Figure 6: Interruption of FT2β activity by artificial miRNA results in early flowering.

(a) Schematic diagram of amiRFT2β. (b) Northern blotting analysis of artificial miRNA accumulation in amiRFT2β transgenic plants. U6 was used as a loading control for RNA gel blot. (c) qRT-PCR analysis of FT2β expression in wild-type Bd21-3 and the indicated amiRFT2β transgenic plants. UBC18 was used as an internal control for normalization of qRT-PCR results. Three six-week-old B. distachyon plants were pooled and collected for RNA isolation. Each qRT-PCR analysis were performed in three biological replicates with similar results. The point represents the mean value of three technical replicates in a representative biological experiment. Error bars indicate s.d. Student’s t-test, *P<0.05 (d) qRT-PCR analysis of FT2α expression in wild-type and the indicated amiRFT2β transgenic plants. (e) Representative photograph of flowering phenotypes in amiRFT2β transgenic and wild-type Bd21-3 plants. White arrows point to spikes. Scale bar, 2 cm. (f) Flowering time of wild-type and the indicated two lines of amiRFT2β transgenic plants. Error bars indicate s.d. (n=10). Student’s t-test, *P<0.05 (g) qRT-PCR analysis of flowering downstream gene VRN1 expression in wild-type and the indicated amiRFT2β transgenic plants. Total RNA from 5-week-old whole-plant tissues, including leaves and shoot apex, were used for VRN1 examination. Student’s t-test, **P<0.01, *P<0.05.