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. 2017 Jun 21;8:1093. doi: 10.3389/fpls.2017.01093

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Copyright © 2017 Pilati, Bagagli, Sonego, Moretto, Brazzale, Castorina, Simoni, Tonelli, Guella, Engelen, Galbiati and Moser.

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) or licensor 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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Agrobacterium-mediated transient trans-activation assay in N. benthamiana. qPCR analysis of GUS expression in leaves of N. benthamiana co-transfected with the activating construct CaMV35Spro:VvABF2 and the following target constructs: (A) VvNAC17pro:GUS, (B) ARMADILLO-likepro:GUS, (C) VvMYB143pro:GUS, (D) XERICO-likepro:GUS, (E) HB5pro:GUS. Leaves co-infiltrated with the pK7WG2 vector devoid of the VvABF2 gene (empty vector) were used as a negative control for the trans-activation assay. Forty-eight hours after the first infiltration, leaves were infiltrated with 50 μM ABA or with a mock solution and samples were collected after 15 min, 1, and 3 h. Relative GUS expression was determined using GUS-specific primers and normalized to the expression of the tobacco EF-1α gene. Data are means of four biological replicates ± SD. (F) Distribution of the ABRE motifs in the five 1 kb promoters analyzed.