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Measuring Nitrate in Plant Cells by in Vivo NMR Using Gd3+ as a Shift Reagent
Shachar-Hill1, Pfeffer, Ratcliffe
1U.S. Department of Agriculture, Agricultural Research Service, Eastern Region Research Center, 600 E. Mermaid Lane, Philadelphia, Pennsylvania, 19118
Journal of Magnetic Resonance. Series B
|April 1, 1996
Summary
Gadolinium (Gd3+) can distinguish nitrate signals inside and outside plant cells using NMR. However, Gd3+ impacts plant growth, leading researchers to explore safer chelated forms for studying nitrate transport.
Area of Science:
- Plant Physiology
- Biochemistry
- Analytical Chemistry
Background:
- Nuclear Magnetic Resonance (NMR) is crucial for studying plant nitrate.
- Distinguishing intracellular and extracellular nitrate signals in plants is challenging.
- Gadolinium (Gd3+) has shown potential as a shift reagent for nitrate NMR.
Purpose of the Study:
- To evaluate Gd3+ as a shift reagent for resolving intracellular and extracellular nitrate signals in plants.
- To assess the physiological effects of Gd3+ on plant nitrate uptake and growth.
- To identify a suitable gadolinium-based agent for studying nitrate transport in plant roots.
Main Methods:
- Utilized Nuclear Magnetic Resonance (NMR) spectroscopy with 14N and 15N isotopes.
- Employed Gadolinium (Gd3+) as a paramagnetic shift reagent.
- Conducted time-course experiments on plant cell suspensions and root material.
- Investigated various chelated forms of gadolinium, including Gd(DTPA-BMA).
Main Results:
- Gd3+ successfully resolved distinct intracellular and extracellular nitrate signals in plant tissues.
- Extended monitoring of nitrate levels was possible with Gd3+.
- Adverse effects of Gd3+ on plant growth and nitrate uptake were observed.
- Gd(DTPA-BMA) emerged as a promising shift reagent with fewer physiological impacts.
Conclusions:
- Gd3+ is an effective shift reagent for differentiating nitrate pools in plants via NMR.
- Free Gd3+ exhibits toxicity, limiting its use in long-term physiological studies.
- Chelated gadolinium, specifically Gd(DTPA-BMA), offers a viable alternative for studying nitrate transport in plant roots under physiological conditions.