Skip to main content
Log in

Strain specificity in transformation of alfalfa by Agrobacterium tumefaciens

  • Original Research Paper
  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

Production of transgenic alfalfa plants by Agrobacterium-mediated transformation requires Agrobacterium infection and regeneration from tissue culture. Variation in alfalfa (Medicago sativa L.) germplasm for resistance to oncogenic and disarmed strains of A. tumefaciens (Smith & Townsend) Conn was tested in plant populations representing the nine distinct sources of alfalfa germplasm introduced into North America and used to develop modern varieties. For each of the virulent strains there was a positive correlation (p=0.05) of resistance to tumorigenesis with the trait for fall dormancy. There was also a significant correlation between plants selected for ineffective nodulation and resistance to tumorigenesis suggesting that the genetic loci required for successful symbiosis are also involved in tumorigenesis. Tissue explants of seedlings from the nine diversity groups were tested for transformation by three disarmed strains containing a plasmid with the scorable marker β-glucuronidase. The strong correlation between dormancy and resistance to oncogenic strains was not observed with disarmed strains. However, there was a strong germplasm-strain interaction or transformation and embryogenesis in a highly embryogenic genotype. Thus, transformation at the whole plant level is germplasm dependent while in tissue culture the bacterial strain used is the critical variable for successful transformation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

pTi:

tumor-inducing plasmid

GUS:

β-glucuronidase

References

  • Austin S, Bingham ET, Mathews DE, Shahan MN, Will J & Burgess RR (1995) Production and field performance of transgenic alfalfa (Medicago sativa L.) expressing alpha-amylase and manganese-dependent lignin peroxidase. Euphytica (in press)

  • Bechtold N, Ellis J & Pelletier G (1983) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C. R. Acad. Sci. Paris. Life Sciences 316: 1194–1199

    Google Scholar 

  • Bingham ET, Hurley LV, Kaatz DM & Saunders JW (1975) Breeding alfalfa which regenerates from callus tissue in culture. Crop Sci. 15: 719–721

    Google Scholar 

  • Bingham ET (1991) Registration of alfalfa hybrid Regen-SY germplasm for tissue culture and transformation research. Crop Sci. 31: 1098

    Google Scholar 

  • Brown DCW & Atanassov A (1985) Role of genetic background in somatic embryogenesis in Medicago. Plant Cell Tiss Org. Cult. 4: 111–122

    Google Scholar 

  • Chabaud M, Passiatore JE, Cannon F & Buchanan-Wollaston V (1988) Parameters affecting the frequency of kanamycin resistant alfalfa obtained by Agrobacterium tumefaciens mediated transformation. Plant Cell Rep. 7: 512–516

    Google Scholar 

  • Frosheiser FI & Barnes DK (1973) Field and greenhouse selection for Phytophthora root rot resistance in alfalfa. Crop Sci. 13: 735–738

    Google Scholar 

  • Frosheiser FI & Barnes DK (1978) Field reaction of artificially inoculated afalfa populations to the Fusarium and bacterial wilt pathogens alone in combination. Phytopath. 68: 943–946

    Google Scholar 

  • Hill KK, Jarvis-Eagan N, Halk EL, Krahn KJ, Liao LW, Mathewson RS, Merlo DT, Nelson SE, Rashka KE & Loesch-Fries LS (1991) The development of virus-resistant alfalfa, Medicago sativa L. Bio/Technology 9: 373–377

    Google Scholar 

  • Horsch R, Fry J, Hoffman N, Eichholtz D, Rogers S & Fraley R (1985) A simple and general method for transferring genes into plants. Science 227: 1229–1231

    Google Scholar 

  • Jefferson RA, Kavanagh TA & Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907

    Google Scholar 

  • Kielly GA & Bowley SR (1992) Genetic control of somatic embryogenesis in alfalfa. Genome 35: 474–477

    Google Scholar 

  • Koncz C & Schell J (1986) The promoter of TL-DNA gene 5 controls the tissue-specific expression of chimeric genes carried by novel type of Agrobacterium binary vector. Mol. Gen. Genet. 204: 383–396

    Google Scholar 

  • Laszo GR, Stein PA, & Ludwig RA (1991) A transformation competent Arabidopsis genomic library in Agrobacterium. Bio/Technology 9: 963–967

    Google Scholar 

  • Mariotti D, Davey MR, Draper J, Freeman JP & Cocking EC (1984) Crown gall tumorigenesis in the forage legume Medicago sativa L. Plant Cell Physiol. 25: 473–482

    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 

  • Ooms G, Hooykaas PJJ, Van Veen RJM, V Beelen P Regensburg-Tuink TJG & Schilperoort RA (1982) Octopine Ti-plasmid deletion mutants of Agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid 7: 15–29

    Google Scholar 

  • Peterson MA & Barnes DK (1981) Inheritance of ineffective nodulation and non-nodulation traits in alfalfa. Crop Sci. 21: 611–616

    Google Scholar 

  • Reisch B & Bingham ET (1980) The genetic control of bud formation from callus cultures of diploid alfalfa. Plant Sci. Lett. 20: 71–77

    Google Scholar 

  • Saunders JW & Bingham ET (1972) Production of alfalfa plants from callus tissue. Crop Sci. 12: 804–808

    Google Scholar 

  • Shahin EA, Spielmann A, Sukhapinda K, Simpson RB & Yashar M (1986) Transformation of cultivated alfalfa using disarmed Agrobacterium tumefaciens. Crop Sci. 26: 1235–1239

    Google Scholar 

  • Viands DR, Barnes DK, Stucker RE & Frosheiser FI (1979) Inheritance of resistance to bacterial wilt in two alfalfa gene pools: Response to selection and quantitative analysis. Crop Sci. 19: 711–714

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Samac, D.A. Strain specificity in transformation of alfalfa by Agrobacterium tumefaciens . Plant Cell Tiss Organ Cult 43, 271–277 (1995). https://doi.org/10.1007/BF00039955

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00039955

Key words

Navigation