Figure 4. qRT-PCR analysis of the abundance of miR1885 and its
targets under cold stress in B. napus.
(A) qRT-PCR analysis of the abundance of miR1885 in B.
napus under cold stress. U6 was used as endogenous reference
gene. (B) Alignment of miR1885 mature sequence between B.
rapa and B. napus . Short black line represents the perfect
matches. (C) Validation of miR1885 target genes and cleavage
sites using 5′-RACE PCR. Predicted TIR, NBS, and LRR domains encoded by
target genes are labeled (Left). Reverse complementary matches of
miR1885 and their target sites were showed and vertical arrows indicated
5 termini of miRNA-guided cleavage products, as identified by 5′ RACE,
with frequency of clones shown (Right). (D ) The transcript
levels of Bn.TIR.A09 and Bn.TNL.A03 in B. napusunder cold treatment. “D” represents days. (E) Relative
transcript levels of cold-related genes in wild-type B. napusunder cold treatment. 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.