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Labeling Maize (Zea mays L.) Leaves with 15 NH4 + and Monitoring Nitrogen Incorporation into Amino Acids by GC/MS Analysis
- Source :
- Current Protocols in Plant Biology. 3:e20073
- Publication Year :
- 2018
- Publisher :
- Wiley, 2018.
-
Abstract
- The human body contains approximately 3.2% nitrogen (N), mainly present as protein and amino acids. Although N exists at a high concentration (78%) in the air, it is not readily available to animals and most plants. Plants are however able to take up both nitrate (NO3- ) and ammonium (NH4+ ) ions from the soil and convert them to amino acids and proteins, which are excellent sources for all animals. Most N is available as the stable isotope 14 N, but a second form, 15 N, is present in very low concentrations. 15 N can be detected in extracts of plants by gas chromatography followed by mass spectrometry (GC/MS). In this protocol, the methods are described for tracing the pathway by which plants are able to take up 15 N-labeled nitrate and ammonium and convert them into amino acids and proteins. A protocol for extracting and quantifying amino acids and 15 N enrichment in maize (Zea mays L.) leaves labeled with 15 NH4+ is described. Following amino acid extraction, purification, and separation by GC/MS, a calculation of the 15 N enrichment of each amino acid is carried out on a relative basis to identify any differences in the dynamics of amino acid accumulation. This will allow a study of the impact of genetic modifications or mutations on key reactions involved in primary nitrogen and carbon metabolism. © 2018 by John Wiley & Sons, Inc.
- Subjects :
- 0106 biological sciences
0301 basic medicine
chemistry.chemical_classification
Chromatography
Extraction (chemistry)
food and beverages
chemistry.chemical_element
General Medicine
Mass spectrometry
01 natural sciences
Nitrogen
Amino acid
03 medical and health sciences
chemistry.chemical_compound
030104 developmental biology
chemistry
Nitrate
Ammonium
Gas chromatography
Gas chromatography–mass spectrometry
010606 plant biology & botany
Subjects
Details
- ISSN :
- 23798068
- Volume :
- 3
- Database :
- OpenAIRE
- Journal :
- Current Protocols in Plant Biology
- Accession number :
- edsair.doi...........e40984eb5449173431f4f3acddf033d1