A simple method for mitochondrial respiration and calcium uptake assessment in pollen tubes

Key mitochondrial processes are known to be widely conserved throughout the eukaryotic domain. However, the scarce availability of working materials may restrict the assessment of such mitochondrial activities in several working models. Pollen tube mitochondrial studies represent one example of this, where tests have been often restricted due the physical impossibility of performing experiments with isolated mitochondria in enough quantities. Here we detail a method to measure in situ mitochondrial respiratory chain activity and calcium transport in tobacco pollen tubes. • Digitonin-mediated plasmalemma permeabilization allows efficient assessment of mitochondrial respiration and calcium uptake. • This method allows quick, reliable and portable measurements from low to high cellular densities, versus methods requiring intracellular calcium reporters.