Skip to main content
SpringerLink
Log in

Acute salt stress differentially modulates nitrate reductase expression in contrasting salt responsive rice cultivars

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

Abstract

Salt stress response includes alteration in the activity of various important enzymes in plants. Nitrate reductase (NR) is one of the known enzyme affected by salt stress. In this study, contrasting salt responsive cultivars (CVS) (IR64-sensitive and CSR 36-tolerant) were considered to study the regulation of NR genes under salt stress conditions. Using Arabidopsis genes Nia1 and Nia2, three different NR genes were identified in rice and their expression study was conducted. Under stress condition, salt-sensitive CVS (IR64) showed a decrease in NR activity under in vitro and in vivo conditions, whereas tolerant CVS showed an increase in NR activity. Different trends for NR activity in contrasting genotype are explained by the variable number of GATA element in the upstream region of the NR gene. This variation of NR activity in contrasting CVS further co-relates with the transcript level of NR genes. The transcript level of three different NR genes also evidenced the effect of CREs in gene regulation. Promoter (1-kb upstream region) of different NR genes contained different abiotic stress-responsive CREs, which explain the differential behavior of these genes towards the abiotic stress. Overall, this study concludes the role of CREs in the regulation of NR gene and indicates the importance of transcriptional control of NR activity under stress condition. This is the first type of report that highlights the role of the regulatory mechanism of NR genes under salt stress condition.

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

Similar content being viewed by others

Abbreviations

NR:

Nitrate reductase

CVS:

Cultivars

CREs:

Cis-regulatory elements

References

  • Apse MP, Blumwald E (2002) Engineering salt tolerance in plants. Curr Opin Biotechnol 13(2):146–150

    Article  CAS  PubMed  Google Scholar 

  • Arshi A, Abdin M, Iqbal M (2002) Growth and metabolism of senna as affected by salt stress. Biol Plant 45(2):295–298

    Article  Google Scholar 

  • Baki G, Siefritz F, Man HM, Weiner H, Kaldenhoff R, Kaiser W (2000) Nitrate reductase in Zea mays L. under salinity. Plant Cell Environ 23(5):515–521

    Article  Google Scholar 

  • Besson-Bard A, Pugin A, Wendehenne D (2008) New insights into nitric oxide signaling in plants. Annu Rev Plant Biol 59(1):21–39. https://doi.org/10.1146/annurev.arplant.59.032607.092830

    Article  CAS  PubMed  Google Scholar 

  • Bi Y-M, Zhang Y, Signorelli T, Zhao R, Zhu T, Rothstein S (2005) Genetic analysis of Arabidopsis GATA transcription factor gene family reveals a nitrate-inducible member important for chlorophyll synthesis and glucose sensitivity. Plant J 44(4):680–692. https://doi.org/10.1111/j.1365-313X.2005.02568.x

    Article  CAS  PubMed  Google Scholar 

  • Carillo P, Mastrolonardo G, Nacca F, Fuggi A (2005) Nitrate reductase in durum wheat seedlings as affected by nitrate nutrition and salinity. Funct Plant Biol 32(3):209–219

    Article  CAS  PubMed  Google Scholar 

  • Cazetta JO, Villela LCV (2004) Nitrate reductase activity in leaves and stems of tanner grass (Brachiaria radicans napper). Sci Agric 61(6):640–648

    Article  CAS  Google Scholar 

  • Chandrasekaran P, Sivakumar R, Nandhitha G, Vishnuveni M, Boominathan P, Senthilkumar M (2017) Impact of PPFM and PGRs on seed germination, stress tolerant index and catalase activity in tomato (Solanum lycopersicum L) under drought. Int J Curr Microbiol App Sci 6(6):540–549

    Article  CAS  Google Scholar 

  • Chaves MM, Flexas J, Pinheiro C (2009) Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot 103(4):551–560

    Article  CAS  PubMed  Google Scholar 

  • Correia MJ, Fonseca F, Azedo-Silva J, Dias C, David MM, Barrote I, Osório ML, Osório J (2005) Effects of water deficit on the activity of nitrate reductase and content of sugars, nitrate and free amino acids in the leaves and roots of sunflower and white lupin plants growing under two nutrient supply regimes. Physiol Plant 124(1):61–70

    Article  CAS  Google Scholar 

  • Cramer MD, Lips SH (1995) Enriched rhizosphere CO2 concentrations can ameliorate the influence of salinity on hydroponically grown tomato plants. Physiol Plant 94(3):425–432

  • Daniel-Vedele F, Filleur S, Caboche M (1998) Nitrate transport: a key step in nitrate assimilation. Curr Opin Plant Biol 1(3):235–239

    Article  CAS  PubMed  Google Scholar 

  • Evans HJ, Nason A (1953) Pyridine nucleotide-nitrate reductase from extracts of higher plants. Plant Physiol 28(2):233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ferrario-Méry S, Valadier M-H, Foyer CH (1998) Overexpression of nitrate reductase in tobacco delays drought-induced decreases in nitrate reductase activity and mRNA. Plant Physiol 117(1):293–302

    Article  PubMed  PubMed Central  Google Scholar 

  • Flores P, Botella M, Martinez V, Cerdá A (2000) Ionic and osmotic effects on nitrate reductase activity in tomato seedlings. J Plant Physiol 156(4):552–557

    Article  CAS  Google Scholar 

  • Flores P, Angeles Botella M, Martínez V, Cerdá A (2002) Response to salinity of tomato seedlings with a split-root system: nitrate uptake and reduction. J Plant Nutr 25(1):177–187

    Article  CAS  Google Scholar 

  • Foyer C, Zhang H (2011) Annual plant reviews, nitrogen metabolism in plants in the post-genomic era, vol 42. John Wiley & Sons

  • Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48(12):909–930. https://doi.org/10.1016/j.plaphy.2010.08.016

    Article  CAS  PubMed  Google Scholar 

  • Gouia H, Ghorbal MH, Touraine B (1994) Effects of NaCl on flows of N and mineral ions and on NO3-reduction rate within whole plants of salt-sensitive bean and salt-tolerant cotton. Plant Physiol 105(4):1409–1418

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hasegawa PM, Bressan RA, Zhu J-K, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu Rev Plant Biol 51(1):463–499

    Article  CAS  Google Scholar 

  • Jamil A, Riaz S, Ashraf M, Foolad M (2011) Gene expression profiling of plants under salt stress. Crit Rev Plant Sci 30(5):435–458

    Article  Google Scholar 

  • Jaworski EG (1971) Nitrate reductase assay in intact plant tissues. Biochem Biophys Res Commun 43(6):1274–1279

    Article  CAS  PubMed  Google Scholar 

  • Katiyar S, Dubey R (1992) Influence of NaCl salinity on behaviours of nitrate reductase and nitrite reductase in rice seedlings differing in salt tolerance. J Agron Crop Sci 169(5):289–297

    Article  CAS  Google Scholar 

  • Kleinhofs A, Chao S, Sharp P (1988) Mapping of nitrate reductase genes in barley and wheat

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25(4):402–408

    Article  CAS  PubMed  Google Scholar 

  • López-Ochoa L, Acevedo-Hernández G, Martínez-Hernández A, Argüello-Astorga G, Herrera-Estrella L (2007) Structural relationships between diverse cis-acting elements are critical for the functional properties of a rbcS minimal light regulatory unit. J Exp Bot 58(15–16):4397–4406

    Article  PubMed  CAS  Google Scholar 

  • Loussaert DF, O'neill D, Simmons CR, Wang H (2010) Nitrate reductases from red algae, compositions and methods of use thereof. Google Patents,

  • Lutts S, Kinet J, Bouharmont J (1996) NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann Bot 78(3):389–398

    Article  CAS  Google Scholar 

  • Meloni DA, Gulotta MR, Martínez CA, Oliva MA (2004) The effects of salt stress on growth, nitrate reduction and proline and glycinebetaine accumulation in Prosopis alba. Braz J Plant Physiol 16(1):39–46

    Article  CAS  Google Scholar 

  • Misra N, Dwivedi U (1990) Nitrogen assimilation in germinating Phaseolus aureus seeds under saline stress. J Plant Physiol 135(6):719–724

    Article  CAS  Google Scholar 

  • Nath M, Yadav S, Kumar Sahoo R, Passricha N, Tuteja R, Tuteja N (2016) PDH45 transgenic rice maintain cell viability through lower accumulation of Na(+), ROS and calcium homeostasis in roots under salinity stress. J Plant Physiol 191:1–11. https://doi.org/10.1016/j.jplph.2015.11.008

    Article  CAS  PubMed  Google Scholar 

  • Ouyang B, Yang T, Li H, Zhang L, Zhang Y, Zhang J, Ye Z (2007) Identification of early salt stress response genes in tomato root by suppression subtractive hybridization and microarray analysis. J Exp Bot 58(3):507–520

  • Passricha N, Saifi S, Ansari MW, Tuteja N (2016a) Prediction and validation of cis-regulatory elements in 5′ upstream regulatory regions of lectin receptor-like kinase gene family in rice. Protoplasma. https://doi.org/10.1007/s00709-016-0979-6

  • Passricha N, Saifi S, Khatodia S, Tuteja N (2016b) Assessing zygosity in progeny of transgenic plants: current methods and perspectives. J Biol Methods 3(3):e46

    Article  PubMed  PubMed Central  Google Scholar 

  • Passricha N, Saifi S, Khatodia S, Tuteja N (2016c) Assessing zygosity in progeny of transgenic plants: current methods and perspectives. J Biol Methods; Vol 3, No 3 (2016)

  • Passricha N, Saifi SK, Gill SS, Tuteja R, Tuteja N (2019a) Chapter 3 - role of plant helicases in imparting salinity stress tolerance to plants. In: Tuteja R (ed) Helicases from All Domains of Life. Academic Press, pp 39–52. https://doi.org/10.1016/B978-0-12-814685-9.00003-8

  • Passricha N, Saifi SK, Kharb P, Tuteja N (2019b) Marker-free transgenic rice plant overexpressing pea LecRLK imparts salinity tolerance by inhibiting sodium accumulation. Plant Mol Biol. https://doi.org/10.1007/s11103-018-0816-8

  • Peuke AD, Glaab J, Kaiser WM, Jeschke WD (1996) The uptake and flow of C, N and ions between roots and shoots in Ricinus communis L. IV. Flow and metabolism of inorganic nitrogen and malate depending on nitrogen nutrition and salt treatment. J Exp Bot 47(3):377–385

    Article  CAS  Google Scholar 

  • PLAUT Z (1974) Nitrate reductase activity of wheat seedlings during exposure to and recovery from water stress and salinity. Physiol Plant 30(3):212–217

    Article  CAS  Google Scholar 

  • Raghuram N, Chandok MR, Sopory SK (1999) Light regulation of nitrate reductase gene expression in maize involves a G-protein. Mol Biol Res Commun 2(2):86–90

    Article  CAS  Google Scholar 

  • Rao RK, Gnanam A (1990) Inhibition of nitrate and nitrite reductase activities by salinity stress in Sorghum vulgare. Phytochemistry 29(4):1047–1049

    Article  CAS  Google Scholar 

  • Redinbaugh MG, Campbell WH (1981) Purification and characterization of NAD (P) H: nitrate reductase and NADH: nitrate reductase from corn roots. Plant Physiol 68(1):115–120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sagi M, Savidov N, L'vov N, Lips S (1997) Nitrate reductase and molybdenum cofactor in annual ryegrass as affected by salinity and nitrogen source. Physiol Plant 99(4):546–553

    Article  CAS  Google Scholar 

  • Saifi SK, Passricha N, Swain DM, Tuteja N (2017) Prediction of cis-regulatory elements for a detailed insight of RuvB family genes from Oryza sativa. Oryza 54(2):135–147

    Article  Google Scholar 

  • Saifi SK, Passricha N, Tuteja R, Tuteja N (2018) Stress-induced Oryza sativa RuvBL1a is DNA-independent ATPase and unwinds DNA duplex in 3′ to 5′ direction. Protoplasma 255(2):669–684. https://doi.org/10.1007/s00709-017-1178-9

    Article  CAS  PubMed  Google Scholar 

  • Sazegari S, Niazi A, Ahmadi FS (2015) A study on the regulatory network with promoter analysis for Arabidopsis DREB-genes. Bioinformation 11(2):101

    Article  PubMed  PubMed Central  Google Scholar 

  • Sepehr MF, Ghorbanli M, Amini F (2012) The effect of water stress on nitrate reductase activity and nitrogen and phosphorus contents in Cuminum cyminum L. Pak J Bot 44(3):899–903

    CAS  Google Scholar 

  • Shabala S (2013) Learning from halophytes: physiological basis and strategies to improve abiotic stress tolerance in crops. Ann Bot 112(7):1209–1221

    Article  PubMed  PubMed Central  Google Scholar 

  • Shahbaz M, Ashraf M (2013) Improving salinity tolerance in cereals. Crit Rev Plant Sci 32(4):237–249

    Article  Google Scholar 

  • Shrivastava P, Kumar R (2015) Soil salinity: a serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi J Biol Sci 22(2):123–131. https://doi.org/10.1016/j.sjbs.2014.12.001

    Article  CAS  PubMed  Google Scholar 

  • Srivastava H (1980) Regulation of nitrate reductase activity in higher plants. Phytochemistry 19(5):725–733

    Article  CAS  Google Scholar 

  • Teakle GR, Manfield IW, Graham JF, Gilmartin PM (2002) Arabidopsis thaliana GATA factors: organisation, expression and DNA-binding characteristics. Plant Mol Biol 50(1):43–56

    Article  CAS  PubMed  Google Scholar 

  • Viégas RA, de Melo AB, da Silveira JG (1999) Nitrate reductase activity and proline accumulation in cashew in response to NaCl salt shock. Revista Brasileira de Fisiologia Vegetal (Brazil)

  • Wang W-X, Pelah D, Alergand T, Shoseyov O, Altman A (2002) Characterization of SP1, a stress-responsive, boiling-soluble, homo-oligomeric protein from aspen. Plant Physiol 130(2):865–875

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wilson I, Bright J, Hancock J, Harrison J, Neill S, Ribeiro D, Barros R, Desikan R, Morris P (2008) Nitric oxide, stomatal closure, and abiotic stress. J Exp Bot 59(2):165–176. https://doi.org/10.1093/jxb/erm293

    Article  CAS  PubMed  Google Scholar 

  • Yadav S, Gill SS, Passricha N, Gill R, Badhwar P, Anjum NA, Francisco J-BJ, Tuteja N (2018) Genome-wide analysis and transcriptional expression pattern-assessment of superoxide dismutase (SOD) in rice and Arabidopsis under abiotic stresses. Plant Gene. https://doi.org/10.1016/j.plgene.2018.10.001

  • Yamaguchi T, Blumwald E (2005) Developing salt-tolerant crop plants: challenges and opportunities. Trends Plant Sci 10(12):615–620

    Article  CAS  PubMed  Google Scholar 

  • You J, Zhang L, Song B, Qi X, Chan Z (2015) Systematic analysis and identification of stress-responsive genes of the NAC gene family in Brachypodium distachyon. PLoS One 10(3):e0122027

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Zhao C-R, Ikka T, Sawaki Y, Kobayashi Y, Suzuki Y, Hibino T, Sato S, Sakurai N, Shibata D, Koyama H (2009) Comparative transcriptomic characterization of aluminum, sodium chloride, cadmium and copper rhizotoxicities in Arabidopsis thaliana. BMC Plant Biol 9(1):32. https://doi.org/10.1186/1471-2229-9-32

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This research work is financially supported by a fellowship grant from the University Grant Commission-Basic Scientific Research (UGC BSR) (F.25-1/2013-14 (BSR)/7-371/2012) to Ms. Pooja Rohilla and the University Grant Commission- Special Assistance Programme (UGC-SAP) (F.20/2012(SAP-II)) grant from UGC, New Delhi. Authors are also thankful to Dr. Nishat Passricha, ICGEB, New Delhi, for the help in the conducting of qPCR experiments.

Author information

Authors and Affiliations

  1. Department of Genetics, Maharshi Dayanand University, Rohtak, Haryana, 124001, India

    Pooja Rohilla & Jaya Parkash Yadav

Authors
  1. Pooja Rohilla
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jaya Parkash Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

RP has contributed to the idea and designed the experiments. YJP is a mentor and edited the manuscript. Both authors thoroughly read and approved the manuscript.

Corresponding author

Correspondence to Jaya Parkash Yadav.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Handling Editor: Néstor Carrillo

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Key message: Under salinity stress Nitrate reductase regulated by transcriptional machinery with the help of CREs

Electronic supplementary material

ESM 1

(DOCX 17 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rohilla, P., Yadav, J.P. Acute salt stress differentially modulates nitrate reductase expression in contrasting salt responsive rice cultivars. Protoplasma 256, 1267–1278 (2019). https://doi.org/10.1007/s00709-019-01378-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00709-019-01378-y

Keywords

Search

Navigation