Measuring Ion Channel Activity During Polar Growth of Pollen Tubes

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Abstract

Ion transporters like H+ATPases, K+channels, and Ca2+channels probably play an essential role during pollen tube tip growth by mediating ion fluxes that are involved in regulation of tube growth, e. g. Ca2+conducting channels localized in the tube tip plasma membrane allow a Ca2+influx that is responsible for the growth speed and the growth direction. However, no Ca2+channels have been identified in the plasma membrane of pollen grains and tubes, so far; secondly, use of conventional patch-clamp techniques requires enzymatic removal of the cell wall to gain access to the protoplast. This approach, in turn, causes loss of the cell polarity. We, therefore, used laser microsurgery to isolate protoplasts from the very tip of pollen tubes while the tube’s polarity is maintained. Using the patch- clamp technique it is then possible to investigate the ion channels in this tip protoplast.