Summary
A protocol is described for the simple, rapid and efficient production of transgenic Arabidopsis plants. The procedure was developed using growth regulator regimes that promote adventitious embryogenesis during or immediately following Agrobacterium mediated transformation. Both the RLD and Columbia genotypes of Arabidopsis were transformed using slightly different growth regulator regimes. For the Columbia genotype two modifications of the protocol were identified which substantially improved regeneration. Cold treatment of the plants used as a source of root explants resulted in a three-fold increase in the number of morphogenic sectors produced. A more important modification was the inclusion of 25 mg/l silver nitrate (an inhibitor of ethylene action) to the medium used for shoot regeneration. This provided a ten-fold increase in the number of shoots produced. These procedures made it possible to obtain over 100 putative transformants of RLD or Columbia from a single 10 cm petri dish, within 2 or 4 weeks after exposure of root explants to the bacteria. When these were transferred to rooting media containing antibiotics, approximately 20% were able to root after kanamycin selection and 80% after hygromycin selection. All the rooted plantlets tested were shown to contain integrated donor DNA as determined by Southern blot analysis.
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Bayliss, MW (1980) Int. Rev. Cytol. Suppl. 11A 113–114
Chang G, Bowman JL, DeJohn AW, Lander ES, Meyerowitz EM (1988) Proc. Natl. Acad. Sci. USA 85: 6856–6860
Chaudbury AM, Singer ER (1980) Plant Cell Reports 8: 368–369
Estelle MA, Somerville CR (1986) Trends in Genetics 2: 89–93
Fromm ME, Taylor LP, Walbot V (1986) Nature 319: 791–793
Gamborg OL, Miller RA, Ojima K (1968) Expt. Cell. Res. 50: 151–158
Hoekema A, Hirsch PR, Hooykaas PJJ, Schiperoort RA (1983) Nature 303: 179–180
Koncz CS, Martini N, Mayerhofer R, Koncz-Kálmán ZS, Körber H, Rédei GP, Schell J (1989) Proc. Natl. Acad. Sci. USA 86: 8467–8471
Koncz CS, Mayerhofer R, Koncz-Kálmán Zs, Nawrath C, Reiss B, Rédei GP, Schell J (1990) EMBO J. 9: 1337–1346
Lloyd AM, Barnason AR, Rogers SG, Byrne MC, Fraley RT, Horsch RB (1986) Science 234: 464–466
Márton L, Wullems GJ, Molendijk L, Schilperoort RA (1979) Nature 277: 129–131
Márton L (1984) In: Cell culture and somatic cell genetics of plants, ed. Vasil IK, Academic Press, New York, Vol. 1, pp 514–521
Meyerowitz E (1989) Cell 56: 263–269
Murashige T, Skoog F (1962) Physiol Plant 15: 473–497
Negrutiu I, Beeflink F, Jacobs M (1975) Plant Sci. Lett. 5: 293–304
Negrutiu I, Jacobs (1978) Z. Pflanzenphysiol. 90: 423–430
Purnhauser L, Dix PJ, Medgyesy P, Czakó M, Márton L (1987) Plant Cell Reports 6: 1–4
Rédei GP (1975) Ann. Rev. Gent. 9: 111–127
Rédei GP, Koncz CS, Schell J (1988) In: Chromosome structure and function eds. Gustavson JP and Appels K, Plenum, New York, pp 175–200
Rogers SO, Bendich AJ (1985) Plant Mol. Biol. 5: 69–76
Schmidt R, Willmitzer (1988) Plant Cell Rep. 7: 583–586
Sheikholeslam SN, Weeks DP (1987) Plant Mol. Biol. 8: 291–298
Valvekens D, Van Montagu M, Van Lijsebettens M (1988) Proc. Natl. Acad. Sci. USA 85: 5536–5540
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Communicated by J. M. Widholm
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Márton, L., Browse, J. Facile transformation of Arabidopsis . Plant Cell Reports 10, 235–239 (1991). https://doi.org/10.1007/BF00232565
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DOI: https://doi.org/10.1007/BF00232565