Figure 3. Metal tolerance complementation assays from TM1 to TM6 of OsMTP8.1 with random and site-directed mutagenesis.

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.