Plant Cell Reports

, Volume 18, Issue 1–2, pp 159–165 | Cite as

Plant regeneration from protoplasts isolated from friable embryogenic callus of cassava

  • E. Sofiari
  • C. J. J. M. Raemakers
  • J. E. M. Bergervoet
  • E. Jacobsen
  • R. G. F. Visser

Abstract

Protoplasts were isolated from friable embryogenic callus (FEC) and from suspensions derived from FEC of cassava genotype TMS60444. Suspensions yielded the highest number of protoplasts (1.5×106 protoplasts/g fresh weight). Protoplasts plated at a density of 105–106/ml in a medium supplemented with 0.5 mg/l α-naphthaleneacetic acid and 1 mg/l zeatin began dividing after 3 days, and after 30 days this resulted in an absolute plating efficiency as high as 2.5%. After 2 months of culture, 60% of the developed calli were highly friable and in appearance identical to the original FEC. The protoplast derived FEC was first purified through two rounds of selection of 3 weeks each before beeing cultured for regeneration of plants. This was done by culturing the protoplast-derived FEC for 11 weeks on maturation medium, yielding a maximum of 184 organized embryos per 10.000 initially cultured protoplasts. Most of the organized embryos were torpedo shaped and matured after they had been isolated from the calli and transferred to fresh medium. Mature embryos were multiplied by secondary somatic embryogenesis at high efficiency (>90%) on a medium supplemented with 8 mg/l 2,4-dichlorophenoxyacetic acid. About 30% of the mature secondary somatic embryos developed into shoots after transfer to a medium supplemented with 1 mg/l N6-benzylaminopurine (BAP). Shoots rooted readily on a medium without BAP.

Key words Cassava Protoplast culture Friable embryogenic callus 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • E. Sofiari
    • 1
  • C. J. J. M. Raemakers
    • 1
  • J. E. M. Bergervoet
    • 1
  • E. Jacobsen
    • 1
  • R. G. F. Visser
    • 1
  1. 1.Graduate School of Experimental Plant Sciences, Department of Plant Breeding, Wageningen Agricultural University, P. O. Box 386, 6700 AJ Wageningen, The NetherlandsNL

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