubus cotyledons andin vitro leaves. Plant Cell Tiss. Org. Cult. 20: 223–228.

    Google Scholar 

  • Graham J, McNicol RJ & Kumar A (1990) Use of the GUS gene as a selectable marker forAgrobacterium-mediated transformation ofRubus. Plant Cell Tiss. Org. Cult. 20: 35–39

    Article  Google Scholar 

  • James DJ, Passey AJ, Barbara DJ & Bevan M (1989) Genetic transformation of apple (Malus pumila) using a disarmed Ti-binary vector. Plant Cell Rep. 7: 658–661

    Google Scholar 

  • James DJ, Passey AJ & Barbara DJ (1990)Agrobacterium-mediated transformation of the cultivated strawberry (Fragaria × ananassa Duch.) using disarmed binary vectors. Plant Sci. 69: 79–94

    Article  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405

    Google Scholar 

  • Maniatis T, Fritsch EF & Sambrook J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Lab, Cold Spring Harbor, NY

    Google Scholar 

  • Mattews H & Litz RE (1990) Kanamycin sensitivity of mango somatic embryos. HortScience 25: 965–966

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497

    Google Scholar 

  • Nehra NS, Chibbar N, Kartha KK, Datla RSS, Crosby WL & Stushnoff C (1990) Genetic transformation of strawberry byAgrobacterium tumefaciens using a leaf regeneration system. Plant Cell Rep. 9: 293–298

    Google Scholar 

  • Otten L & Schilperoort RA (1978) A rapid microscale method for the detection of lysopine and nopaline dehydrogenase activities. Biochem. Biophys. Acta 517: 497–500

    Google Scholar 

  • Smigocki AM & Hammerschlag FA (1991) Regeneration of plants from peach embryo cells infected with a shooty mutant strain ofAgrobacterium. J. Amer. Soc. Hort. Sci. 116: 1092–1097

    Google Scholar 

  • Swartz HJ, Bors RH, Mohamed FA & Naess SK (1990) The effect ofin vitro pretreatment on subsequent shoot organogenesis from excisedRubus andMalus leaves. Plant Cell Tiss. Org. Cult. 21: 179–184

    Article  Google Scholar 

  • Watson JC & Thompson WF (1988) Purification and restriction endonuclease analysis of plant nuclear DNA. In: Weissbach A & Weissbach H (eds) Methods for Plant Molecular Biology (pp 57–75). Academic Press, Orlando, FL

    Google Scholar 

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Authors and Affiliations

  1. Department of Horticulture, University of Maryland, 20742, College Park, MD, USA

    Mahmoud A. Hassan & Harry Jan Swartz

  2. US Department of Agriculture, Plant Hormone Lab, Beltsville Agricultural Research Center-West, 20705, Beltsville, MD, USA

    Gordon Inamine

  3. John Innes Institute, Norwich-Colney Lane, Norwich, UK

    Phillip Mullineaux

Authors
  1. Mahmoud A. Hassan
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  2. Harry Jan Swartz
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  3. Gordon Inamine
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  4. Phillip Mullineaux
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Hassan, M.A., Swartz, H.J., Inamine, G. et al. Agrobacterium tumefaciens-mediated transformation of severalRubus genotypes and recovery of transformed plants. Plant Cell Tiss Organ Cult 33, 9–17 (1993). https://doi.org/10.1007/BF01997592

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  • DOI: https://doi.org/10.1007/BF01997592

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