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In Vitro Cellular & Developmental Biology

, Volume 26, Issue 1, pp 85–90 | Cite as

Artificial seeds of alfalfa (Medicago sativa L.). Induction of desiccation tolerance in somatic embryos

  • Tissa Senaratna
  • Bryan D. McKersie
  • Stephen R. Bowley
Plant Cellular and Development Biology

Summary

The use of somatic embryos from cell culture systems in the clonal propagation of plants would be greatly facilitated if the somatic embryos could be dried and stored in a dormant state similar to true seeds. A cell culture system was developed for alfalfa (Medicago sativa L.) line RL34 which gave high yields of somatic embryos in an approximately synchronized pattern. These somatic embryos were treated with abscisic acid (ABA) at the cotyledonary stage of development to induce desiccation tolerance. With no visual preselection, approximately 60% of the dried embryos converted into plants upon reimbibition. When high quality embryos were selected prior to drying, 90 to 100% conversion rates were observed. The timing of the application of ABA in terms of embryo development was critical with an optimum being at cotyledonary stage spanning approximately 4 days; thus, synchronized embryo development is required for optimal expression in bulk samples. The vigor of the seedlings from dried somatic embryos was greater than those from embryos which had not been dried, but remained substantially lower than those from true seeds.

Key words

artificial seeds alfalfa somatic embryogenesis desiccation tolerance 

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References

  1. Ackerson, R. C. Synthesis and movement of abscisic acid in water-stressed cotton leaves. Plant Physiol. 69:609; 1982.PubMedGoogle Scholar
  2. Atanassov, A.; Brown, D. C. W. Plant regeneration from suspension cultures and mesophyll protoplasts ofMedicago sativa L. Plant Cell Tissue Org. Culture 3:149–162; 1984.CrossRefGoogle Scholar
  3. Bewley, J. D.; Black, M. Physiology and Biochemistry of Seeds, New York: Springer-Verlag; 1982.Google Scholar
  4. Bingham, E. T.; Hurley, L. V.; Kaatz, D. M., et al. Breeding alfalfa which regenerates from callus tissue culture. Crop. Sci. 15:719–721; 1975.CrossRefGoogle Scholar
  5. Chen, T. H. H.; Gusta, L. V. Abscisic acid-induced freezing resistance in cultured plant cells. Plant Physiol. 73:71–75; 1983.PubMedGoogle Scholar
  6. Crowe, J. H.; Crowe, L. M.; Chapman, D. Preservation of Membranes in Anhydrobiotic Organisms: The Role of Trehalose. Science 223:701–703; 1984.CrossRefPubMedGoogle Scholar
  7. Finkelstein, R. R.; Crouch, M. L. Hormonal and osmotic effects on developmental potential of maturing rapeseed. HortScience 22:780–797; 1987.Google Scholar
  8. Gamborg, O.; Miller, R.; Ojima, K. Nutrient requirements of suspension cultures of soybean roots cells. Exp. Cell Res. 50:151–158; 1968.PubMedCrossRefGoogle Scholar
  9. Gray, D. J. Quiescence in monocotyledonous and dicotyledonous somatic embryos induced by dehydration. HortScience 22:810–814; 1987.Google Scholar
  10. Gray, D. J.; Conger, B. V.; Songstad, D. D. Desiccated quiescent somatic embryos of orchardgrass for use as synthetic seeds. In Vitro Cell. Dev. Biol. 23:29–33; 1987.Google Scholar
  11. Kermode, A. R.; Bewley, J. D. The role of maturation drying in the transition from seed development to germination I. Acquisition of desiccation tolerance and germinability during development of Ricinus communis L. seeds. J. Exptl. Bot. 36:1906–1915; 1985.CrossRefGoogle Scholar
  12. Keith, C. N.; McKersie, B. D. Acclimation of callus cultures of Lotus corniculatus L. to freezing stress. Plant Physiol. 80:766–770; 1986.PubMedGoogle Scholar
  13. Kitto, C. L.; Janick, J. Hardening treatments increase survival of synthetically-coated asexual embryos of carrot. J. Amer. Soc. Hort. Sci. 110:283–286; 1985.Google Scholar
  14. Mohapatra, S. S.; Poole, R. S.; Dhindsa, R. S. Abscisic acidregulated gene expression in relation to freezing tolerance in alfalfa. Plant Physiol. 87:468; 1988.PubMedGoogle Scholar
  15. Nitzsche, W. One year storage of dried carrot callus. Z. Pflanzenphysiol. 100:269–271; 1980.Google Scholar
  16. Priestley, D. A. Seed Aging. Cornell University Press. 1986.Google Scholar
  17. Redenbaugh, K.; Slade, D.; Viss, P., et al. Encapsulation of somatic embryos in synthetic seed coats. HortScience 22:803–809; 1987.Google Scholar
  18. Rikin, A.; Atsman, D.; Gitler, C. Chilling injury in cotton (Grossypium hirsutum L.): prevention by abscisic acid. Plant Cell Physiol. 20:1537–1546; 1979.Google Scholar
  19. Senaratna, T.; McKersie, B. D.; Stinson, R. H. Simulation of dehydration injury to membranes from soybean axes by free radicals. Plant Physiol. 77:472–474; 1985a.PubMedGoogle Scholar
  20. Senaratna, T.; McKersie, B. D.; Stinson, R. H. Antioxidant levels in germinating soybean seed axes in relation to free radical and dehydration tolerance. Plant Physyiol. 78:168–171; 1985b.Google Scholar
  21. Zeevaart, J. A. D. Changes in the levels of abscisic acid and its metabolites in excised leaf blades of Xanthium strumarium during and after water stress. Plant Physiol. 66:672–678; 1980.PubMedCrossRefGoogle Scholar

Copyright information

© Tissue Culture Association 1990

Authors and Affiliations

  • Tissa Senaratna
    • 1
  • Bryan D. McKersie
    • 1
  • Stephen R. Bowley
    • 1
  1. 1.Crop Science DepartmentUniversity of GuelphGuelphCanada

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