A phospholipase Dα gene ( AnPLDα ) was cloned from xerophytic desert plant Ammopiptanthus nanus and its overexpression enhanced salt tolerance of a PLDα1 deficient Arabidopsis mutant. Phospholipase Dα (PLDα) hydrolyzes phosphatidylcholine to produce phosphatidic acid, and plays crucial role in plant tolerance to abiotic stress. In this study, a phospholipase Dα gene (AnPLDα) was cloned from xerophyte Ammopiptanthus nanus by the methods of homologous cloning and rapid amplification of cDNA ends, and evaluated for its function in stress tolerance. The full-length cDNA was 2832 bp long, containing an open reading frame of 2427 bp that encodes 808 amino acids. The putative protein was predicted to be localized to the cytoplasm and this was confirmed by transient expression of a fluorescent fusion protein. The endogenous expression of the AnPLDα gene was induced by high salt, dehydration, cold and abscisic acid. The heterologous expression of the AnPLDα gene improved salt tolerance of an Arabidopsis pldα1 knocked out mutant, and positively regulated the expression of the AtABI, AtNCED, AtRD29A, AtRD29B and AtADH genes. Therefore, the AnPLDα gene was concluded to be involved in response to abiotic stress. The AnPLDα gene is a hopeful candidate for transgenic application to improve stress tolerance of commercial crops.
Keywords: Abiotic stress; Crops; Gene cloning; Phospholipase; Stress tolerance.