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Plant and Cell Physiology Advance Access originally published online on May 15, 2007
Plant and Cell Physiology 2007 48(6):856-865; doi:10.1093/pcp/pcm061
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Alternative Oxidase Involvement in Cold Stress Response of Arabidopsis thaliana fad2 and FAD3+ Cell Suspensions Altered in Membrane Lipid Composition

Ana Rita Matos1,2,*, Cécile Hourton-Cabassa1, Dominique Ciçek1, Nathalie Rezé1, Joao Daniel Arrabaça2, Alain Zachowski1 and François Moreau1

1Université Pierre et Marie Curie-Paris 6, CNRS, UMR 7180 Physiologie Cellulaire et Moléculaire des Plantes, Le Raphaël, 3 Rue Galilée, F-94200 Ivry sur Seine, France
2Centro de Engenharia Biológica, Secção de Fisiologia e Bioquímica Vegetais, Faculdade de Ciências da Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal

*Corresponding author: armatos{at}fc.ul.pt; +351217500166.


  Abstract

To investigate how the fatty acid composition of membrane lipids influences cell growth and mitochondrial respiration, in particular the expression and capacity of alternative oxidase (AOX), under cold stress, we used the Arabidopsis thaliana fad2 knockout and FAD3+ -overexpressing cultured cells lines affected in extrachloroplastic fatty acid desaturation activities. At 22°C, fad2 mitochondria exhibited a low polyunsaturated fatty acid content and low protein to lipid ratio, while mitochondria from FAD3+ were enriched in linolenic acid and in total membrane protein. As a consequence, both mutants showed a higher membrane microviscosity than the wild type. After exposure to 9°C, FAD3+ mitochondria exhibited lower microviscosity and lower rigidification upon a temperature downshift than fad2. Furthermore, the extent of reduction of cell growth and respiratiory rates in the phosphorylating state was positively related to the cold sensitivity of each cell line, being more pronounced in fad2 that in the wild type, whereas the stability of those parameters reflected the cold resistance of FAD3+. In contrast, an increase in AOX capacity was observed in the three cell lines at 9°C. These inductions were correlated to AOX protein amounts and seem to result from an accumulation of AOX1c transcripts in the three cell lines and of AOX1a transcripts in wild-type and fad2 cells. The fact that there is no direct relationship between the degree of cold tolerance of each cell line and their ability to enhance their AOX capacity suggests that the participation of AOX in the response of Arabidopsis cells to cold stress does not necessarily favor cold tolerance.

Keywords: Alternative oxidase - Arabidopsis thaliana - Fatty acid desaturase - Low temperature - Mitochondria

Abbreviations: AOX, alternative oxidase; BSA, bovine serum albumin; COX, cytochrome c oxidase; DBI, double bond index; DTT, dithiothreitol; FAD, fatty acid desaturase; PUFA, polyunsaturated fatty acid; ROS, reactive oxygen species; RT–PCR, reverse transcription–PCR; WT, wild type.

(Received April 14, 2007; Accepted May 9, 2007)
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