Advertisement

Molecular Breeding

, Volume 10, Issue 3, pp 143–152 | Cite as

Preservation of transgenic silver birch (Betula pendula Roth) lines by means of cryopreservation

  • Leena Ryynänen
  • Maarit Sillanpää
  • Sari Kontunen-Soppela
  • Heidi Tiimonen
  • Jaakko Kangasjärvi
  • Elina Vapaavuori
  • Hely Häggman
Article

Abstract

The aim of the study was to develop a preservation method for transgenic silver birch (Betula pendula Roth) lines based on cryopreservation. Specific attention was paid to transgene stability and functioning. Vegetative buds collected from one- or two-year-old silver birches representing four transgenic lines and two wild-type lines were used as explants. Generally, the average regeneration of either transgenic or wild-type, cryoperserved and non-cryopreserved control buds was excellent, and varied from 72 to 100 percent. The regeneration percentage of cryopreserved buds was, however, significantly lower than that of non-cryopreserved control buds when estimated two weeks after thawing, but the differences were no longer significant four weeks after thawing. Growth of the plants in the greenhouse was more dependent on the clone than on the cryopreservation treatment. The studied transgenic lines have three (line E/5) to nine (line R/3.2) copies of transferred neomycin phosphotransferase genes that were also found to be stable after cryopreservation. In general, the neomycin phosphotransferase transcript levels did not change due to cryopreservation. The results indicate that it is possible to apply the cryopreservation technique to preserve valuable transgenic lines of a forest tree, silver birch. The method presented here leads to high regeneration percentages combined with transgene stability and functioning.

Betula pendula Roth Cryopreservation Neomycin phosphotransferase Transgenic silver birch 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aronen T. and Häggman H. 1995. Differences in Agrobacterium infections in silver birch and Scots pine. Eur. J. For Path. 25: 197–213.Google Scholar
  2. Aronen T., Krajnakova J., Häggman H. and Ryynänen L. 1999. Genetic fidelity of cryopreserved embryogenic cultures of open pollinated Abies cephalonica. Plant Sci. 142: 163–172.Google Scholar
  3. Benson E.E. and Hamill J.D. 1991. Cryopreservation of post freeze molecular and biosynthetic stability in transformed roots of Beta vulgaris and Nicotiana rustica. Plant Cell Tiss. Org. Cult. 24: 1163–1172.Google Scholar
  4. Blakesley D., Pask N., Henshaw G.G. and Fay M.F. 1996. Biotechnology and the conservation of forest genetic resources: in vitro strategies and cryopreservation. Plant Growth Regul. 20: 11–16.Google Scholar
  5. Chang S., Puryear J. and Cairney C. 1993. A simple and efficient method for isolating RNA from pine trees. Plant Mol. Biol. Rep. 11: 113–116.Google Scholar
  6. Elleuch H., Gazeau C., David H. and David A. 1998. Cryopreservation does not effect the expression of a foreign sam gene in transgenic Papaver somniferum cells. Plant Cell Rep. 18: 94–98.Google Scholar
  7. Forsline P.L., Towill L.E., Waddell J.W., Stushnoff C., Lamboy W.F. and McFerson J.R. 1998. Recovery and longevity of cryopreserved dormant apple buds. J. Amer. Soc. Hort. Sci. 123: 365–370.Google Scholar
  8. Fretz A. and Lörz H. 1995. Cryopreservation of in vitro cultures of barley (Hordeum vulgare L. and H. murinum L.) and transgenic cells of wheat (Triticum aestivum L.). J. Plant Physiol. 146: 489–496.Google Scholar
  9. Gazeau C.M.B., Elleuch H., David A. and Morisset C. 1998. Cryopreservation of transformed Papaver somniferum cells. Cryo-Letters 19: 147–159.Google Scholar
  10. Hobbs S.L.A.K., Kpodar P. and DeLong C.M.O. 1990. The effect of T-DNA copy number, position and methylation on reporter gene expression in tobacco transformants. Plant Mol. Biol. 15: 851–864.Google Scholar
  11. Huhtinen O. and Yahyaoglu Z. 1974. Das frühe Blühen von aus Kalluskulturen herangezogenen Pflänzchen bei der Birke (Betula pendula Roth). Silvae Genet. 23: 32–34.Google Scholar
  12. Häggman H., Ryynänen L. and Aronen T. 2001. Cryopreservation of forest tree germplasm. Acta Horticult. 560: 121–124.Google Scholar
  13. Jones J.D.G., Dunsmuir P. and Bedbrook J. 1985. High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J. 4: 2411–2418.Google Scholar
  14. Jones J.D.G., Gilbert D.E., Grady K.L. and Jorgensen R.A. 1987. T-DNA structure and gene expression in petunia plants transformed by Acrobacterium tumefaciens C58 derivates. Mol. Gen. Genet. 207: 478–485.Google Scholar
  15. Junttila O. 1980. Effect of photoperiod and temperature on apical growth cessation in two ecotypes of Salix and Betula. Physiol. Plant 48: 347–352.Google Scholar
  16. Keinonen-Mettälä K., Pappinen A. and von Weissenberg K. 1998. Comparisons of the efficiency of some promoters in silver birch (Betula pendula). Plant Cell Rep. 17: 356–361.Google Scholar
  17. Koski V. 1985. The timing of hardening and dehardening of forest trees. Acta Horticult. 168: 117–124.Google Scholar
  18. Koski V. and Tallquist P. 1978. Results of long-time measurements of the quantity of flowering and seed crop of forest trees. Folia For. 364: 1–60.Google Scholar
  19. Kurtén U., Nuutila A.-M., Kauppinen V. and Rousi M. 1990. Somatic embryogenesis in cell cultures of birch (Betula pendula Roth). Plant Cell Tiss. Org. Cult. 23: 101–105.Google Scholar
  20. Lemmetyinen J., Keinonen-Mettälä K., Lännenpää M., von Weissenberg K. and Sopanen T. 1998. Activity of the CaMV 35S promoter in various parts of transgenic early flowering birch clones. Plant Cell Rep. 18: 243–248.Google Scholar
  21. Lepistö M. 1973. Accelerated birch breeding in plastic greenhouses. For. Chron. 49: 1–2.Google Scholar
  22. Lloyd G. and McCown B. 1980. Commercially-feasible micropropagation of Mountain Laurel, Kalmia latifolia, by use of shoottip culture. Proc. Int. Plant Prop. Soc. 30: 421–427.Google Scholar
  23. Lodhi M.A., Ye G.-N., Weeden N.F. and Reisch B.I. 1994. A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol. Biol. Rep. 12: 6–13.Google Scholar
  24. Matzke M.A., Primig M., Trnovsky J. and Matzke A.J.M. 1989. Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants. EMBO J. 8: 643–649.Google Scholar
  25. Murashige T. and Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant 15: 473–497.Google Scholar
  26. Oka S., Yakuwa H., Sato K. and Niino T. 1991. Survival and shoot formation in vitro of pear winter buds cryopreserved in liquid nitrogen. HortScience 26: 65–66.Google Scholar
  27. Rinne P., Tuominen H. and Junttila O. 1994. Seasonal changes in bud dormancy in relation to bud morphology, water and starch content, and abscisic acid concentration in adult trees of Betula pubescens. Physiol Plant 90: 1–13.Google Scholar
  28. Romberger J.A. 1963. Meristems, Growth, and Development in Woody Plants. An Analytical Review of Anatomical, Physiological, and Morphogenic Aspects. Technical Bulletin No. 1293. U.S. Department of Agriculture, Forest Service.Google Scholar
  29. Ryynänen L. 1996. Survival and regeneration of dormant silver birch buds stored at super-low temperatures. Can. J. For. Res. 26: 617–623.Google Scholar
  30. Ryynänen L. 1996. Cold hardening and slow cooling: tools for succesful cryopreservation and recovery of in vitro shoot tips of silver birch. Can. J. For. Res. 26: 2015–2022.Google Scholar
  31. Ryynänen L. 1998. Effect of abscisic acid, cold hardening, and photoperiod on recovery of cryopreserved in vitro shoot tips of silver birch. Cryobiology 36: 32–39.Google Scholar
  32. Ryynänen L. 1999. Effect of early spring birch bud type on postthaw regrowth after prolonged cryostorage. Can. J. For. Res. 29: 47–52.Google Scholar
  33. Ryynänen L. and Ryynänen M. 1986. Propagation of adult curlybirch succeeds with tissue culture. Silva Fenn. 20: 139–147.Google Scholar
  34. Ryynänen L. and Häggman H. 1999. Substitution of ammonium ions during cold hardening and post-thaw cultivation enhances recovery of cryopreserved shoot tips of Betula pendula Roth. J. Plant Physiol. 154: 735–742.Google Scholar
  35. Sakai A. and Nishiyama Y. 1978. Cryopreservation of winter vegetative buds of hardy fruit trees in liquid nitrogen. HortScience 13: 225–227.Google Scholar
  36. Sambrook J., Maniatis T. and Fritsch E.F. 1989. Molecular Cloning. 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.Google Scholar
  37. Stushnoff C. and Junttila O. 1986. Seasonal development of cold stress resistance in several plant species at a coastal and a continental location in North Norway. Polar Biol. 5: 129–133.Google Scholar
  38. Susi A. 1997. The effect of transferred chitinase gene on the pathogen resistance of silver birch (Betula pendula). Lic. thesis, University of Joensuu, Joensuu, pp. 60.Google Scholar
  39. Tu H.M., Godfrey L.W. and Sun S.S.M. 1998. Expression of the Brasil nut methionine-rich protein and mutants with increased methionine in transgenic tobacco. Plant Mol. Biol. 37: 829–838.Google Scholar
  40. Tyler N.J. and Stushnoff C. 1988. The effects of prefreezing and controlled dehydration on cryopreservation of dormant vegetative apple buds. Can. J. Plant Sci. 68: 1163–1167.Google Scholar
  41. Töpfer R., Schell J. and Steinbiss H.-H. 1988. Versatile cloning vectors for transient gene expression and direct gene transfer in plant cells. Nucleic Acids Res. 16: 8725.Google Scholar
  42. Valjakka M., Aronen T., Kangasjärvi J., Vapaavuori E. and Häggman H. 2000. Genetic transformation of silver birch (Betula pendula) by particle bombardment. Tree Physiol. 20: 607–613.Google Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Leena Ryynänen
    • 1
  • Maarit Sillanpää
    • 1
  • Sari Kontunen-Soppela
    • 1
  • Heidi Tiimonen
    • 1
  • Jaakko Kangasjärvi
    • 2
  • Elina Vapaavuori
    • 3
  • Hely Häggman
    • 1
  1. 1.Punkaharju Research StationFinnish Forest Research InstitutePunkaharjuFinland
  2. 2.Institute of Biotechnology, The Viikki BiocenterUniversity of HelsinkiHelsinkiFinland
  3. 3.Suonenjoki Research StationFinnish Forest Research InstituteSuonenjokiFinland

Personalised recommendations