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Over-expression of the MxIRT1 gene increases iron and zinc content in rice seeds

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Abstract

Iron and zinc are essential in plant and human nutrition. Iron deficiency has been one of the causes of human mortality, especially in developing countries with high rice consumption. MxIRT1 is a ferrous transporter that has been screened from an iron-efficient genotype of the apple tree, Malus xiaojinensis Cheng et Jiang. In order to produce Fe-biofortified rice with MxIRT1 to solve the Fe-deficiency problem, plant expression vectors of pCAMBIA1302-MxIRT1:GFP and pCAMBIA1302-anti MxIRT1:GFP were constructed that led to successful production of transgenic rice. The transgenic plant phenotypes showed that the expression of endogenous OsIRT1 was suppressed by anti-MxIRT1 in antisense lines that acted as an opposing control, while sense lines had a higher tolerance under Zn- and Fe-deficient conditions. The iron and zinc concentration in T3 seeds increased by three times in sense lines when compared to the wild type. To understand the MxIRT1 cadmium uptake, the MxIRT1 cadmium absorption trait was compared with AtIRT1 and OsIRT1 in transgenic rice protoplasts, and it was found that MxIRT1 had the lowest Cd uptake capacity. MxIRT1 transgenic tobacco-cultured bright yellow-2 (BY-2) cells and rice lines were subjected to different Fe conditions and the results from the non-invasive micro-test technique showed that iron was actively transported compared to cadmium as long as iron was readily available in the environment. This suggests that MxIRT1 is a good candidate gene for plant Fe and Zn biofortification.

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References

  • An P, Wu Q, Wang H, Guan Y, Mu MD, Liao YJ, Zhou DZ, Song PK, Wang CR, Meng LP, Man QQ, Li LX, Zhang JZ, Wang FD (2012) TMPRSS6, but not TF, TFR2 or BMP2 variants are associated with increased risk of iron-deficiency anemia. Hum Mol Genet 21:2124–2131

    Article  CAS  PubMed  Google Scholar 

  • Anuradha K, Agarwal S, Rao YV, Rao KV, Viraktamath BC, Sarla N (2012) Mapping QTLs and candidate genes for iron and zinc concentrations in unpolished rice of Madhukar X Swarna RILs. Gene 508:233–240

    Article  CAS  PubMed  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts: polyphenoloxidase in Beta vulagaris. Plant Physiol 24:1–15

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Azzouzi BE, Tsangaris GT, Pellegrini O, Manuel Y, Benveniste J, Thomas Y (1994) Cadmium induced apoptosis in a human T cell line. Toxicology 88:127–139

    Article  PubMed  Google Scholar 

  • Banuelos GS, Lin ZQ (2008) Development and uses of biofortified agricultural products. CRC Press, Boca Raton

    Book  Google Scholar 

  • Barberon M, Zelazny E, Robert S, Conéjéro G, Curie C, Friml J, Vert G (2011) Monoubiquitin-dependent endocytosis of the iron-regulated transporter 1 (IRT1) transporter controls iron uptake in plants. PNAS 108:E450–E458

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bashir K, Takahashi R, Nakanishi H, Nishizawa NK (2013) The road to micronutrient biofortification of rice: progress and prospects. Front Plant Sci. doi:10.3389/fpls.2013.00015

    Google Scholar 

  • Bughio N, Yamaguchi H, Nishizawa NK, Nakanishi H, Mori S (2002) Cloning an iron regulated metal transporter from rice. J Exp Bot 53:1677–1682

    Article  CAS  PubMed  Google Scholar 

  • Clemens S, Antosiewicz DM, Ward JM, Schachtman DP, Schroeder JI (1998) The plant cDNA LCT1 mediates the uptake of calcium and cadmium in yeast. PNAS 95:12043–12048

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Clemens S, Aarts MGM, Thomine S, Verbruggen N (2013) Plant science: the key to preventing slow cadmium poisoning. Trends Plant Sci 18:92–99

    Article  CAS  PubMed  Google Scholar 

  • Cohen CK, Fox TC, Garvin DF, Kochian LV (1998) The role of iron-deficiency stress responses in stimulating heavy-metal transport in plants. Plant Physiol 116:1063–1072

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Connolly EL, Fett JP, Guerinot ML (2002) Expression of the IRT1 metal transporter is controlled by metals at the levels of transcript and protein accumulation. Plant Cell 14:1347–1357

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ding H, Duan LH, Li J, Yan HF, Zhao M, Zhang FS, Li WX (2010) Cloning and functional analysis of the peanut iron transporter AhIRT1 during iron deficiency stress and intercropping with maize. J Plant Physiol 167:996–1002

    Article  CAS  PubMed  Google Scholar 

  • Drakakaki G, Christou P, Stoger E (2000) Constitutive expression of soybean ferritin cDNA in transgenic wheat and rice results in increased iron levels in vegetative tissues but not in seeds. Transgenic Res 9:445–452

    Article  CAS  PubMed  Google Scholar 

  • Eckhardt U, Marques AM, Buckhout TJ (2001) Two iron regulated cation transporters from tomato complement metal uptake-deficient yeast mutants. Plant Mol Biol 45:437–448

    Article  CAS  PubMed  Google Scholar 

  • Eide D, Broderius M, Fett J, Guerinot MG (1996) A nove liron-regulated metal transporter from plants identified by functional expression in yeast. PNAS 93:5624–5628

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Enomoto Y, Hodoshima H, Shimada H, Shoji K, Yoshihara T, Goto F (2007) Long-distance signals positively regulate the expression of iron uptake genes in tobacco roots. Planta 227:81–89

    Article  CAS  PubMed  Google Scholar 

  • Ghandilyan A, Vreugdenhil D, Aarts MGM (2006) Progress in the genetic understanding of plant iron and zinc nutrition. Plant Physiol 126:407–417

    Article  CAS  Google Scholar 

  • Gitlin JD (2006) Distributing nutrition. Science 314:1252–1253

    Article  CAS  PubMed  Google Scholar 

  • Han ZH, Wang Q, Shen T (1994) Comparison of some physiological and biochemical characteristics between iron-efficient and iron-inefficient species in genus malus. J Plant Nutr 17:1257–1264

    Article  CAS  Google Scholar 

  • Han JH, Song XF, Zhang YP, Li P, Yin LP (2009) Chloroplast localized ZmFDR3 participate in iron transport. Sci China 39:220–227

    Google Scholar 

  • Illing AC, Shawki A, Cunningham CL, Mackenzie B (2012) Substrate profile and metal-ion selectivity of human divalent metal-ion transporter-1. J Biol Chem 287:30485–30496

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ishimaru Y, Suzuki M, Tsukamoto T, Suzuki K, Nakazono M, Kobayashi T, Wada Y, Watanabe S, Matsuhashi S, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2005) Rice plants take up iron as an Fe3+-phytosiderophore and as Fe2+. Plant J 45:335–346

    Article  Google Scholar 

  • Ishimaru Y, Masuda H, Suzuki M, Bashir K, Takahashi M, Nakanishi H, Mori S, Nishizawa NK (2007) Overexpression of the OsZIP4 zinc transporter confers disarrangement of zinc distribution in rice plants. J Exp Bot 58:2909–2915

    Article  CAS  PubMed  Google Scholar 

  • Kanobe MN, Rodermel SR, Bailey T, Scott MP (2013) Changes in endogenous gene transcript and protein levels in maize plants expressing the soybean ferritin transgene. Front Plant Sci. doi:10.3389/fpls.2013.00196

    PubMed Central  PubMed  Google Scholar 

  • Kobayashi T, Nishizawa NK (2012) Iron uptake, translocation, and regulation in higher plants. Annu Rev Plant Biol 63:131–152

    Article  CAS  PubMed  Google Scholar 

  • Korshunova YO, Eide D, Clark WG, Guerinot ML, Pakrasi HB (1999) The IRT1 protein from Arabidopsis thaliana is a metal transporter with a broad substrate range. Plant Mol Biol 40:37–44

    Article  CAS  PubMed  Google Scholar 

  • Lee S, An G (2009) Over-expression of OsIRT1 leads to increased iron and zinc accumulations in rice. Plant Cell Environ 32:408–416

    Article  CAS  PubMed  Google Scholar 

  • Li P, Qi JL, Wang L, Huang QN, Han ZH, Yin LP (2006) Functional expression of MxIRT1, from Malus xiaojinensis, complements an iron uptake deficient yeast mutant for plasma membrane targeting via membrane vesicles trafficking process. Plant Sci 171:52–59

    Article  CAS  Google Scholar 

  • Li S, Zhou X, Huang Y, Zhu L, Zhang S, Zhao Y, Guo J, Chen J, Chen R (2013) Identification and characterization of the zinc-regulated transporters, iron-regulated transporter-like protein (ZIP) gene family in maize. BMC Plant Biol 13:114

    Article  PubMed Central  PubMed  Google Scholar 

  • Lin YF, Liang HM, Yang SY, Boch A, Clemens S, Chen CC, Wu JF, Huang JL, Yeh KC (2009) Arabidopsis IRT3 is a zinc-regulated and plasma membrane localized zinc/iron transporter. New Phytol 182:392–404

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Ma J, He CW, Li XL, Zhang WJ, Xu FS, Lin YJ, Wang LJ (2013) Inhibition of cadmium ion uptake in rice (Oryza sativa) cells by a wall-bound form of silicon. New Phytol. doi:10.1111/nph.12494

    Google Scholar 

  • Lombi E, Tearall KL, Howarth JR, Zhao FJ, Hawkesford MJ, McGrath SP (2002) Influence of iron status on cadmium and zinc uptake by different ecotypes of the hyperaccumulator Thlaspi caerulescens. Plant Physiol 128:1359–1367

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ma G, Jin Y, Li Y, Zhai F, Kok FJ, Jacobsen E, Yang X (2008) Iron and zinc deficiencies in China: what is a feasible and cost-effective strategy? Public Health Nutr 11:632–638

    Article  PubMed  Google Scholar 

  • Maclean JL, Dawe DC, Hardy B, Hettel GP (2002) Rice almanac, third edition, source book for the most important economic activity on earth. CABI Publishing, Oxfordshire

    Google Scholar 

  • Masuda H, Aung MS, Nishizawa NK (2013) Iron biofortification of rice using different transgenic approaches. Rice 6:40

    Article  PubMed Central  PubMed  Google Scholar 

  • Meng FH, Wei YZ (2004) Iron content in rice and research progress of its biological effectiveness. J Northwest Agric For Univ 32:73–78

    Google Scholar 

  • Moulis JM, Thevenod F (2010) New perspectives in cadmium toxicity: an introduction. Biometals 23:763–768

    Article  CAS  PubMed  Google Scholar 

  • Nakanishi H, Ogawa I, Ishimaru Y, Mori S, Nishizawa NK (2006) Iron deficiency enhances cadmium uptake and translocation mediated by the Fe2+ transporters OsIRT1 and OsIRT2 in rice. Soil Sci Plant Nutr 52:464–469

    Article  CAS  Google Scholar 

  • Naren Zhang P, Ma DK, Wang Y, Li S, Yin LP (2012) Overexpression of OsDPR, a novel rice gene highly expressed under iron deficiency, suppresses plant growth. Sci China Life Sci 55:1082–1091

    Article  CAS  PubMed  Google Scholar 

  • Nishida S, Tsuzuki C, Kato A, Aisu A, Yoshida J, Mizuno T (2011) AtIRT1, the primary iron uptake transporter in the root, mediates excess nickel accumulation in Arabidopsis thaliana. Plant Cell Physiol 52:1433–1442

    Article  CAS  PubMed  Google Scholar 

  • Pedas P, Ytting CK, Fuglsang AT, Jahn TP, Schjoerring JK, Husted S (2008) Manganese efficiency in barley: identification and characterization of the metal ion transporter HvIRT1. Plant Physiol 148:455–466

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Plaza S, Tearall KL, Zhao FJ, Buchner P, McGrath SP, Hawkesford MJ (2007) Expression and functional analysis of metal transporter genes in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens. J Exp Bot 58:1717–1728

    Article  CAS  PubMed  Google Scholar 

  • Podar D, Scherer J, Noordally Z, Herzyk P, Nies D, Sanders D (2012) Metal selectivity determinants in a family of transition metal transporters. J Biol Chem 287:3185–3196

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Qu LQ, Yoshihara T, Ooyama A, Goto F, Takaiwa F (2005) Iron accumulation does not parallel the high expression level of ferritin in transgenic rice seeds. Planta 222:225–233

    Article  CAS  Google Scholar 

  • Ramesh SA, Choimes S, Schachtman DP (2004) Over-expression of an Arabidopsis zinc transporter in Hordeum vulgare increases short-term zinc uptake after zinc deprivation and seed zinc content. Plant Mol Biol 54:373–385

    Article  CAS  PubMed  Google Scholar 

  • Rogers EE, Eide DJ, Guerinot ML (2000) Altered selectivity in an Arabidopsis metal transporter. PNAS 97:12356–12360

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Romano N, Macino G (1992) Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences. Mol Microbiol 6:3343–3353

    Article  CAS  PubMed  Google Scholar 

  • Sasaki A, Yamaji A, Yokosho K, Ma JF (2012) Nramp5 is a major transporter responsible for manganese and cadmium uptake in rice. Plant Cell 24:2155–2167

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schikora A, Thimm O, Linke B, Buckhout TJ, Müller M, Schmidt W (2006) Expression, localization, and regulation of the iron transporter LeIRT1 in tomato roots. Plant Soil 284:101–108

    Article  CAS  Google Scholar 

  • Schroeder JI, Delhaize E, Frommer WB, Guerinot ML, Harrison MJ, Herrera-Esrella L, Horie T, Kochian LV, Muns R, Nishizawa NK, Tsay YF, Sanders D (2013) Using membrane transporters to improve crops for sustainable food production. Nature 497:60–66

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sperotto RA (2013) Zn/Fe remobilization from vegetative tissue to rice seeds: should I stay or should I go? Ask Zn/Fe supply! Front Plant Sci. doi:10.3389/fpls.2013.00464

    Google Scholar 

  • Sperotto RA, Ricachenevsky FK, Waldow VA, Fett JP (2012a) Iron biofortification in rice: it’s a long way to the top. Plant Sci 190:24–39

    Article  CAS  PubMed  Google Scholar 

  • Sperotto RA, Vasconcelos MW, Grusak MA, Fett JP (2012b) Effects of different Fe supplies on mineral partitioning and remobilization during the reproductive development of rice (Oryza Sativa L.). Rice 5:27

    Article  PubMed  Google Scholar 

  • UNEP (2008) Draft final review of scientific information on cadmium. http://www.chem.unep.ch/Pb_and_Cd/SR/Draft_final_reviews/Cd_Review/Final_UNEP_Cadmium_review_Nov_2008.pdf

  • Vasconcelos M, Datta K, Oliva N, Khalekuzzaman M, Torrizo L, Krishnan S, Oliveira M, Goto F, Datta SK (2003) Enhanced iron and zinc accumulation in transgenic rice with the ferritin gene. Plant Sci 164:371–378

    Article  CAS  Google Scholar 

  • Vert G, Grotz N, Dedaldechamp F, Gaymard F, Guerinot ML, Briat JF, Curie C (2002) IRT1, an arabidopsis transporter essential for iron uptake from the soil and for plant growth. Plant Cell 14:1223–1233

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Vert G, Barberon M, Zelazny E, Séguéla M, Briat JF, Curie C (2009) Arabidopsis IRT2 cooperates with the high- affinity iron uptake system to maintain iron homeostasis in root epidermal cells. Planta 229:1171–1179

    Article  CAS  PubMed  Google Scholar 

  • Waters BM, Sankaran RP (2011) Moving micronutrients from the soil to the seeds: genes and physiological processes from a biofortification perspective. Plant Sci 180:562–574

    Article  CAS  PubMed  Google Scholar 

  • Waters BM, Lucena C, Romera FJ, Jester GG, Wynn AN, Rojas CL, Alcantara E, Perez-Vicente R (2007) Ethylene involvement in the regulation of the H+-ATPase CsHA1 gene and of the new isolated ferric reductase CsFRO1 and iron transporter CsIRT1 genes in cucumber plants. Plant Physiol Biochem 45:293–301

    Article  CAS  PubMed  Google Scholar 

  • Xiong HC, Guo XT, Kobayashi T, Kakei Y, Nakanishi H, Nozoye T, Zhang LX, Shen HY, Qiu W, Nishizawa NK, Zuo YM (2014) Expression of peanut iron regulated transporter 1 in tobacco and rice plants confers improved iron nutrition. Plant Physiol Biochem 80:83–89

    Article  CAS  PubMed  Google Scholar 

  • Xu Y, Sun T, Yin LP (2006) Application of non-invasive microsensing system to simultaneously measure both H+ and O2 fluxes around the pollen tube. J Integr Plant Biol 48:823–831

    Article  CAS  Google Scholar 

  • Yang G, Ma F, Wang Y, Feng CG, Li P, Xu Y, Zhao WZ, Yin LP (2010) Vesicle-related OsSEC27P enhances H+ secretion in the iron deficient transgenic tobacco root. Chin Sci Bull 55:3298–3304

    Article  CAS  Google Scholar 

  • Yu SM, Kuo YH, Sheu G, Sheu YJ, Liu LF (1991) Metabolic derepression of α-amylase gene expression in suspension-cultured cells of rice. J Biol Chem 266:21131–21137

    CAS  PubMed  Google Scholar 

  • Zhang XN, Han ZH, Yin LL, Kong J, Xu XF, Zhang XZ, Wang Y (2013) Heterologous functional analysis of the Malus xiaojinensis MxIRT1 gene and the His-box motif by expression in yeast. Mol Biol Rep. doi:10.1007/s11033-012-2193-8

    Google Scholar 

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Acknowledgments

This work was supported by the National Natural Science foundation of China (Grant No: 30971856), Beijing Municipal Natural Science Foundation (Grant No: KZ200710028013), Local special project supported by central government-Characteristic key subject-Botany, and the 7th Beijing Municipal Outstanding Teacher Award. This paper has been edited by professional editors at Editage™, a division of Cactus Communications.

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  1. Peng Li

    Present address: School of Life Sciences, Tsinghua University, Beijing, 100083, China

Authors and Affiliations

  1. College of Life Science, Capital Normal University, Beijing, 100048, China

    Song Tan, Rui Han, Peng Li, Guang Yang, Shuang Li, Peng Zhang, Wei-Bing Wang & Li-Ping Yin

  2. School of Mathematical Sciences, Capital Normal University, Beijing, 100048, China

    Wei-Zhong Zhao

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  1. Song Tan
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Tan, S., Han, R., Li, P. et al. Over-expression of the MxIRT1 gene increases iron and zinc content in rice seeds. Transgenic Res 24, 109–122 (2015). https://doi.org/10.1007/s11248-014-9822-z

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