Premium
Analysis by high‐performance liquid chromatography of free amino acids extracted from needles of drought‐stressed and shaded Pinus ponderosa seedlings
Author(s) -
Vance Nan C.,
Zaerr Joe B.
Publication year - 1990
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1990.tb05861.x
Subject(s) - high performance liquid chromatography , amino acid , derivatization , arginine , proline , shading , glutamine , chemistry , chromatography , pinus <genus> , horticulture , botany , biology , biochemistry , art , visual arts
Free amino acid concentrations in needles of drought‐stressed and shaded seedlings of ponderosa pine (Pinus ponderosa Dougl. ex Laws.) were analyzed to determine the influence of low irradiance on a biochemical response to drought stress. A convenient method was developed for separating and quantitating amino acids in needle extracts using derivatization with dansyl chloride and reversed‐phase high‐performance liquid chromatography (HPLC). Twenty‐one amino acids were separated on an Ultrasphere ODS CIS column and detected with a fluorescence spectrophotometer. As little as 10 pmol of each were detected with good peak separation and reproducible retention times. The results of HPLC analysis showed that drought and shading induced an increase in total amino acid concentrations in needles; shading had the greater effect. Arginine and proline concentrations increased most in needles of drought‐stressed seedlings and remained high in unshaded seedlings recovering from drought 48 h after rewatering. Arginine and glutamine increased most in the shaded seedlings, which did not survive severe drought. The large increase in arginine in both drought‐stressed and severely shaded seedlings suggests that sequestering and storage of ammonia are important when stress reduces carbon fixation.