Journal of Plant Research

, Volume 130, Issue 2, pp 373–386 | Cite as

Overexpression of the ascorbate peroxidase gene from eggplant and sponge gourd enhances flood tolerance in transgenic Arabidopsis

  • Chih-Ming Chiang
  • Chiu-Chen Chen
  • Shi-Peng Chen
  • Kuan-Hung Lin
  • Li-Ru Chen
  • Yu-Huei Su
  • His-Cheng Yen
Regular Paper

Abstract

Previously, we found that the flood resistance of eggplant (Solanum melongena) and sponge gourd (Luffa cylindrica) enhanced ascorbate peroxidase (APX) activity under flooding, and consequently, both the SmAPX and LcAPX genes were cloned. In this study, the SmAPX and LcAPX genes were transferred under a ubiquitin promoter to Arabidopsis (At) via Agrobacterium tumefaciens. The expression and amount of APX and APX activities of the SmAPX and LcAPX transgenic lines were significantly higher than those of non-transgenic (NT) plants under a waterlogged condition. Furthermore, the SmAPX, LcAPX, At-sucrose synthases (SUS)-1, phosphoenolpyruvate carboxylase (PEPC), and lactate dehydrogenase (LDH) genes were overexpressed in all transgenic Arabidopsis lines after flooding treatment. Compared to NT plants, the malondialdehyde (MDA) contents and H2O2 accumulation were significantly lower, but germination rates were significantly higher in all transgenic lines with higher APX activity, indicating that the overexpression of SmAPX and LcAPX in Arabidopsis could enhance flood tolerance by eliminating H2O2. Moreover, Arabidopsis seedlings overexpressing SmAPX and LcAPX also displayed greater resistance to flooding and less oxidative injury than NT plants subjected to flooding condition.

Keywords

Flood tolerance Transgenic Arabidopsis Ascorbate peroxidase 

Supplementary material

10265_2016_902_MOESM1_ESM.docx (18 kb)
Supplementary material 1 (DOCX 18 KB)
10265_2016_902_MOESM2_ESM.docx (423 kb)
Supplementary material 2 (DOCX 422 KB)

References

  1. Abdelbagi MI, Evangelina SE, Georgina VV, David JM (2009) Mechanisms associated with tolerance to flooding during germination and early seedling growth in rice (Oryza sativa). Ann Bot 103:197–209CrossRefGoogle Scholar
  2. Bailey-Serres J, Fukao T, Ronald P, Ismail A, Heuer S, Mackill D (2010) Submergence tolerant rice: SUB1’s Journey from landrace to modern cultivar. Rice 3:138–147CrossRefGoogle Scholar
  3. Boris B, Vartapetian A, Michael B, Jackson C (1997) Plant adaptations to anaerobic stress. Ann Bot 79:3–20Google Scholar
  4. Cabello JV, Giacomelli JI, Piattoni CV, Iglesias AA, Chan RL (2016) The sunflower transcription factor HaHB11 improves yield, biomass and tolerance to flooding in transgenic Arabidopsis plants. J Biotech 222:73–83CrossRefGoogle Scholar
  5. Chen W, Yao Q, Patil GB, Agarwal G, Deshmukh RK, Lin L, Wang B, Wang Y, Prince SJ, Song L, Xu D, Yongqiang CA, Valliyodan B, Varshney RK, Nguyen HT (2016) Identification and comparative analysis of differential gene expression in soybean leaf tissue under drought and flooding stress revealed by RNA-sEq. Front Plant Sci 19. doi: 10.3389/fpls.2016.01044
  6. Chiang CM, Chen LFO, Shih SW, Lin KH (2015a) Expression of eggplant ascorbate peroxidase increases the tolerance of transgenic rice plants to flooding stress. J Plant Biochem Biotechnol 24:257–267Google Scholar
  7. Chiang CM, Chien HL, Chen LFO, Hsiung TC, Chiang MC, Chen SP, Lin KH (2015b) Overexpression of the genes coding ascorbate peroxidase from Brassica campestris enhances heat tolerance in transgenic Arabidopsis thaliana. Biol Plant 59:305–315Google Scholar
  8. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefPubMedGoogle Scholar
  9. De Buck S, Windelsb P, De Looseb M, Depicker A (2004) Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable β-glucuronidase accumulation levels. Cell Mol Life Sci 61:2632–2645CrossRefPubMedGoogle Scholar
  10. Diaz-Vivancos P, Faize M, Barba-Espin G, Faize L, Petri C, Hernández JA, Burgos L (2013) Ectopic expression of cytosolic superoxide dismutase and ascorbate peroxidase leads to salt stress tolerance in transgenic plums. Plant Biotechnol J 11:976–985CrossRefPubMedGoogle Scholar
  11. Djanaguiraman M, Prasad PV, Seppanen M (2010) Selenium protects sorghum leaves from oxidative damage under high temperature stress by enhancing antioxidant defense system. Plant Physiol Biochem 48:999–1007CrossRefPubMedGoogle Scholar
  12. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  13. Fukao T, Bailey-Serres J (2008) Ethylene- A key regulator of submergence responses in rice. Plant Sci 175:43–51CrossRefGoogle Scholar
  14. Fukao T, Xu K, Ronald PC, Bailey-Serres J (2006) A variable cluster of ethylene response factor-like genes regulates metabolic and developmental acclimation responses to submergence in rice. Plant Cell 18:2021–2034CrossRefPubMedPubMedCentralGoogle Scholar
  15. Geigenberger P (2003) Response of plant metabolism to too little oxygen. Curr Opin Plant Biol 6:247–256CrossRefPubMedGoogle Scholar
  16. Gibbs DJ, Lee SC, Isa NM, Gramuglia S, Fukao T, Bassel GW, Correia CS, Corbineau F, Theodoulou FL, Bailey-Serres J, Holdsworth MJ (2011) Homeostatic response to hypoxia is regulated by the N-end rule pathway in plants. Nature 479:415–418CrossRefPubMedPubMedCentralGoogle Scholar
  17. Guan Q, Takano T, Liu S (2012) Genetic transformation and analysis of rice OsAPx2 gene in Medicago sativa. Plos One 7:e41233CrossRefPubMedPubMedCentralGoogle Scholar
  18. Hattori Y, Nagai K, Furukawa S, Song XJ, Kawano R, Sakakibara H, Wu J, Matsumoto T, Yoshimura A, Kitano H, Matsuoka M, Mori H, Ashikari M (2009) The ethylene response factors SNORKEL1 and SNORKEL2 allow rice to adapt to deep water. Nature 460:1026–1030CrossRefPubMedGoogle Scholar
  19. Hsing YI, Chern CG, Fan MJ, Lu PC, Chen KT, Lo SF, Sun PK, Ho SL, Lee KW (2007) A rice gene activation/knockout mutant resource for high throughput functional genomics. Plant Mol Biol 63:351–364CrossRefPubMedGoogle Scholar
  20. Hsu FC, Chou MY, Peng HP, Chou SJ, Shih MC (2011) Insights into hypoxic systemic responses based on analyses of transcriptional regulation in Arabidopsis. Plos One 6:e28888CrossRefPubMedPubMedCentralGoogle Scholar
  21. Huynh LN, Vantoai T, Streeter J, Banowetz G (2005) Regulation of flooding tolerance of SAG12: ipt Arabidopsis plants by cytokinin. J Exp Bot 56:1397–1407CrossRefGoogle Scholar
  22. Kim SI, Veena SB, Gelvin SB (2007) Genome-wide analysis of Agrobacterium T-DNA integration sites in the Arabidopsis genome generated under non-selective conditions. Plant J 51:779–791CrossRefPubMedGoogle Scholar
  23. Kim MD, Kim YH, Kwon SY, Yun DJ, Kwak SS, Lee HS (2010) Enhanced tolerance to methyl viologen-induced oxidative stress and high temperature in transgenic potato plants overexpressing the CuZnSOD, APX and NDPK2 genes. Physiol Plant 140:153–162CrossRefPubMedGoogle Scholar
  24. Komatsu S, Nakamura T, Sugimoto Y, Sakamoto K (2014) Proteomic and metabiolomic analyses of soybean root tips under flooding stress. Protein Pept Lett 21:865–884CrossRefPubMedGoogle Scholar
  25. Kosugi H, Kikugawa K (1985) Thiobarbituric acid reaction of aldehydes and oxidized lipids in glacial acetic acid. Lipids 20:915–920CrossRefGoogle Scholar
  26. Kunst A, Draeger B, Ziegenhorn J (1988) Colorimetric methods with glucose oxidase and peroxidase. In: Bergermeyer HU (ed) Methods of enzymatic analysis. VI. Metabolites. I. Carbohydrates. Verlag-Chemie, Weinheim, pp 178–185Google Scholar
  27. Lee SH, Ahsan N, Lee KW, Kim DH, Lee DG, Kwak SS, Kwon SY, Kim TH, Lee BH (2007) Simultaneous overexpression of both CuZn superoxide dismutase and ascorbate peroxidase in transgenic tall fescue plants confers increased tolerance to a wide range of abiotic stresses. J Plant Physiol 164:1626–1638CrossRefPubMedGoogle Scholar
  28. Licausi F, Kosmacz M, Weits DA, Giuntol B, Giorgi F, Voesenek LACJ, Perata P, van Dongen JT (2011) Oxygen sensing in plants is mediated by an N-end rule pathway for protein destabilization. Nature 479:419–422CrossRefPubMedGoogle Scholar
  29. Lin KH, Lo HF, Lin CH, Chan MT (2007) Cloning and expression analysis of ascorbate peroxidase gene from eggplant under flooding stress. Bot Stud 48:25–34Google Scholar
  30. Lin KH, Huang HC, Lin CY (2010) Cloning, expression and physiological analysis of broccoli catalase gene and Chinese cabbage ascorbate peroxidase gene under heat stress. Plant Cell Rep 29:575–593CrossRefPubMedGoogle Scholar
  31. Lin KH, Kuo WS, Chiang CM, Hsiung TC, Chiang MC, Lo HF (2013) Study of sponge gourd ascorbate peroxidase and winter squash superoxide dismutase under respective flooding and chilling stresses. Scientia Hort 162:333–340CrossRefGoogle Scholar
  32. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  33. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–869Google Scholar
  34. Passaia G, Spagnolo FL, Caverzan A, Jardim-Messeder D, Christoff AP, Gaeta ML, de Araujo Mariath JE, Margis R, Margis-Pinheiro M (2013) The mitochondrial glutathione peroxidase GPX3 is essential for H2O2 homeostasis and root and shoot development in rice. Plant Sci 208:93–101CrossRefPubMedGoogle Scholar
  35. Porra R, Thompson W, Kriedelman P (1989) Determination of accurate extraction and simultaneously equation for assaying chlorophyll a and b extracted with different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochem Biophys Acta 975:384–394Google Scholar
  36. Sato Y, Masuta Y, Saito K, Murayama S, Ozawa K (2011) Enhanced chilling tolerance at the booting stage in rice by transgenic overexpression of the ascorbate peroxidase gene, OsAPXa. Plant Cell Rep 30:399–406CrossRefPubMedGoogle Scholar
  37. Setter TL, Waters I, Wallace I, Bhekasut P, Greenway H (1989) Submergence of rice. I. Growth and photosynthetic response to CO2 enrichment of floodwater. Australian. J Plant Physiol 6:251–263Google Scholar
  38. Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Yabuta Y, Youshimura K (2002) Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53:1305–1319CrossRefPubMedGoogle Scholar
  39. Shingaki-Wells RN, Huang S, Taylor NL, Carroll AJ, Zhou W, Millar AH (2011) Differential molecular responses of rice and wheat coleoptiles to anoxia reveal novel metabolic adaptations in amino acid metabolism for tissue tolerance. Plant Physiol 156:1706–1724CrossRefPubMedPubMedCentralGoogle Scholar
  40. Shiono K, Takahashi H, Colmer TD, Nakazono M (2008) Role of ethylene in acclimations to promote oxygen transport in roots of plants in waterlogged soils. Plant Sci 175:52–58CrossRefGoogle Scholar
  41. Tadege M, Dupuis I, Kuhlemeier C (1998) Ethanolic fermentation, new functions for an old pathway. Trends Plant Sci 4:320–325CrossRefGoogle Scholar
  42. Talarczyk A, Krzymowska M, Borucki W, Hennig J (2002) Effect of yeast CTA1 gene expression on response of tobacco plants to tobacco mosaic virus infection. Plant Physiol 129:1032–1048CrossRefPubMedPubMedCentralGoogle Scholar
  43. Vargas WA, Salerno GL (2010) The Cinderella story of sucrose hydrolysis: alkaline/neutral invertases, from cyanobacteria to unforeseen roles in plant cytosol and organelles. Plant Sci 178:1–8CrossRefGoogle Scholar
  44. Varvara PG, Bernard RG (2001) Ethylene and flooding stress in plants. Plant Physiol Biochem 39:1–9CrossRefGoogle Scholar
  45. Wang W, Vignani R, Scali M, Cresti M (2006) An universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis. Electrophoresis 27:2782–2786CrossRefPubMedGoogle Scholar
  46. Wang HS, Yu C, Zhu ZJ, Yu XC (2011) Overexpression in tobacco of a tomato GMPase gene improves tolerance to both low and high temperature stress by enhancing antioxidation capacity. Plant Cell Rep 30:1029–1040CrossRefPubMedGoogle Scholar
  47. Wise AA, Liu Z, Binns AN (2006) Three methods for the introduction of foreign DNA into Agrobacterium. Methods Mol Biol 343:43–53PubMedGoogle Scholar
  48. Yoshiki Y, Kahhara T, Sakabe Y, Yamasaki T (2001) Superoxide and DPPH radical-scavenging activities of soyasponin β related to gallic acid. Biosic Biotechnol Biochem 65:2162–2165CrossRefGoogle Scholar

Copyright information

© The Botanical Society of Japan and Springer Japan 2017

Authors and Affiliations

  • Chih-Ming Chiang
    • 1
  • Chiu-Chen Chen
    • 1
  • Shi-Peng Chen
    • 2
  • Kuan-Hung Lin
    • 3
    • 4
  • Li-Ru Chen
    • 4
  • Yu-Huei Su
    • 1
  • His-Cheng Yen
    • 1
  1. 1.Department of BiotechnologyMing Chuan UniversityTaoyuanTaiwan, Republic of China
  2. 2.Graduate Institute of Plant BiologyNational Taiwan UniversityTaipeiTaiwan, Republic of China
  3. 3.Faculty of Applied SciencesTon Duc Thang UniversityHo Chi Minh CityVietnam
  4. 4.Department of Horticulture and BiotechnologyChinese Culture UniversityTaipeiTaiwan, Republic of China

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