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Plant Molecular Biology

, Volume 85, Issue 4–5, pp 473–484 | Cite as

Expression of Arabidopsis sugar transport protein STP13 differentially affects glucose transport activity and basal resistance to Botrytis cinerea

  • Pauline Lemonnier
  • Cécile Gaillard
  • Florian Veillet
  • Jérémy Verbeke
  • Rémi Lemoine
  • Pierre Coutos-Thévenot
  • Sylvain La Camera
Article

Abstract

Botrytis cinerea is the causing agent of the grey mold disease in more than 200 crop species. While signaling pathways leading to the basal resistance against this fungus are well described, the role of the import of sugars into host cells remains to be investigated. In Arabidopsis thaliana, apoplastic hexose retrieval is mediated by the activity of sugar transport proteins (STPs). Expression analysis of the 14 STP genes revealed that only STP13 was induced in leaves challenged with B. cinerea. STP13-modified plants were produced and assayed for their resistance to B. cinerea and glucose transport activity. We report that STP13-deficient plants exhibited an enhanced susceptibility and a reduced rate of glucose uptake. Conversely, plants with a high constitutive level of STP13 protein displayed an improved capacity to absorb glucose and an enhanced resistance phenotype. The correlation between STP13 transcripts, protein accumulation, glucose uptake rate and resistance level indicates that STP13 contributes to the basal resistance to B. cinerea by limiting symptom development and points out the importance of the host intracellular sugar uptake in this process. We postulate that STP13 would participate in the active resorption of hexoses to support the increased energy demand to trigger plant defense reactions and to deprive the fungus by changing sugar fluxes toward host cells.

Keywords

Sugar transporter protein Arabidopsis thaliana Botrytis cinerea Basal resistance Plant–pathogen interactions 

Notes

Acknowledgments

Pauline Lemonnier and Florian Veillet are supported by Grants from the French Ministry of Higher Education and Research, and the Région Poitou–Charentes, respectively. We are grateful to Vincent Lebeurre and Bruno Faure for helping us producing numerous plants used in this study. Antoine Plasseraud Desgranges is acknowledged for his help in the correction of the manuscript. We would like to thank all our colleagues for inspiring discussions. The CNRS, the University of Poitiers and the Région Poitou–Charentes are gratefully acknowledged for their financial support.

Supplementary material

11103_2014_198_MOESM1_ESM.tif (7 mb)
Fig. S1. STP13 mRNA accumulation in STP13OE-6 plants. Plants were sprayed with mock solution or B. cinerea suspension (5.104 conidia.ml−1). Healthy (0 hpi) or treated leaves from at least 3 plants were harvested at indicated time points. The STP13 transcript levels were quantified by RT-qPCR. Data are expressed as normalized expression (no unit) to the plant reference gene At4g26410 expression level (Czechowski et al. 2005). Data are mean ± SE from 2 independent experiments. STP13 expression level of B. cinerea infected WT plants (48 hpi) is indicated. B.c.: B. cinerea. hpi: hours post-inoculation (TIFF 7185 kb)
11103_2014_198_MOESM2_ESM.tif (5.8 mb)
Fig. S2. Disease phenotype of wild-type (WT) and C2-7 plants infected with B. cinerea. A construct including the cDNA corresponding to STP13 mRNA driven by the CaMV35S promoter was introduced into stp13-2. In the resulting plants (named C2-7), leaves of five-week-old plants have been used for inoculation tests. A. Lesion diameters observed on WT and C2-7 plants 3 days after inoculation with 6 µl droplets containing 104 conidia.ml−1. Data represent the mean (± SE) lesion diameter from 4 independent experiments. In each experiment, at least 5 plants per genotype were infected with B. cinerea. No significant difference was determined between WT and C2-7 plants according to a permutation with general scores test (P < 0,05). B. Lesion size distribution observed on WT and C2-7 plants infected with B. cinerea. Plants were drop-inoculated and diameters of necrotic lesions (LD) were measured after 3 days. Lesions were grouped into 3 classes according to their size: small (LD < WT first quartile), medium (LD = WT interquartile range) and large (LD > WT third quartile). The percentage of lesion size distribution from 4 independent experiments is shown. No significant difference between WT and C2-7 was determined by a Chi square test (P < 0,05) (TIFF 5987 kb)
11103_2014_198_MOESM3_ESM.tif (1.4 mb)
Fig. S3. Six week-old wild-type (WT), stp13-2 and STP13OE-6 plants grown on soil (TIFF 1482 kb)
11103_2014_198_MOESM4_ESM.pdf (19 kb)
Table S2 Table reporting the results of the three-way ANOVA test of PDF1.2 and PAD3 expression. Genotype: wild-type, stp13-2 or STP13OE-6 plants. Time: 0, 24, 48 or 72 h post-treatment. Treatment: mock or B. cinerea (**P < 0,01; ***P < 0,001; nsd: not statistically different)(PDF 18 kb)
11103_2014_198_MOESM5_ESM.pdf (24 kb)
Table S1 List of primers used for RT-qPCR analysis (PDF 23 kb)

References

  1. Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R, Gadrinab C, Heller C, Jeske A, Koesema E, Meyers CC, Parker H, Prednis L, Ansari Y, Choy N, Deen H, Geralt M, Hazari N, Hom E, Karnes M, Mulholland C, Ndubaku R, Schmidt I, Guzman P, Aguilar-Henonin L, Schmid M, Weigel D, Carter DE, Marchand T, Risseeuw E, Brogden D, Zeko A, Crosby WL, Berry CC, Ecker JR (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301(5633):653–657PubMedCrossRefGoogle Scholar
  2. Arbelet D, Malfatti P, Simond-Côte E, Fontaine T, Desquilbet L, Expert D, Kunz C, Soulié M-C (2010) Disruption of the Bcchs3a Chitin Synthase gene in Botrytis cinerea is responsible for altered adhesion and overstimulation of host plant immunity. Mol Plant Microbe Interact 23(10):1324–1334PubMedCrossRefGoogle Scholar
  3. Azevedo H, Conde C, Geros H, Tavares RM (2006) The non-host pathogen Botrytis cinerea enhances glucose transport in Pinus pinaster suspension-cultured cells. Plant Cell Physiol 47(2):290–298PubMedCrossRefGoogle Scholar
  4. Bari R, Jones JG (2009) Role of plant hormones in plant defence responses. Plant Mol Biol 69(4):473–488PubMedCrossRefGoogle Scholar
  5. Biemelt S, Sonnewald U (2006) Plant-microbe interactions to probe regulation of plant carbon metabolism. J Plant Physiol 163(3):307–318PubMedCrossRefGoogle Scholar
  6. Boller T, Felix G (2009) A renaissance of elicitors: perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Ann Rev Plant Biol 60(1):379–406CrossRefGoogle Scholar
  7. Bolouri Moghaddam MR, Van den Ende W (2012) Sugars and plant innate immunity. J Exp Bot 63(11):3989–3998PubMedCrossRefGoogle Scholar
  8. Bourque S, Lemoine R, Sequeira-Legrand A, Fayolle L, Delrot S, Pugin A (2002) The elicitor cryptogein blocks glucose transport in tobacco cells. Plant Physiol 130(4):2177–2187PubMedCentralPubMedCrossRefGoogle Scholar
  9. Büttner M (2007) The monosaccharide transporter(-like) gene family in Arabidopsis. FEBS Lett 581(12):2318–2324PubMedCrossRefGoogle Scholar
  10. Büttner M (2010) The Arabidopsis sugar transporter (AtSTP) family: an update. Plant Biol (Stuttg) 12(Suppl 1):35–41CrossRefGoogle Scholar
  11. Büttner M, Truernit E, Baier K, Scholz-Starke J, Sontheim M, Lauterbach C, Huss VAR, Sauer N (2000) AtSTP3, a green leaf-specific, low affinity monosaccharide-H + symporter of Arabidopsis thaliana. Plant, Cell Environ 23(2):175–184CrossRefGoogle Scholar
  12. Choquer M, Fournier E, Kunz C, Levis C, Pradier J-M, Simon A, Viaud M (2007) Botrytis cinerea virulence factors: new insights into a necrotrophic and polyphageous pathogen. FEMS Microbiol Lett 277(1):1–10PubMedCrossRefGoogle Scholar
  13. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16(6):735–743PubMedCrossRefGoogle Scholar
  14. Czechowski T, Stitt M, Altmann T, Udvardi MK, Scheible W-R (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139(1):5–17PubMedCentralPubMedCrossRefGoogle Scholar
  15. Doehlemann G, Molitor F, Hahn M (2005) Molecular and functional characterization of a fructose specific transporter from the gray mold fungus Botrytis cinerea. Fungal Genet Biol 42(7):601–610PubMedCrossRefGoogle Scholar
  16. Dulermo T, Rascle C, Chinnici G, Gout E, Bligny R, Cotton P (2009) Dynamic carbon transfer during pathogenesis of sunflower by the necrotrophic fungus Botrytis cinerea: from plant hexoses to mannitol. New Phytol 183(4):1149–1162PubMedCrossRefGoogle Scholar
  17. Ehness R, Ecker M, Godt DE, Roitsch T (1997) Glucose and stress independently regulate source and sink metabolism and defense mechanisms via signal transduction pathways involving protein phosphorylation. Plant Cell 9(10):1825–1841PubMedCentralPubMedCrossRefGoogle Scholar
  18. Ferrari S, Plotnikova JM, De Lorenzo G, Ausubel FM (2003) Arabidopsis local resistance to Botrytis cinerea involves salicylic acid and camalexin and requires EDS4 and PAD2, but not SID2, EDS5 or PAD4. Plant J 35(2):193–205PubMedCrossRefGoogle Scholar
  19. Fotopoulos V, Gilbert MJ, Pittman JK, Marvier AC, Buchanan AJ, Sauer N, Hall JL, Williams LE (2003) The monosaccharide transporter gene, AtSTP4, and the cell-wall invertase, Atbetafruct1, are induced in Arabidopsis during infection with the fungal biotroph Erysiphe cichoracearum. Plant Physiol 132(2):821–829PubMedCentralPubMedCrossRefGoogle Scholar
  20. Glazebrook J (2005) Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. Ann Rev Phytopathol 43(1):205–227CrossRefGoogle Scholar
  21. Govrin EM, Levine A (2000) The hypersensitive response facilitates plant infection by the necrotrophic pathogen Botrytis cinerea. Curr Biol 10(13):751–757PubMedCrossRefGoogle Scholar
  22. Hall JL, Williams LE (2000) Assimilate transport and partitioning in fungal biotrophic interactions. Funct Plant Biol 27(6):549–560CrossRefGoogle Scholar
  23. Herbers K, Meuwly P, Frommer WB, Metraux JP, Sonnewald U (1996) Systemic acquired resistance mediated by the ectopic expression of invertase: possible hexose sensing in the secretory pathway. Plant Cell 8(5):793–803PubMedCentralPubMedCrossRefGoogle Scholar
  24. Karimi M, Inzé D, Depicker A (2002) GATEWAY™ vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci 7(5):193–195PubMedCrossRefGoogle Scholar
  25. Kliebenstein DJ, Rowe HC, Denby KJ (2005) Secondary metabolites influence Arabidopsis/Botrytis interactions: variation in host production and pathogen sensitivity. Plant J 44(1):25–36PubMedCrossRefGoogle Scholar
  26. La Camera S, L’Haridon F, Astier J, Zander M, Abou-Mansour E, Page G, Thurow C, Wendehenne D, Gatz C, Metraux JP, Lamotte O (2011) The glutaredoxin ATGRXS13 is required to facilitate Botrytis cinerea infection of Arabidopsis thaliana plants. Plant J 68(3):507–519PubMedCrossRefGoogle Scholar
  27. Lemoine R, La Camera S, Atanassova R, Dédaldéchamp F, Allario T, Pourtau N, Bonnemain J-L, Laloi M, Coutos-Thévenot P, Maurousset L, Faucher M, Girousse C, Lemonnier P, Parrilla J, Durand M (2013) Source to sink transport and regulation by environmental factors. Front Plant Sci 4:272 Google Scholar
  28. Mengiste T (2012) Plant immunity to necrotrophs. Ann Rev Phytopathol 50(1):267–294CrossRefGoogle Scholar
  29. Monaghan J, Zipfel C (2012) Plant pattern recognition receptor complexes at the plasma membrane. Curr Opin Plant Biol 15(4):349–357PubMedCrossRefGoogle Scholar
  30. Mukherjee AK, Carp M-J, Zuchman R, Ziv T, Horwitz BA, Gepstein S (2010) Proteomics of the response of Arabidopsis thaliana to infection with Alternaria brassicicola. J Proteomics 73(4):709–720PubMedCrossRefGoogle Scholar
  31. Norholm MH, Nour-Eldin HH, Brodersen P, Mundy J, Halkier BA (2006) Expression of the Arabidopsis high-affinity hexose transporter STP13 correlates with programmed cell death. FEBS Lett 580(9):2381–2387PubMedCrossRefGoogle Scholar
  32. Pieterse CMJ, Van der Does D, Zamioudis C, Leon-Reyes A, Van Wees SCM (2012) Hormonal modulation of plant immunity. Ann Rev Cell Dev Biol 28(1):489–521CrossRefGoogle Scholar
  33. Poschet G, Hannich B, Buttner M (2010) Identification and characterization of AtSTP14, a novel galactose transporter from Arabidopsis. Plant Cell Physiol 51(9):1571–1580PubMedCrossRefGoogle Scholar
  34. Roitsch T (1999) Source-sink regulation by sugar and stress. Curr Opin Plant Biol 2(3):198–206PubMedCrossRefGoogle Scholar
  35. Roitsch T, Gonzalez MC (2004) Function and regulation of plant invertases: sweet sensations. Trends Plant Sci 9(12):606–613PubMedCrossRefGoogle Scholar
  36. Roitsch T, Balibrea ME, Hofmann M, Proels R, Sinha AK (2003) Extracellular invertase: key metabolic enzyme and PR protein. J Exp Bot 54(382):513–524PubMedCrossRefGoogle Scholar
  37. Rolland F, Baena-Gonzalez E, Sheen J (2006) Sugar sensing and signalling in plants: conserved and novel mechanisms. Ann Rev Plant Biol 57:675–709CrossRefGoogle Scholar
  38. Rowe HC, Kliebenstein DJ (2008) Complex genetics control natural variation in Arabidopsis thaliana resistance to Botrytis cinerea. Genetics 180(4):2237–2250PubMedCentralPubMedCrossRefGoogle Scholar
  39. Rowe HC, Walley JW, Corwin J, Chan EKF, Dehesh K, Kliebenstein DJ (2010) Deficiencies in Jasmonate-mediated plant defense reveal quantitative variation in Botrytis cinerea pathogenesis. PLoS Pathog 6(4):e1000861PubMedCentralPubMedCrossRefGoogle Scholar
  40. Ruiz E, Ruffner HP (2002) Immunodetection of Botrytis-specific invertase in infected grapes. J Phytopathol 150(2):76–85CrossRefGoogle Scholar
  41. Schofield RA, Bi YM, Kant S, Rothstein SJ (2009) Over-expression of STP13, a hexose transporter, improves plant growth and nitrogen use in Arabidopsis thaliana seedlings. Plant, Cell Environ 32(3):271–285CrossRefGoogle Scholar
  42. Schuhegger R, Nafisi M, Mansourova M, Petersen BL, Olsen CE, Svatos A, Halkier BA, Glawischnig E (2006) CYP71B15 (PAD3) catalyzes the final step in camalexin biosynthesis. Plant Physiol 141(4):1248–1254PubMedCentralPubMedCrossRefGoogle Scholar
  43. Sherson SM, Hemmann G, Wallace G, Forbes S, Germain V, Stadler R, Bechtold N, Sauer N, Smith SM (2000) Monosaccharide/proton symporter AtSTP1 plays a major role in uptake and response of Arabidopsis seeds and seedlings to sugars. Plant J 24(6):849–857PubMedCrossRefGoogle Scholar
  44. Slewinski TL (2011) Diverse functional roles of monosaccharide transporters and their homologs in vascular plants: a physiological perspective. Mol Plant 4(4):641–662PubMedCrossRefGoogle Scholar
  45. Staats M, van Kan JAL (2012) Genome Update of Botrytis cinerea Strains B05.10 and T4. Eukaryot Cell 11(11):1413–1414PubMedCentralPubMedCrossRefGoogle Scholar
  46. Stadler R, Büttner M, Ache P, Hedrich R, Ivashikina N, Melzer M, Shearson SM, Smith SM, Sauer N (2003) Diurnal and light-regulated expression of AtSTP1 in guard cells of Arabidopsis. Plant Physiol 133(2):528–537PubMedCentralPubMedCrossRefGoogle Scholar
  47. Stefanato FL, Abou-Mansour E, Buchala A, Kretschmer M, Mosbach A, Hahn M, Bochet CG, Métraux J-P, Schoonbeek H-J (2009) The ABC transporter BcatrB from Botrytis cinerea exports camalexin and is a virulence factor on Arabidopsis thaliana. Plant J 58(3):499–510PubMedCrossRefGoogle Scholar
  48. ten Have A, Mulder W, Visser J, van Kan JA (1998) The endopolygalacturonase gene Bcpg1 is required for full virulence of Botrytis cinerea. Mol Plant Microbe Interact 11(10):1009–1016PubMedCrossRefGoogle Scholar
  49. Thomma BPHJ, Eggermont K, Penninckx IAMA, Mauch-Mani B, Vogelsang R, Cammue BPA, Broekaert WF (1998) Separate jasmonate-dependent and salicylate-dependent defense-response pathways in Arabidopsis are essential for resistance to distinct microbial pathogens. Proc Natl Acad Sci USA 95(25):15107–15111PubMedCentralPubMedCrossRefGoogle Scholar
  50. Thomma BPHJ, Penninckx IAMA, Cammue BPA, Broekaert WF (2001) The complexity of disease signalling in Arabidopsis. Curr Opin Immunol 13(1):63–68PubMedCrossRefGoogle Scholar
  51. Thomma BPHJ, Nürnberger T, Joosten MHAJ (2011) Of PAMPs and effectors: the blurred PTI-ETI dichotomy. Plant Cell 23(1):4–15PubMedCentralPubMedCrossRefGoogle Scholar
  52. Truernit E, Schmid J, Epple P, Illig J, Sauer N (1996) The sink-specific and stress-regulated Arabidopsis STP4 gene: enhanced expression of a gene encoding a monosaccharide transporter by wounding, elicitors, and pathogen challenge. Plant Cell 8(12):2169–2182PubMedCentralPubMedCrossRefGoogle Scholar
  53. Tsuda K, Katagiri F (2010) Comparing signalling mechanisms engaged in pattern-triggered and effector-triggered immunity. Curr Opin Plant Biol 13(4):459–465PubMedCrossRefGoogle Scholar
  54. van Bel AJE (2003) The phloem, a miracle of ingenuity. Plant, Cell Environ 26(1):125–149CrossRefGoogle Scholar
  55. van Kan JAL (2006) Licensed to kill: the lifestyle of a necrotrophic plant pathogen. Trends Plant Sci 11(5):247–253PubMedCrossRefGoogle Scholar
  56. Voegele R, Mendgen K (2011) Nutrient uptake in rust fungi: how sweet is parasitic life? Euphytica 179(1):41–55CrossRefGoogle Scholar
  57. Voegele RT, Struck C, Hahn M, Mendgen K (2001) The role of haustoria in sugar supply during infection of broad bean by the rust fungus Uromyces fabae. Proc Natl Acad Sci USA 98(14):8133–8138PubMedCentralPubMedCrossRefGoogle Scholar
  58. Williamson B, Tudzynski B, Tudzynski P, Van Kan JAL (2007) Botrytis cinerea: the cause of grey mould disease. Mol Plant Pathol 8(5):561–580PubMedCrossRefGoogle Scholar
  59. Yamada K, Kanai M, Osakabe Y, Ohiraki H, Shinozaki K, Yamaguchi-Shinozaki K (2011) Monosaccharide absorption activity of Arabidopsis roots depends on expression profiles of transporter genes under high salinity conditions. J Biol Chem 286(50):43577–43586PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Pauline Lemonnier
    • 1
  • Cécile Gaillard
    • 1
  • Florian Veillet
    • 1
  • Jérémy Verbeke
    • 1
  • Rémi Lemoine
    • 1
  • Pierre Coutos-Thévenot
    • 1
  • Sylvain La Camera
    • 1
  1. 1.UMR CNRS 7267 EBI Ecologie et Biologie des Interactions, Equipe “Physiologie Moléculaire du Transport des Sucres chez les végétaux”Université de PoitiersPoitiers Cedex 9France

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