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.