Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

NRT/PTR transporters are essential for translocation of glucosinolate defence compounds to seeds

Nature volume 488, pages 531–534 (2012)Cite this article

Subjects

Abstract

In plants, transport processes are important for the reallocation of defence compounds to protect tissues of high value1, as demonstrated in the plant model Arabidopsis, in which the major defence compounds, glucosinolates2, are translocated to seeds on maturation3. The molecular basis for long-distance transport of glucosinolates and other defence compounds, however, remains unknown. Here we identify and characterize two members of the nitrate/peptide transporter family, GTR1 and GTR2, as high-affinity, proton-dependent glucosinolate-specific transporters. The gtr1 gtr2 double mutant did not accumulate glucosinolates in seeds and had more than tenfold over-accumulation in source tissues such as leaves and silique walls, indicating that both plasma membrane-localized transporters are essential for long-distance transport of glucosinolates. We propose that GTR1 and GTR2 control the loading of glucosinolates from the apoplasm into the phloem. Identification of the glucosinolate transporters has agricultural potential as a means to control allocation of defence compounds in a tissue-specific manner.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Identification of Arabidopsis transporters with glucosinolate uptake activity.
Figure 2: Biophysical and biochemical transport properties of GTRs.
Figure 3: Glucosinolate analysis of gtr1 and gtr2 knockout mutants.
Figure 4: Localization of GTR1 and GTR2.

Similar content being viewed by others

References

  1. Feeny, P. in Biochemical Interaction Between Plants and Insects (eds Wallace, J. W. & Mansel, R. L. ) 1–40 (Plenum, 1976)

    Book  Google Scholar 

  2. Halkier, B. A. & Gershenzon, J. Biology and biochemistry of glucosinolates. Annu. Rev. Plant Biol. 57, 303–333 (2006)

    Article  CAS  Google Scholar 

  3. Nour-Eldin, H. & Halkier, B. Piecing together the transport pathway of aliphatic glucosinolates. Phyochemistry Rev. 8, 53–67 (2009)

    Article  CAS  Google Scholar 

  4. McKey, D. Adaptive patterns in alkaloid physiology. Am. Nat. 108, 305–320 (1974)

    Article  ADS  Google Scholar 

  5. Züst, T., Joseph, B., Shimizu, K. K., Kliebenstein, D. J. & Turnbull, L. A. Using knockout mutants to reveal the growth costs of defensive traits. Proc. Biol. Sci. 278, 2598–2603 (2011)

    Article  Google Scholar 

  6. Ohnmeiss, T. E. & Baldwin, I. T. Optimal defense theory predicts the ontogeny of an induced nicotine defense. Ecology 81, 1765–1783 (2000)

    Article  Google Scholar 

  7. Wink, M. Wounding-induced increase of quinolizidine alkaloid accumulation in lupin leaves. Z. Naturforschung 38, 905–909 (1983)

    Article  Google Scholar 

  8. Brown, P. D., Tokuhisa, J. G., Reichelt, M. & Gershenzon, J. Variation of glucosinolate accumulation among different organs and developmental stages of Arabidopsis thaliana. Phytochemistry 62, 471–481 (2003)

    Article  CAS  Google Scholar 

  9. Townsend, B. J. & Llewellyn, D. J. Reduced terpene levels in cottonseed add food to fiber. Trends Biotechnol. 25, 239–241 (2007)

    Article  CAS  Google Scholar 

  10. Mailer, R., McFadden, A., Ayton, J. & Redden, B. Anti-nutritional components, fibre, sinapine and glucosinolate content, in Australian canola (Brassica napus L.) meal. J. Am. Oil Chem. Soc. 85, 937–944 (2008)

    Article  CAS  Google Scholar 

  11. Wu, S. & Chappell, J. Metabolic engineering of natural products in plants; tools of the trade and challenges for the future. Curr. Opin. Biotechnol. 19, 145–152 (2008)

    Article  CAS  Google Scholar 

  12. Enneking, D. & Wink, M. in Proceedings of the Third International Food Legumes Research Conference, Linking Research and Marketing Opportunities for Pulses in the 21st Century (ed. Knight, R. ) 671–683 (Kluwer, 2000)

    Google Scholar 

  13. Chen, S., Petersen, B. L., Olsen, C. E., Schulz, A. & Halkier, B. A. Long-distance phloem transport of glucosinolates in Arabidopsis. Plant Physiol. 127, 194–201 (2001)

    Article  CAS  Google Scholar 

  14. Ellerbrock, B. L., Kim, J. H. & Jander, G. Contribution of glucosinolate transport to Arabidopsis defense responses. Plant Signal. Behav. 2, 282–283 (2007)

    Article  Google Scholar 

  15. Nour-Eldin, H. H., Norholm, M. H. & Halkier, B. A. Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes. Plant Methods 2, 17–25 (2006)

    Article  Google Scholar 

  16. Wang, Y. Y. & Tsay, Y. F. Arabidopsis nitrate transporter NRT1.9 is important in phloem nitrate transport. Plant Cell 23, 1945–1957 (2011)

    Article  CAS  Google Scholar 

  17. Fan, S. C., Lin, C. S., Hsu, P. K., Lin, S. H. & Tsay, Y. F. The Arabidopsis nitrate transporter NRT1.7, expressed in phloem, is responsible for source-to-sink remobilization of nitrate. Plant Cell 21, 2750–2761 (2009)

    Article  CAS  Google Scholar 

  18. Bednarek, P. et al. A glucosinolate metabolism pathway in living plant cells mediates broad-spectrum antifungal defense. Science 323, 101–106 (2009)

    Article  ADS  CAS  Google Scholar 

  19. Koroleva, O. A. et al. Identification of a new glucosinolate-rich cell type in Arabidopsis flower stalk. Plant Physiol. 124, 599–608 (2000)

    Article  CAS  Google Scholar 

  20. Balbi, V. & Devoto, A. Jasmonate signalling network in Arabidopsis thaliana: crucial regulatory nodes and new physiological scenarios. New Phytol. 177, 301–318 (2008)

    Article  CAS  Google Scholar 

  21. Foster, R., Williamson, C. S. & Lunn, J. Culinary oils and their health effects. Nutrition Bulletin 34, 4–47 (2009)

    Article  Google Scholar 

  22. Snowdon, R., Luhs, W. & Friedt, W. in Genome Mapping and Molecular Breeding in Plants (ed. Kole, C. ) 55–114 (Springer, 2007)

    Google Scholar 

  23. Wittkop, B., Snowdon, R. & Friedt, W. Status and perspectives of breeding for enhanced yield and quality of oilseed crops for Europe. Euphytica 170, 131–140 (2009)

    Article  Google Scholar 

  24. Clarke, D. B. Glucosinolates, structures and analysis in food. Anal. Methods 2, 310–325 (2010)

    Article  CAS  Google Scholar 

  25. Zhang, J. et al. Metabolite profiling of Arabidopsis seedlings in response to exogenous sinalbin and sulfur deficiency. Phytochemistry 72, 1767–1778 (2011)

    Article  CAS  Google Scholar 

  26. Gottwald, J. R., Krysan, P. J., Young, J. C., Evert, R. F. & Sussman, M. R. Genetic evidence for the in planta role of phloem-specific plasma membrane sucrose transporters. Proc. Natl Acad. Sci. USA 97, 13979–13984 (2000)

    Article  ADS  CAS  Google Scholar 

  27. Vert, G. et al. IRT1, an Arabidopsis transporter essential for iron uptake from the soil and for plant growth. Plant Cell 14, 1223–1233 (2002)

    Article  CAS  Google Scholar 

  28. Nour-Eldin, H. H., Hansen, B. G., Norholm, M. H. H., Jensen, J. K. & Halkier, B. A. Advancing uracil-excision based cloning towards an ideal technique for cloning PCR fragments. Nucleic Acids Res. 34, e122 (2006)

    Article  Google Scholar 

  29. Geu-Flores, F., Nour-Eldin, H. H., Nielsen, M. T. & Halkier, B. A. USER fusion: a rapid and efficient method for simultaneous fusion and cloning of multiple PCR products. Nucleic Acids Res. 35, e55 (2007)

    Article  Google Scholar 

  30. Nørholm, M. A mutant Pfu DNA polymerase designed for advanced uracil-excision DNA engineering. BMC Biotechnol. 10, 21–27 (2010)

    Article  Google Scholar 

Download references

Acknowledgements

We thank M. Palmgren, A. Schulz and D. Kliebenstein for comments on the manuscript and H. K. S. Jepsen, L. B. Møller and A. Kraus for technical assistance. We thank D. Klærke and technicians T. K. Soland, B. L. Christensen, M. Olsen and C. Derrer for providing X. laevis oocytes. We thank the Salk, Stanford, Plant Gene Expression Center (SSP) Consortium and the RIKEN Genome Science Center for providing full-length cDNAs. Imaging data were achieved at the Center for Advanced Bioimaging, University of Copenhagen. H.H.N-.E. was supported by the Danish Research Council for Technology and Production (FTP) grant 09-065827/274-08-0354. M.B. was supported by a Marie Curie fellowship (grant PIEF-GA-2008-221236). I.D. was supported by a Marie-Curie Career Integration Grant of the European Union (FP7-PEOPLE-2011-CIG No. 303674 (Regopoc)). D.G. was supported by the Deutsche Forschungsgemeinschaft grant GE2195/1-1 and R.H. by Deutsche Forschungsgemeinschaft grants within FOR 1061. B.A.H. is partner of The VKR Research Centre for Pro-Active Plants funded by the Villum Kann Rasmussen Foundation. Funding for DynaMo Center of Excellence is provided by the Danish National Research Foundation.

Author information

Author notes

  1. Hussam Hassan Nour-Eldin and Tonni Grube Andersen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Plant and Environmental Sciences, VKR Research Centre for Pro-Active Plants and DynaMo Center of Excellence, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark,

    Hussam Hassan Nour-Eldin, Tonni Grube Andersen, Meike Burow, Svend Roesen Madsen, Morten Egevang Jørgensen, Carl Erik Olsen & Barbara Ann Halkier

  2. Centre for Plant Biotechnology and Genomics, Universidad Politécnica de Madrid, Campus de Montegancedo, Carretera M-40, E-28223 Pozuelo de Alarcón (Madrid), Spain,

    Ingo Dreyer

  3. Julius-von-Sachs Institute, Molecular Plant Physiology and Biophysics, University of Würzburg Julius-von-Sachs-Platz 2, Würzburg, D-97082, Germany

    Rainer Hedrich & Dietmar Geiger

Authors
  1. Hussam Hassan Nour-Eldin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tonni Grube Andersen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Meike Burow
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Svend Roesen Madsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Morten Egevang Jørgensen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Carl Erik Olsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ingo Dreyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Rainer Hedrich
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dietmar Geiger
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Barbara Ann Halkier
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

H.H.N-.E. identified the GTRs, designed the study, performed the phylogenetic analyses, contributed to biochemical characterization, data analysis, confocal localization experiments and analyses of metabolite profiling data and wrote the paper. T.G.A. performed the confocal localization experiments, generated and characterized glucosinolate in the gtr knockout mutants and contributed to biochemical characterization, study design and preparation of the manuscript. M.B. purified glucosinolates necessary for biochemical characterization and contributed to study design, data analysis and preparation of the manuscript. S.R.M. generated and analysed GTR-promoter-β-glucuronidase lines, contributed to confocal localization experiments and performed germination experiments. M.E.J. performed metabolite profiling of seeds and analysed data and performed seed morphology and yield analyses. C.E.O. performed LC–MS analyses for the transporter library screen and for metabolite profiling. I.D. performed theoretical discussion on GTR kinetics. R.H. contributed to the study design and data analyses. D.G. performed the biophysical characterization of the GTR transport mechanism and contributed to the theoretical discussion on GTR kinetics, study design and preparation of the manuscript. B.A.H. contributed to and supervised the study design and contributed to data analyses and preparation of the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Barbara Ann Halkier.

Ethics declarations

Competing interests

The subject matter of this publication forms the basis of a patent application USSN 61/362,390 and EP10075299.7.

Supplementary information

Supplementary Information 1

This file contains Supplementary Figures 1, 3-7 and legend for Supplementary Figure 2 (see separate file for Supplementary Figure 2), additional references, Supplementary Materials and Methods, Supplementary References, Supplementary Table 1, a Supplementary Discussion and full legends for Supplementary Movies 1-4. (PDF 1294 kb)

Supplementary Information 2

This file contains Supplementary Figure 2 (see Supplementary Information file for legend). (PDF 2312 kb)

Supplementary Movie 1

Representative movie of z-stack spanning 27.0 μm of five-week-old leaves from stable Arabidopsis lines transformed with pGTR1(2kb)-GTR1(genomic fragment)-YFP(venus)-GTR1_3’UTR(0.3kb) (seven independent lines), showing plasma membrane localization of GTR1. (MOV 1303 kb)

Supplementary Movie 2

This movie shows background YFP fluorescence in non-transformed plants. (MOV 737 kb)

Supplementary Movie 3

Representative movie of a z-stack spanning 16.4 μm of five-week-old leaves from stable Arabidopsis lines transformed with pGTR2(2kb)-GTR2 (genomic fragment)-mOrange-GTR2_3'UTR(0.3kb) (22 independent lines), showing plasma membrane localization of GTR2. (MOV 862 kb)

Supplementary Movie 4

This movie shows background mOrange fluorescence in non-transformed plants. (MOV 275 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nour-Eldin, H., Andersen, T., Burow, M. et al. NRT/PTR transporters are essential for translocation of glucosinolate defence compounds to seeds. Nature 488, 531–534 (2012). https://doi.org/10.1038/nature11285

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature11285

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing