Plant Cell, Tissue and Organ Culture

, Volume 61, Issue 1, pp 51–58 | Cite as

Somatic embryogenesis and plant regeneration from litchi protoplasts isolated from embryogenic suspensions

  • Changhe YuEmail author
  • Zhenguang Chen
  • Liuxin Lu
  • Jinwen Lin


High yields of protoplasts were isolated from litchi embryogenic suspensions, which were maintained by alternative culture in liquid and on solid media containing silver thiosulfate. Protoplasts in liquid culture and agarose beads were unable to divide sustainedly, whereas embedding of protoplasts in Ca-alginate supported cell division to microcalli and the direct formation of somatic embryos from protoplasts. Nurse cells of litchi further enhanced the culture efficiency when protoplasts were cultured in Ca-alginate beads. White non-hyperhydric somatic embryos were developed from protoplast-derived microcalli or proembryos, and 33.1% of white somatic embryos regenerated into plantlets.

calcium alginate nurse cells silver thiosulfate 


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  1. Damm B & Willmitzer L (1988) Regeneration of fertile plants from protoplasts of different Arabidopsis thaliana genotypes. Mol. Gen. Genet. 213: 15–20CrossRefGoogle Scholar
  2. Frearson EM, Power JB & Cocking EC (1973) The isolation, culture, and regeneration of Petunia leaf protoplasts. Dev. Biol. 33: 130–137PubMedCrossRefGoogle Scholar
  3. Fu L & Tang D (1983) Introduction pollen plants of litchi tree (Litchi chinensis Sonn.). Acta Genetica Sinica 10: 369–374Google Scholar
  4. Gamborg OL, Miller RA & Ojima K (1968) Plant cell cultures. I. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158PubMedCrossRefGoogle Scholar
  5. Huancaruna-Perales E & Schieder O (1993) Plant regeneration form leaf protoplasts of apple. Plant Cell. Tiss. Org. Cult. 34: 71–76.CrossRefGoogle Scholar
  6. Kantharajah, AS, McConchie CA & Dodd WA (1992) In vitro embryo culture and induction of multiple shoots in lychee (Litchi chinensis Sonn.). Ann. Bot. 70: 153–156Google Scholar
  7. Kao KN & Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126: 105–110CrossRefGoogle Scholar
  8. Li ZY, Tanner GJ & Larkin PJ (1990) Callus regeneration from Trifolium subterraneum protoplasts and enhanced protoplast di58 vision by low-voltage treatment and nurse cells. Plant Cell Tiss. Org. Cult. 21: 67–73CrossRefGoogle Scholar
  9. Li ZY, Xia GM & Chen HM (1992) Somatic embryogenesis and plant regeneration from protoplasts isolated from embryogenic cell suspensions of wheat (Triticum aestivum L.). Plant Cell Tiss. Org. Cult. 28: 79–85CrossRefGoogle Scholar
  10. Menzel CM (1985) Propagation of lychee: A review. Sci. Hortic. 25: 31–48CrossRefGoogle Scholar
  11. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioasays with tobacco tissue cultures. Physiol. Plant. 15: 473–497CrossRefGoogle Scholar
  12. Niedz RP (1993) Culturing embryogenic protoplasts of 'Hamlin' sweet orange in calcium alginate beads. Plant Cell Tiss. Org. Cult. 34: 19–25CrossRefGoogle Scholar
  13. Nyman M & Wallin A (1992) Improved culture technique for strawberry (Fragaria x anannassa Duch.) protoplasts and the determination of DNA content in protoplast derived plants. Plant Cell Tiss. Org. Cult. 30: 127–133CrossRefGoogle Scholar
  14. Schnabl H, Youngman RJ & Zimmerman U (1983) Maintenance of plant cell membrane integrity and function by the immobilization of protoplasts in alginate matrices. Planta 158: 392–397CrossRefGoogle Scholar
  15. Teasdale RD & Richards DK (1991) Study of a factor produced by suspension cultured Pinus radiata cells which enhances cell growth at low inoculum densities. Plant Cell Tiss. Org. Cult. 26: 53–59CrossRefGoogle Scholar
  16. Walters TW & Earle ED (1990) A simple, versatile feeder layer system for Brassica oleracea protoplast culture. Plant Cell Rep. 9: 316–319CrossRefGoogle Scholar
  17. Witjaksono, Litz RE & Grosser JW (1998) Isolation, culture and regeneration of avocado (Persea americana Mill.) protoplasts. Plant Cell Rep. 18: 235–242CrossRefGoogle Scholar
  18. Yu CH (1997) Studies on protoplast culture of litchi (Litchi chinensis). PhD dissertation, Fujian Agricultural University, Fuzhou, ChinaGoogle Scholar
  19. Yu CH & Chen ZG (1997) Induction of litchi embryogenic calli by immature embryos and anthers culture in vitro. J. Fujian Agri. Univ. 26: 168–172 (in Chinese with English abstract)Google Scholar
  20. Yu CH & Chen ZG (1998) Embryogenic suspension culture and protoplast isolation in litchee. Chinese J Trop Crops 19: 16–20 (in Chinese with English abstract)Google Scholar
  21. Zhou LN, Kuang ZS, Ma XJ, Chen JQ & Cao J (1996) The study of factors affecting somatic embryogenesis in young embryo culture of Litchi chinensis. J Agric Biotech. 4: 161–165 (in Chinese with English abstract)Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Changhe Yu
    • 1
    Email author
  • Zhenguang Chen
    • 1
  • Liuxin Lu
    • 1
  • Jinwen Lin
    • 1
  1. 1.Department of HorticultureFujian Agricultural UniversityFuzhouChina

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