Plant Cell, Tissue and Organ Culture

, Volume 32, Issue 2, pp 205–212 | Cite as

High frequency somatic embryogenesis and plant regeneration from papaya hypocotyl callus

  • Maureen M. M. Fitch
Research Article

Abstract

High frequency somatic embryogenesis in papaya (Carica papaya L.) tissue cultures was achieved by culturing hypocotyl sections from ten-day-old seedlings on half-strength Murashige and Skoog salts (MS) medium containing modified MS vitamins, 2.3 to 112.5 μM 2,4-dichlorophenoxyacetic acid (2,4-d), 400 mg l-1 glutamine, and 6% sucrose. Four hermaphroditic Hawaiian cultivars produced embryogenic calluses after ten to 14 weeks of culture at 27°C in the dark. Efficiency in embryogenic response of genotypes differed, ‘Kapoho’ > ‘Sunset’ > ‘Sunrise’ > ‘Waimanalo’. The frequency of embryogenesis in induction medium containing 4.5 μM 2,4-d was lowest with 3% sucrose and highest with 7% sucrose. Somatic embryos developed directly from embryogenic calluses on induction medium, or, more often, they differentiated from calluses subcultured on a medium devoid of growth regulators. Between 50 and 500 embryos were produced from each 2-mm hypocotyl section after at least two months on induction medium and two months on maturation medium. Embryos subsequently developed into normal-looking plants on MS medium. Shoot cuttings from germinated embryos and micropropagated plants were rooted with 5.0 μM indole-3-butyric acid (IBA), grown in the greenhouse, and transferred to the field.

Key words

Carica papaya L. 2,4-dichiorophenoxyacetic acid embryogenesis tissue culture 

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References

  1. Ammirato PV (1983) Embryogenesis. In: Evans DA, Sharp WR, Ammirato PV & Yamada Y (Eds) Handbook of Plant Cell Culture, Vol 1 (pp 82–123). MacMillan, New York.Google Scholar
  2. Arora IK & Singh RN (1978) In vitro plant regeneration in papaya. Curr. Sci. 47: 867–868.Google Scholar
  3. Chen FC & Kuo MH (1988) Ovule and ovary culture in papaya and somatic embryogenesis. In: Ma SS, Shii CT, Chang WC & Chang TL (Eds) Proceedings of a Symosium on Tissue Culture of Horticultural Crops. National Taiwan University, Taipei, March 8–9, 1988 (pp 50–61). Department of Horticulture, National Taiwan University, 1991 (in Chinese).Google Scholar
  4. Chen MH (1988a) Tissue culture and ringsport resistant breeding of papaya. In: Ma SS, Shii CT, Chang WC & Chang TL (Eds) Proceedings of a Symposium on Tissue Culture of Horticultural Crops. National Taiwan University, Taipei, March 8–9, 1988 (pp 62–72). Department of Horticulture, National Taiwan University. 1991 (in Chinese).Google Scholar
  5. Chen MH (1988b) Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants. In: Ma SS, Shii CT, Chang WC & Chang TL (Eds) Proceedings of a Symposium on Tissue Culture of Horticutural Crops. National Taiwan University, Taipei, March 8–9, 1988 (pp 230–233), Department of Horticulture, National Taiwan University, 1991.Google Scholar
  6. Chen MH, Wang PJ & Maeda E (1987) Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants. Plant Cell Rep. 6: 348–351.Google Scholar
  7. Chen MH, Chen CC, Wang DN & Chen FC (1991) Somatic embryogenesis and plant regeneration from immature embryos of Carica papaya × Carica cauliflora cultured in vitro. Can. J. Bot. 69: 1913–1918.Google Scholar
  8. Close KR & Ludeman LA (1987) The effect of auxin-like plant growth regulators and osmotic regulation on induction of somatic embryogenesis from elite maize inbreds. Plant Sci. 52: 81–89.Google Scholar
  9. Close TJ, Kortt AA & Chandler PC (1989) A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. Plant Molec. Biol. 13: 95–108.Google Scholar
  10. De Bruijne E, De Langhe E & van Rijck R (1974) Actions of hormones and embryoid formation in callus cultures of Carica papaya. In: International Symposium on Crop Protection, Fytopharmacie en Fytiatrie Rijkslandsbouwhoogeschool Medelelinge 39: 637–645.Google Scholar
  11. Finer JJ (1987) Direct somatic embryogenesis and plant regeneration from immature embryos of hybrid sunflower (Helianthus annuus L.) on a high sucrose-containing medium. Plant Cell Rep. 6: 372–374.Google Scholar
  12. Finer JJ & McMullen MD (1990) Transformation of cotton (Gossypium hirsutum L.) via particle bombardment. Plant Cell Rep. 8: 586–589.Google Scholar
  13. Fitch MMM (1991) Development of Genetic Transformation Systems for Papaya. Ph.D. dissertation, University of Hawaii, Honolulu, Hawaii, August 1991 (p. 277).Google Scholar
  14. Fitch MMM & Manshardt RM (1990) Somatic embryogenesis and plant regeneration from immature zygotic embryos of papaya (Carica papaya L.). Plant Cell Rep. 9: 320–324.Google Scholar
  15. Fitch MMM, Manshardt RM, Gonsalves D, Slightom JL & Sanford JC (1990) Stable transformation of papaya via microprojectile bombardment. Plant Cell Rep. 9: 189–194.Google Scholar
  16. Fromm ME, Morrish F, Armstrong C, Williams R. Thomas J & Klein TM (1990) Inheritance and expression of chimeric genes in the progeny of transgenic maize plants. Bio/Technology 8: 833–839.Google Scholar
  17. Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brian JV, Chambers SA, Adams WR, Willetts NG, Rice TR, Mackey CJ, Krueger RW, Kausch AP & Lemaux PG (1990) Transformation of maize cells and regeneration of fertile transgenic plants. The Plant cell 2: 603–618.Google Scholar
  18. Jordan M, Cortes I & Montenegro G (1982) Regeneration of plantlets by embryogenesis from callus cultures of Carica candamarcensis. Plant Sci. Lett. 28: 321–326.Google Scholar
  19. Kamada H & Harada H (1979a) Studies on the organogenesis in carrot tissue cultures I. Effects of growth regulators on somatic embryogenesis and root formation. Z. Pflanzenphysiol. 91: 255–266.Google Scholar
  20. Kamada H & Harada H (1979b) Studies on the organogenesis in carrot tissue cultures II. Effects of amino acids and inorganic nitrogenous compounds on somatic embryogenesis. Z. Pflanzenphysiol. 91: 453–463.Google Scholar
  21. Larkin PJ & Scowcroft WR (1981) Somaclonal variation-a novel source of variability from cell cultures for plant improvement. Theor. App. Genet. 60: 197–214.Google Scholar
  22. Lee TSG (1987) Micropropagation of sugarcane (Saccharum spp.). Plant Cell Tiss. Org. Cult. 10: 47–55.Google Scholar
  23. Linsmaier EM & Skoog F (1965) Organic growth factor requirements of tabacco tissue culture. Physiol. Plant. 18: 100–127.Google Scholar
  24. Litz RE (1986) Effect of osmotic stress on somatic embryogenesis in Carica suspension cultures. J. Amer. Soc. Hort. Sci. 111: 969–972.Google Scholar
  25. Litz RE & Conover RA (1980) Somatic embryogenesis in cell cultures of Carica stipulata. HortScience 15: 733–735.Google Scholar
  26. Litz RE & Conover RA (1981) In vitro polyembryony in Carica papaya L. ovules. Z. Pflanzenphysiol. 104: 285–288.Google Scholar
  27. Litz RE & Conover RA (1982) In vitro somatic embryogenesis and plant regeneration from Carica papaya L. ovular callus. Plant Sci. Lett. 26: 153–158.Google Scholar
  28. Litz RE & Conover RA (1983) High frequency somatic embryogenesis from Carica suspension cultures. Ann. Bot. 51: 683–686.Google Scholar
  29. Litz RE, O'Hair SK & Conover RA (1983) In vitro growth of Carica papaya L. cotyledons. Sci. Hort. 19: 287–293.Google Scholar
  30. Manshardt RM & Wenslaff TF (1989a) Zygotic polyembryony in interspecific hybrids of Carica papaya and C. cauliflora. J. Amer. Soc. Hort. Sci. 114: 684–689.Google Scholar
  31. Manshardt RM & Wenslaff TF (1989b) Interspecific hybridization of papaya with other Carica species. J. Amer. Soc. Hort. Sci. 114: 689–694.Google Scholar
  32. McGranahan GH, Leslie CA, Uratsu SL, Martin LA & Dandekar AM (1988) Agrobacterium-mediated transformation of walnut somatic embryos and regeneration of transgenic plants. Bio/Technology 6: 800–804.Google Scholar
  33. Mehdi AA & Hogan L (1979) In vitro growth and development of papaya (Carica papaya L.) and date-palm (Phoenix dactylifera L.). HortScience 14: 422.Google Scholar
  34. Miller RM & Drew RA (1990) Effect of explant type on proliferation of Carica papaya L. in vitro. Plant Cell Tiss. Org. Cult. 21: 39–44.Google Scholar
  35. Moore GA & Litz RE (1984) Biochemical markers of Carica papaya, C. cauliflora, and plants from somatic embryos of their hybrid. J. amer. Soc. Hort. Sci. 109: 213–218.Google Scholar
  36. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.Google Scholar
  37. Nagao M & Furutani S (1986) Improving germination of papaya seed by density separation, potassium nitrate, and gibberellic acid. HortSciene 21: 1439–1440.Google Scholar
  38. Orton TJ (1985) Genetic instability during embryogenic cloning of celery. Plant Cell Tiss. Org. Cult. 4: 159–169.Google Scholar
  39. Pang SZ & Sanford JC (1988) Agrobacterium-mediated gene transfer in papaya. J. Amer. Soc. Hort. Sci. 113: 287–291.Google Scholar
  40. Sanford JC (1990) Biolistic plant transformation. Physiol. Plant. 79: 206–209.Google Scholar
  41. Yamamoto H & Tabata M (1989) Correlation of papain-like enzyme production with laticifer formation in somatic embryos of papaya. Plant Cell Rep. 8: 251–254.Google Scholar
  42. Yamamoto H, Tanaka S, Fukui H & Tabata M (1986) Enzymatic difference between laticifers and cultured cells of papaya. Plant Cell Rep. 5: 269–272.Google Scholar
  43. Yie S & Liaw SI (1977) Plant regeneration from shoot tips and callus of papaya. In Vitro 13: 564–568.Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Maureen M. M. Fitch
    • 1
  1. 1.Department of HorticultureUniversity of HawaiiHonolulu
  2. 2.Experiment Station HSPAUSDA, ARSAiea

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