Fig. 2
FTL9 is a promoter of flowering under SDs in B. distachyon. a Interaction analysis between FTL9 and FD1 in yeast cells. Yeast cells growing on selective media without Leu, Trp, His and Ade (-LTHA) represent positive interactions. b Relative quantification of FTL9 and FD1 interactions by β-Gal activity in yeast two-hybrid assays. Bars indicate s.d. c BiFC assays for determination of FTL9 and FD1 interactions in N. benthamiana leaf cells. Scale bar, 20 μm. d Firefly luciferase complementation imaging (FLCI) assays of FTL9 and FD1 interactions in N. benthamiana leaves. e CRISPR/Cas9-mediated target mutagenesis of FTL9. Top: a schematic diagram of the FTL9 bearing the CRISPR-Cas9 target sites. Exons and introns are represented by black boxes and lines, respectively. The lower sequences show the alignment of wild-type Bd21, ftl9-1 and ftl9-2 sequences containing the CRISPR-Cas9 target sites. The ftl9-1 and ftl9-2 mutants contain a 2-bp and 5-bp deletion, respectively. The black underlined sequences mean protospacer adjacent motifs. f Representative phenotypes of two ftl9 mutants under SDs. White arrows point to spikes. Scale bar, 5 cm. g Flowering time of wild-type (WT) Bd21 and the ftl9 mutants under SDs. At least 10 plants for each line were scored. (Student’s t test, ***P < 0.001). h qPCR analysis of flowering downstream gene VRN1 expression in eight-week-old wild-type and the indicated ftl9 mutants under SDs. Error bars indicate s.d. i qPCR analysis of flowering downstream gene FUL2 expression in eight-week-old wild-type and two ftl9 mutants under SDs. Error bars indicate s.d. j Representative phenotypes of three FTL9 over-expressing plants under SDs. White arrows point to spikes. Scale bar, 5 cm. k Flowering time of wild-type Bd21 and the FTL9-OE plants under SDs. At least 10 plants for each line were scored. (Student’s t test, ***P < 0.001). l qPCR analysis of flowering downstream gene VRN1 expression in eight-week-old wild-type and the FTL9-OE plants under SDs. Error bars indicate s.d. m qPCR analysis of flowering downstream gene FUL2 expression in eight-week-old wild-type and the FTL9-OE plants under SDs. Error bars indicate s.d.
