Journal Article

Changes in freezing tolerance, plasma membrane H<sup>+</sup>-ATPase activity and fatty acid composition in <i>Pinus resinosa</i> needles during cold acclimation and de-acclimation

Françoise Martz, Marja-Liisa Sutinen, Sari Kiviniemi and Jiwan P. Palta

in Tree Physiology

Volume 26, issue 6, pages 783-790
Published in print June 2006 | ISSN: 0829-318X
Published online June 2006 | e-ISSN: 1758-4469 | DOI: http://dx.doi.org/10.1093/treephys/26.6.783
Changes in freezing tolerance, plasma membrane H+-ATPase activity and fatty acid composition in Pinus resinosa needles during cold acclimation and de-acclimation

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It has previously been suggested that plasma membrane ATPase (PM H+-ATPase, EC 3.6.1.3.) is a site of incipient freezing injury because activity increases following cold acclimation and there are published data indicating that activity of PM H+-ATPase is modulated by changes in lipids associated with the enzyme. To test and extend these findings in a tree species, we analyzed PM H+-ATPase activity and the fatty acid (FA) composition of glycerolipids in purified plasma membranes (PMs) prepared by the two-phase partition method from current-year needles of adult red pine (Pinus resinosa Ait.) trees. Freezing tolerance of the needles decreased from –56 °C in March to –9 °C in May, and increased from –15 °C in September to –148 °C in January. Specific activity of vanadate-sensitive PM H+-ATPase increased more than twofold following cold acclimation, despite a concurrent increase in protein concentration. During de-acclimation, decreases in PM H+-ATPase activity and freezing tolerance were accompanied by decreases in the proportions of oleic (18:1) and linoleic (18:2) acids and increases in the proportions of palmitic (16:0) and linolenic (18:3) acids in total glycerolipids extracted from the plasma membrane fraction. This pattern of changes in PM H+-ATPase activity and the 18:1, 18:2 and 18:3 fatty acids was reversed during cold acclimation. In the PM fractions, changes in FA unsaturation, expressed as the double bond index (1 × 18:1 + 2 × 18:2 + 3 × 18:3), were closely correlated with changes in H+-ATPase specific activity (r2 = 0.995). Changes in freezing tolerance were well correlated with DBI (r2 = 0.877) and ATPase specific activity (r2 = 0.833) in the PM fraction. Total ATPase activity in microsomal fractions also closely followed changes in freezing tolerance (r2 = 0.969). We conclude that, as in herbaceous plants, simultaneous seasonal changes in PM H+-ATPase activity and fatty acid composition occur during cold acclimation and de-acclimation in an extremely winter hardy tree species under natural conditions, lending support to the hypothesis that FA-regulated PM H+-ATPase activity is involved in the cellular response underlying cold acclimation and de-acclimation.

Keywords: fatty acid unsaturation; frost hardiness; plasma membrane; polar lipids; red pine; seasonal changes

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Subjects: Plant Sciences and Forestry

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