Validation of stress-responsive miRNAs. (a) Validation of phosphate-starvation inducible Bdi-miR399 and Bdi-miR827 by splinted ligation-mediated miRNA detection. Bdi-miR168 was used as a control. (b) Expression pattern of Bdi-miR5200 during submergence treatment and subsequent recovery. Three-week-old seedlings were subjected to submergence stress by completely immersing the plants in water, and subsequent recovery upon removal from the water. Shoots were collected before submergence treatment (0 d), after one-day (1 d) or two-day (2 d) treatments, and one (3 d) or three (5 d) days after recovery. Then 20 μg of total RNA were used to detect Bdi-FTL1 using an RNA blot. Bdi-miR168 and rRNAs were used as controls. miRNA, microRNA; Recov., recovery; rRNA, ribosomal RNA; Submer., submergence.