Isolation of novel indole-3-acetic acid conjugates by immunoaffinity extraction.
An analytical protocol for the isolation and quantification of indole-3-acetic acid (IAA) and its amino acid conjugates was developed. IAA is an important phytohormone and formation of its conjugates plays a crucial role in regulating IAA levels in plants. The developed protocol combines a highly specific immunoaffinity extraction with a sensitive and selective LC-MS/MS analysis. By using internal standards for each of the studied compounds, IAA and seven amino acid conjugates were analyzed in quantities of fresh plant material as low as 30 mg. In seeds of Helleborus niger, physiological levels of these compounds were found to range from 7.5 nmol g(-1) fresh weight (IAA) to 0.44 pmol g(-1) fresh weight (conjugate with Ala). To our knowledge, the identification of IAA conjugates with Gly, Phe and Val from higher plants is reported here for the first time.
Pencík
Ales
A
Faculty of Science, Palacký University, Olomouc, Czech Republic.
Rolcík
Jakub
J
Novák
Ondrej
O
Magnus
Volker
V
Barták
Petr
P
Buchtík
Roman
R
Salopek-Sondi
Branka
B
Strnad
Miroslav
M
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D005998:Glycine / Q000737:chemistry; D039322:Helleborus / Q000737:chemistry; D007210:Indoleacetic Acids / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D010649:Phenylalanine / Q000737:chemistry; D010937:Plant Growth Regulators / Q000276:immunology / Q000302:isolation & purification / Q000592:standards; D012015:Reference Standards; D012639:Seeds / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D053719:Tandem Mass Spectrometry; D014633:Valine / Q000737:chemistry
D000911:Antibodies, Monoclonal; D007210:Indoleacetic Acids; D010937:Plant Growth Regulators; D010649:Phenylalanine; C030737:indoleacetic acid; D014633:Valine; D005998:Glycine