Copper and ectopic expression of the Arabidopsis transport protein COPT1 alter iron homeostasis in rice (Oryza sativa L.)
- 683 Downloads
Copper deficiency and excess differentially affect iron homeostasis in rice and overexpression of the Arabidopsis high-affinity copper transporter COPT1 slightly increases endogenous iron concentration in rice grains.
Higher plants have developed sophisticated mechanisms to efficiently acquire and use micronutrients such as copper and iron. However, the molecular mechanisms underlying the interaction between both metals remain poorly understood. In the present work, we study the effects produced on iron homeostasis by a wide range of copper concentrations in the growth media and by altered copper transport in Oryza sativa plants. Gene expression profiles in rice seedlings grown under copper excess show an altered expression of genes involved in iron homeostasis compared to standard control conditions. Thus, ferritin OsFER2 and ferredoxin OsFd1 mRNAs are down-regulated whereas the transcriptional iron regulator OsIRO2 and the nicotianamine synthase OsNAS2 mRNAs rise under copper excess. As expected, the expression of OsCOPT1, which encodes a high-affinity copper transport protein, as well as other copper-deficiency markers are down-regulated by copper. Furthermore, we show that Arabidopsis COPT1 overexpression (C1 OE ) in rice causes root shortening in high copper conditions and under iron deficiency. C1 OE rice plants modify the expression of the putative iron-sensing factors OsHRZ1 and OsHRZ2 and enhance the expression of OsIRO2 under copper excess, which suggests a role of copper transport in iron signaling. Importantly, the C1 OE rice plants grown on soil contain higher endogenous iron concentration than wild-type plants in both brown and white grains. Collectively, these results highlight the effects of rice copper status on iron homeostasis, which should be considered to obtain crops with optimized nutrient concentrations in edible parts.
KeywordsCopper Iron Metal transport Oryza sativa COPT1 OsIRO2
This work has been supported by grants BIO2011-24848 and BIO2014-56298-P from the Spanish Ministry of Economy and Competitiveness, and by FEDER funds from the European Union. We acknowledge Àngela Carrió-Seguí (Universitat de València) and Kiranmayee Pamidimukkala for their technical help with this manuscript. We also acknowledge the Servei Central d’Instrumentació Científica (Universitat Jaume I) and SCSIE (Universitat de València) for the ICP-MS and atomic absorption spectrophotometry determinations and greenhouse facilities.
SP and LP conceived the idea and wrote the manuscript. AA-B, FA and CD perform the rice microarray and transformation experiments. AA-B, AG-M and AP-G perform the physiological and molecular experiments in rice plants.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Andrés-Colás N, Perea-García A, Mayo de Andrés S, Garcia-Molina A, Dorcey E, Rodríguez-Navarro S, Pérez-Amador MA, Puig S, Peñarrubia L, Mayo de Andres S, García-Molina A, Dorcey E, Rodriguez-Navarro S, Perez-Amador MA, Puig S, Peñarrubia L (2013) Comparison of global responses to mild deficiency and excess copper levels in Arabidopsis seedlings. Metallomics 5:1234–1246. doi: 10.1039/c3mt00025g CrossRefPubMedGoogle Scholar
- Bernal M, Casero D, Singh V, Wilson GT, Grande A, Yang H, Dodani SC, Pellegrini M, Huijser P, Connolly EL, Merchant SS, Krämer U (2012) Transcriptome sequencing identifies SPL7-regulated copper acquisition genes FRO4/FRO5 and the copper dependence of iron homeostasis in Arabidopsis. Plant Cell 24:738–761. doi: 10.1105/tpc.111.090431 CrossRefPubMedPubMedCentralGoogle Scholar
- Di Rienzo JA, Casanoves F, Balzarini MG, Gonzalez L, Tablada M, Robledo CW (2011) InfoStat.Google Scholar
- Garcia-Molina A, Andrés-Colás N, Perea-García A, Del Valle-Tascõn S, Peñarrubia L, Puig S (2011) The intracellular Arabidopsis COPT5 transport protein is required for photosynthetic electron transport under severe copper deficiency. Plant J 65:848–860. doi: 10.1111/j.1365-313X.2010.04472.x CrossRefPubMedGoogle Scholar
- Garcia-Molina A, Andrés-Colás N, Perea-García A, Neumann U, Dodani SC, Huijser P, Peñarrubia L, Puig S (2013) The Arabidopsis COPT6 transport protein functions in copper distribution under copper-deficient conditions. Plant Cell Physiol 54:1378–1390. doi: 10.1093/pcp/pct088 CrossRefPubMedGoogle 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 (2006) Rice plants take up iron as an Fe3+-phytosiderophore and as Fe2+. Plant J 45:335–346. doi: 10.1111/j.1365-313X.2005.02624.x CrossRefPubMedGoogle Scholar
- Ishimaru Y, Masuda H, Bashir K, Inoue H, Tsukamoto T, Takahashi M, Nakanishi H, Aoki N, Hirose T, Ohsugi R, Nishizawa NK (2010) Rice metal-nicotianamine transporter, OsYSL2, is required for the long-distance transport of iron and manganese. Plant J 62:379–390. doi: 10.1111/j.1365-313X.2010.04158.x CrossRefPubMedGoogle Scholar
- Johnson AAT, Kyriacou B, Callahan DL, Carruthers L, Stangoulis J, Lombi E, Tester M (2011) Constitutive overexpression of the OsNAS gene family reveals single-gene strategies for effective iron- and zinc-biofortification of rice endosperm. PLoS One 6:e24476. doi: 10.1371/journal.pone.0024476 CrossRefPubMedPubMedCentralGoogle Scholar
- Lee SSJ, Jeon US, Lee SSJ, Kim Y-K, Persson DP, Husted S, Schjørring JK, Kakei Y, Masuda H, Nishizawa NK, An G (2009) Iron fortification of rice seeds through activation of the nicotianamine synthase gene. Proc Natl Acad Sci 106:22014–22019. doi: 10.1073/pnas.0910950106 CrossRefPubMedPubMedCentralGoogle Scholar
- Marschner P (2012) Marschner’s Mineral Nutrition of Higher Plants, 3rd edn. Elsevier, AmsterdamGoogle Scholar
- Oliva N, Chadha-Mohanty P, Poletti S, Abrigo E, Atienza G, Torrizo L, Garcia R, Dueñas C, Poncio MA, Balindong J, Manzanilla M, Montecillo F, Zaidem M, Barry G, Hervé P, Shou H, Slamet-Loedin IH (2014) Large-scale production and evaluation of marker-free indica rice IR64 expressing phytoferritin genes. Mol Breed 33:23–37. doi: 10.1007/s11032-013-9931-z CrossRefPubMedGoogle Scholar
- Perea-García A, Garcia-Molina A, Andrés-Colás N, Vera-Sirera F, Pérez-Amador MA, Puig S, Peñarrubia L (2013) Arabidopsis copper transport protein COPT2 participates in the crosstalk between iron deficiency responses and low phosphate signaling. Plant Physiol 162:180–194. doi: 10.1104/pp.112.212407 CrossRefPubMedPubMedCentralGoogle Scholar
- Rodrigo-Moreno A, Andrés-Colás N, Poschenrieder C, Gunsé B, Peñarrubia L, Shabala S (2013) Calcium- and potassium-permeable plasma membrane transporters are activated by copper in Arabidopsis root tips: linking copper transport with cytosolic hydroxyl radical production. Plant Cell Environ 36:844–855. doi: 10.1111/pce.12020 CrossRefPubMedGoogle Scholar
- Smyth GK (2005) Limma: linear models for microarray data. Springer, New YorkGoogle Scholar
- Stevens GA, Finucane MM, De-Regil LM, Paciorek CJ, Flaxman SR, Branca F, Peña-Rosas JP, Bhutta ZA, Ezzati M (2013) Global, regional, and national trends in haemoglobin concentration and prevalence of total and severe anaemia in children and pregnant and non-pregnant women for 1995–2011: a systematic analysis of population-representative. Lancet Glob Heal 1:e16–e25. doi: 10.1016/S2214-109X(13)70001-9 CrossRefGoogle Scholar
- Trijatmiko KR, Dueñas C, Tsakirpaloglou N, Torrizo L, Arines FM, Adeva C, Balindong J, Oliva N, Sapasap MV, Borrero J, Rey J, Francisco P, Nelson A, Nakanishi H, Lombi E, Tako E, Glahn RP, Stangoulis J, Chadha-Mohanty P, Johnson AAT, Tohme J, Barry G, Slamet-Loedin IH (2016) Biofortified indica rice attains iron and zinc nutrition dietary targets in the field. Sci Rep 6:19792. doi: 10.1038/srep19792 CrossRefPubMedPubMedCentralGoogle Scholar
- Tsukamoto T, Nakanishi H, Uchida H, Watanabe S, Matsuhashi S, Mori S, Nishizawa NK (2009) 52Fe translocation in barley as monitored by a positron-emitting tracer imaging system (PETIS): evidence for the direct translocation of Fe from roots to young leaves via phloem. Plant Cell Physiol 50:48–57. doi: 10.1093/pcp/pcn192 CrossRefPubMedGoogle Scholar