The rice “fruit-weight 2.2-like” gene family member OsFWL4 is involved in the translocation of cadmium from roots to shoots
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Heterogeneous expression of the rice genes “fruit-weight 2.2-like” (OsFWL) affects Cd resistance in yeast, and OsFWL4 mediates the translocation of Cd from roots to shoots.
Cadmium (Cd) induces chronic and toxic effects in humans. In a previous study (Xu et al. in Planta 238:643–655, 2013), we cloned the rice genes, designated OsFWL1-8, homologous to the tomato fruit-weight 2.2. Here, we show that expression of genes OsFWL3-7 in yeast confers resistance to Cd. The Cd contents of OsFWL3-, -4-, -6- and -7-transformed Cd(II)-sensitive yeast mutant ycf1 cells were strongly decreased compared with those of empty vector, with the strongest resistance to Cd observed in cells expressing OsFWL4. Evaluation of truncated and site-directed mutation derivatives revealed that the CCXXG motifs near the second transmembrane region of OsFWL4 are involved in Cd resistance in yeast. Real-time PCR analysis showed that OsFWL4 expression was induced by CdCl2 stress in rice seedlings. Compared with WT plants, the Cd contents in the shoots of RNAi mediated OsFWL4 knockdown plants were significantly decreased, and Cd translocation from roots to shoots was reduced. According to bimolecular fluorescence complementation, yeast two-hybrid and Western-blotting assays, the OsFWL4 protein forms homo-oligomers. These results suggest that OsFWL4 might act directly as a transporter and is involved in the translocation of Cd from roots to shoots in rice.
KeywordsCCXXG motif Cd resistance Cd translocation Heavy metals OsFWL
Heavy metal ATPase
Low-affinity cation transporter 1
Natural resistance-associated macrophage protein
Oryza sativa FW2.2-like
We thank Dr. Dennis J Thiele (Duke University School of Medicine) for kindly providing the Cd-sensitive ycf1 mutant and Professor Jian Xu (National University of Singapore) for providing the pM999 vectors. We also thank Dr. Tao Luo (Nanchang University) and Tingting Fan (Humboldt-Universitat zu Berlin) for technical assistance. This work was supported by the National Natural Science Foundation of China (Grant no. 31671268).
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