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.
Similar content being viewed by others
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
References
Ahuja MR (1983) Silvae Genetica 32:225–227
Ahuja MR (1984) Silvae Genetica 33:32–37
Butt AD (1985) Plant Sci 42:55–59
Evans DA, Bravo JE (1983) In: Evans DA, Sharp WR, Ammirato PV, Yamada Y (eds) Handbook of Plant Cell Culture, vol I, MacMillan Publ. Co., New York, pp. 124–176
Haberlach GT, Cohen BA, Reichert NA, Baer MA, Towill LE, Helgeson JP (1985) Plant Sci 39:67–74
Lloyd GB, McCown BH (1980) Proc Intl Plant Prop Soc 30:421–427
Murashige T, Skoog F (1962) Physiol Plant 15:473–497
Patniak G, Wilson D, Cocking EC (1981) Z Pflanzenphysiol 102:199–205
Russell JA, McCown BH (1986) submitted
Schenk RU, Hildebrandt AC (1969) Crop Sci 9:629–631
Smith MAL (1984) PhD Thesis, University of Wisconsin, Madison, Wisconsin p105
Smith MAL, McCown BH (1982/83) Plant Sci Lett 28:149–156
Verma DC, Wann SH (1983) In: Potrykus I, Harms CT, Hinnen A, Hutter R, King PJ, Shillito RD (eds) 6th Intl Protoplast Symp, Basel, Experentia Suppl 45:10
Widholm JM (1972) Stain Technology 47:189–194
Wright DC (1985) Plant Cell Tissue Organ Culture 4:95–100
Author information
Authors and Affiliations
Additional information
Communicated by G. C. Phillips
Rights and permissions
About this article
Cite this article
Russell, J.A., McCown, B.H. Techniques for enhanced release of leaf protoplasts in Populus . Plant Cell Reports 5, 284–287 (1986). https://doi.org/10.1007/BF00269823
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00269823