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<p>Nitrogen (N) is a key macronutrient that influences plant growth and development, but its availability in the soil is a major limiting factor for crop productivity. Wheat (<i>Triticum aestivum</i> L.) utilizes only about one-third of the applied nitrogenous fertilizer for its growth and development, leading to a significant economic and environmental loss by unutilized applied N. Nitrate (NO<sub>3</sub><sup>−</sup>) is the major available form of nitrogen source which wheat plants uptake and transport by different nitrate transporters (NRTs). Among NRTs, <i>NRT1.5</i> in <i>Arabidopsis</i> is involved in the nitrate root-to-shoot translocation. The present study undertakes characterization of ~ 2&#xa0;kb promoter regions of three homeologs of the <i>TaNPF7.1</i> gene (A, B, and D sub-genomes) homologous to <i>NRT1.5</i> in an efficient NUE cultivar K9107. Our results demonstrated significant variations in cis-regulatory elements (CREs) and transcription factor (TF) binding sites among the homeologs and between K9107 and the Chinese Spring genotype. Functional validation using <i>Arabidopsis</i> transgenic reporter lines (<i>pTaNPF7.1-A/-B/-D</i>::GUS) under various conditions, including nutrient (nitrate, potassium) and abiotic stress, as well as hormonal treatment, revealed distinct promoter level regulation. All three promoters exhibited nitrate-inducibility in general, but the A and D sub-genomes showed potassium-responsive GUS expression, indicating crosstalk between the two major macronutrients and their coordinated signalling. <i>pTaNPF7.1A</i> showed the highest responsiveness to ABA, <i>pTaNPF7.1B</i> to drought, and <i>pTaNPF7.1D</i> to salt stress, highlighting divergence in the regulation of different stresses. These findings highlight the regulatory divergence of <i>TaNPF7.1</i> through neo-functionalization, enabling better adaptation in various nutritional and abiotic conditions, and offer novel insights into nitrogen use efficiency and stress resilience in wheat.</p>

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Functional validation and regulatory analysis of three homeologous upstream sequences of the wheat low-affinity nitrate transporter gene TaNPF7.1: differential homeolog-specific expression under nutrient and abiotic stress conditions

  • 期刊论文

  • 发表日期：

    2026年5月11日

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  • V. Muhammed Shamnas,
  • Samrita Garnaik,
  • Anuj Kumar,
  • Subodh Kumar Sinha

摘要

Nitrogen (N) is a key macronutrient that influences plant growth and development, but its availability in the soil is a major limiting factor for crop productivity. Wheat (Triticum aestivum L.) utilizes only about one-third of the applied nitrogenous fertilizer for its growth and development, leading to a significant economic and environmental loss by unutilized applied N. Nitrate (NO3−) is the major available form of nitrogen source which wheat plants uptake and transport by different nitrate transporters (NRTs). Among NRTs, NRT1.5 in Arabidopsis is involved in the nitrate root-to-shoot translocation. The present study undertakes characterization of ~ 2 kb promoter regions of three homeologs of the TaNPF7.1 gene (A, B, and D sub-genomes) homologous to NRT1.5 in an efficient NUE cultivar K9107. Our results demonstrated significant variations in cis-regulatory elements (CREs) and transcription factor (TF) binding sites among the homeologs and between K9107 and the Chinese Spring genotype. Functional validation using Arabidopsis transgenic reporter lines (pTaNPF7.1-A/-B/-D::GUS) under various conditions, including nutrient (nitrate, potassium) and abiotic stress, as well as hormonal treatment, revealed distinct promoter level regulation. All three promoters exhibited nitrate-inducibility in general, but the A and D sub-genomes showed potassium-responsive GUS expression, indicating crosstalk between the two major macronutrients and their coordinated signalling. pTaNPF7.1A showed the highest responsiveness to ABA, pTaNPF7.1B to drought, and pTaNPF7.1D to salt stress, highlighting divergence in the regulation of different stresses. These findings highlight the regulatory divergence of TaNPF7.1 through neo-functionalization, enabling better adaptation in various nutritional and abiotic conditions, and offer novel insights into nitrogen use efficiency and stress resilience in wheat.