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Fig. 5 | BMC Plant Biology

Fig. 5

From: Simultaneous gene expression and multi-gene silencing in Zea mays using maize dwarf mosaic virus

Fig. 5

Comparison of insert stability in MDMV OH5 constructs at 7, 14, and 21 dpi. Across three replicates of 20 plants per construct, plants infected with pWX27, pWX68, and pWX56 were assessed for integrity of insertion sequences using RT-PCR with primers flanking the insertion sites. Five plants inoculated with pWX6, the cloned wild type virus, were included for each replicate. For each sample, amplicons were scored in five categories (examples from pWX70 shown in panel C); as full length insert a, full length insert plus smaller bands detected b, multiple bands with no full length insert c, single band of less than full-length insert usually near-wild type virus amplicon d, and no band detected e (example from pWX6 sample shown), compared to plasmid controls pWX70, pWX27 and pWX6 (WT). The number of samples in which bands were scored in categories a and d (panel A and B, respectively) were compared statistically for each construct. At fixed assessment times, constructs designated "ns" were not significantly different from pWX27 GFP at P ≤ 0.05 and constructs with * were significantly different from pWX27 GFP at P ≤ 0.05 based on the pairwise comparisons of least squares mean on the arcsine-square root scale. Samples were taken from the youngest leaves of 7, 14 and 21 dpi of plants for RT-PCR analysis using primers: WX112/WX111 (pWX27); WX317/WX315 (pWX68); WX358/WX367 for GFP insertion (pWX56), and primers WX317/WX315 for VIGS insertion (pWX56). Example test samples shown in panel C are from 14 dpi inoculations with pWX70; L = GeneRuler 100 bp Plus DNA ladder (ThermoFisher Scientific, USA)

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