Skip to main content
Log in

Cloning and Characterization of Functional Trehalose-6-Phosphate Synthase Gene in Maize

  • Original Research
  • Published:
Journal of Plant Biology Aims and scope Submit manuscript

Abstract

Trehalose is a non-reducing disaccharide of glucose that functions as a compatible solute in the stabilization of biological structures under heat and desiccation stress in bacteria, fungi, and some “resurrection plants”. In the plant kingdom, trehalose is biosynthesized by trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Over-expression of exogenous and endogenous genes encoding TPS and TPP is reported to be effective for improving abiotic stress tolerance in tobacco, potato, tomato, rice, and Arabidopsis. On the basis of bioinformatics prediction, we cloned a fragment containing an open reading frame of 2,820 bp from maize, which encodes a protein of 939 amino acids. Phylogenetic analysis showed that this gene belongs to the class I subfamily of the TPS gene family. Analysis of conserved domains revealed the presence of a TPS domain and a TPP domain. Yeast complementation with TPS and TPP mutants demonstrated that this protein has the activity of trehalose-6-phosphate synthase. Semi-quantitative RT-PCR and real-time quantitative PCR indicated that the expression of this gene is upregulated in response to both salt and cold stress.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Adams RP, Kendall E, Kartha KK (1990) Comparison of free sugars in growing and desiccated plants of Selaginella lepidophylla. Biochem Syst Ecol 18:107–110

    Article  CAS  Google Scholar 

  • Almeida AM, Santos M, Villalobos E, Araújo SS, van Dijck P, Leyman B, Cardoso LA, Santos D, Fevereiro PS, Torné JM (2007) Immunogold localization of trehalose-6-phosphate synthase in leaf segments of wild-type and transgenic tobacco plants expressing the AtTPS1 gene from Arabidopsis thaliana. Protoplasma 230:41–49

    Article  CAS  PubMed  Google Scholar 

  • Avonce N, Leyman B, Mascorro-Gallardo JO, Van Dijck P, Thevelein JM, Iturriaga G (2004) The Arabidopsis trehalose-6-P synthase AtTPS1 gene is a regulator of glucose, abscisic acid, and stress signaling. Plant Physiol 136:3649–3659

    Article  CAS  PubMed  Google Scholar 

  • Avonce N, Mendoza-Vargas A, Morett E, Iturriaga G (2006) Insights on the evolution of trehalose biosynthesis. BMC Evol Biol 6:109

    Article  PubMed  Google Scholar 

  • Becker A, Schloeder P, Steele JE, Wegener G (1996) The regulation of trehalose metabolism in insects. Experientia 52:433–439

    Article  CAS  PubMed  Google Scholar 

  • Blázquez MA, Santos E, Flores CL, Martínez-Zapater JM, Salinas J, Gancedo C (1998) Isolation and molecular characterization of the Arabidopsis TPS1 gene, encoding trehalose-6-phosphate synthase. Plant J 13:685–689

    Article  PubMed  Google Scholar 

  • Chary SN, Hicks GR, Choi YG, Carter D, Raikhel NV (2008) Trehalose-6-phosphate synthase/phosphatase regulates cell shape and plant architecture in Arabidopsis. Plant Physiol 146:97–107

    Article  CAS  PubMed  Google Scholar 

  • Cortina C, Culiáñez-Macià FA (2005) Tomato abiotic stress enhanced tolerance by trehalose biosynthesis. Plant Sci 169:75–82

    Article  CAS  Google Scholar 

  • Crowe JH, Crowe LM, Carpenter JF, Aurell Wistrom C (1987) Stabilization of dry phospholipid bilayers and proteins by sugars. Biochem J 15:1–10

    Google Scholar 

  • De Virgilio C, Bürckert N, Bell W, Jenö P, Boller T, Weimken A (1993) Disruption of TPS2, the gene encoding the 100-kDa sub-unit of the trehalose-6-phosphate synthase/phosphatase complex in Saccharomyces cerevisiae, causes accumulation of trehalose-6-phosphate and loss of trehalose-6-phosphate phosphatase activity. Eur J Biochem 212:315–323

    Article  PubMed  Google Scholar 

  • Elble R (1992) A simple and efficient procedure for transformation of yeasts. BioTechniques 13:18–20

    CAS  PubMed  Google Scholar 

  • Garcia AB, de Almeida Engler J, Iyer S, Gerats T, Van Montagu M, Caplan AB (1997) Effects of osmoprotectants upon NaCl stress in rice. Plant Physiol 115:159–169

    CAS  PubMed  Google Scholar 

  • Garg AK, Kim JK, Owens TG, Ranwala AP, Choi YD, Kochian LV, Wu RJ (2002) Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses. Proc Natl Acad Sci USA 99:15898–15903

    Article  CAS  PubMed  Google Scholar 

  • Ge LF, Chao DY, Shi M, Zhu MZ, Gao JP, Lin HX (2008) Overexpression of the trehalose-6-phosphate phosphatase gene OsTPP1 confers stress tolerance in rice and results in the activation of stress responsive genes. Planta 228:191–201

    Article  CAS  PubMed  Google Scholar 

  • Geelen D, Royackers K, Vanstraelen M, De Bus M, Inze D', Van Dijck P, Thevelein JM, Leyman B (2007) Trehalose-6-P synthase AtTPS1 high molecular weight complexes in yeast and Arabidopsis. Plant Sci 173:426–437

    Article  CAS  Google Scholar 

  • Gibson RP, Turkenburg JP, Charnock SJ, Lloyd R, Davis GJ (2002) Insights into trehalose synthesis provided by the structure of the retaining glucosyltransferase OtsA. Chem Biol 9:1337–1346

    Article  CAS  PubMed  Google Scholar 

  • Goddijn OJ, Verwoerd TC, Voogd E, Krutwagen RW, de Graaf PT, Poels J, van Dun K, Ponstein AS, Damm B, Pen J (1997) Inhibition of trehalase activity enhances trehalose accumulation in transgenic plants. Plant Physiol 113:181–190

    Article  CAS  PubMed  Google Scholar 

  • Hohmann S, Neves MJ, de Koning W, Alijo R, Ramos J, Thevelein JM (1993) The growth and signalling defects of the ggs1 (fdp1/byp1) deletion mutant on glucose are suppressed by a deletion of the gene encoding hexokinase PII. Curr Genet 23:281–289

    Article  CAS  PubMed  Google Scholar 

  • Holmström KO, Mäntylä E, Welin B, Mandal A, Palva ET, Tunnela OE, Londesborough J (1996) Drought tolerance in tobacco. Nature 379:683–684

    Article  Google Scholar 

  • Hottiger T, De Virgilio C, Hall MN, Boller T, Wiemken A (1994) The role of trehalose synthesis for the acquisition of thermotolerance in yeast II. Physiological concentrations of trehalose increase the thermal stability of proteins in vitro. Eur J Biochem 219:187–193

    Article  CAS  PubMed  Google Scholar 

  • Jang IC, Oh SJ, Seo JS, Choi WB, Song SI, Kim CH, Kim YS, Seo HS, Choi YD, Nahm BH, Kim JK (2003) Expression of a bifunctional fusion of the Escherichia coli genes for trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase in transgenic rice plants increases trehalose accumulation and abiotic stress tolerance without stunting growth. Plant Physiol 131:516–524

    Article  CAS  PubMed  Google Scholar 

  • Karim S, Aronsson H, Ericson H, Pirhonen M, Leyman B, Welin B, Mäntylä E, Palva ET, Van Dijck P, Holmström KO (2007) Improved drought tolerance without undesired side effects in transgenic plants producing trehalose. Plant Mol Biol 64:371–386

    Article  CAS  PubMed  Google Scholar 

  • Leyman B, Van Dijck P, Thevelein JM (2001) An unexpected plethora of trehalose biosynthesis genes in Arabidopsis thaliana. Trends in Plant Sci 11:510–513

    Google Scholar 

  • Miranda JA, Avonce N, Suárez R, Thevelein JM, Van Dijck P, Iturriaga G (2007) A bifunctional TPS-TPP enzyme from yeast confers tolerance to multiple and extreme abiotic-stress conditions in transgenic Arabidopsis. Planta 226:1411–1421

    Article  CAS  PubMed  Google Scholar 

  • Müller J, Boller T, Wiemken A (1995a) Trehalose and trehalase in plants: recent developments. Plant Sci 112:1–9

    Article  Google Scholar 

  • Müller J, Boller T, Wiemken A (1995b) Effects of validamycin A, a potent trehalase inhibitor, and phyto-hormones on trehalose metabolism in roots and nodules of soybean and cowpea. Planta 197:362–368

    Google Scholar 

  • Müller J, Aeschbacher RA, Wingler A, Boller T, Wiemken A (2001) Trehalose and trehalase in Arabidopsis. Plant Physiol 125:1086–1093

    Article  PubMed  Google Scholar 

  • Neves MJ, Hohmann S, Bell W, Dumortier F, Luyten K, Ramos J, Cobbaert P, de Koning W, Kaneva Z, Thevelein JM (1995) Control of glucose influx into glycolysis and pleiotropic effects studied in different isogenic sets of Saccharomyces cerevisiae mutants in trehalose biosynthesis. Curr Genet 27:110–122

    Article  CAS  PubMed  Google Scholar 

  • Romero C, Bellés JM, Vayá JL, Serrano R, Culiáñez-Macià FA (1997) Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. Planta 201:293–297

    Article  CAS  PubMed  Google Scholar 

  • Satoh-Nagasawa N, Nagasawa N, Malcomber S, Sakai H, Jackson D (2006) A trehalose metabolic enzyme controls inflorescence architecture in maize. Nature 441:227–230

    Article  CAS  PubMed  Google Scholar 

  • Schluepmann H, Pellny T, van Dijken A, Smeekens S, Paul M (2003) Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana. Proc Natl Acad Sci USA 100:6849–6854

    Article  CAS  PubMed  Google Scholar 

  • Schluepmann H, van Dijken A, Aghdasi M, Wobbes B, Paul M, Smeekens S (2004) Trehalose mediated growth inhibition of Arabidopsis seedlings is due to trehalose-6-phosphate accumulation. Plant Physiol 135:879–890

    Article  CAS  PubMed  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  • Thevelein JM, Hohmann S (1995) Trehalose synthase: guard to the gate of glycolysis in yeast? Trends Biochem Sci 20:3–10

    Article  CAS  PubMed  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882

    Article  CAS  PubMed  Google Scholar 

  • Zentella R, Mascorro-Gallardo JO, Van Dijck P, Folch-Mallol J, Bonini B, Van Vaeck C, Gaxiola R, Covarrubias AA, Nieto-Sotelo J, Thevelein JM, Iturriaga G (1999) A Selaginella lepidophylla trehalose-6-phosphate synthase complements growth and stress-tolerance defects in a yeast tps1 mutant. Plant Physiol 119:1473–1482

    Article  CAS  PubMed  Google Scholar 

  • Van Aelst L, Hohmann S, Bulaya B, de Koning W, Sierkstra L, Neves MJ, Luyten K, Alijo R, Ramos J, Coccetti P (1993) Molecular cloning of a gene involved in glucose sensing in the yeast Saccharomyces cerevisiae. Mol Microbiol 8:927–943

    Article  PubMed  Google Scholar 

  • Van Dijck P, Mascorro-Gallardo JO, De Bus M, Royackers K, Iturriaga G, Thevelein JM (2002) Truncation of Arabidopsis thaliana and Selaginella lepidophyll trehalose-6-phosphate synthase unlocks high catalytic activity and supports high trehalose levels on expression in yeast. Biochem J 366:63–71

    PubMed  Google Scholar 

  • Van Laere A (1989) Trehalose, reserve and/or stress metabolite? FEMS Microbiol 63:201–210

    Article  Google Scholar 

  • Vogel G, Aeschbacher RA, Müller J, Boller T, Wiemken A (1998) Trehalose-6-phosphate phosphatases from Arabidopsis thaliana: identification by functional complementation of the yeast tps2 mutant. Plant J 13:673–683

    Article  CAS  PubMed  Google Scholar 

  • Yeo ET, Kwon HB, Han SE, Lee JT, Ryu JC, Byun MO (2000) Genetic engineering of drought resistant potato plants by introduction of the trehalose-6-phosphate synthase (TPS1) gene from Saccharomyces cerevisiae. Mol Cells 10:263–268

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Prof. Johan M. Thevelein at Katholieke University for the gift of yeast strains W303-1A, YSH290 and YSH450, and plasmid AtTPS1-pRS6. Financial support from the Projects of Development Plan of the State Key Fundamental Research (973 Project; 2009CB118400), the National Natural Science Foundation of China (30671309), and the National Key Science and Technology Special Project (2008ZX08003-004) are sincerely appreciated.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Wan-Chen Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jiang, W., Fu, FL., Zhang, SZ. et al. Cloning and Characterization of Functional Trehalose-6-Phosphate Synthase Gene in Maize. J. Plant Biol. 53, 134–141 (2010). https://doi.org/10.1007/s12374-010-9098-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12374-010-9098-7

Keywords

Navigation