title: A reliable method for spectrophotometric determination of glycine betaine in cell suspension and other systems.
abstract: 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.
authors.affiliation: Campus Montecillo, Colegio de Postgraduados, C.P. 56230, Montecillo, Estado de México, Mexico.
authors.forename: Ma Guadalupe
authors.initials: MG
authors.lastname: Valadez-Bustos
authors.affiliation: Unidad de Biotecnología, Campo Experimental Bajío-Inifap, C.P. 38010, Celaya, Guanajuato, Mexico.
authors.forename: Gerardo Armando
authors.initials: GA
authors.lastname: Aguado-Santacruz
authors.affiliation: Laboratorio de Metabolómica y Fisiología Molecular, Cinvestav Unidad Irapuato, C.P. 36821, Irapuato, Guanajuato, Mexico.
authors.forename: Axel
authors.initials: A
authors.lastname: Tiessen-Favier
authors.affiliation: Campus Montecillo, Colegio de Postgraduados, C.P. 56230, Montecillo, Estado de México, Mexico.
authors.forename: Alejandrina
authors.initials: A
authors.lastname: Robledo-Paz
authors.affiliation: Campus Montecillo, Colegio de Postgraduados, C.P. 56230, Montecillo, Estado de México, Mexico.
authors.forename: Abel
authors.initials: A
authors.lastname: Muñoz-Orozco
authors.affiliation: Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, C.P. 31170, Chihuahua, Chihuahua, Mexico.
authors.forename: Quintin
authors.initials: Q
authors.lastname: Rascón-Cruz
authors.affiliation: Campus Montecillo, Colegio de Postgraduados, C.P. 56230, Montecillo, Estado de México, Mexico. Electronic address: asvarela@colpos.mx.
authors.forename: Amalio
authors.initials: A
authors.lastname: Santacruz-Varela
mesh_terms: D001622:Betaine / Q000032:analysis*; D018929:Cell Culture Techniques; D006109:Poaceae / Q000737:chemistry* / Q000166:cytology*; D013053:Spectrophotometry / Q000379:methods*
chemical_list: D001622:Betaine
keywords: Bouteloua; Cell cultures; Glycine betaine; Quaternary ammonium compounds; Stress