Cloning, identification, expression analysis and phylogenetic relevance of two NADP-dependent malic enzyme genes from hexaploid wheat.

The NADP-dependent malic enzyme (NADP-ME; EC1.1.1.40) found in many metabolic pathways catalyzes the oxidative decarboxylation of L-malate, producing pyruvate, CO(2) and NADPH. The NADP-MEs have been well studied in C4 plants but not well in C3 plants. In this study, we identified the NADP-ME isoforms from hexaploid wheat (Triticum aestivum L). Two different NADP-ME transcripts were first identified in this C3 plant. The first is named TaNADP-ME1 [NCBI: EU170134] and encodes a putative plastidic isoform, while the second is named TaNADP-ME2 [NCBI: EU082065] and encodes a cytosolic counterpart. Sequence alignment shows that the two NADP-ME isoforms share an identity of 73.26% in whole amino acids and 64.08% in nucleotide sequences. The phylogenetic analysis deciphers the two NADP-MEs as belonging to the monocots (Group II), which closely resemble OschlME6 and OscytME2, respectively. Tissue-specific analyses indicate that the two NADP-ME genes are both expressed in root, stem and leaf, and that TaNADP-ME1 is a leaf-abundant isoform. Semi-quantitative RT-PCR analysis show that the two NADP-ME transcripts in wheat leaves respond differently to low temperature, salt, dark and drought stresses stimuli and to exogenous abscisic acid (ABA) and salicylic acid (SA). Our results demonstrate that exogenous hormones (ABA and SA), as well as salt, low temperature, dark and drought stresses can regulate the expressions of TaNADP-ME1 and TaNADP-ME2 in wheat. This indicates that the two NADP-ME genes may play an important role in the response of wheat to ABA, SA, low temperature, salt, dark and drought stress.