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Plant Molecular Biology

, Volume 87, Issue 3, pp 219–233 | Cite as

Cucumber ECERIFERUM1 (CsCER1), which influences the cuticle properties and drought tolerance of cucumber, plays a key role in VLC alkanes biosynthesis

  • Wenjiao Wang
  • Yan Zhang
  • Chong Xu
  • Jiaojiao Ren
  • Xiaofeng Liu
  • Kezia Black
  • Xinshuang Gai
  • Qian Wang
  • Huazhong Ren
Article

Abstract

Most land plants have a wax layer which covers their aerial parts to protect them from environmental stresses, such as drought, UV radiation, and pathogenic invasion. The wax biosynthesis has been well studied previously in Arabidopsis, but it still remains elusive in cucumber. Here, we isolated a CER1 homolog CsCER1 in cucumber, and we found that the expression of CsCER1 in the cucumber line 3401 which shows waxy fruit phenotype is much higher than that in the cucumber line 3413 which displays glossy fruit phenotype. Spatial and temporal expression analyses revealed that CsCER1 is specifically expressed in the epidermis where waxes are synthesized, and sub-cellular location showed that CsCER1 protein is localized to the endoplasmic reticulum. The expression of CsCER1 can be induced by low temperature, drought, salt stress and abscisic acid. In addition, abnormal expressions of CsCER1 in transgenic cucumber plants have dramatic effects on very-long-chain (VLC) alkanes biosynthesis, cuticle permeability, and drought resistance. Our data suggested that CsCER1 plays an important role in VLC alkanes biosynthesis in cucumber.

Keywords

Cucumber Wax CsCER1 Alkane biosynthesis Drought resistance 

Notes

Acknowledgments

We thank Professor Jianmin Wan (Institute of Crop Sciences, Chinese Academy of Agricultural Sciences) for providing the ER marker: mCherry-HDEL. Also we thank members of the Professor Ren lab for helpful discussions and technical assistance. WW, HR, and QW designed the experiments, WW performed most of the experiments and wrote the paper along with YZ and KB, XL and JR helped with the in situ hybridization, XG helped with the qRT-PCR, XL helped the subcellular localization, and CX helped with the data analysis. This work was supported by National science & technology research projects of China (2013BAD20B01), National public service sectors (agriculture) project of China (201203003), National high technology research and development program (863) of China (2012AA100103), Innovational Team Program of Beijing Industrial Technology System for Fruit-vegetables (GCTDZJ2014033008) and Scientific & Technological Research Project of Beijing (D131100000713001).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11103_2014_271_MOESM1_ESM.docx (20 kb)
Supplementary material 1 (DOCX 19 kb)

References

  1. Aarts M, Keijzer CJ, Stiekema WJ, Pereira A (1995) Molecular characterization of the CER1 gene of Arabidopsis involved in epicuticular wax biosynthesis and pollen fertility. Plant Cell 7:2115–2127PubMedCentralPubMedCrossRefGoogle Scholar
  2. Aharoni A, Dixit S, Jetter R, Thoenes E, van Arkel G, Pereira A (2004) The SHINE clade of AP2 domain transcription factors activates wax biosynthesis, alters cuticle properties, and confers drought tolerance when overexpressed in Arabidopsis. Plant Cell 16:2463–2480PubMedCentralPubMedCrossRefGoogle Scholar
  3. Andre C, Kim SW, Yu XH, Shanklin J (2013) Fusing catalase to an alkane-producing enzyme maintains enzymatic activity by converting the inhibitory byproduct H2O2 to the cosubstrate O2. PNAS 110:3191–3196PubMedCentralPubMedCrossRefGoogle Scholar
  4. Barrs HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci 15:413–428Google Scholar
  5. Bernard A, Domergue F, Pascal S, Jetter R, Renne C, Faure J-D, Haslam RP, Napier JA, Lessire R, Joubès J (2012) Reconstitution of plant alkane biosynthesis in yeast demonstrates that Arabidopsis ECERIFERUM1 and ECERIFERUM3 are core components of a very-long-chain alkane synthesis complex. Plant Cell 24:3106–3118PubMedCentralPubMedCrossRefGoogle Scholar
  6. Bourdenx B, Bernard A, Domergue F, Pascal S, Léger A, Roby D, Pervent M, Vile D, Haslam RP, Napier JA, Lessire R, Joubès J (2011) Overexpression of Arabidopsis ECERIFERUM1 promotes wax very-long-chain alkane biosynthesis and influences plant response to biotic and abiotic stresses. Plant Physiol 156:29–45PubMedCentralPubMedCrossRefGoogle Scholar
  7. Buschhaus C, Jetter R (2011) Composition differences between epicuticular and intracuticular wax substructures: how do plants seal their epidermal surfaces? J Exp Bot 62:841–853PubMedCrossRefGoogle Scholar
  8. Chen X, Goodwin SM, Boroff VL, Liu X, Jenks MA (2003) Cloning and characterization of the WAX2 gene of Arabidopsis involved in cuticle membrane and wax production. Plant Cell 15:1170–1185PubMedCentralPubMedCrossRefGoogle Scholar
  9. Das D, Eser BE, Han J, Sciore A, Marsh ENG (2011) Oxygen-independent decarbonylation of aldehydes by cyanobacterial aldehyde decarbonylase: a new reaction of diiron enzymes. Angew Chem Int Edit 50:7148–7152CrossRefGoogle Scholar
  10. Fukuda S, Satoh A, Kasahara H, Matsuyama H, Takeuchi Y (2008) Effects of ultraviolet-B irradiation on the cuticular wax of cucumber (Cucumis sativus) cotyledons. J Plant Res 121:179–189PubMedCrossRefGoogle Scholar
  11. Girard A-L, Mounet F, Lemaire-Chamley M, Gaillard C, Elmorjani K, Vivancos J, Runavot J-L, Quemener B, Petit J, Germain V, Rothan C, Marion D, Bakan B (2012) Tomato GDSL1 is required for cutin deposition in the fruit cuticle. Plant Cell 24:3119–3134PubMedCentralPubMedCrossRefGoogle Scholar
  12. Gray JE, Holroyd GH, van der Lee FM, Bahrami AR, Sijmons PC, Woodward FI, Schuch W, Hetherington AM (2000) The HIC signalling pathway links CO2 perception to stomatal development. Nature 408:713–716PubMedCrossRefGoogle Scholar
  13. Gülz P-G, Müller E, Schmitz K, Marner F-J, Güth S (1992) Chemical composition and surface structures of epicuticular leaf waxes of Ginkgo biloba, Magnolia grandiflora and Liriodendron tulipifera. Z Naturforsch C 47:516–526Google Scholar
  14. Heredia A (2003) Biophysical and biochemical characteristics of cutin, a plant barrier biopolymer. BBA Gen Subjects 1620:1–7CrossRefGoogle Scholar
  15. Islam MA, Du H, Ning J, Ye H, Xiong L (2009) Characterization of Glossy1-homologous genes in rice involved in leaf wax accumulation and drought resistance. Plant Mol Biol 70:443–456PubMedCrossRefGoogle Scholar
  16. Jenks MA, Joly RJ, Peters PJ, Rich PJ, Axtell JD, Ashworth EN (1994) Chemically induced cuticle mutation affecting epidermal conductance to water vapor and disease susceptibility in Sorghum bicolor (L.) Moench. Plant Physiol 105:1239–1245PubMedCentralPubMedGoogle Scholar
  17. Jenks MA, Tuttle HA, Eigenbrode SD, Feldmann KA (1995) Leaf epicuticular waxes of the eceriferum mutants in Arabidopsis. Plant Physiol 108:369–377PubMedCentralPubMedGoogle Scholar
  18. Jetter R, Klinger A, Schäffer S (2002) Very long-chain phenylpropyl and phenylbutyl esters from Taxus baccata needle cuticular waxes. Phytochemistry 61:579–587PubMedCrossRefGoogle Scholar
  19. Kim KS, Park SH, Jenks MA (2007a) Changes in leaf cuticular waxes of sesame (Sesamum indicum L.) plants exposed to water deficit. J Plant Physiol 164:1134–1143PubMedCrossRefGoogle Scholar
  20. Kim KS, Park SH, Kim DK, Jenks MA (2007b) Influence of water deficit on leaf cuticular waxes of soybean (Glycine max [L.] Merr.). Int J Plant Sci 168:307–316CrossRefGoogle Scholar
  21. Kosma DK, Bourdenx B, Bernard A, Parsons EP, Lü S, Joubès J, Jenks MA (2009) The impact of water deficiency on leaf cuticle lipids of Arabidopsis. Plant Physiol 151:1918–1929PubMedCentralPubMedCrossRefGoogle Scholar
  22. Kunst L, Samuels A (2003) Biosynthesis and secretion of plant cuticular wax. Prog Lipid Res 42:51–80PubMedCrossRefGoogle Scholar
  23. Kunst L, Samuels L (2009) Plant cuticles shine: advances in wax biosynthesis and export. Curr Opin Plant Biol 12:721–727PubMedCrossRefGoogle Scholar
  24. Lee SB, Jung SJ, Go YS, Kim HU, Kim JK, Cho HJ, Park OK, Suh MC (2009) Two Arabidopsis 3-ketoacyl CoA synthase genes, KCS20 and KCS2/DAISY, are functionally redundant in cuticular wax and root suberin biosynthesis, but differentially controlled by osmotic stress. Plant J 60:462–475PubMedCrossRefGoogle Scholar
  25. Leide J, Hildebrandt U, Reussing K, Riederer M, Vogg G (2007) The developmental pattern of tomato fruit wax accumulation and its impact on cuticular transpiration barrier properties: effects of a deficiency in a β-ketoacyl-coenzyme A synthase (LeCER6). Plant Physiol 144:1667–1679PubMedCentralPubMedCrossRefGoogle Scholar
  26. Lolle SJ, Berlyn GP, Engstrom EM, Krolikowski KA, Reiter W-D, Pruitt RE (1997) Developmental regulation of cell interactions in the Arabidopsis fiddlehead-1 mutant: a role for the epidermal cell wall and cuticle. Dev Biol 189:311–321PubMedCrossRefGoogle Scholar
  27. Luo X, Bai X, Sun X, Zhu D, Liu B, Ji W, Cai H, Cao L, Wu J, Hu M, Liu X, Tang L, Zhu Y (2013) Expression of wild soybean WRKY20 in Arabidopsis enhances drought tolerance and regulates ABA signalling. J Exp Bot 64:2155–2169PubMedCrossRefGoogle Scholar
  28. Mao B, Cheng Z, Lei C, Xu F, Gao S, Ren Y, Wang J, Zhang X, Wang J, Wu F, Guo X, Liu X, Wu C, Wang H, Wan J (2012) Wax crystal-sparse leaf2, a rice homologue of WAX2/GL1, is involved in synthesis of leaf cuticular wax. Planta 235:39–52PubMedCrossRefGoogle Scholar
  29. Markstädter C, Federle W, Jetter R, Riederer M, Hölldobler B (2000) Chemical composition of the slippery epicuticular wax blooms on Macaranga (Euphorbiaceae) ant-plants. Chemoecology 10:33–40CrossRefGoogle Scholar
  30. McNevin JP, Woodward W, Hannoufa A, Feldmann KA, Lemieux B (1993) Isolation and characterization of eceriferum (cer) mutants induced by T-DNA insertions in Arabidopsis thaliana. Genome 36:610–618PubMedCrossRefGoogle Scholar
  31. Nawrath C (2006) Unraveling the complex network of cuticular structure and function. Curr Opin Plant Biol 9:281–287PubMedCrossRefGoogle Scholar
  32. Pollard M, Beisson F, Li Y, Ohlrogge JB (2008) Building lipid barriers: biosynthesis of cutin and suberin. Trends Plant Sci 13:236–246PubMedCrossRefGoogle Scholar
  33. Post-Beittenmiller D (1996) Biochemistry and molecular biology of wax production in plants. Annu Rev Plant Biol 47:405–430CrossRefGoogle Scholar
  34. Riederer M (2006) Thermodynamics of the water permeability of plant cuticles: characterization of the polar pathway. J Exp Bot 57:2937–2942PubMedCrossRefGoogle Scholar
  35. Riederer M, Schreiber L (2001) Protecting against water loss: analysis of the barrier properties of plant cuticles. J Exp Bot 52:2023–2032PubMedCrossRefGoogle Scholar
  36. Rowland O, Zheng H, Hepworth SR, Lam P, Jetter R, Kunst L (2006) CER4 encodes an alcohol-forming fatty acyl-coenzyme A reductase involved in cuticular wax production in Arabidopsis. Plant Physiol 142:866–877PubMedCentralPubMedCrossRefGoogle Scholar
  37. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  38. Samuels L, Kunst L, Jetter R (2008) Sealing plant surfaces: cuticular wax formation by epidermal cells. Plant Biol 59:683–707CrossRefGoogle Scholar
  39. Schirmer A, Rude MA, Li X, Popova E, Del Cardayre SB (2010) Microbial biosynthesis of alkanes. Science 329:559–562PubMedCrossRefGoogle Scholar
  40. Schneider-Belhaddad F, Kolattukudy P (2000) Solubilization, partial purification, and characterization of a fatty aldehyde decarbonylase from a higher plant, Pisum sativum. Arch Biochem Biophys 377:341–349PubMedCrossRefGoogle Scholar
  41. Seo PJ, Lee SB, Suh MC, Park M-J, Go YS, Park C-M (2011) The MYB96 transcription factor regulates cuticular wax biosynthesis under drought conditions in Arabidopsis. Plant Cell 23:1138–1152PubMedCentralPubMedCrossRefGoogle Scholar
  42. Sui X, Meng F, Wang H, Wei Y, Li R, Wang Z, Hu L, Wang S, Zhang Z (2012) Molecular cloning, characteristics and low temperature response of raffinose synthase gene in Cucumis sativus L. J Plant Physiol 169:1883–1891PubMedCrossRefGoogle Scholar
  43. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739PubMedCentralPubMedCrossRefGoogle Scholar
  44. Taton M, Husselstein T, Benveniste P, Rahier A (2000) Role of highly conserved residues in the reaction catalyzed by recombinant ∆7-sterol-C5(6)-desaturase studied by site-directed mutagenesis. Biochemistry 39:701–711PubMedCrossRefGoogle Scholar
  45. Uppalapati SR, Ishiga Y, Doraiswamy V, Bedair M, Mittal S, Chen J, Nakashima J, Tang Y, Tadege M, Ratet P, Chen R, Schultheiss H, Mysore KS (2012) Loss of abaxial leaf epicuticular wax in Medicago truncatula irg1/palm1 mutants results in reduced spore differentiation of anthracnose and nonhost rust pathogens. Plant Cell 24:353–370PubMedCentralPubMedCrossRefGoogle Scholar
  46. Varagona MJ, Schmidt RJ, Raikhel NV (1992) Nuclear localization signal(s) required for nuclear targeting of the maize regulatory protein Opaque-2. Plant Cell 4:1213–1227PubMedCentralPubMedCrossRefGoogle Scholar
  47. von Wettstein-Knowles P (1982) Elongase and epicuticular wax biosynthesis. Physiol Veg 20:797–809Google Scholar
  48. Warui DM, Li N, Nørgaard H, Krebs C, Bollinger JM Jr, Booker SJ (2011) Detection of formate, rather than carbon monoxide, as the stoichiometric co-product in conversion of fatty aldehydes to alkanes by a cyanobacterial aldehyde decarbonylase. J Am Chem Soc 133(10):3316–3319PubMedCentralPubMedCrossRefGoogle Scholar
  49. Yang J, Isabel Ordiz M, Jaworski JG, Beachy RN (2011) Induced accumulation of cuticular waxes enhances drought tolerance in Arabidopsis by changes in development of stomata. Plant Physiol Biochem 49:1448–1455PubMedCrossRefGoogle Scholar
  50. Zhang JY, Broeckling CD, Blancaflor EB, Sledge MK, Sumner LW, Wang ZY (2005) Overexpression of WXP1, a putative Medicago truncatula AP2 domain-containing transcription factor gene, increases cuticular wax accumulation and enhances drought tolerance in transgenic alfalfa (Medicago sativa). Plant J 42:689–707PubMedCrossRefGoogle Scholar
  51. Zhang X, Zhou Y, Ding L, Wu Z, Liu R, Meyerowitz EM (2013) Transcription repressor HANABA TARANU controls flower development by integrating the actions of multiple hormones, floral organ specification genes, and GATA3 family genes in Arabidopsis. Plant Cell 25:83–101PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Wenjiao Wang
    • 1
    • 2
  • Yan Zhang
    • 3
  • Chong Xu
    • 1
    • 2
  • Jiaojiao Ren
    • 1
    • 2
  • Xiaofeng Liu
    • 1
    • 2
  • Kezia Black
    • 1
    • 2
    • 4
  • Xinshuang Gai
    • 1
    • 2
  • Qian Wang
    • 1
    • 2
  • Huazhong Ren
    • 1
    • 2
  1. 1.Department of Vegetable Science, College of Agronomy and Bio-technologyChina Agricultural UniversityBeijingPeople’s Republic of China
  2. 2.Department of Vegetable Science, Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable CropsChina Agricultural UniversityBeijingPeople’s Republic of China
  3. 3.Department of Vegetable Science, College of HorticultureNorthwest Agriculture and Forestry UniversityYanglingPeople’s Republic of China
  4. 4.Department of Food Production, Faculty of Food and AgricultureThe University of the West IndiesSt AugustineTrinidad

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