A reliable method for spectrophotometric determination of glycine betaine in cell suspension and other systems
| Autor: | Gerardo Armando Aguado-Santacruz, Axel Tiessen-Favier, Amalio Santacruz-Varela, Ma. Guadalupe Valadez-Bustos, Alejandrina Robledo-Paz, Quintín Rascón-Cruz, Abel Muñoz-Orozco |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Cell Culture Techniques Biophysics Polyethylene glycol Poaceae 01 natural sciences Biochemistry 03 medical and health sciences chemistry.chemical_compound Betaine Spectrophotometry medicine Denaturation (biochemistry) Ammonium Molecular Biology Chromatography medicine.diagnostic_test Chemistry Extraction (chemistry) Cell Biology 030104 developmental biology Osmolyte Glycine 010606 plant biology & botany |
| Zdroj: | Analytical Biochemistry. 498:47-52 |
| ISSN: | 0003-2697 |
| Popis: | Glycine betaine is a quaternary ammonium compound that accumulates in a large variety of species in response to different types of stress. Glycine betaine counteracts adverse effects caused by abiotic factors, preventing the denaturation and inactivation of proteins. Thus, its determination is important, particularly for scientists focused on relating structural, biochemical, physiological, and/or molecular responses to plant water status. In the current work, we optimized the periodide technique for the determination of glycine betaine levels. This modification permitted large numbers of samples taken from a chlorophyllic cell line of the grass Bouteloua gracilis to be analyzed. Growth kinetics were assessed using the chlorophyllic suspension to determine glycine betaine levels in control (no stress) cells and cells osmotically stressed with 14 or 21% polyethylene glycol 8000. After glycine extraction, different wavelengths and reading times were evaluated in a spectrophotometer to determine the optimal quantification conditions for this osmolyte. Optimal results were obtained when readings were taken at a wavelength of 290 nm at 48 h after dissolving glycine betaine crystals in dichloroethane. We expect this modification to provide a simple, rapid, reliable, and cheap method for glycine betaine determination in plant samples and cell suspension cultures. |
| Databáze: | OpenAIRE |
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