Figure 5. Transgenic B. napus plants overexpressing
miR1885.
(A) Construct with bra-MiR1885 under control of AA6
promoter. pCaMV35S, CaMV35S promoter; Hyg R, hygromycin-resistance gene;
pAA, AA6 promoter; LB, left border of T-DNA; t35S, CaMV35S terminator;
tAA6, AA6 terminator. Eco RI, Nco I and Bst EII are
restriction sites. (B) Plants of T1 transgenic
lines and wild type in the field. Red scale bar = 10 cm. X2–X16,
transgenic plants. (C) Detection of transgene in rapeseed
transgenic lines by PCR using specific primer spanning AA6 promoter andMiR1885 fragment. M, DNA marker. (D) Real-time PCR
showing expression of pri-miR1885 in transgenic lines. ACT7 was
used as endogenous reference gene. (E) Relative abundance of
mature miR1885 in transgenic lines. U6 was used as endogenous
reference gene. (F) Northern blot analysis of
accumulation of mature miR1885 in MIR1885 -OE lines. U6 was
used as endogenous reference gene. (G-H) Relative transcript
levels of Bn.TIR.A09 (G) and Bn.TNL.A03(H) in miR1885-OE lines as compared to WT. ACT7 was used
as endogenous reference gene. Error bar represents mean ± SE; n =
3. * indicates P < 0.05; ** indicates P< 0.01; Student’s t-test.