Abstract
Due to the important role of cotton drought-tolerant varieties and the reported involvement in this trait of trehalose-6-phosphate-synthase, the respective gene (TPS) was isolated and characterized from cultivated cotton, Gossypium hirsutum (ZETA 2 cultivar), using a chromosome-walking technique. TPS has three exons comprising the coding region. Southern blot analysis indicated that the Gossypium genomes (A and D) contain a single copy of TPS per genome. In addition, the expression of this gene was studied in different plant tissues. Plants of the Australian cotton variety Siokra L23, known for its drought tolerance, were subjected to drought stress (using PEG 6000 solution, for 4 h during the dark period of the day and for four consecutive days); leaves, stems and roots were collected after the end of the stress period. Total extracted RNA was examined for the presence of transcripts, in the above-mentioned tissues of stressed and well-watered plants, by reverse transcription-polymerase chain reaction (RT-PCR). The expression levels, determined semi-quantitatively, indicated that the gene was expressed in all plant tissues under both water availability conditions. However, increased expression levels of TPS were observed mainly in stressed leaves and roots compared to those of the well-watered control. This finding is in agreement with the fact that TPS participates in trehalose biosynthesis, known for its participation in stress signal transduction in higher plants.
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Abbreviations
- ABA:
-
Abscisic acid
- TPS :
-
Trehalose-6-phosphate-synthase gene
- TPP :
-
Trehalose-6-phosphate-phosphatase gene
- T6P:
-
Trehalose-6-phosphate
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
References
Adams KL, Wendel JF (2004) Exploring the genomic mysteries of polyploidy in cotton. Biol J Linn Soc 82:573–581
Adams RP, Kendall E, Kartha KK (1990) Comparison of free sugars in growing and dessicated plants of Selaginella lepidophylla. Biochem Syst Ecol 18:107–110
Altshul F, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Avonce N, Leyman B, Gallardo-Mascorro 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
Bajaj S, Targolli J, Liu L-F, Ho T-HD, Wu R (1999) Transgenic approaches to increase dehydration-stress tolerance in plants. Mol Breed 5:493–503
Blazquez MA, Santos E, Flores CL, Martinez-Zapater JM, Salinas J, Gancedo C (1998) Isolation and molecular characterization of the Arabidopsis thaliana TPS1 gene, encoding trehalose-6-phosphate synthase. Plant J 13:685–689
Crowe JH, Crowe LM, Chapman D (1984) Preservation of membranes in anhydrobiotic organisms. The role of trehalose. Science 223:701–703
Crowe JH, Carpenter JF, Crowe LM (1998) The role of vitrification in anhydrobiosis. Annu Rev Physiol 60:73–103
Eastmond PJ, Van Dijken AJH, Spielman M, Kerr A, Tissier AF, Dickinson HG, Jones JDG, Smeekens SC, Graham IA (2002) Trehalose-6-phosphate synthase 1, which catalyses the first step in trehalose synthesis, is essential for Arabidopis embryo maturation. Plant J 29:225–235
Eastmond PJ, Li Y, Graham IA (2003) Is trehalose-6-phosphate a regulator of sugar metabolism in plants? J Exp Bot 54:533–537
Elbein AD, Pan YT, Pastuszak I, Carroll D (2003) New insights on trehalose: a multifunctional molecule. Glycobiology 13:17R–27R
Fritzius T, Aeschbacher RA, Wiemken A, Wingler A (2001) Induction of ApL3 expression by trehalose complements the starch-deficient arabidopsis mutant adg2-1 lacking ApL1, the large subunit of ADP-glucose pyrophosphorylase. Plant Physiol 126:883–889
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
Gibson RP, Turkenburg JP, Charnock SJ, Lloyd R, Davies GJ (2002) Insights into trehalose synthesis provided by the structure of the retaining glucosyltranferase OtsA. Chem Biol 9:1337–1346
Gibson RP, Tarling CA, Roberts S, Withers SG, Davies GJ (2004) The donor subsite of trehalose-6-phosphate synthase. J Biol Chem 279:1950–1955
Goddijn O, Smeekens S (1998) Sensing trehalose biosynthesis in plants. Plant J 14:143–146
Goddijn O, Van Dun K (1999) Trehalose metabolism in plants. Trends Plant Sci 4:315–319
Goddijn O, Verwoerd TC, Voogd E, Krutwagen RWHH, De Graaf PTHM, 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
Higgins D, Thompson J, Gibson T, Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680
Holmström K, Mäntylä E, Welln B, Mandal A, Palva ET (1996) Drought tolerance in tobacco. Nature 379:683–684
Ingram J, Bartels D (1996) The molecular basis of dehydration tolerance in plants. Annu Rev Plant Physiol Plant Mol Biol 47:377–403
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 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
Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163
Leyman B, Dijck P, Thevelein JM (2001) An unexpected plethora of trehalose biosynthesis genes in Arabidopsis thaliana. Trends Plant Sci 6:510–513
Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163:16–20
Müller J, Aeschbacher RA, Sprenger N, Boller T, Wiemken A (2000) Disaccharide-mediated regulation of sucrose: fructan-6-fructosyltransferase, a key enzyme of fructan synthesis in barley leaves. Plant Physiol 123:265–273
Nepomuceno AL, Oosterhuis DM, Stewart JM (1998) Physiological responses of cotton leaves and roots to water deficit induced by polyethylene glycol. Environ Exp Bot 40:29–41
Nepomuceno AL, Oosterhuis DM, Stewart JM, Turley R, Neumaier N, Farias JRB (2002) Expression of heat shock protein and trehalose-6-phosphate synthase homologues induced during water deficit in cotton. Brazilian J Plant Physiol 14:11–20
Nicholas KB, Nicholas HB Jr, Deerfield DW II (1997) GeneDoc: analysis and visualization of genetic variation. EMBNEW.NEWS 4:14
Pellny T, Ghannoum O, Conroy JP, Schluepmann H, Smeekens S, Andralojc J, Krause KP, Goddijn O, Paul MJ (2004) Genetic modification of photosynthesis with E. coli genes for trehalose synthesis. Plant Biotechnol J 2:71–82
Pilon-Smits EAH, Terry N, Sears T, Kim H, Zayed A, Hwang S, van Dun K, Voogd E, Verwoerd TC, Krutwagen RW (1998) Trehalose-producing transgenic tobacco plants show improved growth performance under drought stress. J Plant Physiol 152:525–532
Romero C, Bellés JM, Vayá JL, Serrano R, Culiáñez-Maciá A (1997) Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. Planta 201:293–297
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Schluepmann H, Pellny T, 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
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
Schowalter DB, Sommer SS (1989) The generation of radiolabeled DNA and RNA probes with polymerase chain reaction. Anal Biochem 177:90–94
Stewart Jr CN, Via LE (1993) Rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. Biotechniques 14:748–750
Stiller WN, Constable GA (2001) Influence of cultivar maturity and leaf type on the agronomic water use efficiency of raingrown cotton. In: Proceedings of the 10th Australian agronomy conference. Australian Society of Agronomy, Toowoomba, Hobart. Available online at http://www.regional.org.au./au/asa/2001/1/b/stiller.htm (verified 18 September 2002)
Stiller WN, Eveleigh RR, Constable GA, Reid PE (1998) Varietal characteristics for adaptation of cotton (Gossypium hirsutum L.) to raingrown conditions. In: DL Michalk DL and Pratley JE (eds) Proceedings of the 9th Australian agronomy conference. Australian Society of Agronomy, Toowoomba, Wagga Wagga, pp359–362
Thaller MC, Schippa S, Rossolini GM (1998) Conserved sequence motifs among bacterial, eukaryotic and archaeal phosphatases that define a new phosphohydrolase superfamily. Protein Sci 7:1647–1652
Van Dijck P, Mascorro-Gallardo JO, De Bus M, Royackers K, Iturriaga G, Thevelein JM (2002) Truncation of Arabidopsis thaliana and Selaginella lepidophylla trehalose-6-phosphate synthase unlocks high catalytic activity and supports high trehalose levels on expression in yeast. Biochem J 366:63–71
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
Vogel G, Fiehn O, Bressel LJR, Boller T, Wiemken A, Aeschbaccher A, Wingler A (2001) Trehalose metabolism in Arabidopsis: occurrence of trehalose and molecular cloning and characterization of trehalose-6-phosphate synthase homologues. J Exp Bot 52:1817–1826
Voloudakis AE, Kosmas SA, Tsakas S, Eliopoulos E, Loukas M, Kosmidou K (2002) Expression of selected drought-related genes and physiological response of Greek cotton varieties. Funct Plant Biol 29:1237–1245
Wendel JF (1989) New World tetraploid cottons contain Old World cytoplasm. Proc Natl Acad Sci USA 86:4132–4136
Wendel JF, Cronn RC (2003) Polyploidy and the evolutionary history of cotton. Adv Agron 78:139–186
Wiemken A (1990) Trehalose in yeast, stress protectant rather than reserve carbohydrate. Antonie van Leeuwenhoek 58:209–217
Wingler A (2002) The function of trehalose biosynthesis in plants. Phytochemistry 60:437–440
Wingler A, Fritzius T, Wiemken A, Boller T, Aeschbacher RA (2000) Trehalose induces the ADP-glucose pyrophosphorylase gene, ApL3, and starch synthesis in arabidopsis. Plant Physiol 124:105–114
Zentella R, Mascorro-Gallardo JO, Dijck PV, Foich-Mallol J, Bonini B, Vaeck CV, 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
Acknowledgements
The authors wish to thank Dr. G. Constable (CSIRO Plant Industry) who provided seeds of the Siokra L23 variety, Prof. J.M. Stewart (University of Arkansas) who provided seeds of the diploids species G. herbaceum and G. raimondii. Mr. S. Kosmas is a recipient of a scholarship from the Greek State Scholarships Foundation and is on educational leave from the Hellenic Ministry of Rural Development and Food.
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Kosmas, S.A., Argyrokastritis, A., Loukas, M.G. et al. Isolation and characterization of drought-related trehalose 6-phosphate-synthase gene from cultivated cotton (Gossypium hirsutum L.). Planta 223, 329–339 (2006). https://doi.org/10.1007/s00425-005-0071-5
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DOI: https://doi.org/10.1007/s00425-005-0071-5