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A laser microsurgical method of cell wall removal allows detection of large-conductance ion channels in the guard cell plasma membraneApplication of patch clamp techniques to higher-plant cells has been subject to the limitation that the requisite contact of the patch electrode with the cell membrane necessitates prior enzymatic removal of the plant cell wall. Because the wall is an integral component of plant cells, and because cell-wall-degrading enzymes can disrupt membrane properties, such enzymatic treatments may alter ion channel behavior. We compared ion channel activity in enzymatically isolated protoplasts of Vicia faba guard cells with that found in membranes exposed by a laser microsurgical technique in which only a tiny portion of the cell wall is removed while the rest of the cell remains intact within its tissue environment. "Laser-assisted" patch clamping reveals a new category of high-conductance (130 to 361 pS) ion channels not previously reported in patch clamp studies on plant plasma membranes. These data indicate that ion channels are present in plant membranes that are not detected by conventional patch clamp techniques involving the production of individual plant protoplasts isolated from their tissue environment by enzymatic digestion of the cell wall. Given the large conductances of the channels revealed by laser-assisted patch clamping, we hypothesize that these channels play a significant role in the regulation of ion content and electrical signalling in guard cells.
Document ID
20040088814
Document Type
Reprint (Version printed in journal)
Authors
Miedema, H. (University Park United States)
Henriksen, G. H.
Assmann, S. M.
Evans, M. L.
Date Acquired
August 21, 2013
Publication Date
January 1, 1999
Publication Information
Publication: Protoplasma
Volume: 209
Issue: 2-Jan
ISSN: 0033-183X
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: MCB-9416039
CONTRACT_GRANT: BIR9-303734
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Plant Biology