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Nitrate signals determine the sensing of nitrogen through differential expression of genes involved in nitrogen uptake and assimilation in finger millet

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Abstract

In order to understand the molecular basis of high nitrogen use efficiency of finger millet, five genes (EcHNRT2, EcLNRT1, EcNADH-NR, EcGS, and EcFd-GOGAT) involved in nitrate uptake and assimilation were isolated using conserved primer approaches. Expression profiles of these five genes along with the previously isolated EcDof1 was studied under increased KNO3 concentrations (0.15 to 1,500 μM) for 2 h as well as at 1.5 μM for 24 h in the roots and shoots of 25 days old nitrogen deprived two contrasting finger millet genotypes (GE-3885 and GE-1437) differing in grain protein content (13.76 and 6.15 %, respectively). Time kinetics experiment revealed that, all the five genes except EcHNRT2 in the leaves of GE-3885 were induced within 30 min of nitrate exposure indicating that there might be a greater nitrogen deficit in leaves and therefore quick transportation of nitrate signals to the leaves. Exposing the plants to increasing nitrate concentrations for 2 h showed that in roots of GE-3885, NR was strongly induced while GS was repressed; however, the pattern was found to be reversed in leaves of GE-1437 indicating that in GE-3885, most of the nitrate might be reduced in the roots but assimilated in leaves and vice-versa. Furthermore, compared with the low-protein genotype, expression of HNRT2 was strongly induced in both roots and shoots of high-protein genotype at the least nitrate concentration supplied. This further indicates that GE-3885 is a quick sensor of nitrogen compared with the low-protein genotype. Furthermore, expression of EcDof1 was also found to overlap the expression of NR, GS, and GOGAT indicating that Dof1 probably regulates the expression of these genes under different conditions by sensing the nitrogen fluctuations around the root zone.

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Abbreviations

NRT:

Nitrate transporter

GS:

Glutamine synthetase

GOGAT:

Glutamine oxoglutarate aminotransferase

Dof:

DNA binding with one finger

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Acknowledgments

The authors wish to acknowledge the Department of Biotechnology, Government of India for providing financial support in the form of Program Mode Support for research and development in Agricultural Biotechnology at G.B. Pant University of Agriculture and Technology, Pantnagar (grant No. BT/PR7849/AGR/02/374/2006). Alok Kumar Gupta and Vikram Singh Gaur’s work was supported by the Department of Biotechnology (DBT) and Department of Science and Technology (DST) in the form of fellowships. The support provided by Director, Experiment Station, G.B. Pant University of Agriculture and Technology, Pantnagar is also thankfully acknowledged.

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Authors and Affiliations

  1. Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, GB Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, 263145, India

    Alok Kumar Gupta, Vikram Singh Gaur & Anil Kumar

  2. Department of Biotechnology, SBSPGI, Walawala, Dehradun, Uttarakhand, India

    Sanjay Gupta

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  1. Alok Kumar Gupta
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  2. Vikram Singh Gaur
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  4. Anil Kumar
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Correspondence to Anil Kumar.

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Gupta, A.K., Gaur, V.S., Gupta, S. et al. Nitrate signals determine the sensing of nitrogen through differential expression of genes involved in nitrogen uptake and assimilation in finger millet. Funct Integr Genomics 13, 179–190 (2013). https://doi.org/10.1007/s10142-013-0311-x

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