Molecular Strategies for Improving Mineral Density and Bioavailability in Rice

  • Rajinder K. Jain
  • Jitender Kumar
  • Sunita Jain
  • Vijay K. Chowdhury


Globally, micronutrient malnutrition has become a major health problem affecting over three billion people. Of the various micronutrients, problems (anemia, mental retardation, stunted growth, decreased immune function, and increased mortality) resulting from iron and zinc deficiencies are most prevalent and devastating in the developing countries. Rice serves as a staple food for more than half of the world population, but it has insufficient levels of the key micronutrients (Fe and Zn) to meet daily dietary requirements. Biofortification, which refers to the breeding of plants/crops with high bioavailable micronutrient content using conventional breeding, genetic engineering, and molecular and genomic approaches, has the potential to provide coverage for remote rural populations, where supplementation and fortification programs may not reach, and it inherently targets the poor who consume high levels of staple food and little else. Biofortified rice can be an effective solution to combat micronutrient malnutrition in developing countries with limited resources. The facts that substantial genetic variation for Fe and Zn contents exists in rice and that traits for high nutrient content can be combined with superior agronomic characteristics and high yield have allowed many scientists to use conventional breeding approaches to develop micronutrient-rich rice genotypes. Alternatively, genomic, transformation, and molecular tools have been used to improve our understanding of factors regulating micronutrient contents/bioavailability and rapid discovery of genes involved in iron uptake and storage in target tissues and consequently to develop novel high-Fe and/or high-Zn transgenic plants in rice. At CCS Haryana Agricultural University, Hisar, we have assessed variability for iron and zinc in a collection of 220 rice genotypes and identified several iron- and zinc-rich genotypes which have been used subsequently to raise mapping population and used for identification of QTLs for minerals in brown rice. Material is being used to select mineral-rich high-yielding rice genotypes.


Phytic Acid Brown Rice Transgenic Rice Plant Rice Genotype International Rice Research Institute 


  1. Anuradha K, Agarwal S, Batchu AK, Babu AP, Swamy BPM, Longvah T, Sarla N (2012) Evaluating rice germplasm for iron and zinc concentration in brown rice and seed dimensions. J Phytol 4(1):19–25Google Scholar
  2. Ballot D, Baynes RD, Bothwell TH, Gillooly M, Macfarlane BJ, Macphail AP, Lyons G, Derman DP, Bezwoda WR, Torrance JD, Bothwell JE, Mayet F (1987) The effects of fruit juices and fruits on the absorption of iron from a rice meal. Br J Nutr 57:331–343PubMedCrossRefGoogle Scholar
  3. Banziger M, Long J (2000) The potential of increasing the iron and zinc density of maize through plant breeding. Food Nutr Bull 21:397–400Google Scholar
  4. Bhullar NK, Gruissem W (2013) Nutritional enhancement of rice for human health: the contribution of biotechnology. Biotechnol Adv 31:50–57PubMedCrossRefGoogle Scholar
  5. Bohn L, Meyers AS, Rasmussen K (2008) Phytate: impact on environment and human nutrition. A challenge for molecular breeding. J Zhejiang Univ Sci B 9:165–191PubMedCentralPubMedCrossRefGoogle Scholar
  6. Bouis HE (2002) Plant breeding: a new tool for fighting micronutrient malnutrition. J Nutr 132:491S–494SPubMedGoogle Scholar
  7. Bouis HE, Chassy B, Ochanda JO (2003) Genetically modified food crops and their contribution to human nutrition and food quality. Trends Food Sci Technol 14:191–209CrossRefGoogle Scholar
  8. Brar B, Jain S, Singh R, Jain RK (2011) Genetic diversity for iron and zinc contents in a collection of 220 rice (Oryza sativa L.) genotypes. Indian J Genet 71(1):67–73Google Scholar
  9. Brinch-Pedersen H, Sorensen LD, Holm PB (2002) Engineering crop plants: getting a handle on phosphate. Trends Plant Sci 7:118–125PubMedCrossRefGoogle Scholar
  10. Brinch-Pedersen H, Borg S, Tauris B, Holm PB (2007) Molecular genetic approaches to increasing mineral availability and vitamin content of cereals. J Cereal Sci 46:308–326CrossRefGoogle Scholar
  11. Bughio N, Yamaguchi H, Nishizawa NK, Nakanishi H, Mori S (2002) Cloning an iron-regulated metal transporter from rice. J Exp Bot 53:1677–1682PubMedCrossRefGoogle Scholar
  12. Chandel G, Samuel P, Dubey M, Meena R (2011) In silico expression analysis of QTL specific candidate genes for grain micronutrient (Fe/Zn) content using ESTs and MPSS signature analysis in rice (Oryza sativa L.). J Plant Genet Transgenic 2(1):11–22Google Scholar
  13. Chen H, Siebenmorgen TJ (1997) Effect of rice thickness in degree of milling and associated optical measurements. Cereal Chem 74:821–825CrossRefGoogle Scholar
  14. Chen H, Siebenmorgen TJ, Griffin K (1998) Quality characteristics of long-grain rice milled in two commercial systems. Cereal Chem 75:560–565CrossRefGoogle Scholar
  15. Cheng L, Wang F, Shou H, Huang F, Zheng L, He F, Li J, Zhao FJ, Ueno D, Ma JF, Wu P (2007) Mutation in nicotianamine aminotransferase stimulated the Fe(II) acquisition system and led to iron accumulation in rice. Plant Physiol 145:1647–1657PubMedCentralPubMedCrossRefGoogle Scholar
  16. Curie C, Alonso JM, Le Jean M, Ecker JR, Briat JF (2000) Involvement of NRAMP1 from Arabidopsis thaliana in iron transport. Biochem J 347:749–755PubMedCrossRefGoogle Scholar
  17. Curie C, Cassin G, Couch D, Divol F, Higuchi K, Jean ML (2009) Metal movement within the plant: contribution of nicotianamine and yellow stripe 1-like transporters. Ann Bot 103:1–11PubMedCrossRefGoogle Scholar
  18. Eide D, Broderius M, Fett J, Guerinot ML (1996) A novel iron-regulated metal transporter from plants identified by functional expression in yeast. Proc Natl Acad Sci USA 93:5624–5628PubMedCrossRefGoogle Scholar
  19. Frossard E, Bucher M, Mächler F, Mozafar A, Hurrell R (2000) Potential for increasing the content and bioavailability of Fe, Zn and Ca in plants for human nutrition. J Sci Food Agric 80:861–879CrossRefGoogle Scholar
  20. Galera SG, Rojas E, Sudhakar D, Zhu C, Pelacho AM, Capell T, Christou P (2010) Critical evaluation of strategies for mineral fortification of staple food crops. Transgenic Res 19:165–180CrossRefGoogle Scholar
  21. Garcia-Casal MN, Layrisse M, Solano L, Baron MA, Arguello F, Llovera D, Ramirez J, Leets I, Tropper E (1998) Vitamin A and beta-carotene can improve non-heme iron absorption from rice, wheat and corn by humans. J Nutr 128:646–650PubMedGoogle Scholar
  22. Garcia-Oliveira AL, Tan L, Fu Y, Sun C (2009) Genetic identification of quantitative trait loci for contents of mineral nutrients in rice grain. J Plant Biol 51(1):84–92CrossRefGoogle Scholar
  23. Gibson GR, Beatty ER, Wang X, Cummings JH (1995) Selective stimulation of bifidobacteria in the human colon by oligofructose and inulin. Gastroenterology 108:975–982PubMedCrossRefGoogle Scholar
  24. Gillooly M, Bothwell TH, Torrance JD, Macphail AP, Derman DP, Bezwoda WR, Mills W, Charlton RW, Mayet F (1983) The effects of organic acids, phytates and polyphenols on the absorption of iron from vegetables. Br J Nutr 49:331–342PubMedCrossRefGoogle Scholar
  25. Goto F, Yoshihara T (2001) Improvement of micronutrient contents by genetic engineering – development of high iron content crops. Plant Biotechnol 18:7–15CrossRefGoogle Scholar
  26. Goto FT, Yoshihara T, Shigemoto N, Toki S, Takaiwa F (1999) Iron fortification of rice seed by the soybean Ferritin gene. Nat Biotechnol 17:282–286PubMedCrossRefGoogle Scholar
  27. Gowda SJM, Randhawa GJ, Bisht IS, Firke PK, Singh AK, Abraham Z, Dhillon BS (2012) Morpho-agronomic and simple sequence repeat-based diversity in colored rice (Oryza sativa L.) germplasm from peninsular India. Genet Resour Crop Evol 59:179–189CrossRefGoogle Scholar
  28. Graham RD, Senadhira D, Beebe S, Iglesias C, Monasterio I (1999) Breeding for micronutrient density in edible portions of staple food crops: conventional approaches. Field Crops Res 60:57–80CrossRefGoogle Scholar
  29. Gregorio GB (2002) Progress in breeding for trace elements in staple crops. J Nutr 132:500S–502SPubMedGoogle Scholar
  30. Gregorio GB, Senadhira D, Htut T, Graham RD (1999) Improving iron and zinc value of rice for human nutrients. Agric Dev 23(9):68–87Google Scholar
  31. Gregorio GB, Senadhira D, Htut T, Graham RD (2000) Breeding for trace mineral density in rice. Food Nutr Bull 21:382–386Google Scholar
  32. Grotz N, Guerinot ML (2006) Molecular aspects of Cu, Fe and Zn homeostasis in plants. Biochem Biophys Acta 1763:595–608PubMedCrossRefGoogle Scholar
  33. Guerinot ML (2007) It’s elementary: enhancing Fe3+ reduction improves rice yields. Proc Natl Acad Sci USA 104:7311–7312PubMedCrossRefGoogle Scholar
  34. Gura T (1999) New genes boost rice nutrients. Science 285:994–995PubMedCrossRefGoogle Scholar
  35. Guttieri M, Bowen D, Dorsch JA, Raboy V, Souza E (2003) Identification and characterization of a low phytic acid wheat. Crop Sci 44:418–424CrossRefGoogle Scholar
  36. Harper JF, Surowy TK, Sussman MR (1989) Molecular cloning and sequence of cDNA encoding the plasma membrane proton pump (H+-ATPase) of Arabidopsis thaliana. Proc Natl Acad Sci USA 86:1234–1238PubMedCrossRefGoogle Scholar
  37. 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 36:366–381PubMedCrossRefGoogle Scholar
  38. 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 104:7373–7378PubMedCrossRefGoogle Scholar
  39. Islam FMA, Basford KE, Jara C, Redden RJ, Beebe SE (2002) Seed compositional and disease resistance differences among gene pools in cultivated common bean. Genet Resour Crop Evol 49:285–293. doi: 10.1023/A:1015510428026 CrossRefGoogle Scholar
  40. 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(9):e24476. doi: 10.1371/journal.pone.0024476 PubMedCentralPubMedCrossRefGoogle Scholar
  41. Kaiyang L, Lanzhi L, Zheng X, Zhihong T, Zhonglt H (2008) Quantitative trait loci controlling Cu, Ca, Zn, Mn and Fe content in rice grains. Indian Acad Sci 87:305–310Google Scholar
  42. Khush GS, Brar DS (2002) Biotechnology for rice breeding: progress and impact. In: Sustainable rice production for food security. Proceedings of the 20th session of the international rice commission, Bangkok, Thailand, 23–26 July 2002Google Scholar
  43. Kumar J, Chawla A, Kumar P, Jain RK (2012) Iron and zinc variability in twenty rice (Oryza sativa L.) genotypes. Ann Biol 28(2):90–92Google Scholar
  44. Kuwano M, Ohyama A, Tanaka Y, Mimura T, Takaiwa F, Yoshida KT (2006) Molecular breeding for transgenic rice with low-phytic-acid phenotype through manipulating myo-inositol 3-phosphate synthase gene. Mol Breed 18:263–272CrossRefGoogle Scholar
  45. Larson SR, Rutger JN, Young KA, Raboy V (2000) Isolation and genetic mapping of a non-lethal rice (Oryza sativa L.) low phytic acid 1mutation. Crop Sci 40:1397–1405CrossRefGoogle Scholar
  46. Lee S, An G (2009) Over-expression of OsIRT1 leads to increased iron and zinc accumulations in rice. Plant Cell Environ 32:408–416PubMedCrossRefGoogle Scholar
  47. Li ZK (2001) QTL mapping in rice: a few critical considerations. In: Brar DS, Hardy B, Khush GS (eds) Rice genetics IV. Science Publisher, EnfieldGoogle Scholar
  48. Liu ZC, Yu SX (1997) Nutrition and food sanitation, 2nd edn. Demotic Sanitation Publisher, BeijingGoogle Scholar
  49. Lopez HW, Leenhardt F, Coudray C, Remesy C (2002) Minerals and phytic acid interactions: is it a real problem for human nutrition? Int J Food Sci Technol 37:727–739CrossRefGoogle Scholar
  50. Lu T, Huang X, Zhu C, Huang T, Zhao Q, Xie K (2008) RICD: a rice indica cDNA database resource for rice functional genomics. BMC Plant Biol 8:118PubMedCentralPubMedCrossRefGoogle Scholar
  51. Lucca P, Hurell R, Potrykus I (2001) Genetic engineering approaches to improve the bioavailability and the level of iron in rice grains. Theor Appl Genet 102:392–397CrossRefGoogle Scholar
  52. Mares-Perlman JA, Subar AF, Block G, Greger JL, Luby MH (1995) Zinc intake and sources in the US adult population: 1976–1980. J Am Coll Nutr 14:349–357PubMedCrossRefGoogle Scholar
  53. Maziya-Dixon B, Kling JG, Menkir A, Dixon A (2000) Genetic variation in total carotene, iron and zinc contents of maize and cassava genotypes. Food Nutr Bull 21:419–422Google Scholar
  54. Monasterio I, Graham RD (2000) Breeding for trace minerals in wheat. Food Nutr Bull 21:392–396Google Scholar
  55. Norton GJ, Deacon CM, Xiong L, Huang S, Meharg AA, Price AH (2010) Genetic mapping of the rice ionome in leaves and grain: identification of QTLs for 17 elements including arsenic, cadmium, iron and selenium. Plant Soil 329:139–153CrossRefGoogle Scholar
  56. Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, Hinchliffe E, Adams JL, Silverstone AL, Rachel DR (2005) Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nat Biotechnol 23:482–487PubMedCrossRefGoogle Scholar
  57. Palmer CM, Guerinot ML (2009) Facing the challenges of Cu, Fe and Zn homeostasis in plants. Nat Chem Biol 5:333–340PubMedCentralPubMedCrossRefGoogle Scholar
  58. Pfeiffer WH, McClafferty B (2007) Harvest Plus: breeding crops for better nutrition. Crop Sci 47(S3):S88–S105Google Scholar
  59. Pflieger S, Lefebvre V, Causse M (2001) The candidate gene approach in plant genetics: a review. Mol Breed 7(4):275–291CrossRefGoogle Scholar
  60. Puig S, Andres-Colas N, Garcia-Molina A, Penarrubia L (2007) Copper and iron homeostasis in Arabidopsis: responses to metal deficiencies, interactions and biotechnological applications. Plant Cell Environ 30:271–290PubMedCrossRefGoogle Scholar
  61. Rabbani GH, Ali M (2009) New ideas and concepts, rice bran: a nutrient-dense mill-waste for human nutrition. The ORION Med J 32(3):694–701Google Scholar
  62. Raboy V (2001) Seeds for a better future: ‘low phytate’, grains help to overcome malnutrition and reduce pollution. Trends Plant Sci 6:458–462PubMedCrossRefGoogle Scholar
  63. Raboy V (2002) Progress in breeding low phytate crops. J Nutr 132:503S–505SPubMedGoogle Scholar
  64. Robinson NJ, Procter CM, Connolly EL, Guerinot ML (1999) A ferric-chelate reductase for iron uptake from soils. Nature 397:694–697PubMedCrossRefGoogle Scholar
  65. Scholz-Ahrens KE, Schrezenmeir J (2002) Inulin, oligofructose and mineral metabolism—experimental data and mechanism. Br J Nutr 87:S179–S186PubMedCrossRefGoogle Scholar
  66. Sellappan K, Datta K, Parkhi V, Datta SK (2009) Rice caryopsis structure in relation to distribution of micronutrients (iron, zinc, β-carotene) of rice cultivars including transgenic indica rice. Plant Sci 177:557–562CrossRefGoogle Scholar
  67. Seymour J (1996) Hungry for a new revolution. New Sci 149(2023):32–37Google Scholar
  68. Shaw JG, Friedman JF (2011) Iron deficiency anemia: focus on infectious diseases in lesser developed countries. Anemia. Review Article 2011:10. doi: 10.1155/2011/260380
  69. Sperotto RA, Boffa T, Duartea GL, Santosb LS, Grusakc MA, Fett JP (2010) Identification of putative target genes to manipulate Fe and Zn concentrations in rice grains. J Plant Physiol 167:1500–1506PubMedCrossRefGoogle Scholar
  70. Stangoulis JCR, Huynh BL, Welch RM, Choi EY, Graham RD (2007) Quantitative trait loci for phytate in rice grain and their relationship with grain micronutrient content. Euphytica 154:289–294CrossRefGoogle Scholar
  71. Stein AJ, Sachdev HPS, Qaim M (2008) Genetic engineering for the poor: golden rice and public health in India. World Dev 3:144–158CrossRefGoogle Scholar
  72. Takahashi M (2003) Overcoming Fe deficiency by a transgenic approach in rice. Plant Cell Tissue Organ Cult 72:211–220CrossRefGoogle Scholar
  73. Takahashi M, Terada Y, Nakai I, Nakanishi H, Yoshimura E, Mori S, Nishizawa NK (2003) Role of nicotianamine in the intracellular delivery of metals and plant reproductive development. Plant Cell 15:1263–1280PubMedCentralPubMedCrossRefGoogle Scholar
  74. Tang G, Hu Y, Yin S, Wang Y, Dallal GE, Grusak MA, Russell RM (2012) β-carotene in golden rice is as good as β-carotene in oil at providing vitamin A to children. Am J Clin Nutr 96:658–664PubMedCrossRefGoogle Scholar
  75. The World Health Organization (WHO) (2011) Micronutrient deficiencies: iron deficiency anemia. Available from:
  76. Thomine S, Wang R, Ward JM, Crawford NM, Schroeder JI (2000) Cadmium and iron transport by members of a plant transporter gene family in Arabidopsis with homology to NRAMP genes. Proc Natl Acad Sci USA 97:4991–4996PubMedCrossRefGoogle Scholar
  77. Thorup GL, Kearsey FD (2000) The principles of QTL analysis (a minimal mathematics approach). J Exp Bot 49:1619–1623Google Scholar
  78. Tuberosa R, Salvi S (2007) From QTLs to genes controlling root traits in maize: scale and complexity in plant systems research. Gene-Plant-Crop Relat 2:15–24Google Scholar
  79. Ullah AHJ, Mullaney EJ (1996) Disulfide bonds are necessary for structure and activity in Aspergillus ficuum phytase. Biochem Biophys Res Commun 227:311–317PubMedCrossRefGoogle Scholar
  80. United Nations Children’s Fund (UNICEF) (2009) Vitamin A deficiency: the challenge. Available from:
  81. Vasconcelos M, Datta K, Oliva N, Khalekuzzaman M, Torrizo L, Krishnan S, Margarida O, Goto F, Datta S (2003) Enhanced iron and zinc accumulation in transgenic rice with the ferritin gene. Plant Sci 164:371–378CrossRefGoogle Scholar
  82. Vasconcelos M, Eckert H, Arahana V, Graef G, Grusak MA, Clemente T (2006) Molecular and phenotypic characterization of transgenic soybean expressing the Arabidopsis ferric chelate reductase gene, FRO2. Planta 224:1116–1128PubMedCrossRefGoogle Scholar
  83. Velu G, Rai KN, Muralidharan V, Kulkarni VN, Longvah T, Raveendran TS (2007) Prospects of breeding biofortified pearl millet with high grain iron and zinc content. Plant Breed 126:182–185CrossRefGoogle Scholar
  84. Welch RM, Graham RD (2004) Breeding for micronutrients in staple food crops from a human nutrition perspective. J Exp Bot 55:353–364PubMedCrossRefGoogle Scholar
  85. White PJ, Broadley MR (2005) Biofortifying crops with essential mineral elements. Trends Plant Sci 10:586–593PubMedCrossRefGoogle Scholar
  86. Wirth J, Poletti S, Aeschlimann B, Yakandawala N, Drosse B, Osorio S (2009) Rice endosperm iron biofortification by targeted and synergistic action of nicotianamine synthase and ferritin. Plant Biotechnol J 7:631–644PubMedCrossRefGoogle Scholar
  87. Ye X, Babili AS, Kloeti A, Zhang J, Lucca P, Beyer P, Potrykus I (2000) Engineering the provitamin A (β-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287:303–305PubMedCrossRefGoogle Scholar
  88. Zhu C, Naqvi S, Gomez-Galera S, Pelacho AM, Capell T, Christou P (2007) Transgenic strategies for the nutritional enhancement of plants. Trends Plant Sci 12:548–555, Accessed 24 July 2013PubMedCrossRefGoogle Scholar

Copyright information

© Springer India 2013

Authors and Affiliations

  • Rajinder K. Jain
    • 1
  • Jitender Kumar
    • 1
  • Sunita Jain
    • 2
  • Vijay K. Chowdhury
    • 1
  1. 1.Department of Molecular Biology and BiotechnologyCCS Haryana Agricultural UniversityHisarIndia
  2. 2.Bioinformatics SectionCCS Haryana Agricultural UniversityHisarIndia

Personalised recommendations