biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 44:25-31, 2001 | DOI: 10.1023/A:1017957902042

Effect of Heat Shock Treatment on Hordeum Vulgare Protoplast Transformation Mediated by Polyethylene Glycol

V.K. Tiwari1, J. Zhang1, T.J. Golds1, E.C. Cocking1, M.R. Davey1, J.B. Power1
1 Plant Genetic Manipulation Group, Department of Life Science, University of Nottingham, University Park, Nottingham, UK

The stable transformation of barley protoplasts has been observed in cv. Dissa by PEG-mediated pGL2 plasmid DNA uptake containing hygromycin phosphotransferase (hph) gene under the control of 35S transcript of CaMV. Hygromycin resistant colonies were obtained, when protoplasts were incubated with hygromycin B (25 μg cm-3) at 3 - 4 cell stage. Heat shock treatment, prior to polyethylene glycol (PEG) treatment, markedly increased the protoplast transformation frequency. The presence of active hph gene product, and integration of hph gene was confirmed by hygromycin assay and Southern blotting, respectively. Analysis indicated that rearrangement had possibly occurred during the integration of plasmid DNA into genomic DNA of barley callus. These transformed calli showed no morphogenic response onto the regeneration medium.

Keywords: barley; gene transfer; hygromycin phosphotransferase resistance
Subjects: antibiotics, in vitro culture; auxins; barley; Hordeum vulgare; hygromycin, heat shock, gene transfer; hygromycin phosphotransferase (hph) gene; polyethylene glycol, protoplast transformation; protoplast transformation mediated by polyethylene glycol

Published: March 1, 2001  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Tiwari, V.K., Zhang, J., Golds, T.J., Cocking, E.C., Davey, M.R., & Power, J.B. (2001). Effect of Heat Shock Treatment on Hordeum Vulgare Protoplast Transformation Mediated by Polyethylene Glycol. Biologia plantarum, 44(1), 25-31. doi: 10.1023/A:1017957902042
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Ahokas, H.: Transfection of germinating barley seed electrophoretically with exogenous DNA.-Theor. appl. Genet. 77: 469-472, 1989. Go to original source...
    2. Armstrong, C.L., Petersen, W.L., Buchholz, W.Z., Bowen, B.A., Sulc, S.L.: Factors affecting PEG-mediated stable transformation of maize protoplasts.-Plant Cell Rep. 9: 335-339, 1990. Go to original source...
    3. Cheng, M., Fry, J.E., Pang, S., Zhou, H., Hironaka, C.M., Duncan, D.R., Conner, T.W., Wan, Y.: Genetic transformation of wheat mediated by Agrobacterium tumefaciens.-Plant Physiol. 115: 971-980, 1997. Go to original source...
    4. Criessen, G., Caroline, S., Francis, R., Reynolds, H., Mullineaux, P.: Agrobacterium and microprojectile-mediated viral DNA delivery into barley microspore-derived cultures.-Plant Cell Rep. 8: 680-683, 1990. Go to original source...
    5. Datta, S.K., Datta, K., Soltanifar, N., Donn, G., Potrykus, I.: Herbicide-resistant indica-rice plants from IRRI breeding line IR72 after PEG-mediated transformation of protoplasts.-Plant mol. Biol. 20: 619-629, 1992. Go to original source...
    6. Datta, S.K., Peterhans, A., Datta, K., Potrykus, I.: Genetically engineered fertile indica-rice recovered from protoplasts.-Bio/Technology 8: 736-740, 1990. Go to original source...
    7. Frearson, E.M., Power, J.B., Cocking, E.C.: The isolation, culture and regeneration of Petunia leaf protoplasts.-Dev. Biol. 33: 130-137, 1973. Go to original source...
    8. Fromm, M.E., Taylor, L.P., Walbot, V.: Expression of genes transferred into monocot and dicot plant cells by electroporation.-Proc. nat. Acad. Sci. USA 82: 5824-5828, 1985. Go to original source...
    9. Funatsuki, H., Lörz, H., Lazzeri, P.A.: Use of feeder cells to improve barley protoplast culture and regeneration.-Plant Sci. 85: 179-187, 1992. Go to original source...
    10. Gilmour, D.M., Golds, T.J., Davey, M.R.: Medicago protoplasts: Fusion, culture and plant regeneration.-In: Bajaj, Y.P.S. (ed.): Biotechnology in Agriculture and Forestry. Plant Protoplasts and Genetic Engineering I. Vol. 8. Pp. 370-380. Springer-Verlag, Berlin-Heidelberg-New York 1989. Go to original source...
    11. Gold, T.J., Babczinsky, J., Mordhorst, A., Koop, H.-U.: Protoplast preparation without centrifugation: plant regeneration of barle6 (Hordeum vulgare L).-Plant Cell Rep. 13: 188-192, 1994. Go to original source...
    12. Hänisch ten Cate, C. H., Sree Ramulu, K., Dijkhuis, P., Groot, B.de.: Genetic stability of cultured hairy roots induced by Agrobacterium rhizogenes on tuber discs of potato cv. Bintje.-Plant Sci. 49: 217-222, 1987. Go to original source...
    13. Horn, M.E., Shillito, R.D, Conger, B.V., Harms, C.T.: Transgenic plants of orchardgrass (Dactylis glomerata L.) from protoplasts.-Plant Cell Rep. 7: 469-472, 1988. Go to original source...
    14. Jähne, A., Lazzeri, P.A., Jäger-Gussen, M., Lörz, H.: Plant regeneration from embryogenic cell suspension derived from anther cultures of barley (Hordeum vulgare L).-Theor. appl. Genet. 82: 74-80, 1991. Go to original source...
    15. Junker, B., Zimny, J., Lührs, R., Lörz, H.: Transient expression of chimeric genes in dividing and non-dividing cereal protoplasts after PEG induced DNA uptake.-Plant Cell Rep. 6: 329-332, 1987. Go to original source...
    16. Kao, K.N., Michayluk, M.R.: A method for frequency intergeneric fusion of plant protoplasts.-Planta 115: 355-367, (1974). Go to original source...
    17. Kao, K.N., Michayluk, M.R.: Nutritional requirements for growth of Vicia hajastane cells and protoplasts at very low population density in liquid media.-Planta 126: 105-110, 1975. Go to original source...
    18. Kartha, K.K., Chibbar, R.N., Georges, F., Leung, N., Caswell, K., Kendall, E., Quereshi, J.: Transient expression of chloramphenicol acetyl transferase (CAT) gene in barley cell cultures and immature embryos through microprojectile bombardment.-Plant Cell Rep. 8: 429-432, 1989. Go to original source...
    19. Krens, F.A., Molendijk, L., Wullems, G.J., Schilperoort, R.A.: In vitro transformation of plant protoplasts with Ti-plasmid DNA.-Nature 296: 72-74, 1982. Go to original source...
    20. Lazzeri, P.A., Brettschneider, R., Lührs, R., Lörz, H.: Stable transformation of barley via PEG-induced direct DNA uptake into protoplasts.-Theor. appl. Genet. 81: 437-444, 1991. Go to original source...
    21. Lazzeri, P.A., Lörz, H.: Regenerable suspension and protoplast cultures of barley and stable transformation via DNA uptake into protoplasts.-In: Lycett, G.W., Grierson, D. (ed.): Genetic Engineering of Crop Plants. Pp. 231-238. Butterworth, London 1990. Go to original source...
    22. Lee, B.T., Murdoch, K., Topping, J., Kreis, M., Jones, M.G.K.: Transient gene expression in aleurone protoplasts isolated from developing caryopses of barley and wheat.-Plant mol. Biol. 13: 21-29, 1989. Go to original source...
    23. Lyznik, L.A., Kamo, K.K., Grimes, H.D., Ryan, R., Chang, K.L., Hodges, T.K.: Stable transformation of maize: the impact of feeder cells on protoplast growth and transformation efficiency.-Plant Cell Rep. 8: 292-295, 1989a. Go to original source...
    24. Lyznik, L.A., Ryan, R.D., Ritchie, S.W., Hodges, T.K.: Stable co-transformation of maize protoplasts with gus and neo genes.-Plant mol. Biol. 13: 151-161, 1989b. Go to original source...
    25. Maniatis, T., Fritsch, E.F., Sambrook, J.: Molecular Cloning. A Laboratory Manual.-Cold Spring Harbor Laboratory Press, New York 1982.
    26. Matsuki, R., Onodera, H., Yamauchi, T., Uchimiya, H.: Tissue-specific expression of the rolC promoter of Ti plasmid in transgenic rice plants.-Mol. gen. Genet. 220: 12-16, 1989. Go to original source...
    27. Mendel, R.R., Müller, B., Schulze, J., Kolesnikov, V., Zelenin, A.: Delivery of foreign gene to intact barley cells by high-velocity micro-projectiles.-Theor. appl. Genet. 78: 31-34, 1989. Go to original source...
    28. Meyer, P., Walgenbach, E., Bussmann, K., Hombrecher, G., Saedler, H.: Synchronized tobacco protoplasts are efficiently transformed by DNA.-Mol. gen. Genet. 201: 513-518, 1985. Go to original source...
    29. Negrutiu, I., Shillito, R., Potrykus, I., Biasini, G., Sala, F.: Hybrid genes in the analysis of transformation conditions. I. Setting up a simple method for direct gene transfer in plant protoplasts.-Plant mol. Biol. 8: 863-873, 1987. Go to original source...
    30. Neuhaus, G., Spangenberg, G., Mittelsten-Scheid, O. Schweiger, H.-G.: Transgenic rapeseed plants obtained by the microspore-derived embryoids.-Theor. appl. Genet. 75: 30-36, 1987. Go to original source...
    31. Paszkowski, J., Pisan, B., Shillito, R.D., Potrykus, I.: Genetic transformation of Brassica campestris var. rapa protoplasts with an engineered cauliflower mosaic virus genome.-Plant mol. Biol. 6: 303-312, 1986. Go to original source...
    32. Paszkowski, J., Shillito, R.D., Saul, M., Mandak, V., Hohn, T., Holn, B., Potrykus, I. Direct gene transfer to plants.-EMBO J. 3: 2717-2722, 1984. Go to original source...
    33. Peng, J.Y., Lyznik, L.A., Lee, L., Hodges, T.K.: Co-transformation of indica-rice protoplasts with gus A and neo genes.-Plant Cell Rep. 9: 168-172, 1990. Go to original source...
    34. Potrykus, I.: Gene transfer to cereals: an assessment.-Bio/Technology 8: 535-542, 1990. Go to original source...
    35. Potrykus, I., Harms, C.P., Lörz, H.: Callus formation from cell culture protoplasts of corn (Zea mays L.).-Theor. appl. Genet. 54: 209-214, 1979. Go to original source...
    36. Rhodes, C.A., Pierce, D.A., Mettler, I.J., Mascerenhas, D., Detmer, J.J.: Genetically transformed maize plants from protoplasts.-Science 240: 204-207, 1988b. Go to original source...
    37. Ritala, A., Aspegren, K., Kurten, U., Salmenkallio-Marttila, M., Mannonen, L., Hannus, R., Kauppinen, V., Teeri, T.H., Enari, T.M.: Fertile transgenic barley by particle bombardment of immature embryos.-Plant mol. Biol. 24: 317-325, 1994. Go to original source...
    38. Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A., Allard, R.W.: Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics.-Proc. nat. Acad. Sci. USA 81: 8014-8018, 1984. Go to original source...
    39. Shillito, R.D., Saul, N.W., Paszkowski, J., Müller, M., Potrykus, I.: High efficiency direct gene transfer to plants.-Bio/Technology 3: 274-276, 1985. Go to original source...
    40. Teeri, T.H., Patel, G.H., Aspegren, A., Kauppinen, V.: Chloroplast targeting of neomycin phosphotransferase II with a pea transient peptide in electroporated barley mesophyll protoplasts.-Plant Cell Rep. 8: 187-190, 1989. Go to original source...
    41. Thompson, J.A., Abdullah, R., Chen, W.H., Gartland, K.M.A.: Enhanced protoplast division in rice (Oryza sativa L.) following heat shock treatment.-J. Plant Physiol. 127: 367-370, 1987. Go to original source...
    42. Tingay, S., McElory, D., Kalla, R., Fieg, S.: Agrobacterium tumefaciens-mediated barley transformation.-Plant J. 11: 1369-1376, 1997. Go to original source...
    43. Tiwari, V.K.: Tissue culture and transformation studies of barley.-PhD. Thesis. University of Nottingham, Nottingham 1992.
    44. Töpfer, R., Gronenbom, B., Schell, J., Steinbiss, H.H.: Uptake and transient expression of chimeric genes in seed-derived embryos.-Plant Cell 1: 133-139, 1989. Go to original source...
    45. Toriyama, K., Aimoto, Y., Uchimiya, H., Hinata, K.: Transgenic rice plants after direct gene transfer into protoplasts.-Bio/Technology 6: 1072-1074, 1988. Go to original source...
    46. Uchimiya, H., Fushimi, T., Hashimota, H., Harada, H., Syono, K., Sugawara, Y.: Expression of a gene in callus derived from DNA-treated protoplasts in rice (Oryza sativa L.).-Mol. gen. Genet. 204: 204-207, 1986. Go to original source...
    47. Wan, Y.C., Lemaux, P.G.: Generation of large numbers of independently transformed fertile barley plants.-Plant Physiol. 104: 37-48, 1994. Go to original source...
    48. Wullems, G.L., Molendijk, L., Schilperoort, R.A.: The expression of tumour markers in interspecific somatic hybrids of normal and crown gall cells from Nicotina tabacum.-Theor. appl. Genet. 56: 203-208, 1980. Go to original source...
    49. Yan, Q.S., Zhang, Q.H., Shi, J.B., Li, J.M.: Green plant regeneration from protoplasts of barley (Hordeum vulgare L.).-Chin. Soc. Bull. 36: 932-935, 1990.
    50. Zhang, J., Tiwari, V.K., Golds, T.J., Blackhall, N.W., Cocking, E.C., Mulligan, B.J., Power, J.B., Davey, M.R.: Parameters influencing transient and stable transformation of barley (Hordeum vulgare L.) protoplasts.-Plant Cell Tissue Organ Cult. 41: 125-138, 1995. Go to original source...
    51. Zhang, W., Wu, R.: Efficient regeneration of transgenic plants from rice protoplasts and correctly regulated expression of foreign genes in plants.-Theor. appl. Genet. 76: 835-840, 1988. Go to original source...

    Return to the content