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The influence of freezing on phospholipase D (PLD) was studied in Chorispora bungeana Fisch. & C. A. Mey., which is a naturally cold-tolerant species. During the freezing treatment (-4 °C), PLD activities in both microsomal and mitochondrial membranes increased at day 3, remained at a high level at day 6 and then declined to a moderate level. The RT-PCR analyses showed that PLD activity partially corresponded to the CbPLD gene transcript level. The freezing treatment resulted in increases in the Km and Vmax for microsomal and mitochondrial PLD, respectively. Freezing injury, as measured by electrolyte leakage and malondialdehyde content, peaked at day 6 and then gradually decreased. Alleviation of freezing injury was related to a decreased content of membrane-associated Ca2+. We suggest that the specific mechanism of cold resistance of C. bungeana is linked with PLD. © 2012 Springer Science+Business Media B.V.

Characterization of phospholipase D from Chorispora bungeana callus in response to freezing stress