Cloning a glutathione peroxidase gene from Nelumbo nucifera and enhanced salt tolerance by overexpressing in rice
- 427 Downloads
A full-length cDNA clone encoding an 866 bp-length glutathione peroxidase protein (NnGPX) was isolated from lotus (Nelumbo nucifera L.). The deduced amino acid sequence of the NnGPX gene had significant homology with ATGPX6. A 3D structural model of the NnGPX was constructed by homology modeling. The cloned NnGPX gene was expressed in Escherichia coli, and a fusion protein of about 40 kDa was detected after isopropyl thiogalactoside induction. Under different concentrations of Na2SeO3 treatments, NnGPX was found to be an enzyme that does not contain selenium. Real-time PCR analysis showed that the NnGPX gene was expressed in all organs of lotus, and its high expression mainly occurred in organs with active metabolisms. NnGPX transcript increased remarkably in response to cold, heat, mechanical damage, and salt treatment. Subsequently, the NnGPX gene was introduced in Oryza sativa cv. Yuetai B. PCR results verified the integration of this gene into the genome of rice and reverse transcription-PCR verified that this gene had been expressed in transgenic rice. The transgenic plants were significantly more tolerant to salt stress compared with the wild-type.
KeywordsNelumbo nucifera Glutathione peroxidase (GPX) Reactive oxygen species (ROS) Rice Transformation Salt stress
This research was supported by National Key Technologies R&D Program (No. 2012BAD27B01). The authors thank the anonymous referees for their critical comments that contributed toward improving this manuscript.
- 7.Huang SZ, Tang XJ, Zhang L, Fu JR (2003) Thermotolerance and activity of antioxidative enzymes in lotus seeds. J Plant Physiol Mol Biol 29:421–424Google Scholar
- 9.Li Y, Zhang Y, Wei S, Liu L, Chen Y (2012) Recovery of antioxidant gene expression in sacred lotus (Nelumbo nucifera Gaertn) embryonic axes enhances tolerance to extreme high temperature. Afr J Biotechnol 11:12011–12019Google Scholar
- 16.Liu Y, Teng Z, Wang S, He G (2008) Effects of high temperature stress on soluble sugar and membrane protective enzyme of rice. J Southwest Uni 30:59–63Google Scholar
- 17.Zhang DZ, Wang PH, Zhao HX (1990) Determination of the content of free proline in wheat leaves. Plant Physiol Commun 94:462–465Google Scholar
- 20.Utomo A, Jiang XZ, Furuta S, Yun J, Levin DS, Wang YCJ, Desai KV, Green JE, Chen PL, Lee WH (2004) Identification of a novel putative non-selenocysteine containing phospholipid hydroperoxide glutathione peroxidase (NPGPx) essential for alleviating oxidative stress generated from polyunsaturated fatty acids in breast cancer cells. J Biol Chem 279:43522–43529CrossRefPubMedGoogle Scholar
- 25.Jung BG, Lee KO, Lee SS, Chi YH, Jang HH, Kang SS, Lee K, Lim D, Yoon SC, Yun DJ, Inoue Y, Cho MJ, Lee SY (2002) A Chinese cabbage cDNA with high sequence identity to phospholipid hydroperoxide glutathione peroxidases encodes a novel isoform of thioredoxin-dependent peroxidase. J Biol Chem 277:12572–12578CrossRefPubMedGoogle Scholar
- 26.Nicolas N, Vale´rie C, Jose´ G, Eric G, Masakazu H, Pascal R, David BK, Emmanuelle Issakidis, Jean-Pierre Jacquot, Nicolas Rouhier (2006) Plant glutathione peroxidases are functional peroxiredoxins distributed in several subcellular compartments and regulated during biotic and abiotic stresses. Plant Physiol 142:1364–1379CrossRefGoogle Scholar