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. 2023 Jan 23;24(7):711–724. doi: 10.1111/mpp.13297

FIGURE 2.

FIGURE 2

Identification of ZmSIZ1a/1b double knockout mutants. (a) Schematic diagram of two Cas9 targets on ZmSIZ1a and ZmSIZ1b. (b) Sequence analysis of the target sites (red) in two Zmsiz1a/1b double knockout lines. The wild‐type sequence (WT) is shown at the top, knockouts (KO) below. The protospacer‐adjacent motifs (PAM) are underlined. Deletions are indicated by dashes and the length of the sequence gap is shown. (c) The heat shock‐induced accumulation of SUMO conjugates was impaired in Zmsiz1a/1b double mutant seedlings. The immunoblot was probed with an anti‐AtSUMO1 antibody. Ponceau S‐stained RuBisCO large subunit (rbcL) bands are shown as a loading control. The numbers below the gel blot refer to the quantification of SUMO conjugates relative to the rbcL loading control and then normalizing to the WT without heat shock. CRI(−), the corresponding unedited control plants. (d) The morphologic phenotype of WT, CRI (−), and Zmsiz1a/1b double mutant lines. Bar = 50 cm. (e) Quantification of plant height and the ear height as in (d). Mean ± SD, n = 10, **p < 0.01, Student's t test