, Volume 218, Issue 1, pp 42–49

Expression of ENOD40 during tomato plant development

  • Ingrid Vleghels
  • Jan Hontelez
  • Ana Ribeiro
  • Paul Fransz
  • Ton Bisseling
  • Henk Franssen
Original Article


In legumes, ENOD40 expression is increased upon interaction of plants with rhizobia. Little is known of the expression pattern of ENOD40 during other stages of the plant life cycle. Studies of ENOD40 expression in non-legume development may give an indication of the function of the gene. To investigate the ENOD40 expression pattern during plant development, a fusion between the β-glucuronidase (GUS) reporter gene and 150 bp of the 5′ untranslated region plus 3,000 bp of 5′ untranscribed tomato ENOD40 sequence was constructed and introduced into Lycopersicon esculentum Miller. Based on the observed GUS expression patterns in transgenic tomato we speculate that ENOD40 in tomato has a role in counteracting ethylene-provoked responses.


ENOD40 Ethylene Lycopersicon Promoter analysis Plant development (tomato) 





fluorescence in situ hybridisation


rapid amplification of cDNA ends


restriction fragment length polymorphism


  1. Becker JD, Moreira LM, Kapp D, Frosch SC, Puhler A, Perlic AM (2001) The nodulin vfENOD18 is an ATP-binding protein in infected cells of Vicia faba L. nodules. Plant Mol Biol 47:749–759CrossRefPubMedGoogle Scholar
  2. Charon C, Sousa C, Crespi M, Kondorosi A (1999) Alteration of enod40 expression modifies Medicago truncatula root nodule development induced by Sinorhizobium meliloti. Plant Cell 11:1953–1965CrossRefPubMedGoogle Scholar
  3. Corich G, Goormachtig S, Lievens S, Van Montagu M, Holsters M (1998) Patterns of ENOD40 gene expression in stem-borne nodules of Sesbania rostrata. Plant Mol Biol 37:67–76PubMedGoogle Scholar
  4. Endre G, Kereszt A, Kevel Z, Milhacea S, Kaló P, Kiss GB (2002) A receptor kinase gene regulating symbiotic nodule development. Nature 417:962–966CrossRefPubMedGoogle Scholar
  5. Fang Y, Hirsch AM (1998) Studying early nodulin gene ENOD40 expression and induction by nodulation factor and cytokinin in transgenic alfalfa. Plant Physiol 116:53–68PubMedGoogle Scholar
  6. Flemetakis E, Kavroulakis N, Quaedvlieg NE, Spaink HP, Dimou M, Roussis A, Katinakis P (2000) Lotus japonicus contains two distinct ENOD40 genes that are expressed in symbiotic, nonsymbiotic, and embryonic tissues. Mol Plant Microbe Interact 13:987–994PubMedGoogle Scholar
  7. Gualtieri G, Kulikova O, Limpens E, Kim DJ, Cook DR, Bisseling T, Geurts R (2002) Microsynteny between pea and Medicago truncatula in the SYM2 region. Plant Mol Biol 50:225–235Google Scholar
  8. Guinel FC, LaRue TA (1991) Light microscopy study of nodule initiation in Pisum sativum L. cv Sparkle and in its low-nodulating mutant E2 (sym 5). Plant Physiol 97:1206–1211Google Scholar
  9. Heidstra R, Yang WC, Yalcin Y, Peck S, Emons AM, Van Kammen A, Bisseling T (1997) Ethylene provides positional information on cortical cell division but is not involved in Nod factor-induced root hair tip growth in Rhizobium–legume interaction. Development 124:1781–1787PubMedGoogle Scholar
  10. Khalil MK (1992) Nature of growth regulators effects on Nicotiana tabacum seed germination. Angew Bot 66:106–108Google Scholar
  11. Kosugi S, Ohashi Y, Nakajima K, Arai Y (1990) An improved assay for beta-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous beta-glucuronidase activity. Plant Sci 70:133–140Google Scholar
  12. Kouchi H, Takane K, So RB, Ladha JK, Reddy PM (1999) Rice ENOD40: isolation and expression analysis in rice and transgenic soybean root nodules. Plant J 18:121–129CrossRefPubMedGoogle Scholar
  13. Kulikova O, Gualtieri G, Geurts R, Kim DJ, Cook DR, Huguet T, de Jong H, Franz PF, Bisseling T (2001) Integration of the FISH-pachytene and genetic maps of Medicago truncatula. Plant J 27:49–58PubMedGoogle Scholar
  14. Larsen PB, Woltering EJ, Woodson WR (1993) Ethylene and interorgan signalling in flowers following pollination. In: Schultz JC, Raskin I (eds) Plant signals in interactions with other organisms. American Society for Plant Physiology, Rockville, MD, pp 171–181Google Scholar
  15. Leubner-Metzger G, Petruzzelli L, Waldvogel R, Vögeli-Lange R, Meins F Jr (1998) Ethylene-responsive element binding protein (EREBP) expression and the transcriptional regulation of class I β-1,3-glucanase during tobacco seed germination. Plant Mol Biol 38:785–795PubMedGoogle Scholar
  16. Matvienko, van de Sande K, Yang WC, van Kammen A, Bisseling T, Franssen H (1994) Comparison of soybean and pea ENOD40 cDNA clones representing genes expressed during both early and late stages of nodule development. Plant Mol Biol 26:487–493PubMedGoogle Scholar
  17. Matvienko M, van de Sande K, Pawlowski K, van Kammen A, Franssen H, Bisseling T (1996) Nicotiana tabacum SR1 contains two ENOD40 homologs. In: Stacey G, Mullin B, Gresshoff PM (eds) Biology of plant–microbe interactions. International Society for Molecular Plant–Microbe Interactions, St. Paul, MN, USA,pp 387–391Google Scholar
  18. Papadopoulou K, Roussis A, Katinakis P (1996) Phaseolus ENOD40 is involved in symbiotic and non-symbiotic organogenetic processes: expression during nodule and lateral root development. Plant Mol Biol 30:403–417PubMedGoogle Scholar
  19. Pawlowski K, Kunze R, De Vries S, Bisseling T (1994) Isolation of total, poly(A) and polysomal RNA from plant tissues. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer, Dordrecht, The Netherlands, pp 1–13Google Scholar
  20. Penmetsa RV, Cook RC (1997) A legume ethylene-insensitive mutant hyperinfected by its rhizobial symbiont. Science 275:527–530CrossRefPubMedGoogle Scholar
  21. Reddy PM, Aggarwal RK, Ramos MC, Ladha JK, Brar DS, Kouchi H (1999) Widespread occurrence of the homologues of the early nodulin (ENOD) genes in Oryza species and related grasses. Biochem Biophys Res Commun 258:148–154CrossRefPubMedGoogle Scholar
  22. Roussis A, van de Sande K, Papadopoulou K, Drenth J, Bisseling T, Franssen H, Katinakis P (1995) Characterization of the soybean gene GmENOD40-2. J Exp Bot 46:719–724Google Scholar
  23. Schmidt EDL, van Hengel AJ, de Vries SC (1996) A rapid method for localizing cell-specific transcripts in plant cell cultures. Biochemica 4:25–28Google Scholar
  24. Stam P, Van Ooijen JW (1995) JoinMap (tm) version 2.0: software for the calculation of genetic linkage maps. CPRO-DLO WageningenGoogle Scholar
  25. Stougaard J (2001) Genetics and genomics of root symbiosis. Curr Opin Plant Biol 4:328–335CrossRefPubMedGoogle Scholar
  26. Stracke S, Kistner C, Yoshida S, Mulder L, Sato S, Kaneko T, Tabata S, Sandal N, Stougaard S, Szczyglowski K, Parniske M (2002) A plant receptor-like kinase required for both bacterial and fungal symbiosis. Nature 417:959–962CrossRefPubMedGoogle Scholar
  27. Van de Sande K, Pawlowski K, Czaja I, Wieneke U, Schell J, Schmidt J, Walden R, Matvienko M, Wellink J, van Kammen A, Franssen H, Bisseling T (1996) Modification of phytohormone response by a peptide encoded by ENOD40 of legumes and a nonlegume. Science 273:370–373PubMedGoogle Scholar
  28. Varkonyi-Gasic E, White DWR (2002) The white clover enod40 gene family. Expression patterns of two types of genes indicate a role in vascular function. Plant Physiol 129:1107–1118CrossRefPubMedGoogle Scholar
  29. Woltering EJ, ten Have A, Larsen PB, Woodson WR (1994) Ethylene biosynthetic genes and interorgan signalling during flower senescence. In: Scott RJ, Stead AD (eds) Molecular and cellular aspects of plant reproduction. Cambridge University Press, Cambridge, pp 285–307Google Scholar
  30. Wopereis J, Pajuelo E, Jiang Q, Gresshoff PM, De Bruin FJ, Stougaard J, Szczyglowski K (2000) Short root mutant of Lotus japonicus with a dramatically altered symbiotic phenotype. Plant J 23:97–114CrossRefPubMedGoogle Scholar
  31. Yang WC, Katinakis P, Hendriks P, Smolders A, de Vries F, Spee J, van Kammen A, Bisseling T, Franssen H (1993) Characterization of GmENOD40, a gene showing novel patterns of cell-specific expression during soybean nodule development. Plant J 3:573–585CrossRefPubMedGoogle Scholar
  32. Zhong XB, Fransz PF, Wennekes-van Eden J, Zabel P, van Kammen A, de Jong JH (1996) High resolution mapping by fluorescence in situ hybridisation to pachytene chromosomes and extended DNA fibres. Plant Mol Biol Rep 14:232–242Google Scholar
  33. Zhong XB, Fransz PF, Wennekes-van Eden J, Ramanna MS, van Kammen A, Zabel P, de Jong JH (1998) FISH studies reveal the molecular and chromosomal organisation of individual telomere domains in tomato. Plant J 13:507–517CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Ingrid Vleghels
    • 1
  • Jan Hontelez
    • 1
  • Ana Ribeiro
    • 2
  • Paul Fransz
    • 3
  • Ton Bisseling
    • 1
  • Henk Franssen
    • 1
  1. 1.Department of Molecular BiologyWageningen UniversityWageningenThe Netherlands
  2. 2.Lab. De Bioquimica VegetalInstituto de Tecnologia Quimica e BiologicaOeirasPortugal
  3. 3.Swammerdam Instituut voor LevenswetenschappenAmsterdamThe Netherlands

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