Skip to main content
Log in

Analysis of the two nodulins, sucrose synthase and ENOD2, in transgenic Lotus plants

  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

A partial sucrose synthase clone was isolated from a nodule cDNA library of Lotus japonicus, which had strong homology to sucrose synthase genes from Vicia faba and other plant species. The transcript was 2.7 kb in size and expression was enhanced in nodules compared to roots of L. japonicus. Two overlapping genomic clones containing sucrose synthase sequences were isolated from L. japonicus with a view to mapping and analysing the promoter of this gene.

The early nodulin ENOD2 is a (hydroxy)proline-rich cell wall protein, which is expressed in the inner cortex surrounding the infected region of root nodules. In order to see if this nodulin is involved in the operation of the oxygen diffusion barrier, L. corniculatus was transformed with antisense ENOD2 gene constructs derived from the promoter and coding sequence of soybean. Two of the 20 antisense plant lines had significantly lower dry weight and nitrogenase activity as well as a different O2 profile through the nodules when compared to control plant lines, but there was no difference in ENOD2 transcriptional activity between the control and antisense plants. Possible reasons for this observation are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

kDa:

kilodalton

kb:

kilobases

CAT:

chloramphenicol acetyl transferase

CaMV:

Cauliflower Mosaic Virus

Nos:

Nopaline Synthase

References

  • Bradley D J, Kjellbom P and Lamb C J 1992 Elicitor- and wound-induced oxidative cross-linking of a proline-roch plant cell wall protein: a novel, rapid defense response. Cell 70, 21–30.

    Google Scholar 

  • Franssen H, Thomson D V, Idler K, Kormelink R, van Kammen A and Bisseling T 1987 Nucleotide sequence of two soybean ENOD2 early nodulin genes encoding Ngm-75. Plant Mol. Biol. 14, 103–106.

    Google Scholar 

  • Gonzalez E M, Gordon A J, James C L and Arrese-Igor C 1995 The role of sucrose synthase in the response of soybean nodules to drought. J. Exp. Bot. 46, 1515–1523.

    Google Scholar 

  • Gordon A J 1992 Carbon metabolism in the legume nodule. In Carbon Partitioning within and between Organism. Eds. C J Pollock, J F Farrar and A J Gordon. pp 133–162. BIOS Scientific Publishers, Oxford, UK.

    Google Scholar 

  • Gordon A J, Ougham H J and James C L 1993 Changes in levels of gene transcripts and their corresponding proteins in nodules of soybean plants subjected to dark-induced stress. J. Exp. Bot. 44, 1453–1460.

    Google Scholar 

  • Handberg K and Stougaard J 1992 Lotus japonicus, an autoganous, diploid legume species for classical and molecular genetics. Plant J. 2, 487–496.

    Google Scholar 

  • Hansen J, Jørgensen J-E, Stougaard J and Marcker K A 1989 Hairy roots-a short cut to transgenic root nodules. Plant Cell Rep. 8, 12–15.

    Google Scholar 

  • Hunt S and Layzell D R 1993 Gas exchange of legume nodules and the regulation of nitrogease activity. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44, 483–511.

    Google Scholar 

  • Iannetta P P M, de Lorenzo C, James E K, Fernandez-Pascual M, Sprent J I, Lucas M M Witty J F, de Felipe M R and Minchin F R 1993 Oxygen diffusion in lupin nodules. J. Exp. Bot. 44, 1461–1467.

    Google Scholar 

  • James E K, Sprent J I, Minchin F R and Brewin N J 1991 Intercellular location of glycoprotein in soybean nodules: effect of altered rhizosphere oxygen concentration. Plant Cell Environ. 14, 467–476.

    Google Scholar 

  • Küster H, Fruhling M, Perlick A M and Pühler A 1993 The sucrose synthase gene is predominantly expressed in the root nodule tissue of Vicia faba. Mol. Plant-Microbe Interact. 6, 507–514.

    Google Scholar 

  • Lauridsen P, Franssen H, Stougaard J, Bisseling T and Marcker K A 1993 Conserved regulation of the soybean early nodulin ENOD2 gene promter in determinate and indeterminate transgenic root nodules. Plant J. 3, 483–492.

    Google Scholar 

  • Minchin F R 1996 Regulation of oxygen diffusion in legume nodules. Soil Biol. Biochem. (In press).

  • Minchin F R, Iannetta P P M, Fernandez-Pascual M, de Lorenzo C, Witty J F and Sprent J I 1992 A new procedure for the calculation of oxygen diffusion resistance in legume nodules from flowthrough gas analysis data. Ann. Bot. 70, 283–289.

    Google Scholar 

  • Morell M and Copeland L 1985 Sucrose synthase of soybean nodules. Plant Physiol. 78, 149–154.

    Google Scholar 

  • Salanoubat M and Belliard G 1987 Molecular cloning and sequencing of sucrose synthase cDNA from potato (Solanum tuberosum L.): Preliminary characterization of sucrose synthase mRNA distribution. Gene 60, 47–56.

    Google Scholar 

  • Stougaard J, Marcker K A, Otten L and Schell J 1986 Nodule-specific expression of a chimaeric soybean leghaemoglobin gene in transgenic Lotus corniculatus. Nature 321, 669–674.

    Google Scholar 

  • Thummler F and Verma D P S 1987 Nodulin-100 of soybean is the subunit of sucrose synthase regulated by the availability of free heme in nodules. J. Biol. Chem. 262, 14730–14736.

    Google Scholar 

  • Van de Wiel C, Scheres B, Franssen H, van Lierop M J, van Lammeren A, van Kammen A and Bisseling T 1990 The early nodulin transcript ENOD2 is located in the nodule parenchyma (inner cortex) of pea and soybean root nodules. EMBO J. 9, 1–7.

    Google Scholar 

  • Vanden Bosch K A, Bradley D A, Knox J-P, Perotto S, Butcher G W and Brewin N J 1989 Common components of the infection thread matrix and the intercellular space identified by the immunocyto-chemical analysis of pea nodules and uninfected roots. EMBO J. 8, 335–342.

    Google Scholar 

  • Webb K J, Jones S, Robbins M P and Minchin F R 1990 Characterization of transgenic root cultures of Trifolium repens, Trifolium pratense and Lotus corniculatus and transgenic plants of Lotus corniculatus. Plant Sci. 70, 243–254.

    Google Scholar 

  • Webb K J, Skøt L and Jørgensen B 1994 Plant genes involved in nodule development and nitrogen fixation. In Proceedings of the 1st European Nitrogen Fixation Conference. Eds. G B Kiss and G Endre. pp 234–238. Officina Press, Szeged, Hungary.

    Google Scholar 

  • Werr W, Frommer W B, Maas C and Starlinger P 1985 Structure of the sucrose synthase gene on chromosome 9 of Zea mays L. EMBO J. 4, 1373–1380.

    Google Scholar 

  • Witty J F, Minchin F R, Skøt L and Sheehy J E 1986 Nitrogen fixation and oxygen in legume root nodules. Oxford Surv. Plant Mol. Cell Biol. 3, 274–314.

    Google Scholar 

  • Witty J F, Skøt L and Revsbech N P 1987 Direct evidence for changes in the resistance of legume root nodules to O2 diffusion. J. Exp. Bot. 38, 1129–1140.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Skøt, L., Minchin, F.R., Timms, E. et al. Analysis of the two nodulins, sucrose synthase and ENOD2, in transgenic Lotus plants. Plant Soil 186, 99–106 (1996). https://doi.org/10.1007/BF00035062

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00035062

Key words

Navigation