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. 2019 Oct 25;10:1342. doi: 10.3389/fpls.2019.01342

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Copyright © 2019 Boter, Calleja-Cabrera, Carrera-Castaño, Wagner, Hatzig, Snowdon, Legoahec, Bianchetti, Bouchereau, Nesi, Pernas and Oñate-Sánchez

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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Protein interactions between important regulators of seed germination are conserved in B. napus. Interactions between selected B. napus transcription factor (TF) proteins analyzed in a yeast two-hybrid system. Bait constructs were generated by N-terminal translational fusions between the GAL4-binding domain (BD) coding sequence (CDS) and selected TF CDSs (BD-BnaNAC25, BD-BnaSPT, BD-BnaNF-YC9, and BD-BnaPIL5). Prey constructs were generated by N-terminal translational fusions between the GAL4-activation domain (AD) CDS and selected TF CDSs (AD-BnaRGL2, AD-BnaABI3, and AD-BnaHFR1). The GAL4-AD and the GAL4-AD fused to the CDS of the green fluorescent protein (GFP) were used as negative controls. Yeast strains containing baits were mated to strains containing preys in the combinations indicated in the figure, and the diploid cells obtained were grown on diploid (-L-W) and screening (-L-W-H) plates with or without 3-AT. Cells containing combinations of baits and preys able to interact were able to grow on screening plates at concentrations of 3-AT that blocked the growth of negative controls.