Transformation of Rice via PEG-Mediated DNA Uptake into Protoplasts

  • Karabi Datta
  • Swapan K. Datta
Part of the Methods In Molecular Biology™ book series (MIMB, volume 111)

Abstract

For stable transformation of cereals through PEG-mediated DNA uptake into protoplasts, the two most critical requisites are the ability to isolate and culture protoplasts in large numbers, and the development of an efficient and reliable system for routine plant regeneration from protoplasts (1, 2, 3). Based on early success with mesophyll protoplasts of some dicotyledonous species, extensive efforts were made to induce sustained division in protoplasts isolated from leaves or young shoots of different cereal plants. However, there is still no convincing evidence of sustained divisions in protoplasts isolated from leaves or shoots of any cereal In contrast, protoplasts isolated from embryogenic suspension cultures could be induced to divide in culture (4). Obtaining a fast-growing and highly embryogenic suspension culture is the most important factor for cereal plant regeneration from protoplasts (4, 5, 6, 7, 8). Microspore cultures, and immature or mature embryos may be used to obtain embryogenic calh which can eventually be used to establish embryogenic cell suspensions ECS (2,7).

Keywords

Sucrose Microwave Filtration Mercury Agarose 

References

  1. 1.
    Datta, K, Potrykus, I., and Datta, S. (1992) Efficient fertile plant regeneration from protoplasts of the Indica rice breeding line IR72 (Oryza sativa L) Plant Cell Rep 11,229–233.CrossRefGoogle Scholar
  2. 2.
    Datta, S. K., Peterhans, A, Datta, K., and Potrykus, I. (1990) Genetically engineered fertile Indica-rice plants recovered from protoplasts. Bio/Technology 8, 736–740.CrossRefGoogle Scholar
  3. 3.
    Peterhans, A, Datta, S. K., Datta, K, Goodall, G. J., Potrykus, I., and Paszkowski, J (1990) Recognition efficiency of Dicotyledoneae-specific promoter and RNA processing signals in rice. Mol. Gen Genet 111, 361–368.CrossRefGoogle Scholar
  4. 4.
    Vasil, I. K. (1987) Developing cell and tissue culture systems for the improvement of cereal and grass crops. J Plant Physiol 128, 193–218Google Scholar
  5. 5.
    Datta, S. K., Datta, K., Soltanifar, N, Donn, G., and Potrykus, I. (1992) Herbicide-resistant Indica rice plants from IRRI breeding line IR72 after PEG-mediated transformation of protoplasts Plant Mol Biol 20,619–629.PubMedCrossRefGoogle Scholar
  6. 6.
    Kyozuka, J., Hayashi, Y., and Shimamoto, K. (1987) High frequency plant regeneration form rice protoplasts by novel nurse culture methods. Mol Gen Genet 206,408–413CrossRefGoogle Scholar
  7. 7.
    Datta, S. K, Datta, K., and Potrykus, I (1990) Fertile Indica rice plants regenerated from protoplasts isolated from microspore derived cell suspensions. Plant Cell Rep 9,253–256.CrossRefGoogle Scholar
  8. 8.
    Datta, S. K. (1995) Polyethylene-glycol-mediated direct gene transfer to indica rice protoplasts and regeneration of transgenic plants, in Gene Transfer to Plants (Potrykus, I. and Spangenberg, G, eds), Springer-Verlag, New York, pp 66–74.Google Scholar
  9. 9.
    Datta, S. K. (1996) Genetic transformation of rice from protoplasts of haploid origin, in In Vitro Haploid Production in Higher Plants, 2 (Jain, S. M., Sopory, S K, and Veilleux, R. E, eds.), Kluwer Academic, The Netherlands, pp. 411–423.Google Scholar
  10. 10.
    Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures Physiol Plant. 15, 473–497.CrossRefGoogle Scholar
  11. 11.
    Muller, A. J and Graffe, R. (1978) Isolation and characterization of cell lines of Nicotiana tabacum lacking nitrate reductase. Mol Gen Genet. 161, 67–76.CrossRefGoogle Scholar
  12. 12.
    Ohira, K., Ojima, K., and Fujiwara, A (1973) Studies on the nutrition of rice cell culture. 1. A simple, defined medium for rapid growth in the suspension culture Plant Cell Physiol 14, 1113–1121.Google Scholar
  13. 13.
    Chu, C C, Wang, C. C, Sun, C. S., Hsu, C, Yin, K. C, and Chu, C. Y (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Set. Sin 18,659–668Google Scholar
  14. 14.
    Datta, S. K., Datta, K., and Potrykus, I. (1990) Embryogenesis and plant regeneration from microspores of both “indica” and “japonica” rice (Oryza sativa) Plant Sci. 67, 83–88.CrossRefGoogle Scholar
  15. 15.
    Datta, S K. and Wenzel, G. (1987) Isolated microspore derived plant formation via embryogenesis in Triticum aestivum Plant Sci 48, 49–54.CrossRefGoogle Scholar
  16. 16.
    Shillito, R D, Paszkowski, J., and Potrykus I (1983) Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a number of plant species. Plant Cell Rep. 2, 244–247CrossRefGoogle Scholar
  17. 17.
    Smith, J. A., Green, C. E., and Grengenbach, B. G. (1984) Feeder layer support of low density populations of Zea mays suspension cells. Plant Sci Lett 36, 67–72CrossRefGoogle Scholar
  18. 18.
    Yoshida, S., Forno, D. A., Cock, J. H., Gomez, K. A. (1976) Routine procedure for growing rice plant in culture solution. Laboratory Manual for Physiological Studies of Rice The International Rice Research Institute Los Baños, Philippines. pp 61–66Google Scholar
  19. 19.
    Alam, M. F., Datta, K., Vasquez, A. R., Oliva, N., Khush, G. S., and Datta, S K (1996) Transformation of new plant type lines through biolistic and protoplast methods Rice Genet Newsletter 13, 139–141Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1999

Authors and Affiliations

  • Karabi Datta
    • 1
  • Swapan K. Datta
    • 1
  1. 1.Plant Breeding, Genetics and Biochemistry DivisionIRRIManilaThe Philippines

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