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.
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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
Bingham ET, Hurley LV, Kaatz DM & Saunders JW (1975) Breeding alfalfa which regenerates from callus tissue in culture. Crop Sci. 15: 719–721
Bingham ET (1991) Registration of alfalfa hybrid Regen-SY germplasm for tissue culture and transformation research. Crop Sci. 31: 1098
Brown DCW & Atanassov A (1985) Role of genetic background in somatic embryogenesis in Medicago. Plant Cell Tiss Org. Cult. 4: 111–122
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
Frosheiser FI & Barnes DK (1973) Field and greenhouse selection for Phytophthora root rot resistance in alfalfa. Crop Sci. 13: 735–738
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
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
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
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
Kielly GA & Bowley SR (1992) Genetic control of somatic embryogenesis in alfalfa. Genome 35: 474–477
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
Laszo GR, Stein PA, & Ludwig RA (1991) A transformation competent Arabidopsis genomic library in Agrobacterium. Bio/Technology 9: 963–967
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
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
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
Peterson MA & Barnes DK (1981) Inheritance of ineffective nodulation and non-nodulation traits in alfalfa. Crop Sci. 21: 611–616
Reisch B & Bingham ET (1980) The genetic control of bud formation from callus cultures of diploid alfalfa. Plant Sci. Lett. 20: 71–77
Saunders JW & Bingham ET (1972) Production of alfalfa plants from callus tissue. Crop Sci. 12: 804–808
Shahin EA, Spielmann A, Sukhapinda K, Simpson RB & Yashar M (1986) Transformation of cultivated alfalfa using disarmed Agrobacterium tumefaciens. Crop Sci. 26: 1235–1239
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
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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
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DOI: https://doi.org/10.1007/BF00039955