Abstract
A novel ascorbate peroxidase (APX) cDNA was obtained from Nelumbo nucifera (Elian). The phylogenetic analysis indicated that N. nucifera APX grouped together with chloroplastic APX of high plants. The recombinant protein expressed by PET-30a vector showed APX activity (0.04 mM ascorbate min−1 mg−1 protein). The APX mRNA was expressed in young leaves, roots, terminal buds, and leafstalks. Synergistic expression of N. nucifera APX and MnSOD mRNA was indicated in the short-term response to mechanical wounding.
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Dong C, Li GL, Li ZQ, Zhu HL, Zhou MQ, Hu ZL (2009) Molecular cloning and expression analysis of an MnSOD gene from Nelumbo nucifera. Appl Biochem Biotechnol 158:605–614
Hernandez JA, Olmos E, Corpas FJ, Sevilla F, del Rio LA (1995) Salt-induced oxidative stress in chloroplasts of pea plants. Plant Sci 105:151–167
Koussevitzky S, Suzuki N, Huntington S, Armijo L, Sha W, Cortes D, Shulaev V, Mittler R (2008) Ascorbate peroxidase I plays a key role in the response of Arabidopsis thaliana to stress combination. J Biol Chem 283:34197–34203
Li H, Yang J, Wang YY, Chen ZJ, Tu SS, Feng LL, Zhu YG, Li YS (2009) Expression of a novel OSPGYRP (rice proline-, glycine- and tyrosine-rich protein) gene, which is involved in vesicle trafficking, enhanced cold tolerance in E. coli. Biotechnol Lett 31:905–910
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−△△Ct methods. Methods 25:402–408
Lu ZQ, Takano T, Liu SK (2005) Purification and characterization of two ascorbate peroxidases of rice (Oryza sativa L.) expressed in Escherichia coli. Biotechnol Lett 27:63–67
Mittova V, Theodoulou FL, Kiddle G, Volokita M, Tal M, Foyer CH, Guy M (2004) Comparison of mitochondria ascorbate peroxidase in the cultivated tomato, Lycopersicon esculentum, and its wild, salt-tolerant relative, L. pennellii: a role for matrix isoforms in protection against oxidative damage. Plant Cell Environ 27:237–250
Nishikawa F, Kato M, Wang R, Hyodo H, Ikoma Y, Sugiura M, Yano M (2003) Two APX from broccoli: Identification, expression and characterization of their recombinant proteins. Postharv Biol Technol 27:147–156
Orozco-Cardenas M, Ryan CA (1999) Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway. Proc Natl Acad Sci USA 96:6553–6557
Park SY, Ryu SH, Jang LC, Kwon SY, Kim JG, Kwak SS (2004) Molecular cloning of a cytosolic ascorbate peroxidase cDNA from cell cultures of sweet potato and its expression in response to stress. Mol Genet Genomic 271:339–346
Shi WM, Muramoto Y, Ueda A, Takabe T (2001) Cloning of peroxisomal ascorbate peroxidase gene from barley and enhanced thermotolerance by overexpressing in Arabidopsis thaliana. Gene 273:23–27
Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Takeda T, Yabuta Y, Yoshimura K (2002) Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53:1305–1319
Shin SY, Lee HS, Kwon SY, Kwon ST, Kwak SS (2005) Molecular characterization of a cDNA encoding copper/zinc superoxide dismutase from cultured cells of Manihot esculenta. Plant Physiol Biochem 43:55–60
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599
Teixeira FK, Menezes-Benavente L, Margis R, Margis-Pinheiro M (2004) Analysis the molecular evolutionary history of the ascorbate peroxidase gene family: inferences from the rice genome. J Mol Evol 59:761–770
Varsamos S, Wendelaar Bonga SE, Flik G, Quere R, Commes T (2003) Cloning of a proopiomelanocortin cDNA from the pituitary gland of the sea bass (Dicentrarchus labrax) and assessment of mRNA expression in different tissues by means of real-time PCR. J Endocrinol 176:405–414
Wada K, Tada T, Nakamura Y, Ishikawa T, Yabuta Y, Yoshimura K, Shigeoka S, Nishimura K (2003) Crystal structure of chloroplastic ascorbate peroxidase from tobacco plants and structural insights into its instability. J Biochem 134:239–244
Wang X, Yang HJ, Ruan LW, Liu X, Li F, Xu X (2008) Cloning and characterization of a thermostable superoxide dismutase from thermophilic bacterium Rhodothermus sp. XMH10. J Ind Microbiol Biotechnol 35:133–139
Yoshimura K, Ishikawa T, Nakamura Y, Tamoi M, Takeda T, Nishimura K, Shigeoka S (1998) Comparative study on recombinant chloroplastic and cytosolic ascorbate peroxidase isozymes of spinach. Arch Biochem Biophys 353:55–63
Acknowledgments
This work is financially supported by the Key Grant Project of the Chinese Ministry of Education (no. 307018), National Science and Technology Pillar Program (no. 2007BAD37B06), Natural Science Foundation of Hubei Province (no. 2007ABD003), and National Natural Science Foundation of China (no. 30771375).
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Dong, C., Zheng, X., Li, G. et al. Cloning and Expression of One Chloroplastic Ascorbate Peroxidase Gene from Nelumbo nucifera . Biochem Genet 49, 656–664 (2011). https://doi.org/10.1007/s10528-011-9440-x
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DOI: https://doi.org/10.1007/s10528-011-9440-x