A novel approach for real-time monitoring of leaf wounding responses demonstrates unprecedently fast and high emissions of volatiles from cut leaves

Wounding is a key plant stress that results in a rapid, within seconds to a few minutes, release of ubiquitous stress volatiles and stored volatiles in species with storage structures. Understanding the timing and extent of wound-dependent volatile elicitation is needed to gain an insight into different emission controls, but real-time monitoring of plant emissions through wounding treatments has been hampered by the need to stop the measurements to perform the wounding, slow stabilization of gas flows upon chamber closure and smearing out the signal by large chambers and long sampling lines. We developed a novel leaf cutter that allows to rapidly perform highly precise leaf cuts within the leaf chamber. The cutter was fitted to the standard Walz GFS-3000 portable gas-exchange system leaf chamber and chamber exhaust air for analysis with a proton transfer reaction time-of-flight mass-spectrometer (PTR-TOF-MS) was taken right at the leaf chamber outlet. Wounding experiments in four species of contrasting leaf structure demonstrated significant species differences in timing, extent and blend of emitted volatiles, and showed unprecedently high emission rates of several stress volatiles and stored monoterpenes. In light of the rapid rise of release of de novo synthesized and stored volatiles, the results of this study suggest that past studies have underestimated the rate of elicitation and maximum emission rates of wound-dependent volatiles.