Cloning a glutathione peroxidase gene from Nelumbo nucifera and enhanced salt tolerance by overexpressing in rice.
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.
State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, 430072, People's Republic of China.
Ying
Y
Diao
Huaxue
H
Xu
Guolin
G
Li
Aiqing
A
Yu
Xia
X
Yu
Wanling
W
Hu
Xingfei
X
Zheng
Shaoqing
S
Li
Youwei
Y
Wang
Zhongli
Z
Hu
D000222:Adaptation, Physiological / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D016000:Cluster Analysis; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D018014:Gene Transfer Techniques; D005979:Glutathione Peroxidase / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D031653:Nelumbo / Q000201:enzymology*; D012275:Oryza / Q000235:genetics* / Q000502:physiology; D030821:Plants, Genetically Modified / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D054712:Salinity; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D005979:Glutathione Peroxidase