Adventitious shoot formation and plant regeneration from Pinus pinaster Sol. ex Aiton

  • Filomena Calixto
  • M. Salomé Pais
Miropropagation

Summary

Pinus pinaster plants were regenerated from cotyledons excised from in vitro germinated seeds and axenically cultured on induction medium (GMD). 6-Benzyladenine (2.2 µM) induced the highest frequency of direct bud formation from cotyledons. An average of 13.1 ± 2.1 elongated shoots per cotyledon was obtained. Germination time influenced shoot induction, and the organogenic potential decreased with explant age. Cotyledons remained for 21 d on induction medium, and in order to promote adventitious shoot elongation, they were transferred to Gupta and Durzan’s DCR medium without growth regulators, containing 0.5% (wt/vol) activated charcoal and 3% (wt/vol) sucrose. Rooting was achieved by application of an indole-3-butyric acid, (396.6 µM) pulse (24 h at 4° C), followed by transfer to a sterile mixture of peat plus perlite (1:1 vol/vol). Ninety-eight to 100% of the regenerated plants were successfully acclimatized. All plants have survived after transfer to the field.

Key words

cotyledon micropropagation organogenesis Pinus pinaster rhizogenesis 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Abdullah, A.; Grace, J. Regeneration of calabrian pine juvenile needles. Plant Sci. 53:147–155; 1987.CrossRefGoogle Scholar
  2. 2.
    Aitken, J.; Horgan, K. J.; Thorpe, T. A. Influence of explant selection on the shoot forming capacity of juvenile tissue of Pinus radiata. Can. J. For. Res. 11:112–117; 1981.Google Scholar
  3. 3.
    Aitken-Christie, J.; Sing, A. P.; Horgan, K. J., et al. Explant developmental state and shoot formation in Pinus radiata cotyledons. Bot. Gaz. 146:196–203; 1985.CrossRefGoogle Scholar
  4. 4.
    Aitken-Christie, J.; Thorpe, T. A. Clonal propagation; gymnosperms. In: Vasil, I. K., ed. Cell culture and somatic cell genetics of plants. Vol. I. San Diego: Academic Press, Inc.; 1984:82–95.Google Scholar
  5. 5.
    Bornman, C. H. Possibilities and constraints in the regeneration of trees from cotyledonary needles of Picea abies in vitro. Physiol. Plant. 57:5–16; 1983.CrossRefGoogle Scholar
  6. 6.
    Bornman, C. H. Picea abies. In: Bonga, J. M.; Durzan, D. J., ed. Cell tissue culture in forestry. Vol. 3. Dordrecht: Martinus Nijhoff Publishers; 1987:2–29.Google Scholar
  7. 7.
    David, A. Manifestations de diverses potentialités organogènes et micropropagation végétative chez le pin maritime (Pinus pinaster Sol.) Ann. Resc. Sylvicoles AFOCEL: 57–75; 1979.Google Scholar
  8. 8.
    David, A.; David, H. Manifestations de diverses potentialités organogènes d’organes ou de fragments d’organes de pin maritime (Pinus pinaster Sol.) en culture in vitro. C. R. Acad. Sci. Paris 284:627–630; 1977.Google Scholar
  9. 9.
    David, A.; David, H.; Mateille, T. In vitro adventitious budding on Pinus pinaster cotyledons and needles. Physiol. Plant. 56:102–107; 1982.CrossRefGoogle Scholar
  10. 10.
    David, H.; Isemukali, K.; David, A. Obtention de plants de pin maritime (Pinus pinaster Sol.) à partir des brachiblastes ou d’apex caulinaires de trés jeunes sujets cultivés in vitro. C. R. Acad. Sci. Paris 287:245–248; 1978.Google Scholar
  11. 11.
    Desmetrau, D. X.; Alazard, P.; Chaperon, H. Monographie genetiques de Pinus pinaster. Ann. Forestales 9/4:125–150; 1982.Google Scholar
  12. 12.
    Dumas, E. Micropropagation d’une clone agé de pin maritime en vue de l’obtention de pieds-mères. Ann. Resc. Sylvicoles AFOCEL 95–107; 1987.Google Scholar
  13. 13.
    Flinn, B. S.; Webb, D. T.; Georgis, W. In vitro control of caulogenesis by growth regulators and media components in embryonic explants of eastern white pine (Pinus strobus). Can. J. Bot. 64:1948–1956; 1985.CrossRefGoogle Scholar
  14. 14.
    Franclet, A.; David, A.; David, H., et al. Première mise en évidence morphologique d’un rajeunissement de méristèmes primaires caulinaires de pin maritime (Pinus pinaster Sol.). C. R. Acad. Sci. Paris. 290:927–930; 1980.Google Scholar
  15. 15.
    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.CrossRefGoogle Scholar
  16. 16.
    Jelaska, S. European pines. In: Bonga, J. M.; Durzan, D. J., ed. Cell tissue culture in forestry. Vol. 3. Dordrecht: Martinus Nijhoff Publishers; 1987:2–29.Google Scholar
  17. 17.
    Mohammed, G. H.; Dunstan, D. I.; Thorpe, T. A. Influence of nutrient medium upon shoot initiation on vegetative explants excised from 15-to 18-year-old Picea glauca. N. Z. For. Sci. 16:297–305; 1986.Google Scholar
  18. 18.
    Noh, E. W.; Minocha, S. C.; Riemenschneider, D. E. Adventitious shoot formation from embryonic explants of red pine (Pinus resinosa). Physiol. Plant. 74:119–124; 1988.CrossRefGoogle Scholar
  19. 19.
    Patel, K. R.; Thorpe, T. A. Histochemical examination of shoot initiation in cultured cotyledon explants of radiata pine. Bot. Gaz. 145:312–322; 1984.CrossRefGoogle Scholar
  20. 20.
    Quorin, M.; Lepoivre, P. Étude de milieux adaptés aux cultures in vitro de Prunus. Acta Hortic. 78:437–442; 1977.Google Scholar
  21. 21.
    Rancillac, M. Mise au point d’une méthode de multiplication végétative “in vitro” du pin maritime (Pinus pinaster Sol.) pour la constitution de clones à partir de semences. In: AFOCEL (Editors). Micropropagation d’arbres forestière. AFOCEL et Rech. 12:41–48; 1979.Google Scholar
  22. 22.
    Rancillac, M. Perspectives d’application des culture d’organs in vitro à la multiplication végétative du pin maritime, Pinus pinaster Sol. Ann. Sci. Forestières 38:55–59; 1981.CrossRefGoogle Scholar
  23. 23.
    Rancillac, M. Maritime pine (Pinus pinaster Sol.). In: Bajaj, Y. P. S., ed. Biotechnology in agriculture and forestry. Vol. 16, Trees III. Berlin: Springer-Verlag; 1991:317–338.Google Scholar
  24. 24.
    Rancillac, M.; Faye, M.; David, A. In vitro rooting of cloned shoots in Pinus pinaster. Physiol. Plant. 56:97–101; 1982.CrossRefGoogle Scholar
  25. 25.
    Schenk, R. U.; Hildebrant, A. C. Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell culture. Can. J. Bot. 50:199–204; 1972.Google Scholar
  26. 26.
    Smith, D. R. Radiata pine (Pinus radiata D. Don) In: Bajaj, Y. P. S., ed. Biotechnology in agriculture and forestry. Vol. 1. Berlin: Springer-Verlag; 1986:274–291.Google Scholar
  27. 27.
    Sommer, H. E.; Brown, C. L.; Kormanik, P. P. Differentiation of plantlets in longleaf pine (Pinus palustris Mill.) tissue culture in vitro. Bot. Gaz. 136:196–200; 1975.CrossRefGoogle Scholar
  28. 28.
    Thomas, M. J.; Travan, H. Influence relative de la BAP et l’IBA sur la néoformation de bourgeons et de racine sur les plantules du Biota orientalis (Cupressacées). Physiol. Plant. 56:118–122; 1982.CrossRefGoogle Scholar
  29. 29.
    Thorpe, T. A. Organogenesis in vitro: structural, physiological and biochemical aspects. In: Vasil, I. K., ed. Perspectives in plant cell and tissue culture. Int. Rev. Cytol. Suppl. Vol. 11A. New York: Academic Press; 1980:71–111.Google Scholar
  30. 30.
    Travan, H.; Troton, D.; Calvayrac, R. Morphological, histological and lipid changes during adventitious budding in Pinus pinaster cultured cotyledons. J. Exp. Bot. 204:907–915; 1988.CrossRefGoogle Scholar
  31. 31.
    Von Arnold, S.; Alsterborg, E.; Walles, B. Micromorphological studies of adventitious bud formation of Picea abies embryos treated with cytokinin. Physiol. Plant. 72:248–256; 1988.CrossRefGoogle Scholar
  32. 32.
    Von Arnold, S.; Eriksson, T. C. Induction of adventitious buds of Norway spruce grown in vitro. Physiol. Plant. 44:283–287; 1978.CrossRefGoogle Scholar
  33. 33.
    Von Arnold, S.; Eriksson, T. C. In vitro studies of adventitious shoot formation in Pinus contorta. Can. J. Bot. 59:870–874; 1981.Google Scholar
  34. 34.
    Von Arnold, S.; Eriksson, T. Initial stages in the course of adventitious bud formation on embryos of Picea abies. Physiol. Plant. 64:41–47; 1985.CrossRefGoogle Scholar
  35. 35.
    Yeung, E. C.; Aitken, J.; Biondi, S., et al. Shoot histogenesis in cotyledon explant of radiata pine. Bot. Gaz. 142:494–501; 1981.CrossRefGoogle Scholar

Copyright information

© Society for In Vitro Biology 1997

Authors and Affiliations

  • Filomena Calixto
    • 1
  • M. Salomé Pais
    • 1
  1. 1.Centro de Biotecnologia VegetalFaculdade de Ciências de LisboaLisboaPortugal

Personalised recommendations