NADP-malic enzyme (NADP-ME, EC 188.8.131.52) functions in many different pathways in plants, and has recently been implicated in plant defense such as in responses to wounding and UV-B radiation. In this study, we isolated a complementary DNA (cDNA) clone by using the differential display method and screening of a root cDNA library of rice (Oryza sativa. L) under carbonate (NaHCO3) stress, and identified it as one of the rice NADP-ME genes (we named it NADP-ME 2 , GenBank accession no. AB053295). The 5′ end of NADP-ME 2 was obtained by the 5′-RACE method, and the full-length cDNA had a length of 2217 bp encoding 593 amino acids. Expression of NADP-ME 2 mRNA in roots was induced by stress from carbonates (NaHCO3 and Na2CO3), NaCl, and environmental pH changes. NADP-ME 2 transcripts increased during 72-h exposures to NaHCO3, NaCl, and PEG stresses. Furthermore, NADP-ME activities in leaves and roots of rice seedlings increased by more than 50% in the presence of carbonates (NaHCO3 and Na2CO3), NaCl, and PEG. These results indicate that rice NADP-ME 2 responds to salts and osmotic stresses. Transgenic Arabidopsis plants over-expressing NADP-ME 2 were obtained through transformation, screening, Northern analysis and in situ NADP-ME activity assay. Transgenic Arabidopsis plants over-expressing NADP-ME 2 grew well in 1/2 × MS medium with 100 mM NaCl or 4% mannitol, whereas growth of wild-type (WT) Arabidopsis seedlings was strongly inhibited. In addition, under 125 mM NaCl stress, the root lengths of transgenic lines were about twice as long as those of the WT. These results suggest that NADP-ME 2 has a role in enhancing tolerance of plants to salt and osmotic stress.
Plant Molecular Biology – Springer Journals
Published: Jan 24, 2007
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