Biologia Plantarum

, 36:183 | Cite as

Increase of callus and embryoid production from hypocotyl protoplasts of sunflower (Helianthus annuus L.) by culture in microdrops

  • J. Šamaj
  • A. Okolot
  • M. Bobak
  • Yu. Gleba
Original Papers


Hypocotyls of seedling plants of the sunflower inbred line HNK-81 were used as a source of protoplasts. Optimal conditions of isolation and culture of protoplasts were established. Using the method of individual culture in microdrops, a 77% final plating efficiency was achieved. Cytokinins, especially zeatin, showed a significant influence on the formation of globular embryo-like structures, but these failed to develop into mature embryos. Calli obtained in liquid media containing auxins and cytokinins grew on agar solidified media without growth regulators.


Cellulase Zeatin Cell Coloni Mature Embryo Protoplast Culture 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.





2,4-dichlorophenoxyacetic acid




Murashige-Skoog medium


1-naphthaleneacetic acid




  1. Bohorova, N.E., Cocking, E.C., Power, J.B.: Isolation, culture and callus regeneration of protoplasts of wild and cultivated Helianthus species.—Plant Cell Rep.5: 256–258, 1986.CrossRefGoogle Scholar
  2. Burrus, M., Chanabe, Ch., Alibert, G., Bidney, D.: regeneration of fertile plants from protoplasts of sunflower (Helianthus annuus L.).—Plant Cell Rep.10: 161–166, 1991.CrossRefGoogle Scholar
  3. Dupuis, J.M., Gomiero, M., Plantevin, C., Chagvardieff, P.: Différenciacion de formes embryoides issues de protoplastes de tournesol (Helianthus annuus L.).—Compt. rend. Acad. Sci. Paris307: 465–468, 1988.Google Scholar
  4. Fischer, Ch., Hahne, G.: Sunflower cotyledon protoplasts. Their development in liquid and solidified medium.—Physiol. Plant.82(Suppl.): A112, 1991.Google Scholar
  5. Fischer, Ch., Klethi, P., Hahne, G.: Protoplasts from cytoledon and hypocotyl of sunflower (Helianthus annuus L.)—Shoot regeneration and seed production.—Plant Cell Rep.11: 632–636, 1992.CrossRefGoogle Scholar
  6. Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirement of suspension cultures of soybean root cells.—Exp. Cell Res.50: 151–158, 1968.PubMedCrossRefGoogle Scholar
  7. Gleba, Yu.Yu.: Microdroplet culture: tobacco plants from single mesophyll protoplasts.— Naturwissenschaften65: 158, 1978.CrossRefGoogle Scholar
  8. Guilley, E., Hahne, G.: Callus formation from isolated sunflower (Helianthus annuus) mesophyll protoplasts.—Plant Cell Rep.8: 226–229, 1989.CrossRefGoogle Scholar
  9. Chanabe, Ch., Burrus, M., Alibert, G.: Factors affecting the improvement of colony formation from sunflower protoplasts.—Plant Sci.64: 125–132, 1989.CrossRefGoogle Scholar
  10. Kao, K.N., Michayluk, M.R.: Nutritional requirements for growth ofVicta hajastana cells and protoplasts at a very low population density in liquid media.—Planta126: 105–110, 1975.CrossRefGoogle Scholar
  11. Krasnyanski, S., Menczel, L.: Shoot regeneration from protoplast derived callus of sunflower (Helianthus annuus L.).—Physiol. Plant.82 (Suppl.): A14, 1991.Google Scholar
  12. Lenee, P., Chupeau, Y.: Isolation and culture of sunflower protoplasts (Helianthus annuus L.): Factors influencing the viability of cell colonies derived from protoplasts.—Plant Sci.43: 69–75, 1986.CrossRefGoogle Scholar
  13. Lenee, P., Chupeau, Y.: Development of nitrogen assimilating enzymes during growth of cells derived from protoplasts of sunflower and tobacco.—Plant Sci.59: 109–117, 1989.CrossRefGoogle Scholar
  14. Medgyesy, P., Menczel, L., Maliga, P.: The use of cytoplasmic streptomycin resistance: chloroplast transfer fromNicotiana tabacum intoNicotiana sylvestris, isolation of their somatic hybrids.— Mol. gen. Genet.179: 693–698, 1980.CrossRefGoogle Scholar
  15. Moyne, A.L., Thor, V., Pélissier, B., Bergounioux, C., Freyssinet, G., Gadal, P.: Callus and embryoid formation from protoplasts ofHelianthus annuus.—Plant Cell Rep.7: 437–440, 1988.Google Scholar
  16. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures.—Physiol. Plant.15: 473–497, 1962.CrossRefGoogle Scholar
  17. Nagata, T., Takebe, I.: Planting of isolated tobacco mesophyll protoplasts on agar medium.—Planta99: 12–20, 1971.CrossRefGoogle Scholar
  18. Paterson, K.E., Everett, N.P.: Regeneration ofHelianthus annuus inbred plants from callus.—Plant Sci.42: 125–132, 1985.CrossRefGoogle Scholar
  19. Shillito, R.D., Paszkowski, J., Potrykus, I.: Agarose planting and bead type culture technique enable and stimulate development of protoplast-derived colonies in number of plant species.—Plant Cell Rep.2: 224–247, 1983.CrossRefGoogle Scholar
  20. Schmitz, P., Shnabl, B.: Regeneration and evacuolation of protoplasts from mesophyll, hypocotyl and petioles fromHelianthus annuus L..—J. Plant Physiol.135: 223–227, 1989.Google Scholar
  21. Šamaj, J., Auxtová, O., Bobák, M.: Different regeneration potential of various sunflower (Helianthus annuus L.) genotypes in meristem culture.—Biol. Plant.36: 309–311, 1994.CrossRefGoogle Scholar

Copyright information

© Institute of Experimental Botany 1994

Authors and Affiliations

  • J. Šamaj
    • 1
  • A. Okolot
    • 2
  • M. Bobak
    • 1
  • Yu. Gleba
    • 2
  1. 1.Department of Plant PhysiologyComenius UniversityBratislavaSlovak Republic
  2. 2.Institute of Cell Biology and EngineeringKiev GSP 22Ukraine

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