Fig. 5

Pollen viability and morphology of tomato plants in different SlAMS modification modes. a. PCR Identification of positive plants transformed with the pCRISPR/Cas9-SlAMS vector. Lanes 1–38: PCR products of Npt II; Lane B: no target DNA; Lane N: nontransgenic plant; Lane P: pCRISPR/Cas9-SlAMS plasmid. b. Pollen viability test after SlAMS knockout (CR-T0–2) and overexpression (OV-T0–10). Blue-stained pollen grains are non-viable; colorless pollen grains are viable. c. Scanning electron microscope (SEM) images of tomato pollen grains. Wild type (WT, oval shape); pCRISPR/Cas9-SlAMS (showing shrinkage and diamond-like shapes, from CR-T0–2). pCAMBIA2301-SlAMS (showing shriveled and atrophic shapes, from OV-T0–10); d. Target site mutation examination in CRISPR/Cas9-mediated SlAMS plant. Target shows the editing site sequence. PAM indicates the adjacent motif of the protospacer sequence. CR-ams1 indicated the first type of mutation; CR-ams2 indicated the second type; CR-ams3 indicated the third type