Accessible Requires Authentication Published by De Gruyter May 31, 2013

Sequences of Mn-sod gene from Pyropia haitanensis (Bangiales, Rhodophyta) and its expression under heat shock

Rui Yang, Wei Liu, Xiao-Long Zhang, Mei-Li Shen, Xue Sun and Hai-Min Chen
From the journal Botanica Marina


Manganese superoxide dismutase (Mn-SOD) is an essential enzyme involved in the detoxification of reactive oxygen by dismutation of the superoxide radical anion. In this study, full-length complementary DNA (cDNA) and genomic DNA of Mn-sod genes were cloned from Pyropia haitanensis (phsod), and its expression was monitored using quantitative polymerase chain reaction (qPCR). Scavenging of reactive oxygen species under heat shock was analyzed as well. The full-length cDNA of the phsod gene consists of 935 bp (GenBank accession number: EU715988) with an open reading frame of 675 bp, which encodes 224 amino acids. The length of the genomic sequence is 1416 bp and includes four exons and three introns. A highly conserved Mn-SOD signature (DVWEHAYY) was found. The transit peptide in the N-terminal helps protein targeting to the mitochondria. Results showed that the expression levels of phsod increased 3.4 times compared to controls and then suddenly decreased under a heat shock of 35°C for 30 min. After heat shock exposure and return to normal cultivation temperature, the expression levels of phsod decreased within 1 h of release from the heat shock and then increased within 3 h. SOD activity correlated with the expression of the phsod gene and, as expected, the concentration of ·O2- showed negative correlations with SOD activity.

Corresponding author: Rui Yang, Ministry of Education, Key Laboratory of Applied Marine Biotechnology, Ningbo University, No. 818 Fenghua Road, Ningbo, Zhejiang, 315211, China; and School of Marine Sciences, Ningbo University, No. 818 Fenghua Road, Ningbo, Zhejiang, 315211, China

This research was supported by the National Natural Science Foundation of China No. 40776077, Natural Science Foundation of Zhejiang No. LY12D06003, National Scientific and Technological Support Plan Subject, 2011BAD13B08, Zhejiang Scientific Innovative Team No. 2012R10025-07 and No.2011R09012-06, Ningbo Marine Algae Biotechnology Team No. 2011B81007 and No. 2012A610140 and the Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences. We thank Prof. Matthew J. Dring, Prof. Kirsten Heimann and Dr. Gunda Stöber for their assistance in editing this paper.


Armond, P.A., O. Björkman and L.A. Staehelin. 1980. Dissociation of supramolecular complexes in chloroplast membranes a manifestation of heat damage to the photosynthetic apparatus. BBA-Biomembranes 601: 433–442. Search in Google Scholar

Asada, K. and M. Takahashi. 1987. Production and scavenging of active oxygen in photosynthesis. In: (D.J. Kyle et al., eds.) Photoinhibition (topics in photosynthesis). Vol. 9. Elsevier Science, Amsterdam. pp. 227–287. Search in Google Scholar

Bagnoli, F., D. Giannino, S. Caparrini, A. Camussi, D. Mariotti and M.L. Racchi. 2002. Molecular cloning, characterisation and expression of a manganese superoxide dismutase gene from peach (Prunus persica [L.] Batsch). Mol. Genet. Genomics 267: 321–328. Search in Google Scholar

Bolwell, G.P. 1999. Role of active oxygen species and NO in plant defence responses. Curr. Opin. Plant. Biol. 2: 287–294. Search in Google Scholar

Boveris, A. and B. Chance. 1973. The mitochondrial generation of hydrogen peroxide. General properties and effect of hyperbaric oxygen. Biochem. J. 134: 707–716. Search in Google Scholar

Bowler, C., L. Slooten, S. Vandenbranden, R. De Rycke, J. Botterman, C. Sybesma, M. Van Montagu and D. Inzé. 1991. Manganese superoxide dismutase can reduce cellular damage mediated by oxygen radicals in transgenic plants. EMBO J. 10: 1723–1732. Search in Google Scholar

Brinster, R.L., J.M. Allen, R.R. Behringer, R.E. Gelinas and R.D. Palmiter. 1988. Introns increase transcriptional efficiency in transgenic mice. Proc. Natl. Acad. Sci. USA 85: 836–840. Search in Google Scholar

Buchman, A.R. and P. Berg. 1988. Comparison of intron-dependent and intron-independent gene expression. Mol. Cell. Biol. 8: 4395–4405. Search in Google Scholar

Cadenas, E. and K.J.A. Davies. 2000. Mitochondrial free radical generation, oxidative stress and aging. Free Radic. Biol. Med. 29: 222–230. Search in Google Scholar

Callis, J., M. Fromm and V. Walbot. 1987. Introns increase gene expression in cultured maize cells. Genes Dev. 1: 1183–1200. Search in Google Scholar

Choo, K., P. Snoeijs and M. Pedersén. 2004. Oxidative stress tolerance in the filamentous green algae Cladophora glomerata and Enteromorpha ahlneriana. J. Exp. Mar. Biol. Ecol. 298: 111–123. Search in Google Scholar

Collén, J. and I.R. Davison. 1999a. Stress tolerance and reactive oxygen metabolism in the intertidal red seaweeds Mastocarpus stellatus and Chondrus crispus. Plant Cell. Environ. 22: 1143–1151. Search in Google Scholar

Collén, J. and I.R. Davison. 1999b. Reactive oxygen metabolism in intertidal Fucus spp. (Phaeophyceae). J. Phycol. 35: 62–69. Search in Google Scholar

Collén, J. and I.R. Davison. 1999c. Reactive oxygen production and damage in intertidal Fucus spp. (Phaeophyceae). J. Phycol. 35: 54–61. Search in Google Scholar

Dai, Q., B. Yan, S. Huang, X. Liu, S. Peng, M.L.L. Miranda, A.Q. Chavez, B.S. Vergara and D.M. Olszyk. 1997. Response of oxidative stress defence systems in rice (Oryza sativa) leaves with supplemental UV-B radiation. Physiol. Plantarum 101: 301–308. Search in Google Scholar

Davison, I.R. and G.A. Pearson. 1996. Stress tolerance in intertidal seaweeds. J. Phycol. 32: 197–211. Search in Google Scholar

del Río, L.A., F.J. Corpas, L.M. Sandalio, J.M. Palma, M. Gómez and J.B. Barroso. 2002. Reactive oxygen species, antioxidant systems and nitric oxide in peroxisomes. J. Exp. Bot. 53: 1255–1272. Search in Google Scholar

Duncker, B.P., P.L. Davies and V.K. Walker. 1997. Introns boost transgene expression in Drosophila melanogaster. Mol. Gen. Genet. 254: 291–296. Search in Google Scholar

Eitinger, T. 2004. In vivo production of active nickel superoxide dismutase from Prochlorococcus marinus MIT9313 is dependent on its cognate peptidase. J. Bacteriol. 186: 7821–7825. Search in Google Scholar

Elstner, E.F. and A. Heupet. 1976. Inhibition of nitrite formation from hydroxylarnmoniumchloride: a simple assay for superoxide dismutases. Ann. Biochem. 70: 616–620. Search in Google Scholar

Fink, R.C. and J.G. Scandalios. 2002. Molecular evolution and structure-function relationships of the superoxide dismutase gene families in angiosperms and their relationship to other eukaryotic and prokaryotic superoxide dismutases. Arch. Biochem. Biophys. 399: 19–36. Search in Google Scholar

Fridovich, I. 1986. Superoxide dismutases. In: (A. Meister, ed.) Advances in enzymology and related areas of molecular biology. Wiley-Interscience, New York. pp. 61–97. Search in Google Scholar

Gruss, P., C.J. Lai, R. Dhar and G. Khoury. 1979. Splicing as a requirement for biogenesis of functional 16SmRNA of simian virus 40. Proc. Natl. Acad. Sci. USA 76: 4317–4321. Search in Google Scholar

Hale, A.J., C.A. Smith, L.C. Sutherland, V.E.A. Stoneman, V.L. Longthorne, A.C. Culhane and G.T. Williams. 1996. Apoptosis: molecular regulation of cell death. Eur. J. Biochem. 236: 1–26. Search in Google Scholar

Heinrich, S., K. Valentin, S. Frickenhaus, U. John and C. Wiencke. 2012. Transcriptomic analysis of acclimation to temperature and light stress in Saccharina latissima (Phaeophyceae). PLoS One 7: 1–13. Search in Google Scholar

Kaminaka, H., S. Morita, H. Yohoi, T. Masumura and H. Tanaka. 1997. Molecular cloning and characterization of a cDNA for plastidic copper/zinc-superoxide dismutase in rice (Oryza sativa L.). Plant Cell Physiol. 38: 65–69. Search in Google Scholar

Kreslavski, V.D., R. Carpentier, V.V. Klimov, N. Murata and S.I. Allakhverdiev. 2007. Molecular mechanisms of stress resistance of the photosynthetic apparatus. Biochemistry (Moscow). Suppl. Ser. A: Mem. Cell. Biol. 1: 185–205. Search in Google Scholar

Liu, F. and S.J. Pang. 2010. Performances of growth, photochemical efficiency, and stress tolerance of young sporophytes from seven populations of Saccharina japonica (Phaeophyta) under short-term heat stress. J. Appl. Phycol. 22: 221–229. Search in Google Scholar

Liu, L., M.J. White and T.H. MacRae. 1999. Transcription factors and their genes in higher plants: Functional domains, evolution and regulation. Eur. J. Biochem. 262: 247–257. Search in Google Scholar

Liu, W., Yang R., L. Xu, Zhang X., Wang Y. and Sun X. 2012. Cloning and expression of hsp70 for Porphyra haitanensis. J. Ningbo University (NSEE) 25: 17–25. Search in Google Scholar

Lu, I.F., M.S. Sung and T.M. Lee.2006. Salinity stress and hydrogen peroxide regulation of antioxidant defence system in Ulva fasciata. Mar. Biol. 150: 1–15. Search in Google Scholar

Mahalingam, R. and N. Fedoroff. 2003. Stress response, cell death and signalling: the many faces of reactive oxygen species. Physiol. Plantarum 119: 56–68. Search in Google Scholar

Marais, G. and L. Duret. 2001. Synonymous codon usage, accuracy of translation, and gene length in Caenorhabditis elefans. J. Mol. Evol. 52: 275–280. Search in Google Scholar

Misra, H.P. and I. Fridovich. 1977. Purification and properties of superoxide dismutase from a red alga, Porphyridium cruentum. J. Biol. Chem. 252: 6421–6423. Search in Google Scholar

Mittler, R., S. Vanderauwera, M. Gollery and F.V. Breusegem. 2004. Reactive oxygen gene network of plants. Trends Plant Sci. 9: 490–498. Search in Google Scholar

Miyasaka, H. 2002. Translation initiation AUG context varies with codon usage bias and gene length in Drosophila melanogaster. J. Mol. Evol. 55: 52–64. Search in Google Scholar

Nakamura, Y., N. Sasaki, M. Kobayashi, N. Ojima, M. Yasuike, Y. Shigenobu, M. Satomi, Y. Fukuma, K. Shiwaku, A. Tsujimoto, T. Kobayashi, I. Nakayama, F. Ito, K. Nakajima, M. Sano, T. Wada, S. Kuhara, K. Inouye, T. Gojobori and K. Ikeo. 2013. The first symbiont-free genome sequence of marine red Alga, susabi-nori (Pyropia yezoensis). Plos One 8:e57122. Search in Google Scholar

Palenik, B., B. Brahamsha, F.W. Larimer, M. Land, L. Hauser, P. Chain, J. Lamerdin, W. regala, E.E. Allen, J. McCarren, I. Paulsen, A. Dufresne, F. Partensky, E.A. Webb and J. Waterbury. 2003. The genome of a motile marine Synechococcus. Nature 424: 1037–1042. Search in Google Scholar

Palmiter, R.D., E.P. Sandgren, M.R. Avarbock, D.D. Allen and R.L. Brinster. 1991. Heterologous introns can enhance expression of transgenes in mice. Proc. Natl. Acad. Sci. USA 88: 478–482. Search in Google Scholar

Paulsen, G.M. 1994. High temperature responses of crop plants. In: (K.J. Boote, J.M. Bennett, T.R. Sinclairand and G.M. Paulsen, eds.) Physiology and determination of crop yield. ASA, CSSA and SSSA, Madison, WI. pp. 365–389. Search in Google Scholar

Pinhero, R.G., M.V. Rao, G. Paliyath, D.P. Murr and R.A. Fletcher. 1997. Changes in activities of antioxidant enzymes and their relationship to genetic and paclobutrazol-induced chilling tolerance of maize seedlings. Plant Physiol. 114: 695–704. Search in Google Scholar

Rao, M.V., G. Tamura and S. Shinano. 1996. Ultraviolet-B- and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol. 110: 125–136. Search in Google Scholar

Ross, C. and K.L. Van Alstyne. 2007. Intraspecific variation in stress-induced hydrogen peroxide scavenging by the ulvoid macroalga Ulva lactuca. J. Phycol. 43: 466–474. Search in Google Scholar

Sahoo, D., X. Tang and C. Yarish. 2002. Porphyra: the economic seaweed as a new experimental system. Curr. Sci. 83: 1313–1316. Search in Google Scholar

Sakamoto, A., Y. Nosaka and K. Tanaka. 1993. Cloning and sequence analysis of a complementary DNA for manganese-superoxide dismutase from rice (Oryza sativa L.). Plant Physiol. 103: 1477–1478. Search in Google Scholar

Seel, W.E., G.A.F. Hendry and J.A. Lee. 1992. The combined effects of desiccation and irradiance on mosses from xeric and hydric habitats. J. Exp. Bot. 43: 1023–1030. Search in Google Scholar

Shigeoka, S., T. Ishikawa, M. Tamoi, Y. Miyagawa, T. Takeda, Y. Yabuta and K. Yoshimura. 2002. Regulation and function of ascorbate peroxidase isoenzymes. J. Exp. Bot. 53: 1305–1319. Search in Google Scholar

Sutherland, J.E., S.C. Lindstrom, W.A. Nelson, J. Brodie, M.D.J. Lynch, M.S. Hwang, H.G. Choi, M. Miyata, N. Kikuchi, M.C. Oliveira, T. Farr, C. Neefus, A. Mols-Mortensen, D. Milstein and K.M. Müller. 2011. A new look at an ancient order: generic revision of the Bangiales (Rhodophyta). J. Phycol. 47: 1131–1151. Search in Google Scholar

Vinogradov, A.E. 2001. Intron length and codon usage. J. Mol. Evol. 52: 2–5. Search in Google Scholar

Wang, R., T. Liu, X. Zhou, Y. Zhuang and Y. Mao. 2006. Cloning and sequence analysing of Mn-SOD gene from Porphyra yezoensis Ueda. High Technol. Lett. 16: 522–528. Search in Google Scholar

Wang, Y. and X.X. Tang. 2005. Relationships between antioxidant activities and heat-resistant features of two Laminaria japonica strains. Chin. J. Appl. Ecol. 16: 1507–1512. Search in Google Scholar

Wolfe-Simon, F., D. Grzebyk, O. Schofield and P.G. Falkowski. 2005. The role and evolution of superoxide dismutases in algae. J. Phycol. 41: 453–465. Search in Google Scholar

Wu, G., R.W. Wilen, A.J. Robertson and L.V. Gusta. 1999. Isolation, chromosomal localization, and differential expression of mitochondrial manganese superoxide dismutase and chloroplastic copper/zinc superoxide dismutase genes in wheat. Plant Physiol. 120: 513–520. Search in Google Scholar

Xu, M.J., Y.X. Mao, X.C. Zhang, X.J. Zhou, Z.H. Sui, H.L. Zhou and J.H. Li. 2005. Bioinformatic analysis of expressed sequence tags from sporophyte of Porphyra yezoensis (bagiaeeae, Rhodophyta). Proy. Nat. Sci. 15: 24–34. Search in Google Scholar

Youn, H.D., E.J. Kim, J.H. Roe, Y.C. Hah and S.O. Kang. 1996. A novel nickel-containing superoxide dismutase from Streptomyces spp. Biochem. J. 318: 889–896. Search in Google Scholar

Zhang, X., R. Yang, Q. Yi, X. Sun, X. Wu and Y. Wang. 2010. Molecular cloning and characterization analysis of cytosolic ascorbate peroxidase gene from Porphyra haitanensis. Acta Oceanol. Sin. 32: 165–174. Search in Google Scholar

Zhu, D. and J.G. Scandalios. 1993. Maize mitochondrial manganese superoxide are encoded by a differentially expressed multigene family. Proc. Natl. Acad. Sci. USA 90: 9310–9314. Search in Google Scholar

Received: 2012-7-1
Accepted: 2013-5-10
Published Online: 2013-05-31
Published in Print: 2013-06-01

©2013 by Walter de Gruyter Berlin Boston