Skip to main content
Log in

Nutrient utilization during bioreactor culture, and maturation of somatic embryo cultures ofPicea mariana andPicea glauca-engelmannii

  • Micropropagation
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Summary

Embryogenic cell cultures ofPicea mariana (black spruce) and the species complexPicea glauca-engelmannii (interior spruce) were maintained either as suspensions in liquid medium in 250 or 500-ml-capacity shake-flasks, 7-liter-capacity airlift or mechanically stirred bioreactors, or on agar-solidified medium. Cultures from each of the maintenance conditions were subsequently transferred to agar-solidified LP medium containing 40µM (±) -abscisic acid for maturation into cotyledonary stage embryos. For both species, the highest maturation frequency resulted from cultures grown in the airlift bioreactor. With black spruce cells grown in the airlift bioreactor containing LP medium with 60 mM sucrose, a maximum of 7.1 g·liter−1 dry weight and 2892 embryos·ml−1 were obtained after 15 days. For interior spruce cells, a maximum dry weight of 5.9 g·liter−1 and 2698 embryos·ml−1 were obtained after 21 to 30 days. During culture over 2 wk, ammonia was almost completely utilized by both species, wherease nitrate was depleted to 40% of the initial concentration. Sucrose was rapidly hydrolyzed to glucose and fructose by both species. Black spruce cultures preferentially metabolized glucose, whereas interior spruce preferentially metabolized fructose. Improved growth of interior spruce cells in mechanically stirred bioreactors occurred when cultured in LP medium with 60 mM fructose as the sole carbon source.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ammirato, P. V. Embryogenesis. In: Evans, D. A.; Sharp, W. R.; Ammirato, P. V., et al. eds. Handbook of plant cell culture, vol. 1. New York: Macmillan Publishing Co.; 1983a:82–123.

    Google Scholar 

  • Ammirato, P. V. The regulation of somatic embryo development in plant cell cultures: suspension culture techniques and hormone requirements. Bio/Technology 3:68–74; 1983b.

    Article  Google Scholar 

  • Attree, S. M.; Dunstan, D. I.; Fowke, L. C. White spruce [Picea glauca (Moench) Voss] and black spruce [Picea mariana (Mill) B.S.P.]. In: Bajaj, Y. P. S., ed. Biotechnology in agriculture and forestry, vol 16. Berlin: Springer-Verlag; 1991:423–445.

    Google Scholar 

  • Bapat, V. A.; Fulzele, D. P.; Heble, M. R., et al. Production of sandalwood somatic embryos in bioreactors. Curr. Sci. 59:746–748; 1990.

    Google Scholar 

  • Carrier, D.-J.; Consentino, G.; Neufeld, R., et al. Nutritional and hormonal requirements ofGinkgo biloba embryo-derived callus and suspension cell culture. Plant Cell Rep. 8:635–638; 1990.

    Article  CAS  Google Scholar 

  • Cazzulino, D.; Pedersen, H.; Chin, C.-K. Bioreactors and image analysis for scale-up and plant propagation. In: Vasil, I. K., ed. Cell culture and somatic cell genetics of plants, vol. 8. San Diego, CA: Academic Press; 1991:147–177.

    Google Scholar 

  • Chen, T. H. H.; Thompson, B. G.; Gerson, D. F.In vitro production of alfalfa somatic embryos in fermentation systems. J. Ferment. Technol. 65:353–357; 1987.

    Article  Google Scholar 

  • De-Eknamkul, W.; Ellis, B. E. Effects of macronutrients on growth and rosmarinic acid formation in cell suspension cultures ofAnchusa officinalis. Plant Cell Rep. 4:46–49; 1985.

    Article  CAS  Google Scholar 

  • Dougall, D. K.; Frazier, G. C. Nutrient utilization during biomass and anthocyanin accumulation in suspension cultures of wild carrot cells. Plant Cell Tissue Organ Cult. 18:95–104; 1989.

    Article  CAS  Google Scholar 

  • Dunstan, D. I.; Bekkaoui, F.; Pilon, M., et al. Effects of abscisic acid and analogues on the maturation of white spruce (Picea glauca) somatic embryos. Plant Sci. 58:77–84; 1988.

    Article  CAS  Google Scholar 

  • Durzan, D. J. Ammonia: its analogues, metabolic products and site of action in somatic embryogenesis. In: Bonga, J. M.; Durzan, D. J., eds. Cell and tissue culture in forestry, vol. 2. Dordrecht, The Netherlands: Martinus Nijhoff Publishers; 1987:92–136.

    Google Scholar 

  • Fowler, M. W. Substrate utilization by plant-cell cultures. J. Chem. Technol. Biotechnol. 32:338–346; 1982.

    Article  CAS  Google Scholar 

  • Gérard, J.; Antoine, P.; Lepoivre, P., et al. Uptake kinetics of medium constituents during batch growth of potato cell cultures. Biotechnol. Lett. 13:381–384; 1991.

    Article  Google Scholar 

  • Gupta, P. K.; Timmis, R.; Pullman, G., et al. Development of an embryogenic system for automated propagation of forest trees. In: Vasil, I. K., ed. Scale-up and automation in plant propagation, vol. 8. San Diego, CA: Academic Press; 1991:75–93.

    Google Scholar 

  • Hakman, I.; van Arnold, S. Somatic embryogenesis and plant regeneration from suspension cultures ofPicea glauca (white spruce). Physiol. Plant. 72:579–587; 1988.

    CAS  Google Scholar 

  • Hong, Y. C.; Labuza, T. P.; Harlander, S. K. Growth kinetics of strawberry cell suspension cultures in shake flask, airlift, stirred-jar, and roller bottle bioreactors. Biotech. Prog. 5:137–143; 1989.

    Article  Google Scholar 

  • Kim, D.-I.; Pedersen, H.; Chin, C.-K. Cultivation ofThalictrum rugosum cell suspension in an improved airlift bioreactor: stimulatory effect of carbon dioxide and ethylene on alkaloid production. Biotechnol. Bioeng. 38:331–339; 1991.

    Article  CAS  PubMed  Google Scholar 

  • Kumar, P. P.; Reid, D. M.; Thorpe, T. A. The role of ethylene and carbon dioxide in differentiation of shoot buds in excised cotyledons ofPinus radiata in vitro. Physiol. Plant. 69:244–252; 1987.

    Article  CAS  Google Scholar 

  • Kumar, P. P.; Joy, R. W. IV; Thorpe, T. A. Ethylene and carbon dioxide accumulation, and growth of cell suspension cultures ofPicea glauca (white spruce). J. Plant Physiol. 135:592–596; 1989.

    Google Scholar 

  • Levi, A.; Sink, K. C. Differential effects of sucrose, glucose and fructose during somatic embryogenesis in asparagus. J. Plant Physiol. 137:184–189; 1990.

    CAS  Google Scholar 

  • Lulsdorf, M. M.; Tautorus, T. E.; Kikcio, S. I., et al. Growth parameters of embryogenic suspension cultures of interior spruce (Picea glauca engelmannii complex) and black spruce (Picea mariana Mill.). Plant Sci. 82:227–234; 1992.

    Article  CAS  Google Scholar 

  • McDonald, K. A.; Jackman, A. P. Bioreactor studies of growth and nutrient utilization in alfalfa suspension cultures. Plant Cell Rep. 8:455–458; 1989.

    Article  CAS  Google Scholar 

  • Nigra, H. M.; Alvarez, M. A.; Giulietti, A. M. Effect of carbon and nitrogen sources on growth and solasodine production in batch suspension cultures ofSolanum eleagnifolium Cav. Plant Cell Tissue Organ Cult. 21:55–60; 1990.

    Article  CAS  Google Scholar 

  • Owens, J. N.; Molder, M. The reproductive cycle of interior spruce. Victoria, B.C.: Ministry of Forests; 1984.

    Google Scholar 

  • Preil, W. Application of bioreactors in plant propagation. In: Debergh, P. C.; Zimmerman, R. H., eds. Micropropagation, technology and application. Dordrecht, the Netherlands: Kluwer Academic Publishers; 1991:425–445.

    Google Scholar 

  • Preil, W.; Florek, P.; Wix, U., et al. Towards mass propagation by use of bioreactors. Acta Hortic. 226:99–106; 1988.

    Google Scholar 

  • Roberts, D. R.; Sutton, B. C. S.; Flinn, B. S. Synchronous and high frequency germination of interior spruce somatic embryos following partial drying at high relative humidity. Can. J. Bot. 68:1086–1090; 1990.

    Google Scholar 

  • Scragg, A. H. Large-scale cultivation ofHelianthus annuus cell suspensions. Enzyme Microb. Technol. 12:82–85; 1990.

    Article  Google Scholar 

  • Scragg, A. H.; Ashton, S.; York, A. et al. Growth ofCatharanthus roseus suspensions for maximum biomass and alkaloid accumulation. Enzyme Microb. Technol. 12:292–298; 1990.

    Article  CAS  Google Scholar 

  • Stuart, D. A.; Strickland, S. G.; Walker, K. A. Bioreactor production of alfalfa somatic embryos. HortScience 22:800–803; 1987.

    Google Scholar 

  • Tal, B.; Gressel, J.; Goldberg, I. The effect of medium constituents on growth and diosgen in production byDioscorea deltoidea cells grown in batch culture. Planta Med. 44:111–115; 1982.

    Article  PubMed  CAS  Google Scholar 

  • Taticek, R. A.; Moo-Young, M.; Legge, R. L. Effect of bioreactor configuration on substrate uptake by cell suspension cultures of the plantEschscholtzia californica. Appl. Microb. Biotechnol. 33:280–286; 1990.

    Article  CAS  Google Scholar 

  • Tautorus, T. E.; Attree, S. M.; Fowke, L. C., et al. Somatic embryogenesis from immature and mature zygotic embryos, and embryo regeneration from protoplasts in black spruce (Picea mariana Mill.). Plant Sci. 67:115–124; 1990.

    Article  Google Scholar 

  • Tautorus, T. E.; Fowke, L. C.; Dunstan, D. I. Somatic embryogenesis in conifers. Can. J. Bot. 69:1873–1899; 1991.

    Google Scholar 

  • Tautorus, T. E.; Lulsdorf, M. M.; Kikcio, S. I., et al. Bioreactor culture ofPicea mariana Mill. (black spruce) and the species complexPicea glauca-engelmannii (interior spruce) somatic embryos. Growth parameters. Appl. Microb. Biotechnol. 38:46–51; 1992.

    CAS  Google Scholar 

  • Treat, W. J.; Engler, C. R.; Soltes, E. J. Culture of photomixotrophic soybean and pine in a modified fermentor using a novel impeller. Biotechnol. Bioeng. 34:1191–1202; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Tremblay, L.; Tremblay, F. M. Carbohydrate requirements for the development of black spruce (Picea mariana (Mill.) B.S.P.) and red spruce (P. rubens Sarg.) somatic embryos. Plant Cell Tissue Organ Cult. 27:95–103; 1991.

    Article  CAS  Google Scholar 

  • von Arnold, S.; Eriksson, T.In vitro studies of adventitious shoot formation inPinus contorta. Can. J. Bot. 59:870–874; 1981.

    Google Scholar 

  • Webster, F. B.; Roberts, D. R.; McInnis, S. M., et al. Propagation of interior spruce by somatic embryogenesis. Can. J. For. Res. 20:1759–1765; 1990.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

NRCC no. 36479

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tautorus, T.E., Lulsdorf, M.M., Kikcio, S.I. et al. Nutrient utilization during bioreactor culture, and maturation of somatic embryo cultures ofPicea mariana andPicea glauca-engelmannii . In Vitro Cell Dev Biol - Plant 30, 58–63 (1994). https://doi.org/10.1007/BF02632121

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02632121

Key words

Navigation