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. 2021 Mar 19;17(3):e1009461. doi: 10.1371/journal.pgen.1009461

Fig 2. FRILAIR promotes strawberry ripening through working as a noncanonical target mimic of miR397.

Fig 2

(A) Schematic diagram of FRILAIR showing the target site for miR397. Blue shading indicates the miR397 target site with base-pairing profile expanded below. (B) Schematic view of pFveCas13b. Protospacer sequence of sgRNA for pFveCas13b is exhibited on the bottom panel, and mature miR397 sequence is highlighted in red. (C) qRT-PCR analysis of mature miR397 expression in miR397 knockdown (miR397i) strawberry fruits. Strawberry fruits transformed with pFveCas13b vector without sgRNA were used as control. U6 was used as the internal control. (D) qRT-PCR analysis of FRILAIR expression in miR397i strawberry fruits. GAPDH was used as the internal control. (E) miR397 cleavage sites in FRILAIR determined by RNA ligase-mediated 5’ RACE. Cleavage positions of FRILAIR are indicated by arrows, and numbers of 5’ RACE sequenced clones are shown above the arrows. (F) qRT-PCR analysis of expression of FRILAIR in FRILAIR OE, FRILAIR mut1 and FRILAIR mut2 over-expression fruits. Strawberry fruits transformed with empty vector were used as control. GAPDH was used as the internal control. (G) Schematic diagrams of FRILAIR mut1 and FRILAIR mut2. (H) Phenotypic analyses of fruits from FRILAIR OE, FRILAIR mut1, FRILAIR mut2 and miR397i. Control-1, strawberry fruits transformed with empty vector; Control-2, strawberry fruits transformed with pFveCas13b vector without sgRNA. (I) Total anthocyanin content in fruits from Control-1, Control-2, FRILAIR OE, FRILAIR mut1, FRILAIR mut2 and miR397i. Agrobacterium tumefaciens-mediated transient transformations were performed on immature Falandi fruits at the big green stage. All analyses were conducted five days after infection. Statistically significant differences from control were determined by Student’s t-test: *P <0.05; **P <0.01. Values are means ±SEM of three biological replicates. Scale bar: 1 cm.