Plant Cell Reports

, Volume 5, Issue 4, pp 284–287 | Cite as

Techniques for enhanced release of leaf protoplasts in Populus

  • Julie A. Russell
  • Brent H. McCown
Article

Abstract

Microscopic examination of Populus leaf tissue following enzyme treatment revealed two factors contributing to low protoplast yields: (1) poor penetration of the enzymes into the tissue, and (2) entrapment of protoplasts in leaf debris during protoplast purification procedures. A simple combination of rapid grinding of the tissue in an Omni-mixer prior to enzyme treatment and forceful washing of leaf-debris after digestion provided high exposure of the cells, uniform digestion, and high yields of protoplasts of two Populus clones. Protoplasts exhibited cell wall regeneration and long-term viability in culture. The relative yield advantages of the techniques varied with the inherent digestibility of each clone but could produce up to 600 percent greater protoplast yields in a woody plant species in which protoplast isolation was previously limited. The techniques were also applicable to an herbaceous species, Solanum etuberosum.

Abbreviations

BA

benzyladenine

NAA

naphthalene acetic acid

WPM

Woody Plant Medium, Lloyd and McCown (1980)

MS

Murashige and Skoog Medium (1962)

(NC-XXXX)

North Central Forest Experiment Station accession number assigned to Populus hybrid clones

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Julie A. Russell
    • 1
  • Brent H. McCown
    • 1
  1. 1.Department of HorticultureUniversity of WisconsinMadisonUSA

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