Spin-label EPR study in thylakoid membranes from a new herbicide-resistant D1 mutant from soybean cell cultures deficient in fatty acid desaturation

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Fatty acid desaturation effect on the lipid fluidity in thylakoid membranes isolated from the STR7 mutant was investigated by electron paramagnetic resonance (EPR) using spin label probes. The spectra of both 5- and 16-n-doxylstearic acid probes were measured as a function of the temperature between 10-305 K and compared to those of the wild type. This complete thermal evolution provides a wider picture of the dynamics. The spectra of the 5-n-doxylstearic acid probe were identical in both STR7 mutant and wild type thylakoids as well as their temperature evolution. However, differences were found with the 16-n-doxylstearic acid probe at temperatures between 230-305 K. The differences in the thermal evolution of the EPR spectra can be interpreted as a 5-10 K shift toward higher temperatures of the probe motional rates in the STR7 mutant as compared with that in the wild type. At temperatures below 230 K no differences were observed. The results indicated that the lipid motion in the outermost region of the thylakoids is the same in the STR7 mutant than in the wild type while the fluidity in the inner region of the STR7 mutant membrane decreases. Our data point out a picture of the STR7 thylakoid membrane in which the lipid motion is slower most probably as a consequence of fatty acid desaturation deficiency.
M.A. and I.G.-R. were recipients of a contract and a fellowship, respectively, from the Ministerio de Educación y Cultura of Spain. This work was supported by the Dirección General de Investigación Científica y Técnica (Grant PB98-1632) and by the Diputación General de Aragón (Project P17/98).