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. 2017 Jul 19;8:1274. doi: 10.3389/fpls.2017.01274

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Copyright © 2017 Huang, Lv and Wang.

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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Quantitative PCR assay of the flavonoid pathway genes in transgenic tobacco flowers carrying EsMYB9 gene. The expression levels of 10 structural genes of flavonoid biosynthetic pathway and two bHLH regulatory genes (NtAn1a and NtAn1b) are determined by quantitative PCR assay. Tobacco Tub1 gene is used as an internal control, and comparative Ct method is used to determine the relative expression level. Three EsMYB9 overexpressing tobacco lines (MYB9-4, -14, and -17) and the control plant carrying the empty vector are used for qPCR assay. Each column represents average value with SD bar from three technical replicates. Asterisk represents a significant difference from the control plant at the level of P < 0.05 using LSD statistical analysis.