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Identification and localisation of the rice nicotianamine aminotransferase gene OsNAAT1 expression suggests the site of phytosiderophore synthesis in rice

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Abstract

Rice plants (Oryza sativa L.) take up iron using iron-chelating compounds known as mugineic acid family phytosiderophores (MAs). In the biosynthetic pathway of MAs, nicotianamine aminotransferase (NAAT) catalyses the key step from nicotianamine to the 3′′-keto form. In the present study, we identified six rice NAAT genes (OsNAAT1–6) by screening a cDNA library made from Fe-deficient rice roots and by searching databases. Among the NAAT homologues, OsNAAT1 belongs to a subgroup containing barley functional NAAT (HvNAAT-A and HvNAAT-B) as well as a maize homologue cloned by cDNA library screening (ZmNAAT1). Northern blot and RT-PCR analysis showed that OsNAAT1, but not OsNAAT2–6, was strongly up-regulated by Fe deficiency, both in roots and shoots. The OsNAAT1 protein had NAAT enzyme activity in vitro, confirming that the OsNAAT1 gene encodes functional NAAT. Promoter–GUS analysis revealed that OsNAAT1 was expressed in companion and pericycle cells adjacent to the protoxylem of Fe-sufficient roots. In addition, expression was induced in all cells of Fe-deficient roots, with particularly strong GUS activity evident in the companion and pericycle cells. OsNAAT1 expression was also observed in the companion cells of Fe-sufficient shoots, and was clearly induced in all the cells of Fe-deficient leaves. These expression patterns highly resemble those of OsNAS1, OsNAS2 and OsDMAS1, the genes responsible for MAs biosynthesis for Fe acquisition. These findings strongly suggest that rice synthesises MAs in whole Fe-deficient roots to acquire Fe from the rhizosphere, and also in phloem cells to maintain metal homeostasis facilitated by MAs-mediated long-distance transport.

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Abbreviations

DMA:

2′-Deoxymugineic acid

GUS:

β-Glucuronidase

MAs:

Mugineic acid family phytosiderophores

NA:

Nicotianamine

References

  • Bashir K, Inoue H, Nagasaka S, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2006) Cloning and characterization of deoxymugineic acid synthase genes from graminaceous plants. J Biol Chem 281:32395–32402

    Article  PubMed  CAS  Google Scholar 

  • Colangelo EP, Guerinot ML (2006) Put the metal to petal: metal uptake and transport throughout plants. Curr Opin Plant Biol 9:322–330

    Article  PubMed  CAS  Google Scholar 

  • Curie C, Panaviene Z, Loulergue C, Dellaporta SL, Briat JF, Walker EL (2001) Maize yellow stripe 1 encodes a membrane protein directly involved in Fe(III) uptake. Nature 409:346–349

    Article  PubMed  CAS  Google Scholar 

  • Herbik A, Koch G, Mock HP, Dushkov D, Czihal A, Thielmann J, Stephan UW, Baumlein H (1999) Isolation, characterization and cDNA cloning of nicotianamine synthase from barley. A key enzyme for iron homeostasis in plants. Eur J Biochem 265:231–239

    Article  PubMed  CAS  Google Scholar 

  • Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza Sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282

    Article  PubMed  CAS  Google Scholar 

  • Higuchi K, Kanazawa K, Nishizawa NK, Mori S (1994) Purification and characterization of nicotianamine synthase from Fe deficient barley roots. Plant Soil 165:173–179

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Higuchi K, Suzuki K, Nakanishi H, Yamaguchi H, Nishizawa NK, Mori S (1999) Cloning of nicotianamine synthase genes, novel genes involved in the biosynthesis of phytosiderophores. Plant Physiol 119:471–479

    Article  PubMed  CAS  Google Scholar 

  • Higuchi K, Watanabe S, Takahashi M, Kawasaki S, Nakanishi H, Nishizawa NK, Mori S (2001) Nicotianamine synthase gene expression differs in barley and rice under Fe-deficient conditions. Plant J 25:159–167

    Article  PubMed  CAS  Google Scholar 

  • Inoue H, Higuchi K, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2003) Three rice nicotianamine synthase genes, OsNAS1, OsNAS2, and OsNAS3 are expressed in cells involved in long-distance transport of iron and differentially regulated by iron. Plant J 36:366–381

    Article  PubMed  CAS  Google Scholar 

  • Ishimaru Y, Kim S, Tsukamoto T, Oki H, Kobayashi T, Watanabe S, Matsuhashi S, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2007) Mutational reconstructed ferric chelate reductase confers enhanced tolerance in rice to iron deficiency in calcareous soil. Proc Natl Acad Sci USA 104:7373–7378

    Article  PubMed  CAS  Google Scholar 

  • Kanazawa K, Higuchi K, Nishizawa NK, Fushiya S, Chino M, Mori S (1994) Nicotianamine aminotransferase activities are correlated to the phytosiderophore secretions under Fe-deficient conditions in Gramineae. J Exp Bot 45:1903–1906

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Kawai S, Itoh K, Takagi S, Nomoto K (1988) Studies on phytosiderophore: biosynthesis of mugineic acid and 2′-deoxymugineic acid in Hordeum vulgare L. var Minorimugi. Tetrahyd Lett 29:1053–1056

    Article  CAS  Google Scholar 

  • Kobayashi T, Nakanishi H, Takahashi M, Kawasaki S, Nishizawa NK, Mori S (2001) In vivo evidence that Ids3 from Hordeum vulgare encodes a dioxygenase that converts 2′-deoxymugineic acid to mugineic acid in transgenic rice. Planta 212:864–871

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi T, Nakayama Y, Itai RN, Nakanishi H, Yoshihara T, Mori S, Nishizawa NK (2003) Identification of novel cis-acting elements, IDE1 and IDE2, of the barley IDS2 gene promoter conferring iron-deficiency-inducible, root-specific expression in heterogeneous tobacco plants. Plant J 36:780–793

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi T, Nakayama Y, Takahashi M, Inoue H, Nakanishi H, Yoshihara T, Mori S, Nishizawa NK (2004) Construction of artificial promoters highly responsive to iron deficiency. Soil Sci Plant Nutr 50:1167–1175

    CAS  Google Scholar 

  • Kobayashi T, Suzuki M, Inoue H, Itai RN, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2005) Expression of iron-acquisition-related genes in iron-deficient rice is co-ordinately induced by partially conserved iron-deficiency-responsive elements. J Exp Bot 56:1305–1316

    Article  PubMed  CAS  Google Scholar 

  • Koike S, Inoue H, Mizuno D, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2004) OsYSL2 is a rice metal-nicotianamine transporter that is regulated by iron and expressed in the phloem. Plant J 39:415–424

    Article  PubMed  CAS  Google Scholar 

  • Lytle CM, Jolley VD (1991) Iron deficiency stress response of various C-3 and C-4 grain crop genotypes: strategy II mechanism evaluated. J Plant Nutr 14:341–361

    CAS  Google Scholar 

  • Ma JF, Nomoto K (1993) Two related biosynthetic pathways of mugineic acids in Gramineous plants. Plant Physiol 102:373–378

    Article  PubMed  CAS  Google Scholar 

  • Ma JF, Shinada T, Matsuda C, Nomoto K (1995) Biosynthesis of phytosiderophores, mugineic acids, associated with methionine cycling. J Biol Chem 270:16549–16554

    Article  PubMed  CAS  Google Scholar 

  • Ma JF, Taketa S, Chang YC, Iwashita T, Matsumoto H, Takeda K, Nomoto K (1999) Genes controlling hydroxylations of phytosiderophores are located on different chromosomes in barley (Hordeum vulgare L.). Planta 207:590–596

    Article  CAS  Google Scholar 

  • Mizuno D, Higuchi K, Sakamoto T, Nakanishi H, Mori S, Nishizawa NK (2003) Three nicotianamine synthase genes isolated from maize are differentially regulated by iron nutritional status. Plant Physiol 132:1989–1997

    Article  PubMed  CAS  Google Scholar 

  • Mori S, Nishizawa N (1987) Methionine as a dominant precursor of phytosiderophores in Graminaceae plants. Plant Cell Physiol 28:1081–1092

    CAS  Google Scholar 

  • Mori S, Nishizawa N, Hayashi H, Chino M, Yoshimura E, Ishihara J (1991) Why are young rice plants highly susceptible to iron deficiency? Plant Soil 130:143–156

    Article  CAS  Google Scholar 

  • Murata Y, Ma JF, Yamaji N, Ueno D, Nomoto K, Iwashita T (2006) A specific transporter for iron(III)-phytosiderophore in barley roots. Plant J 46:563–572

    Article  PubMed  CAS  Google Scholar 

  • Nakanishi H, Okumura N, Umehara Y, Nishizawa NK, Chino M, Mori S (1993) Expression of a gene specific for iron deficiency (Ids3) in the roots of Hordeum vulgare. Plant Cell Physiol 34:401–410

    PubMed  CAS  Google Scholar 

  • Nakanishi H, Yamaguchi H, Sasakuma T, Nishizawa NK, Mori S (2000) Two dioxygenase genes, Ids3 and Ids2, from Hordeum vulgare are involved in the biosynthesis of mugineic acid family phytosiderophores. Plant Mol Biol 44:199–207

    Article  PubMed  CAS  Google Scholar 

  • Negishi T, Nakanishi H, Yazaki J, Kishimoto N, Fujii F, Shimbo K, Yamamoto K, Sakata K, Sasaki T, Kikuchi S, Mori S, Nishizawa NK (2002) cDNA microarray analysis of gene expression during Fe-deficiency stress in barley suggests that polar transport of vesicles is implicated in phytosiderophore secretion in Fe-deficient barley roots. Plant J 30:83–94

    Article  PubMed  CAS  Google Scholar 

  • Nozoye T, Inoue H, Takahashi M, Ishimaru Y, Nakanishi H, Mori S, Nishizawa NK (2007) The expression of iron homeostasis-related genes during rice germination. Plant Mol Biol 64:35–47

    Article  PubMed  CAS  Google Scholar 

  • Ogo Y, Itai RN, Nakanishi H, Inoue H, Kobayashi T, Suzuki M, Takahashi M, Mori S, Nishizawa NK (2006) Isolation and characterization of IRO2, a novel iron-regulated bHLH transcription factor in graminaceous plants. J Exp Bot 57:2867–2878

    Article  PubMed  CAS  Google Scholar 

  • Ogo Y, Itai RN, Nakanishi H, Kobayashi T, Takahashi M, Mori S, Nishizawa NK (2007) The rice bHLH protein OsIRO2 is an essential regulator of the genes involved in Fe uptake under Fe-deficient conditions. Plant J 51:366–377

    Article  PubMed  CAS  Google Scholar 

  • Ohata T, Kanazawa K, Mihashi S, Nishizawa NK, Fushiya S, Nozoe S, Chino M, Mori S (1993) Biosynthetic pathway of phytosiderophores in iron-deficient Graminaceous plants. Development of an assay system for the detection of nicotianamine aminotransferase activity. Soil Sci Plant Nutr 39:745–749

    CAS  Google Scholar 

  • Römheld V, Marschner H (1986) Evidence for a specific uptake system for iron phytosiderophore in roots of grasses. Plant Physiol 80:175–180

    PubMed  Google Scholar 

  • Schaaf G, Ludewig U, Erenoglu BE, Mori S, Kitahara T, von Wirén N (2004) ZmYS1 functions as a proton-coupled symporter for phytosiderophore- and nicotianamine-chelated metals. J Biol Chem 279:9091–9096

    Article  PubMed  CAS  Google Scholar 

  • Shojima S, Nishizawa NK, Fushiya S, Nozoe S, Irifune T, Mori S (1990) Biosynthesis of phytosiderophores. In-vitro biosynthesis of 2′-deoxymugineic acid from l-methionine and nicotianamine. Plant Physiol 93:1497–1503

    PubMed  CAS  Google Scholar 

  • Suzuki M, Takahashi M, Tsukamoto T, Watanabe S, Matsuhashi S, Yazaki J, Kishimoto N, Kikuchi S, Nakanishi H, Mori S, Nishizawa NK (2006) Biosynthesis and secretion of mugineic acid family phytosiderophores in zinc-deficient barley. Plant J 48:85–97

    Article  PubMed  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Takagi S, Nomoto K, Takemoto S (1984) Physiological aspect of mugineic acid, a possible phytosiderophore of graminaceous plants. J Plant Nutr 7:469–477

    Article  CAS  Google Scholar 

  • Takahashi M, Yamaguchi H, Nakanishi H, Shioiri T, Nishizawa NK, Mori S (1999) Cloning two genes for nicotianamine aminotransferase, a critical enzyme in iron acquisition (Strategy II) in graminaceous plants. Plant Physiol 121:947–956

    Article  PubMed  CAS  Google Scholar 

  • Takahashi M, Nakanishi H, Kawasaki S, Nishizawa NK, Mori S (2001) Enhanced tolerance of rice to low iron availability in alkaline soils using barley nicotianamine aminotransferase genes. Nat Biotechnol 19:466–469

    Article  PubMed  CAS  Google Scholar 

  • Takizawa R, Nishizawa NK, Nakanishi H, Mori S (1996) Effect of iron deficiency on S-adenosylmethionine synthetase in barley roots. J Plant Nutr 19:1189–1200

    CAS  Google Scholar 

  • von Wirén N, Mori S, Marschner H, Römheld V (1994) Iron inefficiency in maize mutant ys1 (Zea mays L. cv yellow-stripe) is caused by a defect in uptake of iron phytosiderophores. Plant Physiol 106:71–77

    Google Scholar 

  • von Wirén N, Klair S, Bansal S, Briat JF, Khodr H, Shioiri T, Leigh RA, Hider RC (1999) Nicotianamine chelates both Fe(III) and Fe(II). Implications for metal transport in plants. Plant Physiol 119:1107–1114

    Article  Google Scholar 

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Acknowledgement

We thank Mr. Tatsuya Sakamoto for assistance with cloning of ZmNAAT.

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Author notes

  1. Haruhiko Inoue

    Present address: Division of Plant Sciences, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

Authors and Affiliations

  1. Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Haruhiko Inoue, Michiko Takahashi, Takanori Kobayashi, Motofumi Suzuki, Hiromi Nakanishi, Satoshi Mori & Naoko K. Nishizawa

  2. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (JST), Saitama , Japan

    Haruhiko Inoue, Takanori Kobayashi & Naoko K. Nishizawa

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  1. Haruhiko Inoue
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  2. Michiko Takahashi
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Correspondence to Naoko K. Nishizawa.

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Inoue, H., Takahashi, M., Kobayashi, T. et al. Identification and localisation of the rice nicotianamine aminotransferase gene OsNAAT1 expression suggests the site of phytosiderophore synthesis in rice. Plant Mol Biol 66, 193–203 (2008). https://doi.org/10.1007/s11103-007-9262-8

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