title: Techniques for enhanced release of leaf protoplasts in Populus.
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.
authors.affiliation: Department of Horticulture, University of Wisconsin, 1575 Linden Drive, 53706, Madison, WI, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Russell
authors.forename: B H
authors.initials: BH
authors.lastname: McCown