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. 2022 May 3;13:877404. doi: 10.3389/fpls.2022.877404

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Copyright © 2022 He, Zhang, Xu, Chen and Cai.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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BnRALF10 physically interacted with BnFER. (A) BnRALF10 interacts with the extracellular domain of BnFER in yeast. Yeast (strain AH109) co-transformed with pGBKT7 (BD) or the BD-ectoBnFER bait vector and pGADT7 (AD)-BnRALF10 prey vector was grown on SD–Trp–Leu, SD–Trp–Leu–His–Ade plates for interaction confirmation. Only yeast co-transformed with BD-ectoBnFER and AD-BnRALF10 grew on the SD–Trp–Leu–His–Ade medium. BD + AD-BnRALF10 was used as negative control. (B) BiFC assay to detect BnRALF10–BnFER interaction in N. benthamiana. YFP fluorescence was detected in N. benthamiana leaves co-expressing BnRALF10-cYFP and BnFER-nYFP (Lower) but not in the control (cYFP + BnFER-nYFP; Upper). Scale bar: 20 μm. (C) LCI assay to monitor BnRALF10–BnFER interaction in N. benthamiana. LUC activity was detected in N. benthamiana leaves co-expressing BnRALF10-CLuc and BnFER-NLuc but not in the controls (BnRALF10-CLuc + NLuc, CLuc + BnFER-NLuc and CLuc + NLuc).