Expression of an NADP-malic enzyme gene in rice (Oryza sativa. L) is induced by environmental stresses; over-expression of the gene in Arabidopsis confers salt and osmotic stress tolerance.
NADP-malic enzyme (NADP-ME, EC 1.1.1.40) 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-ME2, GenBank accession no. AB053295). The 5' end of NADP-ME2 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-ME2 mRNA in roots was induced by stress from carbonates (NaHCO3 and Na2CO3, NaCl, and environmental pH changes. NADP-ME2 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-ME2 responds to salts and osmotic stresses. Transgenic Arabidopsis plants over-expressing NADP-ME2 were obtained through transformation, screening, Northern analysis and in situ NADP-ME activity assay. Transgenic Arabidopsis plants over-expressing NADP-ME2 grew well in 1/2 x 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-ME2 has a role in enhancing tolerance of plants to salt and osmotic stress.
Liu
Shenkui
S
Alkali Soil Natural Environmental Science Center (ASNESC), Stress Molecular Biology Laboratory, Northeast Forestry University, Harbin 150040, P. R. China.
Cheng
Yuxiang
Y
Zhang
Xinxin
X
Guan
Qingjie
Q
Nishiuchi
Shunsaku
S
Hase
Kenichi
K
Takano
Tetsuo
T
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics* / Q000502:physiology; D002254:Carbonates / Q000494:pharmacology; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D006863:Hydrogen-Ion Concentration; D008291:Malate Dehydrogenase / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D009997:Osmotic Pressure; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D018517:Plant Roots / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D012965:Sodium Chloride / Q000494:pharmacology*; D014867:Water / Q000378:metabolism
D002254:Carbonates; D010940:Plant Proteins; D012333:RNA, Messenger; D014867:Water; D012965:Sodium Chloride; D008291:Malate Dehydrogenase; C031917:malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)