Gene cloning and expression of cytosolic glutathione reductase in rice (Oryza sativa L.).
We have isolated a cDNA (RGRC2) encoding glutathione reductase (GR) from rice (Oryza sativa L.). The comparison of deduced amino acid sequences from RGRC2 and other plant GR cDNAs indicated that RGRC2 encodes a putative cytosolic isoform. The recombinant RGRC2 protein had enzymatic properties comparable to those of GR from rice embryo. Subcellular fractionation showed that the RGRC2 protein is localized primarily in cytosol. mRNA and protein of RGRC2 were observed mainly in roots and calli but little in leaf tissues. Southern blot analysis showed that the RGRC2 gene exists as a single copy gene. Here, we have also isolated a genomic clone completely corresponding to RGRC2. The RGRC2 gene is split into 16 exons spread about 7.4 kb of chromosomal DNA, with coding sequence beginning in the 2nd exon and ending in the 16th exon. From the presence of two ABA-responsive elements in the 5'-flanking region of RGRC2, we examined the expression in rice seedlings treated with ABA and the ABA-related environmental stresses, chilling, drought and salinity. The expression of RGRC2 was strongly induced by all these treatments. We suggest that the expression of the rice cytosolic GR gene is regulated via ABA-mediated signal transduction pathway under environmental stresses.
Laboratory of Genetic Engineering, Faculty of Agriculture, Kyoto Prefectural University, Japan.
H
H
Kaminaka
S
S
Morita
M
M
Nakajima
T
T
Masumura
K
K
Tanaka
D020121:5' Untranslated Regions; D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D003080:Cold Temperature; D003600:Cytosol / Q000201:enzymology*; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005091:Exons; D018628:Gene Dosage; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005980:Glutathione Reductase / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology / Q000235:genetics*; D011994:Recombinant Proteins; D012639:Seeds / Q000201:enzymology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D020121:5' Untranslated Regions; D018076:DNA, Complementary; D007527:Isoenzymes; D011994:Recombinant Proteins; D000040:Abscisic Acid; D005980:Glutathione Reductase