Chloride, glutathiones, and insect-derived elicitors introduced into the xylem trigger electrical signaling

Ricca assays allow the direct introduction of compounds extracted from plants or the organisms that attack them into the leaf vasculature. Using chromatographic fractionation of Arabidopsis (Arabidopsis thaliana) leaf extracts, we found glutamate was the most active low mass elicitor of membrane depolarization. However, other known elicitors of membrane depolarization are generated in the wound response. These include unstable aglycones generated by glucosinolate (GSL) breakdown. None of the aglycone-derived GSL-breakdown products, including nitriles and isothiocyanates, that we tested using Ricca assays triggered electrical activity. Instead, we found that glutathione and the GSL-derived compound sulforaphane glutathione triggered membrane depolarizations. These findings identify a potential link between GSL breakdown and glutathione in the generation of membrane depolarizing signals. Noting that the chromatographic fractionation of plant extracts can dilute or exchange ions, we found that Cl- caused glutamate receptor-like3.3-dependent membrane depolarizations. In summary, we show that, in addition to glutamate, glutathione derivatives as well as chloride ions will need to be considered as potential elicitors of wound-response membrane potential change. Finally, by introducing aphid (Brevicoryne brassicae) extracts or the flagellin-derived peptide flg22 into the leaf vasculature we extend the use of Ricca assays for the exploration of insect/plant and bacteria/plant interactions. Using a century-old method it is possible to introduce diverse molecules of non-plant origin into the vasculature in intact leaves.

Automated summary

Ricca assays allow the direct introduction of compounds extracted from plants or organisms into the leaf vasculature, enabling the study of plant responses to external stimuli. This study identifies glutamate as the most active elicitor of membrane depolarization and reveals a potential link between GSL breakdown and glutathione in the generation of membrane depolarizing signals. It also suggests that chloride ions may play a role in wound-response membrane potential changes.