Letter | Published:

Arabidopsis NITROGEN LIMITATION ADAPTATION regulates ORE1 homeostasis during senescence induced by nitrogen deficiency

Nature Plantsvolume 4pages898903 (2018) | Download Citation


Nitrogen is an important macronutrient in plants and its deficiency induces rapid leaf senescence. Two genes, ORE1 and NITROGEN LIMITATION ADAPTATION (NLA), have been implicated in regulating the senescence process but their relationship is unclear1,2. Here, we show that nla and pho2 (also known as ubc24) plants develop rapid leaf senescence under nitrogen-starvation condition, whereas ore1 and nla/ore1 and pho2 (ubc24)/ore1 plants stay green. These results suggest that ORE1 acts downstream of NLA and PHO2 (UBC24). NLA interacts with ORE1 in the nucleus and regulates its stability through polyubiquitination using PHO2 (UBC24) as the E2 conjugase. Our findings identified ORE1 as a downstream target of NLA/PHO2 (UBC24) and showed that post-translational regulation of ORE1 levels determines leaf senescence during nitrogen deficiency.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Data availability

All data generated or analysed in this study are included in this article and Supplementary Information files. The data are available from the corresponding author upon request.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


  1. 1.

    Kant, S. et al. Genetic regulation by NLA and microRNA827 for maintaining nitrate-dependent phosphate homeostasis in Arabidopsis. PLoS Genet. 7, e1002021 (2011).

  2. 2.

    Kim, J. H. et al. Trifurcate feed-forward regulation of age-dependent cell death involving miR164 in Arabidopsis. Science 323, 1053–1057 (2009).

  3. 3.

    Gan, S. et al. Making sense of senescence (molecular genetic regulation and manipulation of leaf senescence). Plant Physiol. 113, 313–319 (1997).

  4. 4.

    Qiu, K. et al. EIN3 and ORE1 accelerate degreening during ethylene-mediated leaf senescence by directly activating chlorophyll catabolic genes in Arabidopsis. PLoS Genet. 11, e1005399 (2015).

  5. 5.

    Rauf, M. et al. ORE1 balances leaf senescence against maintenance by antagonizing G2-like-mediated transcription. EMBO Rep. 14, 382–388 (2013).

  6. 6.

    Woo, H. R. et al. The delayed leaf senescence mutants of Arabidopsis, ore1, ore3, and ore9 are tolerant to oxidative stress. Plant Cell Physiol. 45, 923–932 (2004).

  7. 7.

    Balazadeh, S. et al. Salt-triggered expression of the ANAC092-dependent senescence regulon in Arabidopsis thaliana. Plant Signal. Behav. 5, 733–735 (2010).

  8. 8.

    Balazadeh, S. et al. A gene regulatory network controlled by the NAC transcription factor ANAC092/AtNAC2/ORE1 during salt-promoted senescence. Plant J. 62, 250–264 (2010).

  9. 9.

    Meng, S. et al. Arabidopsis NRT1.5 mediates the suppression of nitrate starvation-induced leaf senescence by modulating foliar potassium level. Mol. Plant 9, 461–470 (2016).

  10. 10.

    Agüera, E. et al. Induction of leaf senescence by low nitrogen nutrition in sunflower (Helianthus annuus) plants. Physiol. Plant. 138, 256–267 (2010).

  11. 11.

    Schulte auf’m Erley, G. et al. Leaf senescence induced by nitrogen deficiency as indicator of genotypic differences in nitrogen efficiency in tropical maize. J. Plant Nutr. Soil Sci. 170, 106–114 (2007).

  12. 12.

    Matallana-Ramirez, L. P. et al. NAC transcription factor ORE1 and senescence-induced BIFUNCTIONAL NUCLEASE1 (BFN1) constitute a regulatory cascade in Arabidopsis. Mol. Plant 6, 1438–1452 (2013).

  13. 13.

    Yaeno, T. & Iba, K. BAH1/NLA, a RING-type ubiquitin E3 ligase, regulates the accumulation of salicylic acid and immune responses to Pseudomonas syringae DC3000. Plant Physiol. 148, 1032–1041 (2008).

  14. 14.

    Park, B. S. et al. NITROGEN LIMITATION ADAPTATION recruits PHOSPHATE2 to target the phosphate transporter PT2 for degradation during the regulation of Arabidopsis phosphate homeostasis. Plant Cell 26, 454–464 (2014).

  15. 15.

    Peng, M. et al. A mutation in NLA, which encodes a RING-type ubiquitin ligase, disrupts the adaptability of Arabidopsis to nitrogen limitation. Plant J. 50, 320–337 (2007).

  16. 16.

    Xie, Q. et al. SINAT5 promotes ubiquitin-related degradation of NAC1 to attenuate auxin signals. Nature 419, 167–170 (2002).

  17. 17.

    Lin, W. Y. et al. NITROGEN LIMITATION ADAPTATION, a target of microRNA827, mediates degradation of plasma membrane-localized phosphate transporters to maintain phosphate homeostasis in Arabidopsis. Plant Cell 25, 4061–4074 (2013).

  18. 18.

    Hu, C. D. et al. Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Mol. Cell 9, 789–798 (2002).

  19. 19.

    Kerppola, T. K. et al. Bimolecular fluorescence complementation (BiFC) analysis as a probe of protein interactions in living cells. Annu. Rev. Biophys. 37, 465–487 (2008).

  20. 20.

    Zhao, M. et al. Involvement of miR169 in the nitrogen-starvation responses in Arabidopsis. New Phyt. 190, 906–915 (2011).

  21. 21.

    He, H. et al. Two young microRNAs originating from target duplication mediate nitrogen starvation adaptation via regulation of glucosinolate synthesis in Arabidopsis thaliana. Plant Physiol. 164, 853–865 (2014).

  22. 22.

    Hsieh, L. C. et al. Uncovering small RNA-mediated responses to phosphate deficiency in Arabidopsis by deep sequencing. Plant Physiol. 151, 2120–2132 (2009).

  23. 23.

    Pant, B. D. et al. Identification of nutrient-responsive Arabidopsis and rapeseed microRNAs by comprehensive real-time polymerase chain reaction profiling and small RNA sequencing. Plant Physiol. 150, 1541–1555 (2009).

  24. 24.

    Liang, G. et al. Uncovering miRNAs involved in crosstalk between nutrient deficiencies in Arabidopsis. Sci. Rep. 5, 11813 (2015).

  25. 25.

    Bari, R. et al. PHO2, microRNA399, and PHR1 define a phosphate-signaling pathway in plants. Plant Physiol. 141, 988–999 (2006).

  26. 26.

    Kuo, H. F. et al. The role of microRNAs in phosphorus deficiency signalling. Plant Physiol. 156, 1016–1024 (2011).

  27. 27.

    Fujii, H. et al. A miRNA involved in phosphate-starvation response in Arabidopsis. Curr. Biol. 15, 2038–2043 (2005).

  28. 28.

    Aung, K. et al. pho2, a phosphate overaccumulator, is caused by a nonsense mutation in a microRNA399 target gene. Plant Physiol. 141, 1000–1011 (2006).

  29. 29.

    Chiou, T. J. et al. Regulation of phosphate homeostasis by microRNA in Arabidopsis. Plant Cell 18, 412–421 (2006).

  30. 30.

    Sakuraba, Y. et al. The Arabidopsis transcription factor NAC016 promotes drought stress responses by repressing AREB1 transcription through a trifurcate feed-forward regulatory loop Involving NAP. Plant Cell 27, 1771–1787 (2015).

  31. 31.

    Mahmood, K. et al. ANAC032 positively regulates age-dependent and stress-induced senescence in Arabidopsis thaliana. Plant Cell Physiol. 57, 2029–2046 (2016).

  32. 32.

    Millner, P. D. et al. The Beltsville method for soilless production of vesicular-arbuscular mycorrhizal fungi. Mycorrhiza 2, 9–15 (1992).

  33. 33.

    Zhang, X. et al. Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. Nat. Protoc. 1, 641–646 (2006).

  34. 34.

    Liu, T. Y. et al. PHO2-dependent degradation of PHO1 modulates phosphate homeostasis in Arabidopsis. Plant Cell 24, 2168–2183 (2012).

  35. 35.

    Porra, R. J. et al. Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectrometry. Biochim. Biophys. Acta 975, 384–394 (1989).

Download references


This work was supported by an RSSS grant (no. NRF-RSSS-002) from the National Research Foundation, Singapore to N.H.C and by a MEXT KAKENHI grant (no. 25113001), Japan to N.M.

Author information


  1. Temasek Life Sciences Laboratory, National University of Singapore, 117604, Singapore

    • Bong Soo Park
    • , Tao Yao
    • , Jun Sung Seo
    • , Eriko Chi Cheng Wong
    • , Chung-Hao Huang
    •  & Nam-Hai Chua
  2. Plant Gene Regulation Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    • Nobutaka Mitsuda


  1. Search for Bong Soo Park in:

  2. Search for Tao Yao in:

  3. Search for Jun Sung Seo in:

  4. Search for Eriko Chi Cheng Wong in:

  5. Search for Nobutaka Mitsuda in:

  6. Search for Chung-Hao Huang in:

  7. Search for Nam-Hai Chua in:


B.S.P. and N.H.C. designed the experiments. B.S.P., T.Y., J.S.S., E.C.C.W., C.H.H. and N.M. executed the experiments. All of the authors interpreted and discussed the data. B.S.P. and N.H.C. wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Nam-Hai Chua.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–7, Supplementary Tables 1–4.

  2. Reporting Summary

About this article

Publication history




Issue Date



Further reading