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. 2018 Oct 5;178(3):989–1001. doi: 10.1104/pp.18.00828

Figure 5.

Figure 5.

Facilitating transgenic screen of Arabidopsis plants with optimal silencing by an intronic amiRNA-producing fluorescent reporter. A, Schematic diagram of an intronic amiRNA-producing GFP reporter. B, An intronic amiR-CNGC2-producing GFP reporter (GFPamiRCNGC2-m1) facilitates the identification of transgenic plants with optimal CNGC2 silencing. Note that transgenic lines with GFP fluorescence show a cyan color when irradiated by a blue light flashlight in the dark, whereas the wild type (WT) and the cngc2 null mutant show a dark purple color. Bar = 1 cm. C, Comparison of the success rates to obtain optimal silencing plants by different transgenic selection strategies. D, Detection of mature amiR-CNGC2-m1 in representative transgenic plants with GFP fluorescence. Mature amiRNAs were detected using stem-loop RT-PCR, and the PCR amplicons were subjected to 12% PAGE analysis. Arabidopsis SnoR101 served as an internal control.