Expression of nta-miR6019/6020 decreases during tobacco seedling growth.

<p>(A) Northern blot hybridization of sRNAs isolated from TG34 plants at 1-, 3- and 6 WAG. Hybridization probes, miR6019, miR156, miR168 and miR166 are indicated to the left and probe sequences are listed in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006756#ppat.1006756.s009" target="_blank">S3 Table</a>. (B) Maps of the <i>nta-MIR6019/6020a</i> promoter reporter <i>a-3kpro</i>::<i>GUS</i> and <i>nta-MIR6019/6020b</i> promoter reporter <i>b-2kpro</i>::<i>GUS</i> constructs. Open arrows represent the a-3k or b-2k promoter, and blue boxes represent GUS CDS. (C) GUS staining of <i>a-3kpro</i>::<i>GUS</i> and <i>b-2kpro</i>::<i>GUS</i> transgenic plants and SR1 wild-type plants at 1-, 3- and 6 WAG. The number (n) of stained plants and the length of each bar are indicated on the image. (D) Intensity of GUS staining in <i>a-3kpro</i>::<i>GUS</i> and <i>b-2kpro</i>::<i>GUS</i> transgenic seedlings at 1, 3 and 6WAG. GUS activity was quantified using the average gray values that are plotted on the Y-axis. (E) Relative GUS mRNA levels determined by qRT-PCR in <i>a-3kpro</i>::<i>GUS</i> and <i>b-2kpro</i>::<i>GUS</i> transgenic seedlings at 1-, 3- and 6WAG. The data are the means of three replicates with SD. Different letters indicate significant differences between the treatments according to a one-way ANOVA test (P < 0.05). Y axes all indicate relative fold.