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Variation in NRT1.1B contributes to nitrate-use divergence between rice subspecies

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Abstract

Asian cultivated rice (Oryza sativa L.) consists of two main subspecies, indica and japonica. Indica has higher nitrate-absorption activity than japonica, but the molecular mechanisms underlying that activity remain elusive. Here we show that variation in a nitrate-transporter gene, NRT1.1B (OsNPF6.5), may contribute to this divergence in nitrate use. Phylogenetic analysis revealed that NRT1.1B diverges between indica and japonica. NRT1.1B-indica variation was associated with enhanced nitrate uptake and root-to-shoot transport and upregulated expression of nitrate-responsive genes. The selection signature of NRT1.1B-indica suggests that nitrate-use divergence occurred during rice domestication. Notably, field tests with near-isogenic and transgenic lines confirmed that the japonica variety carrying the NRT1.1B-indica allele had significantly improved grain yield and nitrogen-use efficiency (NUE) compared to the variety without that allele. Our results show that variation in NRT1.1B largely explains nitrate-use divergence between indica and japonica and that NRT1.1B-indica can potentially improve the NUE of japonica.

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Figure 1: NRT1.1B variation contributes to differences in nitrate use.
Figure 2: Functional characterization and tissue-localization assay of NRT1.1B.
Figure 3: Variation in NRT1.1B could affect nitrate uptake, nitrate root-to-shoot transport and the expression of nitrate-responsive genes.
Figure 4: Phylogenetic analysis of NRT1.1B.
Figure 5: NRT1.1B-indica introgression improves NUE.
Figure 6: NRT1.1B-indica transgenic plants showed higher NUE than 1B-japonica transgenic plants.

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References

  1. Oka, H.I. Origin of Cultivated Rice (Japan Scientific Societies Press, 1983).

  2. Koutroubas, S.D. & Ntanos, D.A. Genotypic differences for grain yield and nitrogen utilization in Indica and Japonica rice under Mediterranean conditions. Field Crops Res. 83, 251–260 (2003).

    Article  Google Scholar 

  3. Kronzucker, H.J., Glass, A.D.M., Siddiqi, M.Y. & Kirk, G.J.D. Comparative kinetic analysis of ammonium and nitrate acquisition by tropical lowland rice: implications for rice cultivation and yield potential. New Phytol. 145, 471–476 (2000).

    Article  CAS  Google Scholar 

  4. Kirk, G.J. & Kronzucker, H.J. The potential for nitrification and nitrate uptake in the rhizosphere of wetland plants: a modelling study. Ann. Bot. 96, 639–646 (2005).

    Article  CAS  Google Scholar 

  5. Ta, T.C. & Ohira, K. Effects of various environmental and medium conditions on the response of indica and japonica rice plants to ammonium and nitrate nitrogen. Soil Sci. Plant Nutr. 27, 347–355 (1981).

    Article  CAS  Google Scholar 

  6. Tsay, Y.F., Schroeder, J.I., Feldmann, K.A. & Crawford, N.M. The herbicide sensitivity gene Chl1 of Arabidopsis encodes a nitrate-inducible nitrate transporter. Cell 72, 705–713 (1993).

    Article  CAS  Google Scholar 

  7. Scholten, H.J. & Feenstra, W.J. Expression of the mutant character of chlorate-resistant mutants of Arabidopsis thaliana in cell-culture. J. Plant Physiol. 123, 45–54 (1986).

    Article  CAS  Google Scholar 

  8. Oostindiër-Braaksma, F.J. & Feenstra, W.J. Isolation and characterization of chlorate-resistant mutants of Arabidopsis thaliana. Mutat. Res. 19, 175–185 (1973).

    Article  Google Scholar 

  9. Wang, X.M., Scholl, R.L. & Feldmann, K.A. Characterization of a chlorate-hypersensitive, high nitrate reductase Arabidopsis thaliana mutant. Theor. Appl. Genet. 72, 328–336 (1986).

    Article  CAS  Google Scholar 

  10. Teng, S. et al. QTLs and candidate genes for chlorate resistance in rice (Oryza sativa L.). Euphytica 152, 141–148 (2006).

    Article  CAS  Google Scholar 

  11. Plett, D. et al. Dichotomy in the NRT gene families of dicots and grass species. PLoS One 5, e15289 (2010).

    Article  CAS  Google Scholar 

  12. Monden, I., Olsowski, A., Krause, G. & Keller, K. The large cytoplasmic loop of the glucose transporter GLUT1 is an essential structural element for function. Biol. Chem. 382, 1551–1558 (2001).

    Article  CAS  Google Scholar 

  13. Mandal, A. et al. A key structural domain of the Candida albicans Mdr1 protein. Biochem. J. 445, 313–322 (2012).

    Article  CAS  Google Scholar 

  14. Zhao, Y. et al. Crystal structure of the E. coli peptide transporter YbgH. Structure 22, 1152–1160 (2014).

    Article  CAS  Google Scholar 

  15. Liu, K.H., Huang, C.Y. & Tsay, Y.F. CHL1 is a dual-affinity nitrate transporter of Arabidopsis involved in multiple phases of nitrate uptake. Plant Cell 11, 865–874 (1999).

    Article  CAS  Google Scholar 

  16. Ho, C.H., Lin, S.H., Hu, H.C. & Tsay, Y.F. CHL1 functions as a nitrate sensor in plants. Cell 138, 1184–1194 (2009).

    Article  CAS  Google Scholar 

  17. Ho, C.H. & Tsay, Y.F. Nitrate, ammonium, and potassium sensing and signaling. Curr. Opin. Plant Biol. 13, 604–610 (2010).

    Article  CAS  Google Scholar 

  18. Gojon, A., Krouk, G., Perrine-Walker, F. & Laugier, E. Nitrate transceptor(s) in plants. J. Exp. Bot. 62, 2299–2308 (2011).

    Article  CAS  Google Scholar 

  19. Biswas, S. & Akey, J.M. Genomic insights into positive selection. Trends Genet. 22, 437–446 (2006).

    Article  CAS  Google Scholar 

  20. Caicedo, A.L. et al. Genome-wide patterns of nucleotide polymorphism in domesticated rice. PLoS Genet. 3, 1745–1756 (2007).

    Article  CAS  Google Scholar 

  21. Gao, L.Z. & Innan, H. Nonindependent domestication of the two rice subspecies, Oryza sativa ssp. indica and ssp. japonica, demonstrated by multilocus microsatellites. Genetics 179, 965–976 (2008).

    Article  CAS  Google Scholar 

  22. Huang, X. et al. A map of rice genome variation reveals the origin of cultivated rice. Nature 490, 497–501 (2012).

    Article  CAS  Google Scholar 

  23. Moll, R.H., Kamprath, E.J. & Jackson, W.A. Analysis and interpretation of factors which contribute to efficiency of nitrogen-utilization. Agron. J. 74, 562–564 (1982).

    Article  Google Scholar 

  24. Moll, R.H., Kamprath, E.J. & Jackson, W.A. Development of nitrogen efficient prolific hybrids of maize. Crop Sci. 27, 181–186 (1987).

    Article  CAS  Google Scholar 

  25. Bart, R., Chern, M., Park, C.J., Bartley, L. & Ronald, P.C. A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts. Plant Methods 2, 13 (2006).

    Article  Google Scholar 

  26. Almagro, A., Lin, S.H. & Tsay, Y.F. Characterization of the Arabidopsis nitrate transporter NRT1.6 reveals a role of nitrate in early embryo development. Plant Cell 20, 3289–3299 (2008).

    Article  CAS  Google Scholar 

  27. Li, C. et al. A rice plastidial nucleotide sugar epimerase is involved in galactolipid biosynthesis and improves photosynthetic efficiency. PLoS Genet. 7, e1002196 (2011).

    Article  CAS  Google Scholar 

  28. Felsenstein, J. PHYLIP—phylogeny inference package (version 3.2). Cladistics 5, 164–166 (1989).

    Google Scholar 

  29. Zhang, H., Gao, S., Lercher, M.J., Hu, S. & Chen, W.H. EvolView, an online tool for visualizing, annotating and managing phylogenetic trees. Nucleic Acids Res. 40, W569–W572 (2012).

    Article  CAS  Google Scholar 

  30. Kawahara, Y. et al. Improvement of the Oryza sativa Nipponbare reference genome using next generation sequence and optical map data. Rice (NY) 6, 4 (2013).

    Article  Google Scholar 

  31. Edgar, R.C. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 32, 1792–1797 (2004).

    Article  CAS  Google Scholar 

  32. Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol. Biol. Evol. 30, 2725–2729 (2013).

    Article  CAS  Google Scholar 

  33. Jensen, J.D., Thornton, K.R., Bustamante, C.D. & Aquadro, C.E. On the utility of linkage disequilibrium as a statistic for identifying targets of positive selection in nonequilibrium populations. Genetics 176, 2371–2379 (2007).

    Article  Google Scholar 

  34. Alachiotis, N., Stamatakis, A. & Pavlidis, P. OmegaPlus: a scalable tool for rapid detection of selective sweeps in whole-genome datasets. Bioinformatics 28, 2274–2275 (2012).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank H. Xue (Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, China) for providing the nrt1.1b mutant and M. Schläppi (Marquette University, Milwaukee, Wisconsin, USA) and X. Wang (China Agricultural University, Beijing, China) for language editing. This work was supported by grants from the Ministry of Science and Technology of China (2014AA10A602-5, 2015CB755702) and the Chinese Academy of Sciences (XDA08010400).

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Contributions

B.H. performed most of the experiments. J.T. and W.W. constructed the NIL. W.W., Z.Z., Z.P., Q.D. and Y.L. performed the field test. S.O. and H.W. performed the population genetic analysis. H.L. assayed 15N accumulation in rice cultivars. R.C. carried out the in situ hybridization. X.C. and L. Li performed the Xenopus oocyte injection. Y.W., C.L., L. Liu, K.D. and C.X. performed the chlorate-sensitivity assay. C.C., B.H. and L.Z. designed the experiments. B.H., C.C. and S.O. wrote the manuscript.

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Correspondence to Chengcai Chu.

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The authors declare no competing financial interests.

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Supplementary Figures 1–14, Supplementary Tables 1–3 and Supplementary Note (PDF 2815 kb)

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Hu, B., Wang, W., Ou, S. et al. Variation in NRT1.1B contributes to nitrate-use divergence between rice subspecies. Nat Genet 47, 834–838 (2015). https://doi.org/10.1038/ng.3337

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