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Genetically Transformed Maize (Zea mays L.) Plants from Protoplasts

  • C. A. Rhodes
  • D. A. Pierce
  • I. J. Mettler
  • D. Mascarenhas
  • J. J. Detmer
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 9)

Abstract

Agriculturally important cereal crops, including maize, have been difficult to engineer genetically using current techniques for gene insertion. With few exceptions (Graves and Goldman 1986; Grimsley et al. 1987), most of the graminaceous crops are not readily susceptible to infection by Agrobacterium tumefaciens, which is a vector for gene transfer commonly used with many dicot species. Genes can be transferred directly into protoplasts, without an Agrobacterium vector, by methods that permit DNA to cross the plasmalemma (Krens et al. 1982; Shillito et al. 1985; Riggs and Bates 1986; Fromm et al. 1986). Stable transformation of maize cells has been achieved through direct uptake of DNA into protoplasts that had been permeabilized by electroporation (Fromm et al. 1986; Pierce et al. 1987), but until recently (Rhodes et al. 1988 a), no plants had been recovered from maize protoplasts. We now describe regeneration of maize plants derived from protoplasts into which a gene encoding neomycin phosphotransferase II (NPT II) was introduced via electroporation. NPT II permits plant cells to grow on inhibitory levels of the antibiotic kanamycin (Fraley et al. 1983; Fromm et al. 1986) and can be used as a dominant marker to select for transformed cells. Following is the account of our published work on the regeneration of transformed plants from maize protoplasts (Rhodes et al. 1988 b).

Keywords

Maize Plant Maize Streak Virus Relative Specific Activity Maize Protoplast Black Mexican Sweet 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Barker RF, Idler KB, Thompson DV, Kemp JD (1983) Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTi5955. Plant Mol. Biol 2: 335CrossRefGoogle Scholar
  2. Dellaporta S, Wood J, Hicks J (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19CrossRefGoogle Scholar
  3. Edallo S, Zucchinali C, Perenzin M, Salamini F (1981) Chromosomal variation and frequency of spontaneous mutation associated with in vitro culture and plant regeneration in maize. Maydica 26: 39Google Scholar
  4. Fraley R, Rogers S, Horsch R, Sanders P, Flick J, Adams S, Bittner M, Brand L, Fink C, Fry J, Galluppi G, Goldberg S, Hoffman N, Woo S (1983) Expression of bacterial genes in plant cells. Proc Natl Acad Sci USA 80: 4803PubMedCrossRefGoogle Scholar
  5. Fromm M, Taylor LP, Walbot V (1985) Expression of genes electroporated into monocot and dicot plant cells. Proc Natl Acad Sci USA 82: 5824PubMedCrossRefGoogle Scholar
  6. Fromm MF, Taylor LP, Walbot V (1986) Stable transformation of maize after gene transfer by electroporation. Nature (London) 319: 791CrossRefGoogle Scholar
  7. Graves ACF, Goldman S (1986) The transformation of Zea mays seedlings with Agrobacterium tumefaciens: Detection of T-DNA specific enzyme activities. Plant Mol Biol 7: 43CrossRefGoogle Scholar
  8. Grimsley N, Hohn T, Davies JW, Hohn B (1987) Agrobacterium-mediated delivery of infectious maize streak virus into maize plants. Nature (Lond) 325:177Google Scholar
  9. Guilley H, Dudley RK, Jonard G, Balazs E, Richards K (1982) Transcription of cauliflower mosaic virus DNA: Detection of promoter sequences and characterization of transcripts. Cell 30: 763Google Scholar
  10. Krens FH, Molendijk L, Wullems G, Schilperoort RA (1982) In vitro transformation of plant protoplasts with Ti-plasmid DNA. Nature (Lond) 296: 72CrossRefGoogle Scholar
  11. Mettler IJ, Lachmansingh AR, Detmer JJ, Pierce DA, Mascarenhas D (unpublished data)Google Scholar
  12. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473CrossRefGoogle Scholar
  13. Pierce DA, Mettler IJ, Lachmansingh AR, Pomeroy LM, Weck EA, Mascarenhas D (1987) Effect of 35S leader modifications on promoter activity. In: UCLA Symposia on Molecular and Cellular Biology, New Series (Liss, New York), vol. 62, pp 301–310Google Scholar
  14. Reiss B, Sprengel R, Will H, Schaller H (1984) A new and sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30: 211PubMedCrossRefGoogle Scholar
  15. Rhodes CA, Phillips RL, Green CE (1986) Cytogenetic stability of aneuploid maize tissue cultures. Can J Genet Cytol 28: 374Google Scholar
  16. Rhodes C, Lowe K, Ruby K (1988a) Plant regeneration from protoplasts isolated from embryogenic maize cell cultures. Biotechnology 6: 56CrossRefGoogle Scholar
  17. Rhodes C, Pierce I, Mettler D, Mascarenhas D, Detmer J (1988b) Genetically transformed maize plants from protoplasts. Science 240: 204PubMedCrossRefGoogle Scholar
  18. Riggs CD, Bates GW (1986) Stable transformation of tobacco by electroporation: Evidence for plasmid concatenation. Proc Natl Acad Sci USA 83: 5602PubMedCrossRefGoogle Scholar
  19. Schröder G, Schröder J (1982) Hybridization selection and translation of T-DNA encoded mRNAs from octopine tumors. Mol Gen Genet 185: 51CrossRefGoogle Scholar
  20. Shillito RD, Saul MW, Paszkowski J, Muller M, Potrykus I (1985) High efficiency direct gene transfer to plants. Biotechnology 3: 1099CrossRefGoogle Scholar
  21. Southern E (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503PubMedCrossRefGoogle Scholar
  22. Werr W, Frommer WB, Maas C, Starlinger P (1985) Structure of the sucrose synthase gene on chromosome 9 of Zea mays L. EMBO J 4: 1373PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1989

Authors and Affiliations

  • C. A. Rhodes
    • 1
  • D. A. Pierce
    • 2
  • I. J. Mettler
    • 1
  • D. Mascarenhas
    • 3
  • J. J. Detmer
    • 3
  1. 1.Sandoz Crop Protection CorporationPalo AltoUSA
  2. 2.EniChemMonmouth JunctionUSA
  3. 3.BioGrowthRichmondUSA

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