Plant Cell, Tissue and Organ Culture

, Volume 42, Issue 3, pp 299–302 | Cite as

An improved protocol for the culture of cassava leaf protoplasts

  • P. Anthony
  • M. R. Davey
  • J. B. Power
  • K. C. Lowe
Research Notes

Abstract

Viable protoplasts (yield > 1.9 × 107 g−1 fresh weight; mean viability 85±2%, n=5) were isolated from leaves of axenic shoot cultures of Manihot esculenta Crantz. cv. M. Thai 8. Protoplasts were cultured for up to 50 days in liquid, ammonium-free MS medium, overlaying agarose-solidified B5 medium with short glass rods embedded perpendicularly within, and protruding from, the agarose layer. Control protoplasts were cultured identically, but without glass rods. Sustained protoplast division was observed only in the presence of glass rods, where the initial plating efficiency was almost 6-fold greater than control (p < 0.05). The mean final plating efficiency of treated cultures was 1.0±0.2% while, in contrast, significant colony formation was not observed in controls.

Key words

Manihot esculenta two-phase culture system glass rods plating efficiency 

Abbreviations

BA

6-benzyladenine

CPPU

N-(2-chloro-4-pyridyl)-N'-phenylurea

MES

2[N-morpholino]ethane sulphonic acid

MS

Murashige & Skoog (1962)

NAA

α-naphthaleneacetic acid

IPE

initial plating efficiency

FPE

final plating efficiency

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References

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • P. Anthony
    • 1
  • M. R. Davey
    • 1
  • J. B. Power
    • 1
  • K. C. Lowe
    • 1
  1. 1.Department of Life ScienceUniversity of NottinghamNottinghamUK

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