Fig. 8

a Graphical representation of StTm2 and StPHO2 TAS-like loci and the most abundant siRNAs generated from these two loci are shown. Phased siRNAs predicted to be generated from StTm2 and StPHO2. Eight siRNAs off 18, whereas 4 siRNAs off 8, with higher abundance values are shown for both loci, respectively. Black arrows pointing downwards represent predicted miRNA cleavage site in each locus. b Detection of two phased siRNAs (highlightd in bold in panel (a) on sense strand of StTm2 ([+]siR1 and [+]siR3) and two from antisense strand of StPHO2 ([−]siR3 and [−]siR4) by RT-PCR. c 5′ RLM-RACE assay confirms the cleavage of StTm2 by one of the siRNAs generated from itself i.e. (−)siR15. The siRNA cleavage frequency is also shown by downward arrows in black (10/11). Alignment between mature miRNA (bold) and target gene sequence is shown. d Expression analysis of StTm2 and StPHO2 at 4, 7 and 10 d time-points under LD vs SD photoperiod conditions. EIF3e was used as a reference gene for normalization. The expression level of respective target gene at LD4 was considered as 1 for measurement of relative expression at other time-points. Error bars represent standard error of means from two biological replicates. e Expression analysis for two phased siRNAs generated from StTm2 ([+]siR1 and [+]siR3) and two from StPHO2 ([−]siR3 and [−]siR4) loci at 4, 7 and 10 d time-points under LD vs SD photoperiod conditions. U6 was used as a reference gene for normalization. The expression level of respective siRNA at LD4 was considered as 1 for measurement of relative expression at other time-points. Error bars represent standard error of means from two biological replicates. For graphs, a dotted line represents LD condition, whereas a thick line represents SD condition. Asterisks (one, two and three) indicate significant differences at p < 0.05, p < 0.01, p < 0.001, respectively, using a Student’s t-test. ns = not significant at p < 0.05