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Golgi-Localized OsFPN1 is Involved in Co and Ni Transport and Their Detoxification in Rice
Manman Kan1, Toru Fujiwara1, Takehiro Kamiya2,3
1Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Abstract:
Cobalt (Co) and nickel (Ni) are beneficial and essential elements for plants, respectively, with the latter required for urease activity, which hydrolyzes urea into ammonium in plants. However, excess Co and Ni are toxic to plants and their transport mechanisms in rice are unclear. Here, we analyzed an ethyl methanesulfonate (EMS)-mutagenized rice mutant, 1187_n, with increased Co and Ni contents in its brown rice and shoots. 1187_n has a mutation in OsFPN1, which was correlated with a high Co and Ni phenotype in F2 crosses between the parental line and mutant. In addition, CRISPR/Cas9 mutants exhibited a phenotype similar to that of 1187_n, demonstrating that OsFPN1 is the causal gene of the mutant. In addition to the high Co and Ni in brown rice and shoots, the mutant also exhibited high Co and Ni concentrations in the xylem sap, but low concentrations in the roots, suggesting that OsFPN1 is involved in the root-to-shoot translocation of Co and Ni. The growth of 1187_n and CRISPR/Cas9 lines were suppressed under high Co and Ni condition, indicating OsFPN1 is required for the normal growth under high Co and Ni. An OsFPN1-green fluorescent protein (GFP) fusion protein was localized to the Golgi apparatus. Yeast carrying GFP-OsFPN1 increased sensitivity to high Co contents and decreased Co and Ni accumulation. These results suggest that OsFPN1 can transport Co and Ni and is vital detoxification in rice.
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