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Cloning of two contrasting high-affinity sulfate transporters from tomato induced by low sulfate and infection by the vascular pathogen Verticillium dahliae

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Abstract

Two cDNAs, LeST1-1 (AF347613) and LeST1-2 (AF347614), encoding sulfate transporters have been cloned from tomato (Lycopersicon esculentum Mill.) by reverse transcription–polymerase chain reaction and their expression characterised. Sharing 76% identity at the amino acid level, the transporters are phylogenetically associated with the Group-1, high-affinity plant sulfate transporters. Both were shown to have high affinity for sulfate by uptake kinetic analysis using a yeast (Saccharomyces cerevisiae) sulfate-transporter mutant. K m values of 11.5 μM and 9.8 μM were calculated for LeST1-1 and LeST1-2, respectively, the same order of magnitude as those previously reported for several other Group-1 high-affinity sulfate transporters. In situ hybridisation to S-deficient tomato roots showed LeST1-1 to be expressed in the epidermis and pericycle, whereas LeST1-2 expression was located to the epidermis only. Northern analysis shows that the mRNA abundances of both LeST1-1 and LeST1-2 are upregulated in the root in response to sulfate deprivation. LeST1-1 is specifically expressed in root tissue, a characteristic of Group-1 sulfate transporters. LeST1-2, however, was also detected in tomato leaves and stems and is upregulated and expressed to a similar extent in these tissues under conditions of sulfate deprivation. Induction of LeST1-2 expression was also observed in the vascular tissues of a resistant line of tomato infected with the vascular wilt pathogen Verticillium dahliae.

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Abbreviations

APSR:

adenosine 5′-phosphosulfate reductase

dpi:

days post-infection

RT–PCR:

reverse transcription–polymerase chain reaction

S:

sulfur

ST:

sulfate transporter

References

  • Ashikari T, Kiuchi-Goto N, Tanaka Y, Shibano Y, Amachi T, Yoshizumi H (1989) High expression, and efficient secretion of Rhizopus oryzae glucoamylase in the yeast Saccharomyces cerevisiae. Appl Microbiol Biotechnol 30:515–520

    CAS  Google Scholar 

  • Bolchi A, Petrucco S, Tenca P-L, Foroni C, Ottonello S (1999) Coordinate modulation of maize sulfate permease and ATP sulfurylase mRNAs in response to variations in sulfur nutritional status: stereospecific down-regulation by l-cysteine. Plant Mol Biol 39:527–537

    CAS  PubMed  Google Scholar 

  • Cherest H, Davidian J-C, Thomas D, Benes V, Ansorge W, Surdin-Kerjan Y (1997) Molecular characterization of two high-affinity sulfate transporters in Saccharomyces cerevisiae. Genetics 145:627–635

    PubMed  Google Scholar 

  • Cooper RM, Resende MLV, Flood J, Rowan MG, Beale MH, Potter U (1996) Detection and cellular localisation of elemental sulphur in disease-resistant genotypes of Theobroma cacao. Nature 379:159–162

    Article  CAS  Google Scholar 

  • Davidian J-C, Hatzfeld Y, Cathala N, Tagmount A, Vidmar J (2000) Sulfate uptake and transport in plants. In: Brunold C, Rennenberg H, De Kok LJ, Stulen I, Davidian J-C (eds) Sulfur nutrition and sulfur assimilation in higher plants: molecular, biochemical and physiological aspects. Haupt, Berne, pp 19–40

  • Gotor C, Cejudo FJ, Barroso C, Vega JM (1997) Tissue-specific expression of ATCYS-3A, a gene encoding the cytosolic isoform of O-acetylserine(thiol)lyase in Arabidopsis. Plant J 11:347–352

    Article  CAS  PubMed  Google Scholar 

  • Hawkesford MJ (2000) Plant responses to sulphur deficiency and the genetic manipulation of sulphate transporters to improve S-utilization efficiency. J Exp Bot 51:131–138

    CAS  PubMed  Google Scholar 

  • Hawkesford MJ (2003) Transporter gene families in plants: the sulphate transporter gene family — redundancy or specialization? Physiol Plant 117:155–163

    Article  CAS  Google Scholar 

  • Hawkesford MJ, Wray JL (2000) Molecular genetics of sulphur assimilation. Adv Bot Res 33:159–223

    CAS  Google Scholar 

  • Hawkesford MJ, Buchner P, Hopkins L, Howarth JR (2002) Sulphate uptake and transport. In: Abrol YP, Ahmad A (eds) Sulphur in higher plants. Kluwer, Dordrecht, pp 71–86

  • Hell R (1997) Molecular physiology of plant sulfur metabolism. Planta 202:138–148

    CAS  PubMed  Google Scholar 

  • Herschbach C, Pilch B, Tausz M, Rennenberg H, Grill D (2002) Sulphate uptake and xylem loading of young pea (Pisum sativum L.) Plant Soil 242:227–233

    Google Scholar 

  • Lappartient AG, Vidmar JJ, Leustek T, Glass ADM, Touraine B (1999) Inter-organ signaling in plants: regulation of ATP sulfurylase and sulfate transporter genes expression in roots mediated by phloem-translocated compound. Plant J 18:89–95

    PubMed  Google Scholar 

  • McGrath SP, Zhao FJ, Withers PJA (1996) Development of sulphur deficiency in crops and its treatment. Proceedings of the Fertiliser Society No. 379. The Fertiliser Society, Peterborough

  • Poirier Y, Thoma S, Somerville C, Schiefelbein J (1991) A mutant of Arabidopsis thaliana deficient in xylem loading of phosphate. Plant Physiol 97:1087–1093

    CAS  Google Scholar 

  • Resende MLV, Flood J, Ramsden JD, Rowan MG, Beale MH, Cooper RM (1996) Novel phytoalexins including elemental sulphur in the resistance of cocoa (Theobroma cacao L.) to Verticillium wilt (Verticillium dahliae Kleb.). Physiol Mol Plant Pathol 48:347–359

    Article  CAS  Google Scholar 

  • Saito K (2000) Regulation of sulfate transport and synthesis of sulfur-containing amino acids. Curr Opin Plant Biol 3:188–195

    CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Sherman F (1991) Getting started with yeast. Methods Enzymol 194:3–21

    CAS  PubMed  Google Scholar 

  • Shibagaki N, Rose A, McDermott JP, Fujiwara T, Hayashi H, Yomeyama T, Davis JP (2002) Selenate-resistant mutants of Arabidopsis thaliana identify Sultr1;2, a sulfate transporter required for efficient transport of sulfate into roots. Plant J 29:475–486

    Article  CAS  PubMed  Google Scholar 

  • Smith FW, Ealing, PM, Hawkesford MJ, Clarkson DT (1995a) Plant members of a family of sulfate transporters reveal functional subtypes. Proc Natl Acad Sci USA 92:9373–9377

    PubMed  Google Scholar 

  • Smith FW, Hawkesford MJ, Prosser IM, Clarkson DT (1995b) Isolation of a cDNA from Saccharomyces cerevisiae that encodes a high-affinity sulphate transporter at the plasma membrane. Mol Gen Genet 247:709–715

    CAS  PubMed  Google Scholar 

  • Smith FW, Hawkesford MJ, Ealing PM, Clarkson, DT, Vandenberg PJ, Belcher A, Warrilow AGS (1997) Regulation of expression of a cDNA from barley roots encoding a high-affinity sulphate transporter. Plant J 12:875–884

    PubMed  Google Scholar 

  • Takahashi H, Sasakura N, Noji M, Saito K (1996) Isolation and characterization of a cDNA encoding a sulfate transporter from Arabidopsis thaliana. FEBS Lett 392:95–99

    PubMed  Google Scholar 

  • Takahashi H, Yamazaki M, Sasakura N, Watanabe A, Leustek T, de Almeida Engler J, Engler G, van Montagu M, Saito K (1997) Regulation of sulfur assimilation in higher plants: a sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana. Proc Natl Acad Sci USA 94:11102–11107

    CAS  PubMed  Google Scholar 

  • Takahashi H, Asanuma W, Saito K (1999a) Cloning of an Arabidopsis cDNA encoding a chloroplast localizing sulphate transporter isoform. J Exp Bot 50:1713–1714

    CAS  Google Scholar 

  • Takahashi H, Sasakura N, Kimura A, Watanabe A, Saito K (1999b) Identification of two leaf-specific sulfate transporters in Arabidopsis thaliana (Accession No. AB012048 and AB004060). (PGR99-154) Plant Physiol 121:686

    Google Scholar 

  • Takahashi H, Watanabe-Takahashi A, Smith FW, Blake-Kalf M, Hawkesford MJ, Saito K (2000) The roles of three functional sulphate transporters involved in uptake and translocation of sulphate in Arabidopsis thaliana. Plant J 23:171–182

    PubMed  Google Scholar 

  • 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 22:4673–5680

    PubMed  Google Scholar 

  • Verwoerd TC, Dekker BM, Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17: 2362

    PubMed  Google Scholar 

  • Vidmar JJ, Tagmount A, Cathala N, Touraine B, Davidian J-C (2000) Cloning and characterization of a root specific high-affinity sulfate transporter from Arabidopsis thaliana. FEBS Lett 475:65–69

    Article  CAS  PubMed  Google Scholar 

  • Williams JS, Hall SA, Hawkesford MJ, Beale MH, Cooper RM (2002) Elemental sulfur and thiol accumulation in tomato and defense against a fungal vascular pathogen. Plant Physiol 128:150–159

    Article  CAS  PubMed  Google Scholar 

  • Yoshimoto N, Takahashi H, Smith FW, Yamaya T, Saito K (2002) Two distinct high-affinity sulfate transporters with different inducibilities mediate uptake of sulfate in Arabidopsis roots. Plant J 29:465–473

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We acknowledge Jane Williams and Richard Cooper at the University of Bath, UK, for growth and infection of tomato material, Nicole Cathala at BPMP Montpellier for yeast transformation and Peter Buchner, Gordon Forbes, Laura Hopkins, Saroj Parmar and Katie Tearall for their assistance. Rothamsted Research receives grant-aided support from the Biotechnology and Biological Sciences Research Council (BBSRC) of the UK. This research was supported by a BBSRC ROPA award (206/9912300), DEFRA Programme AR09 and the British Council.

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Authors and Affiliations

  1. Rothamsted Research, West Common, Harpenden, Hertfordshire, AL5 2JQ, UK

    Jonathan R. Howarth & Malcolm J. Hawkesford

  2. Biochimie et Physiologie Moléculaire des Plantes, INRA–CNRS, ENSA-M, UM2, 2 Place Viala, 34060, Montpellier, Cedex 1, France

    Pierre Fourcroy & Jean-Claude Davidian

  3. CSIRO Plant Industry, Queensland Bioscience Precinct, 306 Carmody Rd., QLD 4067, St. Lucia, Australia

    Frank W. Smith

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  1. Jonathan R. Howarth
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  2. Pierre Fourcroy
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  3. Jean-Claude Davidian
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  4. Frank W. Smith
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  5. Malcolm J. Hawkesford
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Correspondence to Malcolm J. Hawkesford.

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Howarth, J.R., Fourcroy, P., Davidian, JC. et al. Cloning of two contrasting high-affinity sulfate transporters from tomato induced by low sulfate and infection by the vascular pathogen Verticillium dahliae . Planta 218, 58–64 (2003). https://doi.org/10.1007/s00425-003-1085-5

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