Skip to main content
SpringerLink
Log in

Iron accumulation does not parallel the high expression level of ferritin in transgenic rice seeds

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

To answer the question whether iron accumulation in transgenic rice seeds depends on the expression level of exogenous soybean ferritin, we generated two kinds of ferritin hyper-expressing rice lines by introducing soybean ferritin SoyferH-1 gene under the control of the rice seed storage glutelin gene promoter, GluB-1 and the rice seed storage globulin gene promoter, Glb-1, (GluB-1/SoyferH-1 and Glb-1/SoyferH-1, DF lines), and by introducing the SoyferH-1 gene under the control of Glb-1 promoter alone (Glb-1/SoyferH-1, OF lines). Ferritin expression was restricted to the endosperm in both lines and protein levels determined by western blot analysis were up to 13-fold higher than in a construct previously reported FK22 (GluB-1/SoyferH-1, in genetically Kitaake bachground); however, the maximum iron concentrations in seeds of both of the new lines were only about 30% higher than FK22. The maximum iron concentration in the OF and DF lines was about threefold higher than in the non-transformant. The mean Fe concentration in leaves of ferritin over-expressing lines decreased to less than half of the non-transformant while that the plant biomasses and seed yields of the ferritin-transformed lines were not significantly different from those of the non-transformant, suggesting that accumulation of Fe in seeds of hyper-expression ferritin rice did not always depend on the expression level of exogenous ferritin but may have been limited by Fe uptake and transport. No obvious differences were observed for other divalent-metal concentrations (Ca, Cd, Cu, Mg, Mn and Zn) in the seeds among all experimental lines and non-transformant.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

bp:

Base pair

CTAB:

Cetyl trimethyl ammonium bromide

DF:

Double transformation line with GluB-1/SoyferH-1 and Glb-1/SoyferH-1

NT:

Non-transformant

OF:

Single transformation line with Glb-1/SoyferH-1

PCR:

Polymerase chain reaction

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

References

  • Beard JL, Burton JW, Theil EC (1996) Purified ferritin and soybean meal can be sources of iron for treating iron deficiency in rats. J Nutr 126:154–160

    Google Scholar 

  • Becker D, Kemper E, Schell J, Masterson R (1992) New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol Biol 20:1195–1197

    CAS  PubMed  Google Scholar 

  • Briat JF, Lobréaux S (1997) Iron transport and storage in plants. Trends Plant Sci 2:187–193

    Google Scholar 

  • Cohen KC, Fox TC, Garvin DF, Kochian LV (1998) The role of iron-deficiency stress in stimulating heavy-metal transport in plants. Plant Physiol 116:1063–1672

    Google Scholar 

  • Delhaize E (1996) A metal-accumulator mutant of Arabidopsis thaliana. Plant Physiol 111:549–855

    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

    Google Scholar 

  • Finnegan EJ, Peacock WJ, Dennis ES (2000) DNA methylation, a key regulator of plant development and other processes. Curr Opin Genet Dev 10:217–223

    Google Scholar 

  • Flavell RB (1994) Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc Natl Acad Sci USA 91:3490–3496

    CAS  PubMed  Google Scholar 

  • Gillespie S, Haddad L (2001) Malnutrition in Asia and the Pacific. In: Nutrition and development series, attacking the double burden of malnutrition in Asia and the Pacific. Asian Development Bank, International Food Policy Research Institute, pp 5–14

  • Goto F, Yoshihara T, Saiki H (1998) Iron accumulation in tobacco plants expressing soybean ferritin gene. Transgenic Res 7:173–180

    Google Scholar 

  • Goto F, Yoshihara T, Shigemoto N, Toki S, Takaiwa F (1999) Iron fortification of rice seed by the soybean ferritin gene. Nat Biotech 17:282–286

    Google Scholar 

  • Grusak MA, Welch RM, Kochian LV (1990) Physiological characterization of a single-gene mutant of Pisum sativum L. exhibiting excess iron accumulation. I. Root iron reduction and iron uptake. Plant Physiol 93:976–981

    Google Scholar 

  • Guerinot ML, Yi Y (1994) Iron: nutritious, noxious, and not readily available. Plant Physiol 104:815–820

    Google Scholar 

  • Harrison P, Arosio P (1996) The ferritin: molecular properties iron storage function and cellular regulation. Biochim Biophys Acta 1275:161–203

    CAS  PubMed  Google Scholar 

  • Himelblau E, Amasino RM (2001) Nutrients mobilized from leaves of Ababidopsis thaliana during leaf senescence. J Plant Physiol 158:1317–1323

    Google Scholar 

  • Kokot S, Phuong TD (1999) Elemental content of Vietnamese rice. Part2. Multivariate data analysis. Analyst 124:561–569

    Google Scholar 

  • Laulhere JP, Lescure AM, Briat JF (1988) Purification and characterization of ferritins from maize, pea, and soybean seeds. J Biol Chem 263:10298–10294

    Google Scholar 

  • Lucca P, Hurrell R, Potrykus I (2001) Genetic engineering approaches to improve the bioavailability and the level of iron in rice grains. Theor Appl Genet 102:392–397

    Google Scholar 

  • Lueders T, Friedrich M (2000) Archaeal population dynamics during sequential reduction processes in rice field soil. Appl Environ Microbiol 66:2732–2742

    Google Scholar 

  • Mauk CS, Nooden LD (1992) Regulation of mineral redistribution in pod-bearing soybean explants. J Exp Bot 43:1429–1440

    Google Scholar 

  • Meins FJ (2000) RNA degradation and models for post-transcriptional gene silencing. Plant Mol Biol 43:261–273

    Google Scholar 

  • Meyer P, Saedler H (1996) Homology-dependent gene silencing in plants. Annu Rev Plant Mol Biol 47:23–48

    Google Scholar 

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

    Google Scholar 

  • Murray-Kolb LE, Theil EC, Takaiwa F, Goto F, Yoshihara T, Beard JL (2002) Transgenic rice is a source of iron for iron-depleted rats. J Nutr 132:957–960

    Google Scholar 

  • Proudhon D, Briat JF, Lescure AM (1989) Iron induction of ferritin synthesis in soybean. Plant Physiol 90:586–590

    Google Scholar 

  • Qu LQ, Takaiwa F (2004) Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice. Plant Biotech J 2:113–125

    Google Scholar 

  • Qu LQ, Wei XL, Satoh H, Kumamaru T, Ogawa M, Takaiwa F (2002). Inheritance of alleles for glutelin α-2 subunit genes in rice and identification of their corresponding cDNA clone. Theor Appl Genet 105:1099–1108

    Google Scholar 

  • Qu LQ, Tada Y, Takaiwa F (2003) In situ western hybridization: a new highly sensitive technique to detect protein distribution in seeds. Plant Cell Rep 22:282–285

    Google Scholar 

  • Theil EC (1987) Ferritin: structure, gene regulation and cellular function in animals, plants and microorganisms. Annu Rev Biochem 56:289–315

    Google Scholar 

  • Theil EC (2004) Iron, ferritin, and nutrition. Annu Rev Nutr 24:327–343

    Google Scholar 

  • US Department of Agriculture, US Department of Health and Human Services Nutrition and your health (1995) Dietary guidelines for Americans, 4th edn. US Government Printing Office, Home and Garden Bulletin no. 232, Washington

  • Van Wuytsuwinkel O, Vansuyt G, Grignon N, Fourcroy P, Briat JF (1999) Iron homeostasis alteration in transgenic tobacco overexpressing ferritin. Plant J 17:93–97

    Google Scholar 

  • Vansuyt G, Mench M, Briat JF (2000) Soil-dependent variability of leaf iron accumulation in transgenic tobacco overexpressing ferritin. Plant Physiol Biochem 38:499–506

    Google Scholar 

  • 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

    Google Scholar 

  • Welch RM, Norvell WA, Schaefer SC, Shaff JE, Kochian LV (1993) Induction of iron (III) and copper (II) reduction in pea (Pisum sativum L.) roots by Fe and Cu status: does the root cell plasmalemma Fe (III)-chelate reductase perform a general role in regulating cation uptake? Planta 190:555–561

    Google Scholar 

  • Yoshihara T, Masuda T, Jiang T, Goto T, Mori S, Nishizawa NK (2003) Analysis of some divalent metal concentrations in tobacco expressing the exogenous soybean ferritin gene. J Plant Nutr 26:2253–2265

    Google Scholar 

Download references

Acknowledgements

We thank Ms. K. Tsunokwa and S. Sumino (CRIEPI) and Ms. M. Utsuno, Y. Suzuki and X. Wang (NIAS) for their outstanding technical assistance. This research was supported in part by a research grant from the Ministry of Agriculture, Forestry and Fishery of Japan to F. Takaiwa, Core Research for Evolutional Science and Technology (CREST) and by a Grant-in-aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) to T. Yoshihara.

Author information

Authors and Affiliations

  1. Laboratory of Genetic Engineering, Department of Plant Biotechnology, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan

    Le Qing Qu, Akio Ooyama & Fumio Takaiwa

  2. Plant Molecular Biology Group, Laboratory of Environmental Science, Central Research Institute Electric Power Industry, 1646 Abiko, Chiba, 270-1194, Japan

    Toshihiro Yoshihara & Fumiyuki Goto

  3. Institute of Botany, The Chinese Academy of Sciences, Beijing, 100093, China

    Le Qing Qu

Authors
  1. Le Qing Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Toshihiro Yoshihara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Akio Ooyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fumiyuki Goto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fumio Takaiwa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Toshihiro Yoshihara or Fumio Takaiwa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Qu, L.Q., Yoshihara, T., Ooyama, A. et al. Iron accumulation does not parallel the high expression level of ferritin in transgenic rice seeds. Planta 222, 225–233 (2005). https://doi.org/10.1007/s00425-005-1530-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-005-1530-8

Keywords

Search

Navigation