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Molecular Characterization of GS2 and Fd-GOGAT Homeologues and Their Biased Response to Nitrogen Stress in Bread Wheat (Triticum aestivum L.)

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Abstract

Nitrogen (N) is one of the major limiting elements affecting the growth and productivity of cereal crops including bread wheat. Identification and detailed characterization of the candidate genes involved in the assimilation of N from inorganic to organic form is very important for better N utilization. Glutamine synthetase 2 (GS2) and Ferredoxin-dependent-glutamate synthase (Fd-GOGAT) are the two critical enzymes involved in GS/GOGAT cycle, which are necessary for primary N-assimilation. In the present study, GS2 and Fd-GOGAT genes were cloned from a popular bread wheat variety HD-2967 and their individual homeologues (A, B and D) were characterized under different levels of N-stress. Homeologous cDNAs of GS2 and Fd-GOGAT showed > 93% and > 98% sequence similarity, respectively, in the coding region but were distinguishable at UTRs. The expression patterns and contribution of each homeologue of both the genes in HD-2967 under different N treatments showed downregulation in the shoot and upregulation in the root tissues. However, the contribution of the individual homeologue varied with the tissue, time point and level of N-stress. Further, when the expression pattern and contribution of each homeologue were studied in a diverse set of wheat genotypes, a generalised contribution pattern of each homeologue of wheat was observed. While the transcript ratio of A and D homeologues of both the genes demonstrated to be compensating each other, Fd-GOGAT-B showed the least contribution in all the conditions and at all stages of growth. Substantial variation in cis-regulatory elements was observed among all three homeologues of both the genes. Overall, characterization and spatio-temporal contribution of GS2 and Fd-GOGAT homeologues would provide applied value for further detailed studies as well as their exploration in primary N-assimilation process in wheat.

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Abbreviations

GS2:

Glutamine synthetase

Fd-GOGAT:

Ferredoxin-dependent-glutamate synthase

N:

Nitrogen

NO3 − :

Nitrate

CDS:

Coding sequence

mmol/L:

Millimolar/litre

MS:

Murashige and Skoog

H:

Hour

N:P:K:

Nitrogen:phosphorus:potassium

Kg/h:

Kilogram/hour

RNA:

Ribonucleic acid

cDNA:

Complementary deoxy- ribonucleic acid

dNTPs:

Deoxynucleoside triphosphates

PCR:

Polymerase chain reaction

qPCR:

Quantitative-polymerase chain reaction

Tm:

Melting temperature

°C:

Degree celsius

ORF:

Open reading frame

MW:

Molecular weight

pI:

Isoelectric point

NCBI:

National Center for Biotechnology Information

Μl:

Microlitre

Ng:

Nanogram

Bp:

Basepair

UTR:

Untranslated region

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Acknowledgements

Authors would also like to acknowledge the support and guidance provided by the Project Director, ICAR-National Institute for Plant Biotechnology, New Delhi. The authors also want to thank Puja Mandal for helping in editing the manuscript.

Funding

This research is part of the Indo-UK Virtual Nitrogen Centre project, financially supported by the Department of Biotechnology, Govt. of India (BT/IN/UK-VNC/43/KV/2015-16) under the Indo-UK Centre for Improvement of Nitrogen Use Efficiency in Wheat (INEW) and NIPB institutional in-house project on Biochemical and Molecular Basis of Nitrogen Utilization in Wheat. 

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Authors and Affiliations

  1. ICAR-National Institute for Plant Biotechnology, Pusa Campus, New Delhi, 110012, India

    Gayatri, Subodh Kumar Sinha & Pranab Kumar Mandal

  2. Amity Institute of Biotechnology, Amity University, Sector 125, Noida, UttarPradesh, 201301, India

    Gayatri & Pranita Roy

  3. ICAR- Indian Institute of Wheat and Barley Research, Karnal, 132001, India

    Karnam Venkatesh

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  1. Gayatri
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  2. Karnam Venkatesh
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  3. Subodh Kumar Sinha
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  4. Pranita Roy
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  5. Pranab Kumar Mandal
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Contributions

Conceptualization, PKM and G; Data curation, PKM; Formal analysis, G; Funding acquisition, PKM; Investigation, PKM; Methodology, G; Project administration, PKM; Resources, PKM, KV (9 wheat genotypes); Software, G; Supervision, PKM; Validation, PKM; Visualization, PKM; Writing-original draft, G; Writing-review and editing PKM, KV, SKS, PR. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Pranab Kumar Mandal.

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Handling Editor: Ranjit Riar.

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Gayatri, Venkatesh, K., Sinha, S.K. et al. Molecular Characterization of GS2 and Fd-GOGAT Homeologues and Their Biased Response to Nitrogen Stress in Bread Wheat (Triticum aestivum L.). J Plant Growth Regul 41, 2555–2569 (2022). https://doi.org/10.1007/s00344-021-10433-z

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  • DOI: https://doi.org/10.1007/s00344-021-10433-z

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