Abstract

Plants absorb nitrogen mainly through their roots. Nitrogen sensing is required for the absorption and transport of different nitrogen nutrients. In this study, we constructed biosensors with immobilized Panax notoginseng root tip meristems based on a three-electrode system and successfully determined the kinetics of the interactions between the P. notoginseng root tip meristems and five important nitrogen nutrients, namely, urea, sodium nitrate, sodium glutamate, disodium inosinate, and disodium guanylate. We discovered that the biosensor’s sensing kinetics was similar to the enzyme–substrate kinetics, and the receptor–ligand interconnected allosteric interaction constant Ka (mol/L), analogous to the Michaelis constant, was calculated. The result showed that the root tip meristems of two- to four-year-old P. notoginseng plants had a higher capacity to sense inorganic nitrogen nutrients (sodium nitrate and urea) than the three organic nitrogen nutrients. The ability of the plants to sense inorganic nitrogen nutrients decreased with an increase in plant age. The sensing sensitivity of four-year-old P. notoginseng plants to disodium inosinate and disodium guanylate was 100- to 10,000-fold lower than that of the two- and three-year-old plants. Additionally, the capability to sense sodium glutamate decreased initially and then increased with an increase in plant age. The biosensors reached an ultra-sensitive level ( 1 × 10 − 22 mol/L) in sensing the five nitrogen nutrients and exhibited advantages such as good stability and reproducibility, low cost, a simple structure, and a rapid response, providing a new approach for quantitative determination of the capability of plants to sense different nitrogen nutrients.

Highlights

  • Nitrogen nutrition is closely related to plant growth [1]

  • Nitrate transceptors (NRT) NRT1.1 and NRT2.1 [2,3,4,5] as well as ammonium transporters (AMT) AMT1.1 and AMT1.3 [6, 7] were identified in the root system of the model plant Arabidopsis thaliana

  • Compared with a nanogold receptor sensor [25], which has the advantage that the effect truly caused by the receptor can be exactly known, the biosensor prepared using the root tip meristem could reflect the allostery resulting from the interaction between the extracellular domain of the receptor and the nitrogen nutrient ligand and the change in the on/off state of the ion channels in the cell membrane caused by the intracellular signal transduction and cascade amplification after the binding of the receptor and ligand

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Summary

Introduction

Botanists have made significant progress in understanding the absorption and transport of nitrate nitrogen, ammonium nitrogen, and amino acid nitrogen in plants. Glutamate receptors (GLRs) are related to defense signals in A. thaliana [11, 12]. Most of these studies focused on the transport of nitrogen nutrients and signal transduction in plants. It is relevant to study the nitrogen-sensing receptors of plants, assess the mechanisms of nitrogen absorption and transport, and quantify the capacity of plants to sense different nitrogen nutrients. Plant extracellular sensing, the pathways of intracellular signal amplification and transduction, and nitrogen sensing receptor-related quantification methods have been seldom reported so far

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