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Phaseolus ENOD40 is involved in symbiotic and non-symbiotic organogenetic processes: expression during nodule and lateral root development

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Abstract

ENOD40 is an early nodulin gene, recently isolated from legume species forming nodules either after Rhizobium infection or spontaneously. ENOD40 cDNAs from Phaseolus plants were isolated and nucleotide sequence determination revealed a 85% and 88.5% homology with the reported soybean cDNA clones. The putative polypeptide deduced coincides with the soybean one but a stop codon, almost in the middle of the respective ORF, renders it much shorter. This polypeptide was overexpressed as a fusion protein in Escherichia coli. Although the spatial expression pattern of the gene in the root pericycle and nodule primordium at early stages of development as well as in the pericycle of the vascular bundles and uninfected cells in mature nodules is comparable to the gene's expression pattern in soybean, differences in developmental regulation are evident. We have shown that ENOD40 transcripts are also detected at very early stages of lateral root development, in the dividing pericycle cells of the root stele that give rise to the lateral root primordia. The presence of Rhizobium causes an enhancement of the gene's expression and also induction of the gene in the vascular tissues of developed lateral roots. Interestingly, a discrimination on the gene's expression level in adventious and acropetal incipient lateral root primordia, emerging in infected and uninfected roots, is observed. This indicates that the gene's product may be involved in the hormonal status of the plant and that ENOD40 may be used as a molecular marker in lateral root initiation.

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References

  1. Asad S, Fang Y, Wycoff KL, Hirsch AM: Isolation and characterization of cDNA and genomic clones of MsENOD40; transcripts are detected in meristematic cells of alfalfa. Protoplasma 183: 10–23 (1994).

    Google Scholar 

  2. Bergensen FJ: Root Nodules of Legumes: Structure and Functions. Research Studies Press/Willey, Chishester (1982).

    Google Scholar 

  3. Bisseling T, van de Bos RC, van Kammen A: The effect of ammonium nitrate on the synthesis of nitrogenase and the concentration of leghemoglobin in pea root nodules induced by Rhizobium leguminosarum. Biochim Biophys Acta 539: 1–11 (1978).

    Google Scholar 

  4. Blakely LM, Blakely RM, Elliot DS: Experimental studies on lateral root formation in radish seedling roots. II Analysis of the dose-responce to exogenous auxin. Plant Physiol 87: 414–419 (1988).

    Google Scholar 

  5. Calvert HE, Pence MK, Pierce M, Malik NSA, Bauer WD: Anatomical analysis of the development and distribution of Rhizobium infections in soybean roots. Can J Bot 62: 2375–2384 (1984).

    Google Scholar 

  6. Charlton WA: Lateral root initiation. In: Waisel Y, Wshel A, Kafkafi U (eds) Plant Roots: The Hidden Half, pp. 103–128. New York (1991).

  7. Crespi MD, Jurkevitch E, Poiret M, Aubeton-Carafa Y, Petrovits G, Kondorosi E, Kondorosi A: Enod40, a gene expressed during nodule organogenesis, codes for a nontranslatable RNA involved in plant growth. EMBO J 13: 5099–5112 (1994).

    Google Scholar 

  8. De Block M, Debrouwer D: RNA-RNA in situ hybridization using digoxigenin-labelled probes: the use of high-molecular-weihght polyvinyl alcohol in the alkaline phosphatase indoxyl-nitroblue tetrazolium reaction. Anal Biochem 215: 86–89 (1993).

    Google Scholar 

  9. Franssen HJ, Vijn I, Yang WC, Bisseling T: Developmental aspects of the Rhizobium-legume symbiosis. Plant Mol Biol 19: 89–107 (1992).

    Google Scholar 

  10. Gresshof PM, Delves AC: Plant genetic approach to symbiotic nodulation and nitrogen fixation in legumes. In: Blanstein AD, King PJ (eds) A Genetic Approach to Plant Biochemistry, pp. 159–206. Springer-Verlag, New York (1986).

    Google Scholar 

  11. Hirsch AM, Fang Y: Plant hormones and nodulation: what's the connection? Plant Mol Biol 26: 5–9 (1994).

    Google Scholar 

  12. Klee H, Estelle M: Molecular genetic approaches to plant hormone biology. Annu Rev Plant Physiol Plant Mol Biol 41: 521–551 (1991).

    Google Scholar 

  13. Kouchi H, Hata S: Isolation and characterization of novel nodulin cDNAs representing genes expressed at early stages of soybean nodule development. Mol Gen Genet 238: 106–119 (1993).

    Google Scholar 

  14. Kouchi H, Tsukamoto M, Tajima S: Differential expression of nodule specific (nodulin) genes in infected, uninfected and cortical cells of soybean (Glycine soya) root nodules. J Plant Physiol 135: 608–617 (1989).

    Google Scholar 

  15. Lancelle SA, Torrey JG: Early development of Rhizobium-induced root nodules of Parasporia rigida. II. Nodule morphogenesis and symbiotic development. Can J Bot 63: 25–35 (1984).

    Google Scholar 

  16. Lancelle SA, Torrey JG: Early development of Rhizobium-induced root nodules of Parasporia rigida. I. Infection and early nodule initiation. Protoplasma 123: 26–37 (1984).

    Google Scholar 

  17. Legocki RP, Verma DPS: Identification of ‘nodule-specific’ host proteins (nodulins) involved in the development of Rhizobium-legume symbiosis. Cell 20: 153–163 (1995).

    Google Scholar 

  18. Mac Isaak SM, Sawhney VK, Phorecky Y: Regulation of root formation in lettuce (Lactuca sativa) seeding roots. Interacting effects of a α-naphthaleneacetic acid and kinetin. Physiol Plant 77: 287–293 (1989).

    Google Scholar 

  19. Marvel DJ, Kuldau G, Hirsch AM, Richards E, Torrey JG, Ausubel FM: Conservation of nodulation genes between Rhizobium meliloti and a slow-growing Rhizobium strain that nodulates a non-legume host. Proc Nat Acad Sci USA 82: 5841–5842 (1985).

    Google Scholar 

  20. Matvienko M, van de Sande K, Yang WC, van Kammen A, Bisseling T, Franssen H: Comparison of soybean and pea ENOD40 cDNA clones representing genes expressed during both early and late stages of nodule development. Plant Mol Biol 26: 487–493 (1994).

    Google Scholar 

  21. Miao GH, Verma DPS: Soybean nodulin-26 gene encoding a channel protein is expressed only in infected cells of nodules and is regulated differently in roots of homologous and heterologous plants. Plant Cell 5: 781–794 (1993).

    Google Scholar 

  22. Nap JP, Bisseling T: Developmental biology of plant-procaryote symbiosis: the legume root nodule. Science 250: 948–954 (1990).

    Google Scholar 

  23. Newcomb EH, Kaneko Y, van den Bosch KA: Specialization of the inner cortex for ureide production in soybean root nodules. Protoplasma 150: 150–159 (1989).

    Google Scholar 

  24. Newcomb W, Sippel D, Robertson RL: The early morphogenesis of Glycine max. and Pisum sativum root nodules. Can J Bot 57: 2603–2616 (1979).

    Google Scholar 

  25. Pate JS, Gunning BES, Biarty LG: Ultrastructure and functioning of the transport system of the leguminous root nodule. Planta 85: 11–34 (1969).

    Google Scholar 

  26. Rogers SO, Bendich AJ: Extraction of DNA from plant tissues. In: Gelvin SB, Schilperoort RA, Verma DPS (eds) Plant Molecular Biology Manual, pp. 1–11. Kluwer Academic Publishers, Dordrecht (1988).

    Google Scholar 

  27. Sambrook J, Fritsch T, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  28. Sanchez F, Padilla JE, Perez H, Lara M: Control of nodulin genes in root-nodule development and metabolism. Annu Rev Plant Physiol Plant Mol Biol 42: 507–528 (1991).

    Google Scholar 

  29. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  30. Scheres B, van de Wiel C, Zalensky A, Horvath B, Spaink HP, van Eck H, Zwartkruis F, Wolters A, Gloudemans T, van Kammen A, Bisseling T: The ENOD12 gene product is involved in the infection process during the pea-Rhizobium interaction. Cell 60: 281–294 (1988).

    Google Scholar 

  31. Scheres B, van Engelen F, van der Knaap E, van de Wiel C, van Kammen A, Bisseling T: Sequential induction of nodulin gene expression in developing pea nodules. Plant Cell 2: 687–700 (1990).

    Google Scholar 

  32. Smith DB, Johnson KS: Single-strep purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67: 31–40 (1988).

    Google Scholar 

  33. Staden R: Graphic methods to determine the function of nucleic acid sequences. Nucl Acids Res 12: 521–528 (1984).

    Google Scholar 

  34. Taylor BH, Scheuring CF: A molecular marker for lateral root initiation: The RSI-1 gene of tomato (Lycopersicon esculentum Mill) is activated in early lateral root primordia. Mol Gen Genet 243: 148–157 (1994).

    Google Scholar 

  35. Torrey JG: Root hormones and plant growth. Annu Rev Plant Physiol 27: 435–459 (1976).

    Google Scholar 

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

    Google Scholar 

  37. van Kammen A: Suggested nomenclature for plant genes involved in nodulation and symbiosis. Plant Mol Biol Rep 2: 43–45 (1984).

    Google Scholar 

  38. Vasse J, De Billy F, Camut S, Truchet G: Correlation between ultrastructural differentiation of bacteroids and nitrogen fixation in alfalfa nodules. J Bact 172: 4295–4306 (1990).

    Google Scholar 

  39. Yang WC, Katinakis P, Hendriks P, Smolders A, de Vries F, Spee J, van Kammen A, Bisseling T, Franssen H: Characterization of GmENOD40, a gene showing novel patterns of cell-specific expression during soybean nodule development. Plant J 3: 573–585 (1993).

    Google Scholar 

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Papadopoulou, K., Roussis, A. & Katinakis, P. Phaseolus ENOD40 is involved in symbiotic and non-symbiotic organogenetic processes: expression during nodule and lateral root development. Plant Mol Biol 30, 403–417 (1996). https://doi.org/10.1007/BF00049320

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  • DOI: https://doi.org/10.1007/BF00049320

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