Fig. 6

Validation of the target genes for Hvu-miR396, miR160 and miR159. a: Overlap between Hvu-miR396 targets predicted through psRNA Target software and degradome data analysis; b: Validation of the target genes for Hvu-miR396. Upper row shows the distribution of the degradome tag reads along the full length of the target mRNA sequence. Lower row shows modified 5′ RACE-PCR mapping of AK376404 mRNA cleavage sites. Arrows indicate miR396 cleavage sites. Numbers above depict the fraction of cloned products that terminated at the predicted cleavage site; c-d: Real-time qRT-PCR detection of miR160 (c), miR159 (d) and their target gene levels during seed imbibition. Total RNA was isolated from embryo at different time points in germination stage including dry seed, 12 HAI, 18 HAI, 24 HAI. The expression level in dry seed was set as 1.0. Error bars represent the SD from three independent experiments