The plasma membrane–bound phospholipase Dδ enhances freezing tolerance in Arabidopsis thaliana

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Freezing injury is a major environmental limitation on the productivity and geographical distribution of plants. Here we show that freezing tolerance can be manipulated in Arabidopsis thaliana by genetic alteration of the gene encoding phospholipase Dδ (PLDδ), which is involved in membrane lipid hydrolysis and cell signaling. Genetic knockout of the plasma membrane–associated PLDδ rendered A. thaliana plants more sensitive to freezing, whereas overexpression of PLDδ increased freezing tolerance. Lipid profiling revealed that PLDδ contributed approximately 20% of the phosphatidic acid produced in wild-type plants during freezing, and overexpression of PLDδ increased the production of phosphatidic acid species. The PLDδ alterations did not affect the expression of the cold-regulated genes COR47 or COR78 or alter cold-induced increases in proline or soluble sugars, suggesting that the PLD pathway is a unique determinant of the response to freezing and may present opportunities for improving plant freezing tolerance.

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Figure 1: Decreased freezing tolerance in PLDδ-KO A. thaliana.
Figure 2: Increased freezing sensitivity of cold-acclimated PLDδ-KO seedlings of A. thaliana.
Figure 3: Overexpression of PLDδ (PLDδ-OE) and the effect of cold acclimation and freezing on the PLDδ expression in A. thaliana.
Figure 4: Increased freezing tolerance in PLDδ-overexpressing plants (PLDδ-OE).
Figure 5: Effect of PLDδ-KO and PLDδ-OE on the expression of cold-regulated genes and H2O2-induced cell death.
Figure 6: Changes in phosphatidic acid (PA) species and proline and soluble sugar content after cold treatments of PLDδ-KO, PLDδ-OE and wild-type A. thaliana.


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This work was supported by grants from the National Science Foundation (NSF), US Department of Agriculture, Kansas NSF EPSCoR and the Kansas State University Plant Biotechnology Center. The authors would like to thank Charles Rife for freezing chamber use, Todd Williams for acquisition of the ESI-MS/MS data and Christen Buseman for help with processing of the lipid profiling data. This is contribution 04-265-J from the Kansas Agricultural Experiment Station.

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Correspondence to Xuemin Wang.

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Li, W., Li, M., Zhang, W. et al. The plasma membrane–bound phospholipase Dδ enhances freezing tolerance in Arabidopsis thaliana. Nat Biotechnol 22, 427–433 (2004) doi:10.1038/nbt949

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