Skip to main content
SpringerLink
Log in

Light and abiotic stresses regulate the expression of GDP-l-galactose phosphorylase and levels of ascorbic acid in two kiwifruit genotypes via light-responsive and stress-inducible cis-elements in their promoters

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Ascorbic acid (AsA) plays an essential role in plants by protecting cells against oxidative damage. GDP-l-galactose phosphorylase (GGP) is the first committed gene for AsA synthesis. Our research examined AsA levels, regulation of GGP gene expression, and how these are related to abiotic stresses in two species of Actinidia (kiwifruit). When leaves were subjected to continuous darkness or light, ABA or MeJA, heat, or a hypoxic environment, we found some correlation between the relative levels of GGP mRNA and AsA concentrations. In transformed tobacco plants, activity of the GGP promoter was induced by all of these treatments. However, the degree of inducibility in the two kiwifruit species differed among the GGP promoter deletions. We deduced that the G-box motif, a light-responsive element, may have an important function in regulating GGP transcripts under various light conditions in both A. deliciosa and A. eriantha. Other elements such as ABRE, the CGTCA motif, and HSE might also control the promoter activities of GGP in kiwifruit. Altogether, these data suggest that GGP expression in the two kiwifruit species is regulated by light or abiotic stress via the relative cis-elements in their promoters. Furthermore, GGP has a critical role in modulating AsA concentrations in kiwifruit species under abiotic stresses.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

ABA:

Abscisic acid

AsA:

Ascorbic acid

GGP :

GDP-l-galactose phosphorylase

GUS :

β-Glucuronidase

MeJA:

Methyl jasmonate

RT-qPCR:

Quantitative reverse transcription-polymerase chain reactions

4-MU:

4-Methylumbelliferone

References

  • Agius F, Amaya I, Botella MA, Valpuesta V (2005) Functional analysis of homologous and heterologous promoters in strawberry fruits using transient expression. J Exp Bot 56:37–46

    PubMed  CAS  Google Scholar 

  • Arguello-Astorga GR, Herrera-Estrella LR (1996) Ancestral multipartite units in light-responsive plant promoters have structural features correlating with specific phototransduction pathways. Plant Physiol 112:1151–1166

    Article  PubMed  CAS  Google Scholar 

  • Asif MH, Dhawan P, Nath P (2000) A simple procedure for the isolation of high quality RNA from ripening banana fruit. Plant Mol Biol Rep 18:109–115

    Article  CAS  Google Scholar 

  • Baker SS, Wilhelm KS, Thomashow MF (1994) The 5′-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression. Plant Mol Biol 24:701–713

    Article  PubMed  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Brenner C (2002) Hint, Fhit, and GalT: function, structure, evolution, and mechanism of three branches of the histidine triad superfamily of nucleotide hydrolases and transferases. Biochemistry 41:9003–9014

    Article  PubMed  CAS  Google Scholar 

  • Bulley SM, Rassam M, Hoser D, Otto W, Schünemann N, Wright M, MacRae E, Gleave A, Laing WA (2009) Gene expression studies in kiwifruit and gene over-expression in Arabidopsis indicates that GDP-l-galactose guanyltransferase is a major control point of vitamin C biosynthesis. J Exp Bot 60:765–778

    Article  PubMed  CAS  Google Scholar 

  • Bulley S, Wright M, Rommens C, Yan H, Rassam M, Lin-Wang K, Andre C, Brewster D, Karunairetnam S, Allan AC (2012) Enhancing ascorbate in fruits and tubers through over-expression of the l-galactose pathway gene GDP-l-galactose phosphorylase. Plant Biotechnol J 10:390–397

    Article  PubMed  CAS  Google Scholar 

  • Chaouch S, Queval G, Vanderauwera S, Mhamdi A, Vandorpe M, Langlois-Meurinne M, Van Breusegem F, Saindrenan P, Noctor G (2010) Peroxisomal hydrogen peroxide is coupled to biotic defense responses by isochorismate synthase 1 in a daylength-related manner. Plant Physiol 153:1692–1705

    Article  PubMed  CAS  Google Scholar 

  • Conklin P, Barth C (2004) Ascorbic acid, a familiar small molecule intertwined in the response of plants to ozone, pathogens, and the onset of senescence. Plant Cell Environ 27:959–970

    Article  CAS  Google Scholar 

  • Creelman RA, Mullet JE (1995) Jasmonic acid distribution and action in plants: regulation during development and response to biotic and abiotic stress. Proc Natl Acad Sci USA 92:4114–4119

    Article  PubMed  CAS  Google Scholar 

  • Daraselia ND, Tarchevskaya S, Narita JO (1996) The promoter for tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase gene 2 has unusual regulatory elements that direct high-level expression. Plant Physiol 112:727–733

    Article  PubMed  CAS  Google Scholar 

  • De Tullio M, Arrigoni O (2004) Hopes, disillusions and more hopes from vitamin C. Cell Mol Life Sci 61:209–219

    Article  PubMed  Google Scholar 

  • Diaz-De-Leon F, Klotz KL, Lagrimini LM (1993) Nucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene. Plant Physiol 101:1117–1118

    Article  PubMed  CAS  Google Scholar 

  • Dolferus R, Jacobs M, Peacock WJ, Dennis ES (1994) Differential interactions of promoter elements in stress responses of the Arabidopsis Adh gene. Plant Physiol 105:1075–1087

    Article  PubMed  CAS  Google Scholar 

  • Dowdle J, Ishikawa T, Gatzek S, Rolinski S, Smirnoff N (2007) Two genes in Arabidopsis thaliana encoding GDP-l-galactose phosphorylase are required for ascorbate biosynthesis and seedling viability. Plant J 52:673–689

    Article  PubMed  CAS  Google Scholar 

  • Fink JS, Verhave M, Kasper S, Tsukada T, Mandel G, Goodman RH (1988) The CGTCA sequence motif is essential for biological activity of the vasoactive intestinal peptide gene cAMP-regulated enhancer. Proc Natl Acad Sci USA 85:6662–6666

    Article  PubMed  CAS  Google Scholar 

  • Foster R, Izawa T, Chua NH (1994) Plant bZIP proteins gather at ACGT elements. FASEB J 8:192–200

    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 

  • Gong M, Hay S, Marshall KR, Munro AW, Scrutton NS (2007) DNA binding suppresses human AIF-M2 activity and provides a connection between redox chemistry, reactive oxygen species, and apoptosis. J Biol Chem 282:30331–30340

    Article  PubMed  CAS  Google Scholar 

  • Hartmann U, Valentine WJ, Christie JM, Hays J, Jenkins GI, Weisshaar B (1998) Identification of UV/blue light-response elements in the Arabidopsis thaliana chalcone synthase promoter using a homologous protoplast transient expression system. Plant Mol Biol 36:741–754

    Article  PubMed  CAS  Google Scholar 

  • Huang C, He W, Guo J, Chang X, Su P, Zhang L (2005) Increased sensitivity to salt stress in an ascorbate-deficient Arabidopsis mutant. J Exp Bot 56:3041–3049

    Article  PubMed  CAS  Google Scholar 

  • Ioannidi E, Kalamaki MS, Engineer C, Pateraki I, Alexandrou D, Mellidou I, Giovannonni J, Kanellis AK (2009) Expression profiling of ascorbic acid-related genes during tomato fruit development and ripening and in response to stress conditions. J Exp Bot 60:663–678

    Article  PubMed  CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  • Jimenez A, Creissen G, Kular B, Firmin J, Robinson S, Verhoeyen M, Mullineaux P (2002) Changes in oxidative processes and components of the antioxidant system during tomato fruit ripening. Planta 214:751–758

    Article  PubMed  CAS  Google Scholar 

  • Laing WA, Wright MA, Cooney J, Bulley SM (2007) The missing step of the l-galactose pathway of ascorbate biosynthesis in plants, an l-galactose guanyltransferase, increases leaf ascorbate content. Proc Natl Acad Sci USA 104:9534–9539

    Article  PubMed  CAS  Google Scholar 

  • Larkindale J, Hall JD, Knight MR, Vierling E (2005) Heat stress phenotypes of Arabidopsis mutants implicate multiple signaling pathways in the acquisition of thermotolerance. Plant Physiol 138:882–897

    Article  PubMed  CAS  Google Scholar 

  • Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327

    Article  PubMed  CAS  Google Scholar 

  • Li M, Ma F, Liang D, Li J, Wang Y (2010) Ascorbate biosynthesis during early fruit development is the main reason for its accumulation in Kiwi. PLoS ONE 5(12):e14281

    Article  PubMed  Google Scholar 

  • Linster CL, Clarke SG (2008) l-Ascorbate biosynthesis in higher plants: the role of VTC2. Trends Plant Sci 13:567–573

    Article  PubMed  CAS  Google Scholar 

  • Linster CL, Gomez TA, Christensen KC, Adler LN, Young BD, Brenner C, Clarke SG (2007) Arabidopsis VTC2 encodes a GDP-l-galactose phosphorylase, the last unknown enzyme in the Smirnoff–Wheeler pathway to ascorbic acid in plants. J Biol Chem 282:18879–18885

    Article  PubMed  CAS  Google Scholar 

  • Liu Z-B, Ulmasov T, Shi X, Hagen G, Guilfoyle TJ (1994) Soybean GH3 promoter contains multiple auxin-inducible elements. Plant Cell 6:645–657

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Ma YH, Ma FW, Zhang JK, Li MJ, Wang YH, Liang D (2008) Effects of high temperature on activities and gene expression of enzymes involved in ascorbate–glutathione cycle in apple leaves. Plant Sci 175:761–766

    Article  CAS  Google Scholar 

  • Martínez-Hernández A, López-Ochoa L, Argüello-Astorga G, Herrera-Estrella L (2002) Functional properties and regulatory complexity of a minimal RBCS light-responsive unit activated by phytochrome, cryptochrome, and plastid signals. Plant Physiol 128:1223–1233

    Article  PubMed  Google Scholar 

  • Massot C, Stevens R, Génard M, Longuenesse JJ, Gautier H (2012) Light affects ascorbate content and ascorbate-related gene expression in tomato leaves more than in fruits. Planta 235:153–163

    Article  PubMed  CAS  Google Scholar 

  • Mellidou I, Chagne D, Laing W, Keulemans J, Davey MW (2012) Allelic variation in paralogues of GDP-l-galactose phosphorylase is a major determinant of vitamin C concentrations in apple fruit. Plant Physiol 160:1613–1629

    Article  PubMed  CAS  Google Scholar 

  • Müller-Moulé P (2008) An expression analysis of the ascorbate biosynthesis enzyme VTC2. Plant Mol Biol 68:31–41

    Article  PubMed  Google Scholar 

  • Müller-Moulé P, Golan T, Niyogi KK (2004) Ascorbate-deficient mutants of Arabidopsis grow in high light despite chronic photooxidative stress. Plant Physiol 134:1163–1172

    Article  PubMed  Google Scholar 

  • Nash J, Luehrsen KR, Walbot V (1990) Bronze-2 gene of maize: reconstruction of a wild-type allele and analysis of transcription and splicing. Plant Cell 2:1039–1049

    PubMed  CAS  Google Scholar 

  • Pastuglia M, Roby D, Dumas C, Cock JM (1997) Rapid induction by wounding and bacterial infection of an S gene family receptor-like kinase gene in Brassica oleracea. Plant Cell 9:49–60

    PubMed  CAS  Google Scholar 

  • Phillips BW, Sharma R, Leco PA, Edwards DR (1999) A sequence-selective single-strand DNA-binding protein regulates basal transcription of the murine tissue inhibitor of metalloproteinases-1 (Timp-1) gene. J Biol Chem 274:22197–22207

    Article  PubMed  CAS  Google Scholar 

  • Sparkes IA, Runions J, Kearns A, Hawes C (2006) Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 1:2019–2025

    Article  PubMed  CAS  Google Scholar 

  • Tokunaga T, Esaka M (2007) Induction of a novel XIP-type xylanase inhibitor by external ascorbic acid treatment and differential expression of XIP-family genes in rice. Plant Cell Physiol 48:700–714

    Article  PubMed  CAS  Google Scholar 

  • Urzica EI, Adler LN, Page MD, Linster CL, Arbing MA, Casero D, Pellegrini M, Merchant SS, Clarke SG (2012) Impact of oxidative stress on ascorbate biosynthesis in Chlamydomonas via regulation of the VTC2 gene encoding a GDP-l-galactose phosphorylase. J Biol Chem 287:14234–14245

    Article  PubMed  CAS  Google Scholar 

  • Walker JC, Howard EA, Dennis ES, Peacock WJ (1987) DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene. Proc Natl Acad Sci USA 84:6624–6628

    Article  PubMed  CAS  Google Scholar 

  • Wheeler GL, Jones MA, Smirnoff N (1998) The biosynthetic pathway of vitamin C in higher plants. Nature 393:365–369

    Article  PubMed  CAS  Google Scholar 

  • Wu CY, Suzuki A, Washida H, Takaiwa F (1998) The GCN4 motif in a rice glutelin gene is essential for endosperm-specific gene expression and is activated by Opaque-2 in transgenic rice plants. Plant J 14:673–683

    Article  PubMed  CAS  Google Scholar 

  • Xu W, Yu Y, Ding J, Hua Z, Wang Y (2010) Characterization of a novel stilbene synthase promoter involved in pathogen-and stress-inducible expression from Chinese wild Vitis pseudoreticulata. Planta 231:475–487

    Article  PubMed  CAS  Google Scholar 

  • Yabuta Y, Mieda T, Rapolu M, Nakamura A, Motoki T, Maruta T, Yoshimura K, Ishikawa T, Shigeoka S (2007) Light regulation of ascorbate biosynthesis is dependent on the photosynthetic electron transport chain but independent of sugars in Arabidopsis. J Exp Bot 58:2661–2671

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (30871700) and the Research Fund for the Doctoral Program of Higher Education of China (20090204110009). The authors are grateful to Professor Yuejin Wang for providing us with vector samples and to Priscilla Licht for help in revising our English composition.

Author information

Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Juan Li, Dong Liang, Mingjun Li & Fengwang Ma

Authors
  1. Juan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dong Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mingjun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fengwang Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Mingjun Li or Fengwang Ma.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 52 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, J., Liang, D., Li, M. et al. Light and abiotic stresses regulate the expression of GDP-l-galactose phosphorylase and levels of ascorbic acid in two kiwifruit genotypes via light-responsive and stress-inducible cis-elements in their promoters. Planta 238, 535–547 (2013). https://doi.org/10.1007/s00425-013-1915-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-013-1915-z

Keywords

Search

Navigation