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. 2017 Jul 10;7:5026. doi: 10.1038/s41598-017-04913-0

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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BiFC assay to verify the MKK-MAPK interactions identified by Y2H. Chickpea MKKs were tagged with N-terminal half of YFP and MAPKs were tagged with C-terminal half of YFP. Plasmid DNA of MKK and MAPK clone was precipitated on gold particles and bombarded on onion epidermal cells. Cells were observed for YFP fluorescence under a confocal microscope after 48 hours. YFP and merged (bright field and YFP) photographs are presented here for 20 interaction pairs of chickpea MKK-MAPK. BiFC assay negative controls (not shown here) were also checked. Scale bars, 50 µm.