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Molecular cloning and characterization of SoNCED, a novel gene encoding 9-cis-epoxycarotenoid dioxygenase from sugarcane (Saccharum officinarum L.)

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Abstract

Abscisic acid (ABA) plays important roles in adaptive responses to various environmental stresses. The rate-limiting step in ABA biosynthesis is the oxidative cleavage of cis-epoxycarotenoids, which is catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED). In this experiment, a full-length cDNA encoding NCED gene was cloned by RT-PCR and RACE from sugarcane (Saccharum officinarum L.). The full-length of SoNCED is 2,521 bp with 1,827 bp open reading frame, encoding a peptide of 608 amino acids. The calculated molecular weight of protein was 65.9 kDa with isoelectric point of 6.04. Conserved domains prediction indicated a chloroplast-targeting peptide located at N-terminus of SoNCED. Phylogenetic tree, constructed by Neighbor-Joining method indicated that SoNCED shared high identity with the NCEDs reported from other plant species. Sequence alignment revealed that the basic secondary structure including α-helices, β-strands, β-propeller and His residues coordinating catalytic sites of SoNCED were highly conserved as in the NCEDs from other plants. Tissue specific expression analysis using quantitative real-time PCR showed a significant increase in SoNCED mRNA level and its correlation with O2 – production rate and ABA accumulation in leaves and roots of sugarcane variety GT21 when exposed to water stress. Further, the stimulation of SoNCED mRNA level, O2 – production rate and ABA content after exogenous application of ABA (100 μMol l−1) proved its involvement in pathways providing tolerance to drought stress.

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References

  • Battisti DS, Naylor RL (2009) Historical warnings of future food insecurity with unprecedented seasonal heat. Science 323:240–244

    Article  PubMed  CAS  Google Scholar 

  • Chernys JT, Zeevaart JAD (2000) Characterization of the 9-cis-epoxycarotenoid dioxygenase gene family and the regulation of abcisic acid biosynthesis in avocado. Plant Physiol 124:343–353

    Article  PubMed  CAS  Google Scholar 

  • Cutler SR, Rodriguez PL, Finkelstein RR, Abrams SR (2010) Abscisic acid: emergence of a core signaling network. Annu Rev Plant Biol 61:651–679

    Article  PubMed  CAS  Google Scholar 

  • del Río LA, Sandalio LM, Corpas FJ, Palma JM, Barroso JB (2006) Reactive oxygen species and reactive nitrogen species in peroxisomes. Production, scavenging, and role in cell signaling. Plant Physiol 141:330–335

    Article  PubMed  Google Scholar 

  • Elstner EF, Heupel A (1976) Inhibition of nitrite formation from hydroxylammoniumchloride: a simple assay for superoxide dismutase. Anal Biochem 70:616–620

    Article  PubMed  CAS  Google Scholar 

  • Endo A, Sawada Y, Takahashi H, Okamoto M, Ikegami K, Koiwai H, Seo M, Toyomasu T, Mitsuhashi W, Shinozaki K, Mikio N, Yuji K, Tomokazu K, Eiji N (2008) Drought induction of Arabidopsis 9-cis-epoxycarotenoid dioxygenase occurs in vascular parenchyma cells. Plant Physiol 147:1984–1993

    Article  PubMed  CAS  Google Scholar 

  • Finkelstein R, Reeves W, Ariizumi T, Steber C (2008) Molecular aspects of seed dormancy. Annu Rev Plant Biol 59:387–415

    Article  PubMed  CAS  Google Scholar 

  • Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930

    Article  PubMed  CAS  Google Scholar 

  • Guan LM, Zhao J, Scandalios JG (2000) Cis-elements and trans-factors that regulate expression of maize Cat1 antioxidant gene in response to ABA and osmotic stress: H2O2 is the likely intermediary signaling molecule for the response. Plant J 22:87–95

    Article  PubMed  CAS  Google Scholar 

  • Hu X, Zhang A, Zhang J, Jiang M (2006) Abscisic acid is a key inducer of hydrogen peroxide production in leaves of maize plants exposed to water stress. Plant Cell Physiol 47:1484–1495

    Article  PubMed  CAS  Google Scholar 

  • Ikegami K, Okamoto M, Seo M, Koshiba T (2009) Activation of abscisic acid biosynthesis in the leaves of Arabidopsis thaliana in response to water deficit. J Plant Res 122:235–243

    Article  PubMed  CAS  Google Scholar 

  • Jaspers P, Kangasjärvi J (2010) Reactive oxygen species in abiotic stress signaling. Physiol Plant 138:405–413

    Article  PubMed  CAS  Google Scholar 

  • Jiang M, Zhang J (2001) Effect of abscisic acid on active oxygen species, antioxidative defence system and oxidative damage in leaves of maize seedlings. Plant Cell Physiol 42:1265–1273

    Article  PubMed  CAS  Google Scholar 

  • Jiang M, Zhang J (2002) Water stress-induced abscisic acid accumulation triggers the increased generation of reactive oxygen species and up-regulates the activities of antioxidant enzymes in maize leaves. J Exp Bot 53:2401–2410

    Article  PubMed  CAS  Google Scholar 

  • Kaminaka H, Morita S, Tokumoto M, Yokoyama H, Masumura T, Tanaka K (1999) Molecular cloning and characterization of a cDNA for an iron-superoxide dismutase in rice (Oryza sativa L.). Biosci Biotechnol Biochem 63:302–308

    Article  PubMed  CAS  Google Scholar 

  • Kim TH, Böhmer M, Hu H, Nishimura N, Schroeder JI (2010) Guard cell signal transduction network: advances in understanding abscisic acid, CO2, and Ca2+ signaling. Annu Rev Plant Biol 61:561–591

    Article  PubMed  CAS  Google Scholar 

  • Kloer DP, Schulz GE (2006) Structural and biological aspects of carotenoid cleavage. Cell Mol Life Sci 63:2291–2303

    Article  PubMed  CAS  Google Scholar 

  • Kogan FN (1997) Global drought watch from space. Bull Am Meteorol Soc 78:621–636

    Article  Google Scholar 

  • Kozak M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15:8125–8148

    Article  PubMed  CAS  Google Scholar 

  • Li Q, Li P, Sun L, Wang YP, Ji K, Sun YF, Dai SJ, Chen P, Duan CR, Leng P (2012) Expression analysis of β-glucosidase genes that regulate abscisic acid homeostasis during watermelon (Citrullus lanatus) development and under stress conditions. J Plant Physiol 169:78–85

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Lu SY, Su W, Li HH, Guo ZF (2009) Abscisic acid improves drought tolerance of triploid bermudagrass and involves H2O2- and NO-induced antioxidant enzyme activities. Plant Physiol Bioch 47:132–138

    Article  CAS  Google Scholar 

  • Marris E (2008) Water: more crop per drop. Nature 453:273–277

    Article  Google Scholar 

  • Maurel C, Verdoucq L, Luu DT, Santoni V (2008) Plant aquaporins: membrane channels with multiple integrated functions. Annu Rev Plant Biol 59:595–624

    Article  PubMed  CAS  Google Scholar 

  • Messing SA, Gabelli SB, Echeverria I, Vogel JT, Guan JC, Tan BC, Klee HJ, McCarty DR, Amzel LM (2010) Structural insights into maize Viviparous14, a key enzyme in the biosynthesis of the phytohormone abscisic acid. Plant Cell 22:2970–2980

    Article  PubMed  CAS  Google Scholar 

  • Miller G, Shulaev V, Mittler R (2008) Reactive oxygen signaling and abiotic stress. Physiol Plant 133:481–489

    Article  PubMed  CAS  Google Scholar 

  • Navrot N, Rouhier N, Gelhaye E, Jaquot JP (2007) Reactive oxygen species generation and antioxidant systems in plant mitochondria. Physiol Plant 129:185–195

    Article  CAS  Google Scholar 

  • Qin X, Zeevaart JAD (1999) The 9-cis-epoxycarotenoid cleavage reaction is the key regulatory step of abscisic acid biosynthesis in water-stressed bean. Proc Natl Acad Sci U S A 96:15354–15361

    Article  PubMed  CAS  Google Scholar 

  • Qin X, Zeevaart JAD (2002) Overexpression of a 9-cis-epoxycarotenoid dioxygenase gene in Nicotiana plumbaginifolia increases abscisic acid and phaseic acid levels and enhances drought tolerance. Plant Physiol 128:544–551

    Article  PubMed  CAS  Google Scholar 

  • Qin X, Yang SH, Kepsel AC, Schwartz SH, Zeevaart JAD (2008) Evidence for abscisic acid biosynthesis in Cuscuta reflexa, a parasitic plant lacking neoxanthin. Plant Physiol 147:816–822

    Article  PubMed  CAS  Google Scholar 

  • Qiu YP, Yu DQ (2009) Over-expression of the stress-induced OsWRKY45 enhances disease resistance and drought tolerance in Arabidopsis. Environ Exp Bot 65:35–47

    Article  CAS  Google Scholar 

  • Rodrigo MJ, Alquezar B, Zacarías L (2006) Cloning and characterization of two 9-cis-epoxycarotenoid dioxygenase genes, differentially regulated during fruit maturation and under stress conditions, from orange (Citrus sinensis L. Osbeck). J Exp Bot 57:633–643

    Article  PubMed  CAS  Google Scholar 

  • Schwartz SH, Tan BC, Gage D, Zeevaart JAD, McCarty DR (1997) Specific oxidative cleavage of carotenoids by VP14 of maize. Science 276:1872–1874

    Article  PubMed  CAS  Google Scholar 

  • Schwartz SH, Qin X, Zeevaart JAD (2003) Elucidation of the indirect pathway of abscisic acid biosynthesis by mutants, genes, and enzymes. Plant Physiol 131:1591–1601

    Article  PubMed  CAS  Google Scholar 

  • Shinozaki K, Yamaguchi-Shinozaki K (2007) Gene networks involved in drought stress response and tolerance. J Exp Bot 58:221–227

    Article  PubMed  CAS  Google Scholar 

  • Suhita D, Raghavendra AS, Kwak JM, Vavasseur A (2004) Cytoplasmic alkalization precedes reactive oxygen species production during methyl jasmonate- and abscisic acid-induced stomatal closure. Plant Physiol 134:1536–1545

    Article  PubMed  CAS  Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    Article  PubMed  CAS  Google Scholar 

  • Tan BC, Schwartz SH, Zeevaart JAD, McCarty DR (1997) Genetic control of abscisic acid biosynthesis in maize. Proc Natl Acad Sci U S A 94:12235–12240

    Article  PubMed  CAS  Google Scholar 

  • Tan BC, Cline K, McCarty DR (2001) Localization and targeting of the VP14 epoxy-carotenoid dioxygenase to chloroplast membranes. Plant J 27:373–382

    Article  PubMed  CAS  Google Scholar 

  • Vaahtera L, Brosché M (2011) More than the sum of its parts - How to achieve a specific transcriptional response to abiotic stress. Plant Sci 180:421–430

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Wang Z, Dong J, Wang M, Gao H (2009) Cloning of a 9-cis-epoxycarotenoid dioxygenase gene and the responses of Caragana korshinskii to a variety of abiotic stresses. Genes Genet Syst 84:397–405

    Article  PubMed  CAS  Google Scholar 

  • Yang J, Guo Z (2007) Cloning of a 9-cis-epoxycarotenoid dioxygenase gene (SgNCED1) from Stylosanthes guianensis and its expression in response to abiotic stresses. Plant Cell Rep 26:1383–1390

    Article  PubMed  CAS  Google Scholar 

  • Zhang YM, Tan JL, Guo ZF, Lu SY, He SJ, Shu W, Zhou BY (2009) Increased abscisic acid levels in transgenic tobacco over-expressing 9-cis-epoxycarotenoid dioxygenase influence H2O2 and NO production and antioxidant defences. Plant Cell Environ 32:509–519

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This present study was supported by International Scientific Exchange Program projects (2008DFA30600, 2009DFA30820), Guangxi R & D Research Program projects (Gui Ke Neng 0815011, Gui Ke Chan 1123008-1), Guangxi Natural Science Foundation project (2011GXNSFF018002) and Guangxi Academy of Agricultural Sciences research fund (2011YT01).

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Authors and Affiliations

  1. College of Agriculture, State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, 530004, Guangxi, People’s Republic of China

    Chang-Ning Li, Qian Nong, Li-Tao Yang & Yang-Rui Li

  2. Guangxi Crop Genetic Improvement and Biotechnology Lab, Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture, P. R. China, Guangxi Key Laboratory of Sugarcane Genetic Improvement, Guangxi Academy of Agricultural Sciences, #174, East Da Xue Road, Nanning, 530007, Guangxi, People’s Republic of China

    Chang-Ning Li, Manoj-Kumar Srivastava, Li-Tao Yang & Yang-Rui Li

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  1. Chang-Ning Li
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  2. Manoj-Kumar Srivastava
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  3. Qian Nong
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Correspondence to Yang-Rui Li.

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Li, CN., Srivastava, MK., Nong, Q. et al. Molecular cloning and characterization of SoNCED, a novel gene encoding 9-cis-epoxycarotenoid dioxygenase from sugarcane (Saccharum officinarum L.). Genes Genom 35, 101–109 (2013). https://doi.org/10.1007/s13258-013-0065-9

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