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. 2018 May 7;9:606. doi: 10.3389/fpls.2018.00606

FIGURE 4.

FIGURE 4

Influence of the signature motif substitution of OsNrat1 or OsNramp3 on transport activity for Al and Mn. (A) Effect of mutated OsNrat1 or OsNramp3 on Al tolerance. Yeast strain (BY4741) transformed with empty vector pYES2, OsNrat1, OsNramp3, OsNrat1I240M,I241V,T242M, OsNrat1T242M, OsNramp3M239I,V 240I,M241T, OsNramp3M241T were spotted on LPM without uracil medium (pH 4.2) buffered with 5 mM succinic acid with or without AlCl3 at serial dilutions (from left to right: 10 μl cell suspension with OD 0.2, 0.02, 0.002, and 0.0002) and incubated at 30°C for 3 days. (B) Transport activity of mutated OsNrat1 or OsNramp3 for Al. Yeast cells expressing different mutants were exposed to a solution containing 50 μM AlCl3 (pH 4.2) for 6 h. Data are mean ± SD of three biological replicates. Different letters above the bars indicate significant differences (P < 0.05, Tukey’s test). (C) Complementation of manganese uptake. Transformed smf1 were grown on a medium (pH 6.0) buffered with 50 mM MES in the presence or absence of EGTA. The plates were incubated at 30°C for 3 days.