Fig. 1

Generation of OsNramp5 knockout mutants. (A) Schematic diagram of OsNramp5 gene structure. UTRs, exons, and introns are indicated by grey blocks, black blocks, and lines, respectively. The nucleotide sequences of the CRISPR/Cas9 target together with PAM (GGG) and the gene edited OsNramp5 mutants are shown at the bottom of the gene structure. Deletions and insertions are indicated by dashes and red letters, respectively. (B) Alignment of amino acid sequences of OsNramp5 and its variants. The amino acid sequences were aligned by Clustral W method under the software MegAlign and viewed by software GeneDoc. The identical residues among all aligned proteins were highlighted in black. (C) Grain Cd content of T5105 and OsNramp5 knockout mutants. (D) Grain Mn content of T5105 and OsNramp5 knockout mutants. Asterisks in (C) and (D) indicate a significant difference between T5105 and OsNramp5 mutants at ** P < 0.01 by Student’s t-test. All data are means ± SD of at least three biological replicates