Biologia plantarum 46:205-210, 2003 | DOI: 10.1023/A:1022894409408

Cryopreservation of Embryogenic Culture of Pinus roxburghii

G. Mathur1, V.A. Alkutkar1, R.S. Nadgauda1
1 National Chemical Laboratory, Tissue Culture Pilot Plant, Pune-, India

Embryogenic cultures of chir pine (Pinus roxburghii Sarg.) were cryopreserved successfully in liquid nitrogen. It was found that using sorbitol and dimethyl sulfoxide (DMSO) as cryoprotectants was essential for the survival of the tissue. Among the different concentrations of the cryoprotectants used, the most effective treatment was observed to be 0.3 M sorbitol and 5 % DMSO. On staining the cryopreserved tissue with fluorescein diacetate, it was observed that only a few meristematic embryo heads survived and resumed growth after a very short initial lag phase. The recovered cultures showed normal regrowth on proliferation medium and, it was also observed that washing off the cryoprotectants was necessary for the cultures to survive. The results indicate that cryopreservation can be used for conserving the germplasm, and in maintaining the embryogenic capacity of the tissue.

Keywords: chir pine; cryo-storage; somatic embryogenesis
Subjects: chir pine, in vitro culture, cryopreservation; cryopreservation; cytokinins; 2,4-dichlorophenoxyacetic acid; dimethyl sulfoxide, crypreservation; in vitro culture, nutrient media modification; in vitro culture, somatic embryogenesis; Pinus roxburghii

Prepublished online: March 1, 2003; Published: September 1, 2003Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Mathur, G., Alkutkar, V.A., & Nadgauda, R.S. (2003). Cryopreservation of Embryogenic Culture of Pinus roxburghii. Biologia plantarum, 46(2), 205-210. doi: 10.1023/A:1022894409408.
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. Aronen, T.S., Krajnakova, J., Häggman, H.M., Ryynänen, L.A.: Genetic fidelity of cryopreserved embryogenic cultures of open-pollinated Abies cephalonica.-Plant Sci. 142: 163-172, 1999. Go to original source...
    2. Arya, S., Kalia, R.K., Arya, I.D.: Induction of somatic embryogenesis in Pinus roxburghii Sarg.-Plant Cell Rep. 19: 775-780, 2000. Go to original source...
    3. Attree, S.M., Fowke, L.C.: Embryogeny of gymnosperms: advances in synthetic seed technology of conifers.-Plant Cell Tissue Organ Cult. 35: 1-35, 1993. Go to original source...
    4. Cyr, D.R., Lazaroff, W.R., Grimes, S.M.A., Quan, G., Bethune, T.D., Dunstan, D.I., Roberts, D.R.: Cryopreservation of interior spruce (Picea glauca engelmanni complex) embryogenic cultures.-Plant Cell Rep. 13: 574-577, 1994. Go to original source...
    5. DeVerno, L.L., Park, Y.S., Bonga, J.M., Barrett, J.D.: Somaclonal variation in cryopreserved embryogenic clones of white spruce [Picea glauca (Moench) Voss.].-Plant Cell Rep. 18: 948-953, 1999. Go to original source...
    6. Eastman, P.A.K., Webster, F.B., Pitel, J.A., Roberts, D.R.: Evaluation of somaclonal variation during somatic embryogenesis of interior spruce (Picea engelmannii complex) using culture morphology and isozyme analysis.-Plant Cell Rep. 10: 425-430, 1991. Go to original source...
    7. Find, J.I., Floto, F., Krogstrup, P., Moller, J.D., Norgaard, J.V., Kristensen, M.M.H.: Cryopreservation of an embryogenic suspension culture of Picea sitchensis and subsequent plant regeneration.-Scand. J. Forest Res. 8: 156-162, 1993.
    8. Ford, C.S., Jones, N.B., Van Staden, J.: Cryopreservation and plant regeneration from somatic embryos of Pinus patula.-Plant Cell Rep. 19: 610-615, 2000. Go to original source...
    9. Fourré, J.L., Berger, P., Niquet, L., André, P.: Somatic embryogenesis and somaclonal variation in Norway spruce: morphogenesis, cytogenetic and molecular approaches.-Theor. appl. Genet. 94: 159-169, 1997. Go to original source...
    10. Gupta, P. K., Durzan, D.J.: Shoot multiplication from mature trees of Douglas fir (Pseudotsuga menziesii) and sugar pine (Pinus lambertiana).-Plant Cell Rep. 4: 177-179, 1985. Go to original source...
    11. Gupta, P.K., Durzan, D.J., Finkle, B.J.: Somatic embryogenesis in embryogenic cell masses of Picea abies (Norway spruce) and Pinus taeda (loblolly pine) after thawing from liquid nitrogen.-Can. J. Forest Res. 17: 1130-1134, 1987.
    12. Gupta, P.K., Grob, J.A.: Somatic embryogenesis is conifers.-In: Jain, S.M., Gupta, P.K., Newton, R.J. (ed.): Somatic Embryogenesis in Woody Plants. Vol. 1. Pp. 81-98. Kluwer Academic Publishers, Dordrecht 1995.
    13. Häggman, H.M., Ryynänen, L.A., Aronen, T.S., Krajnakova, J.: Cryopreservation of embryogenic cultures of Scots pine.-Plant Cell Tissue Organ Cult. 54: 45-53, 1998. Go to original source...
    14. Isabel, N., Tremblay, L., Michaud, M., Tremblay, F.M., Bosquet, J.: RAPDs as an aid to evaluate the genetic integrity of somatic embryogenesis-derived populations of Picea mariana.-Theor. appl. Genet. 86: 81-87, 1993. Go to original source...
    15. Kartha, K.K., Fowke, L.C., Leung, N.L., Caswell, K.L., Hakman, I.: Induction of somatic embryos and plantlets from cryopreserved cell cultures of white spruce (Picea glauca).-Plant Physiol. 132: 529-539, 1988. Go to original source...
    16. Klimaszewska, K., Ward, C., Cheliak, W.M.: Cryopreservation and plant regeneration from embryogenic cultures of larch (Larix × eurolepis) and black spruce (Picea mariana).-J. exp. Bot. 43: 73-79, 1992. Go to original source...
    17. Laine, E., Bade, P., David, A.: Recovery of plants from cryopreserved embryogenic cell suspensions of Pinus caribaea.-Plant Cell Rep. 11: 295-298, 1992. Go to original source...
    18. Mathur, G., Von Arnold, S., Nadgauda, R.S.: Studies on somatic embryogenesis from immature zygotic embryos of chir pine (Pinus roxburghii Sarg.).-Curr. Sci. 79: 999-1004, 2000.
    19. Nørgaard, J.V., Baldursson, S., Krogstrup, P.: Genotypic differences in the ability of embryogenic Abies normanniana cultures to survive cryopreservation.-Silvae Genet. 42: 93-97, 1993a.
    20. Nørgaard, J.V., Duran, V., Johnsen, Ø., Krogstrup, P., Baldursson, S., Von Arnold, S.: Variations in cryotolerance of embryogenic Picea abies cell lines and the association to genetic, morphological, and physiological factors.-Can. J. Forest Res. 23: 2560-2567, 1993b.
    21. Tewari, D.N.: A Monograph on Chir Pine (Pinus roxburghii Sarg.).-Vedam Books International, New Delhi 1994.
    22. Timmis, R.: Bioprocessing for the tree production in the forest industry: conifer somatic embryogenesis.-Biotechnol. Progr. 14: 156-166, 1998. Go to original source...
    23. Widholm, J.M.: The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells.-Stain Technol. 47: 189-194, 1972. Go to original source...

    Return to the content