Cloning and Functional Analysis of Thylakoidal Ascorbate Peroxidase (TAPX) Gene in Sugarcane

详细信息    查看全文

• 作者：Sheng Wang ; Kun-Kun Zhang ; Xing Huang ; Yan-Jiao Fan ; Li-Tao Yang ; Yang-Rui Li
• 关键词：Sugarcane ; Thylakoid ; Ascorbate peroxidase (TAPX) ; Expression analysis ; Prokaryotic expression ; Oxidative stress
• 刊名：Sugar Tech
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：17
• 期：4
• 页码：356-366
• 全文大小：2,304 KB
• 参考文献：Asada, K. 1992. Ascorbate peroxidase鈥擜 hydrogen peroxide-scavenging enzyme in plants. Physiologia Plantarum 85(2): 235鈥?41.CrossRef
  Asada, K. 1999. The water-water cycle in chloroplasts: scavenging of active oxygens and dissipation of excess photons. Annual Review of Plant Physiology and Plant Molecular Biology 50: 601鈥?39.CrossRef PubMed
  Badawi, G.H., N. Kawano, Y. Yamauchi, E. Shimada, R. Sasaki, A. Kubo, and K. Tanaka. 2004. Over-expression of ascorbate peroxidase in tobacco chloroplasts enhances the tolerance to salt stress and water deficit. Physiologia Plantarum 121(2): 231鈥?38.CrossRef PubMed
  Bandeo臒lu, E., F. Eyido臒an, M. Y眉cel, and H.A. 脰ktem. 2004. Antioxidant responses of shoots and roots of lentil to NaCl-salinity stress. Plant Growth Regulation 42: 69鈥?7.CrossRef
  Chen, G.X., and K. Asada. 1989. Asorbate peroxidase in tea leaves occurrence of two isoenzymes and their differences in enzymatic and molecular properties. Plant and Cell Physiology 30(7): 987鈥?98.
  Chen, Y., H. Wang, X. Wang, A. Cao, and P. Chen. 2006. Cloning and expression of peroxisomal ascorbate peroxidase gene from wheat. Molecular Biology Reporters 33(3): 207鈥?13.CrossRef
  Chew, O., J. Whelan, and A.H. Millar. 2003. Molecular definition of the ascorbate-glutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants. Journal of Biological Chemistry 278(47): 46869鈥?6877.CrossRef PubMed
  D鈥橝rcy-Lameta, A., R. Ferrari-Iliou, D. Contour-Ansel, P.T. Anh-Thu, and Y. Zuily-Fodil. 2006. Isolation and characterization of four ascorbate peroxidase cDNAs responsive to water deficit in cowpea leaves. Annals of Botany 97(1): 133鈥?40.PubMed Central CrossRef PubMed
  Deng, Z.Y., H.B. Liu, G.M. Zhang, F.X. Fang, H. He, Y.R. Li, R.Z. Yang, M. Li, G.L. Chen, C.X. Liang, and S.L. Bi. 2009. Survey of frost occurred regular harm in sugarcane. Sugar Crops of China 1: 47鈥?0.
  Foyer, C.H., and B. Hailiwell. 1997. Purification and properties of dehydroascorbate reductase from spinach leaves. Phytochemistry 16(9): 1347鈥?350.CrossRef
  Guan, Q.J., L. Li, T. Takano, and S.K. Liu. 2009. Cloning of an ascorbate peroxidase gene from Puccinellia tenuiflora and its expression analysis. Genomics and Applied Biology 28(4): 631鈥?39.
  Hernandez, J.A., E. Olmos, F.J. Corpas, F. Sevilla, and L.A. del Rio. 1995. Salt-induced oxidative stress in chloroplasts of pea plants. Plant Science 105(2): 151鈥?67.CrossRef
  Hou, Y.X., H.R. Tang, Y. Zhang, Y. Luo, and Q. Chen. 2009. Cloning and expression analysis of ascorbate peroxidase gene during fruit development and ripening in Fragaria. World Journal of Agricultural Science 5(6): 675鈥?79.
  Ishikawa, T., and S. Shigeoka. 2008. Recent advances in ascorbate biosynthesis and the physiological significance of ascorbate peroxidase in photosynthesizing organisms. Bioscience, Biotechnology, and Biochemistry 72(5): 1143鈥?154.CrossRef PubMed
  Jiang, M., Z.X. Zhang, and L.G. Yuan. 2012. Cloning and expression analysis of an ascorbate peroxidase gene BoAPX2 from Brassica oleracea var. italic. Aata Phytopathologica Sinica 42(4): 374鈥?80.
  Li, Y.R., and L.T. Yang. 2009. New developments of sugarcane industry and technology in China since 1990s. Southwest China Journal of Agricultural Sciences 22(5): 1469鈥?476.
  Li, Y.R., F.X. Fang, J.M. Wu, X. Li, R.H. Zhang, X.H. Liu, H. He, R.Z. Yang, L.T. Yang, G.L. Chen, S.Q. Sun, J.L. Xie, X.Y. Liu, W.H. Huang, W.X. Duan, W.Z. He, and M. Wang. 2011. Survey and defensive of production cold damage on sugarcane in Guang Xi in 2010/2011. Journal of Southern Agriculture 42(1): 37鈥?2.
  Lin, L., X. Wang, and Y. Wang. 2006. cDNA clone, fusion expression and purification of the novel gene related to ascorbate peroxidase from Chinese wild Vitis pseudoreticulata in E. coli. Molecular Biology Reporters 33(3): 197鈥?06.CrossRef
  Lu, H., R.L. Han, and X.N. Jiang. 2009. Heterologous expression and characterization of a proxidomal ascorbate peroxidase from Populus tomentosa. Molecular Biology Reporters 36(1): 21鈥?7.CrossRef
  Mano, S., K. Yamaguchi, M. Hayashi, and M. Nishimura. 1997. Stromal and thylakoid-bound ascorbate peroxidases are produced by alternative splicing in pumpkin. FEBS Letters 413(1): 21鈥?6.CrossRef PubMed
  Murgia, I., D. Tarantino, C. Vnanini, M. Bracale, S. Carravieri, and C. Soave. 2004. Arabidopsis thaliana plants overexpressing thylakoidal ascorbate peroxidase show increased resistance to Paraquat-induced photooxidative stress and to nitric oxide-induced cell death. Plant Journal 38(6): 940鈥?53.CrossRef PubMed
  Najami, N., T. Janda, W. Barriah, G. Kayam, M. Tal, M. Guy, and M. Volokita. 2008. Ascorbate peroxidase gene family in tomato: its identification and characterization. Molecular Genetics and Genomics 279(2): 171鈥?82.CrossRef PubMed
  Shi, F., R. Yamamoto, S. Shimamura, S. Hiraga, N. Nakayama, T. Nakamura, K. Yukawa, M. Hachinohe, H. Matsumoto, and S. Komatsu. 2008. Cytosolic ascorbate peroxidase 2 (cAPX 2) is involved in the soybean response to flooding. Phytochemistry 69(6): 1295鈥?303.CrossRef PubMed
  Shigeoka, S., T. Ishikawa, M. Tamoi, Y. Miyagawa, T. Takeda, and Y. Yabuta. 2002. Regulation and function of 85 ascorbate peroxidase isoenzymes. Journal of Experimental Botony 53(372): 1305鈥?319.CrossRef
  Stone, J.R. 2004. An assessment of proposed mechanism for sensing hydrogen peroxide in mammalian systems. Archives of Biochemistry and Biophysics 422: 119鈥?24.CrossRef PubMed
  Sun, W.H., X.Y. Chen, B. Du, W. Song, Y. Zhou, and Y. Wang. 2011. Tobacco seedlings overexpressing StAPX enhanced resistance to oxidative stress. Plant Physiology Journal 47: 613鈥?18.
  Sun, W.H., F. Li, D.F. Shu, X.C. Dong, X.M. Yang, and Q.W. Meng. 2009. Tobacco plants transformed with tomato sense LetAPX enhanced salt tolerance. Scientia Agricultura Sinica 42(4): 1165鈥?171.
  Teixeirra, F.K., L. Menezes-Renavente, V.C. Galv茫o, R. Margis, and M. Margis-Pinheiro. 2006. Rice ascorbate peroxidase gene family encodes functionally diverse isoforms localized in different subcellular compartments. Planta 224(2): 300鈥?14.CrossRef
  Wang, J., H. Zhang, and R.D. Allen. 1999. Overexpression of an Arabidopsis peroxisomal ascorbate peroxidase gene in tobacco increases protection against oxidative stress. Plant and Cell Physiology 40(7): 725鈥?32.CrossRef PubMed
  Yamaguchi-Shinozaki, K., and K. Shinozaki. 2006. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annual Review of Plant Biology 57: 781鈥?03.CrossRef PubMed
  Yoshimura, K., Y. Yabuta, T. Ishikawa, and S. Shiqeoka. 2000. Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. Plant Physiology 123(1): 223鈥?34.PubMed Central CrossRef PubMed
  
• 作者单位：Sheng Wang (1)
  Kun-Kun Zhang (1)
  Xing Huang (2)
  Yan-Jiao Fan (1)
  Li-Tao Yang (1) (2)
  Yang-Rui Li (2)
  
  1. Agricultural College, State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, 530004, China
  2. Guangxi Key Laboratory of Sugarcane Genetic Improvement, and Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture, Sugarcane Research Institute of Guangxi Academy of Agricultural Sciences-Sugarcane Research Center of Chinese Academy of Agricultural Sciences, Nanning, 530007, China
  
• 刊物主题：Agriculture;
• 出版者：Springer India
• ISSN：0974-0740

文摘

Ascorbate peroxidase (APX) is a key enzyme for active oxygen scavenging of higher plants, and it plays an important role in plants against abiotic stresses. In this study, a thylakoidal APX (TAPX) gene was cloned from sugarcane leaves by using RT-PCR technique, which was registered in GenBenk with the accession number JQ958327. Bioinformatics analysis showed that the new gene contained a 1,422 bp open reading frame and encoded 473 amino acids, and the predicted molecular weight was 51.1 kDa, containing the typical functional area and heme binding sites of peroxidase gene family. Real-time fluorescence quantitative PCR analysis showed the TAPX gene expressed in the roots, stems and leaves of sugarcane plant, and the highest expression level was found in the leaves. The TAPX gene was expressed in three exogenous stresses, i.e. polyethylene glycol (PEG), NaCl and H₂O₂. The expression pattern varied due to different regulatory mechanisms and the impact of low temperature stress on the expression is not significant. The TAPX gene was successfully expressed in E. coli. The TAPX gene excessive plant expression vector was built and transformed into tobacco with Agrobacterium-mediation, and transgenic tobacco plants were obtained. In conclusion, the TAPX gene was mainly expressed in the green tissue of sugarcane plant. It is presumed that TAPX gene is related with sugarcane resistance to osmotic stress. Further investigation on the biological functions of TAPX in sugarcane are warranted. Keywords Sugarcane Thylakoid Ascorbate peroxidase (TAPX) Expression analysis Prokaryotic expression Oxidative stress