Figure 4. Metal tolerance complementation assays in yeast of N-terminal and C-terminal mutants of OsMTP8.1.

(A) Metal tolerance complementation assays in yeast of N-terminal deletion mutants of OsMTP8.1. (B) Metal tolerance complementation assays in yeast of C-terminal deletion and site-directed mutagenesis of OsMTP8.1. Complementation of S. cerevisiae mutant pmr1 grown on selective media supplemented with 3 mM and 8 mM Mn²⁺. For complementation tests, transformants were pre-cultured in SD-Urea overnight. Pre-cultured cells were diluted to an OD_{600 nm} of 1.0, 10 μL of cell suspensions were spotted on SD-Urea plates supplemented with the indicated concentrations of Mn²⁺. Plates were incubated for 72 h at 30 °C.