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Enhanced tolerance of rice to low iron availability in alkaline soils using barley nicotianamine aminotransferase genes

Nature Biotechnology volume 19, pages 466–469 (2001)Cite this article

Abstract

One of the widest ranging abiotic stresses in world agriculture arises from low iron (Fe) availability due to high soil pH, with 30% of arable land too alkaline for optimal crop production. Rice is especially susceptible to low iron supply, whereas other graminaceous crops such as barley are not. A barley genomic DNA fragment containing two naat genes, which encode crucial enzymes involved in the biosynthesis of phytosiderophores, was introduced into rice using Agrobacterium-mediated transformation and pBIGRZ1. Phytosiderophores are natural iron chelators that graminaceous plants secrete from their roots to solubilize iron in the soil. The two transgenes were expressed in response to low iron nutritional status in both the shoots and roots of rice transformants. Transgenic rice expressing the two genes showed a higher nicotianamine aminotransferase activity and secreted larger amounts of phytosiderophores than nontransformants under iron-deficient conditions. Consequently, the transgenic rice showed an enhanced tolerance to low iron availability and had 4.1 times greater grain yields than that of the nontransformant rice in an alkaline soil.

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Figure 1
Figure 2: Construct of pBIGRZ-NAAT binary vector and molecular analysis of the transformants.
Figure 3: NAAT activity and DMA secretion.
Figure 4: Transgenic rice harboring naat genes is tolerant to low iron availability in an alkaline soil.
Figure 5: Growth features and grain yield of transgenic rice and wild-type rice in an alkaline soil.

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References

  1. Marschner, H. Mineral nutrition of higher plants, Edn. 2. (Academic Press, London, UK; 1995).

    Google Scholar 

  2. Takagi, S. Naturally occurring iron-chelating compounds in oat- and rice-root washing. I. Activity measurement and preliminary characterization. Soil Sci. Plant Nutr. 22, 423–433 (1976).

    Article  CAS  Google Scholar 

  3. Mihashi, S. & Mori, S. Characterization of mugineic-acid-Fe transporter in Fe-deficient barley roots using the multi-compartment transport box method. Bio Metals 2, 146–154 (1989).

    Article  CAS  Google Scholar 

  4. Römheld, V. & Marschner, H. Evidence for a specific uptake system for iron phytosiderophores in roots of grasses. Plant Physiol. 70, 175–180 (1986).

    Article  Google Scholar 

  5. Mori, S. & Nishizawa, N.K. Methionine as a dominant precursor of phytosiderophores in Gramineae plants. Plant Cell Physiol. 28, 1081–1092 (1987).

    CAS  Google Scholar 

  6. Shojima, S., Nishizawa, N.K. & Mori, S. Establishment of a cell-free system for the biosynthesis of nicotianamine. Plant Cell Physiol. 30, 673–677 (1989).

    CAS  Google Scholar 

  7. Shojima, S. et al. Biosynthesis of phytosiderophores: in vitro biosynthesis of 2′-deoxymugineic acid from l-methionine and nicotianamine. Plant Physiol. 93, 1497–1503 (1990).

    Article  CAS  Google Scholar 

  8. Higuchi, K., Kanazawa, K., Nishizawa, N.K. & Mori, S. The role of nicotianamine synthase in response to Fe nutrition status in Gramineae. Plant Soil 178, 171–177 (1996).

    Article  CAS  Google Scholar 

  9. Kanazawa, K. et al. Nicotianamine aminotransferase activities are correlated to the phytosiderophore secretions under Fe-deficient conditions in Gramineae. J. Exp. Bot. 45, 1903–1906 (1994).

    Article  CAS  Google Scholar 

  10. Takahashi, M. et al. Cloning two genes for nicotianamine aminotransferase, a critical enzyme in iron acquisition (strategy II) in graminaceous plants. Plant Physiol. 121, 947–956 (1999).

    Article  CAS  Google Scholar 

  11. Akiyama, K. et al. Development of a system of rice transformation with long genome inserts for their functional analysis for positional cloning. Plant Cell Physiol. Suppl. 38, s94 (1997).

    Google Scholar 

  12. Higuchi, K. et al. Nicotianamine synthase gene expression differs in barley and rice under Fe-deficient conditions. Plant J. 25, 159–168 (2001).

    Article  CAS  Google Scholar 

  13. Mori, S. et al. Why are young rice plants highly susceptible to iron deficiency? Plant Soil 130, 143–156 (1991).

    Article  CAS  Google Scholar 

  14. Kanazawa, K., Higuchi, K., Nishizawa, N.K., Fushiya, S. & Mori, S. Detection of two distinct isozymes of nicotianamine aminotransferase in Fe-deficient barley roots. J. Exp. Bot. 46, 1241–1244 (1995).

    Article  CAS  Google Scholar 

  15. Takizawa, R., Nishizawa, N.K., Nakanishi, H. & Mori, S. Effect of iron deficiency on S-adenosyl-methionine synthetase in barley roots. J. Plant Nutr. 19, 1189–1200 (1996).

    Article  CAS  Google Scholar 

  16. Higuchi, K. et al. Cloning of nicotianamine synthase, novel genes involved in the biosynthesis of phytosiderophores. Plant Physiol. 119, 471–480 (1999).

    Article  CAS  Google Scholar 

  17. Suzuki, K., Higuchi, K., Nakanishi, H., Nishizawa, N-K. & Mori, S. Cloning of nicotianamine synthase genes from Arabidopsis thaliana. Soil Sci. Plant Nutr. 45, 993–1002 (1999).

    Article  CAS  Google Scholar 

  18. Okumura, N. et al. A dioxygenase gene (Ids2) expressed under iron deficiency conditions in the roots of Hordeum vulgare. Plant Mol. Biol. 25, 705–719 (1994).

    Article  CAS  Google Scholar 

  19. Nakanishi, H. et al. Expression of a gene specific for iron deficiency (Ids3) in the roots of Hordeum vulgare. Plant Cell Physiol. 34, 401–410 (1993).

    CAS  PubMed  Google Scholar 

  20. Nakanishi, H., Yamaguchi, H., Sasakuma, T., Nishizawa, N.K. & Mori, S. Two dioxygenase genes, Ids 3 and Ids 2, from Hordeum vulgare are involved in the biosynthesis of mugineic acid family phytosiderophores. Plant Mol. Biol. 44, 199–207 (2000).

    Article  CAS  Google Scholar 

  21. Kobayashi, T. et al. In-vivo evidence that Ids 3 from Hordeum vulgare encodes a dioxygenase that converts 2′-deoxymugineic acid to mugineic acid in transgenic rice. Planta, in press (2001).

  22. Okumura, N., Nishizawa, N.K., Umehara, Y. & Mori, S. An iron deficiency-specific cDNA from barley roots having two homologous cysteine-rich MT domains. Plant Mol. Biol. 17, 531–533 (1991).

    Article  CAS  Google Scholar 

  23. Suzuki, K. et al. Formate dehydrogenase, an enzyme of anaerobic metabolism, is induced by Fe-deficiency in barley roots. Plant Physiol. 116, 725–732 (1998).

    Article  CAS  Google Scholar 

  24. Yamaguchi, H., Nakanishi, H., Nishizawa, N.K., & Mori, S. Induction of the IDI1 gene in Fe-deficient barley roots: a gene encoding a putative enzyme that catalyses the methionine salvage pathway for phytosiderophore production. Soil Sci. Plant Nutr. 46, 1–9 (2000).

    Article  CAS  Google Scholar 

  25. Yamaguchi, H., Nakanishi, H., Nishizawa, N.K. & Mori, S. Isolation and characterization of IDI2, a new Fe-deficiency induced cDNA from barley roots, which encodes a protein related to the α-subunit of eukaryotic initiation factor 2B (eIF2Bα). J. Exp. Bot. 51, 2001–2007 (2000).

    Article  CAS  Google Scholar 

  26. Itai, R. et al. Induced activity of adenine phosphoribosyltransferase (APRT) in iron-deficient barley roots: a possible role for phytosiderophore production. J. Exp. Bot. 51, 1179–1188 (2000).

    Article  CAS  Google Scholar 

  27. Chu, C. The N6 medium and its application to anther culture of cereal crops. In Proceedings of Symposium on Plant Tissue Culture, pp. 43–50. (Science Press,1978).

    Google Scholar 

  28. Kanazawa, K., Higuchi, K., Nakanishi, H., Nishizawa, N.K., & Mori, S. Characterizing nicotianamine aminotransferase: improving its assay system and details of the regulation of its activity by Fe nutrition status. Soil Sci. Plant Nutr. 44, 717–721 (1998).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Dr. Pax Blamey in Queensland University for reading of this manuscript and Mr. Takeo Tanaka for his valuable assistance.

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Authors and Affiliations

  1. Laboratory of Plant Molecular Physiology, The University of Tokyo, Tokyo, 113-8657, Japan

    Michiko Takahashi, Hiromi Nakanishi & Satoshi Mori

  2. CREST, Tsukuba, 305-8602, Japan

    Michiko Takahashi & Satoshi Mori

  3. National Institute of Agrobiological Resources, Tsukuba, 305-8602, Japan

    Shinji Kawasaki

  4. Laboratory of Plant Biotechnology, The University of Tokyo, Tokyo, 113-8657, Japan

    Naoko K. Nishizawa

Authors
  1. Michiko Takahashi
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  2. Hiromi Nakanishi
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  4. Naoko K. Nishizawa
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  5. Satoshi Mori
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Correspondence to Satoshi Mori.

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Takahashi, M., Nakanishi, H., Kawasaki, S. et al. Enhanced tolerance of rice to low iron availability in alkaline soils using barley nicotianamine aminotransferase genes. Nat Biotechnol 19, 466–469 (2001). https://doi.org/10.1038/88143

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