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Techniques for enhanced release of leaf protoplasts in Populus

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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.

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naphthalene acetic acid


Woody Plant Medium, Lloyd and McCown (1980)


Murashige and Skoog Medium (1962)


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


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Communicated by G. C. Phillips

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Russell, J.A., McCown, B.H. Techniques for enhanced release of leaf protoplasts in Populus . Plant Cell Reports 5, 284–287 (1986).

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