Epigenome Editing of Potato by Grafting Using Transgenic Tobacco as siRNA Donor
Fig 2
TGS induction of the endogenous gene StGBSSI in 35SGBpIR transgenic potato.
(A) Schematic diagram of the GBSSI 5'-flanking region in potato ‘Waseshiro’. Black boxes are untranslated regions, and green boxes are exons. Orange bar shows the target region. Green and red bars indicate regions used for methylation level and transcription level analyses, respectively. The construct of 35SGBpIR and CoGBpIR is drawn schematically. (B) Methylation level of the target region in the potato leaves of in vitro sub-culturing shoots. WT: wild type, t33: 35SGBpIR line 33. Asterisks show statistically significant (**; P<0.01 Student's t test) differences relative to 35SGFP. Means and SD of 3 biological replicates are shown. (C) DNA methylation status in the target (orange) and its flanking regions (black) of GBSSI. The leaves were subjected to bisulphite sequencing. The percentages of methylation at individual cytosines are shown. Upper and lower are WT and t33 results, respectively. 5'-flanking region from -621 to -568 couldn't analyze due to lack of appropriate primer sets. The methylation rates of cytosines with different sequence backgrounds are shown in the inset box. (D) Down-regulation of the GBSSI transcript level in t33. (E) Percentage of amylose in the starch of WT and t33 tubers (n = 5) which were grown in a glasshouse.