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Concurrent visualization of gusA and lacZ reporter gene expression

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Lotus japonicus Handbook

Abstract

Here we describe conditions under which two reporter genes, encoding β-glucuronidase (gusA) and β-galactosidase (lacZ), can be used to simultaneously distinguish between the sites of expression of plant and bacterial genes, respectively, in symbiotic tissues. These conditions involve the fixation of cytoplasmic proteins with cold methanol before detecting gusA expression, and the addition of detergents to the incubation solution in order to inactivate endogenous plant β-galactosidases.

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References

  • Bauer P, Ratet P, Crespi MD, Schultze M, and Kondorosi A. (1996) Nod factors and cytokinins induce similar cortical cell divisions, amyloplanst deposition and MsEnod12A expression pattern in alfalfa roots. The Plant Journal 10, 91–96.

    Article  CAS  Google Scholar 

  • Beeckman T, and Engler G. (1994) An easy technique for the clearing of histochemically stained plant tissue. Plant Molecular Biology Reporter 12, 37–42.

    Google Scholar 

  • Jefferson RA, Kavanagh TA, and Bevan MW. (1987) GUS-fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO Journal 6, 3901–3907.

    CAS  PubMed  Google Scholar 

  • Leong SA, Willimans PD, and Ditta GS. (1985) Analysis of the 5′ regulatory region of the gene for δ-aminolevulinic acid synthetase of Rhizobium meliloti. Nucleic Acids Research 13, 5965–5976.

    CAS  PubMed  Google Scholar 

  • Pichon M, Journet E-P, de Billy F, Dedieu A, Huguet T, Truchet G, and Barker DG. (1994a) ENOD12 gene expression as a molecular marker for comparing Rhizobium-dependant and —independent nodulation in alfalfa. Molecular Plant-Microbe Interactions. 7, 740–747.

    CAS  Google Scholar 

  • Pichon M, Journe, E-P, Dedieu A, de Billy F, Truchet G, and Barker DG. (1994b) Rhizobium meliloti elicits transient expression of the early noduline gene ENOD12 in the differentiating root epidermis of transgenic alfalfa. Plant Cell 4, 1199–1211.

    Google Scholar 

  • Scheres B, van de Wiel C, Zalenski A, Horvath B, Spaink H, Zwartkruis F, Wolters A-M, Gloudemans T, van Kammen A, and Bisseling T. (1990) The ENOD12 gene product is involved in the infection process during the pea-Rhizobium interaction. Cell, 60, 281–294.

    Article  CAS  PubMed  Google Scholar 

  • Trinh TH, Ratet P, Kondorosi E, Durand P, Kamaté K, Bauer P, and Kondorosi A. (1998) Rapid and efficient transformation of diploid Medicago truncatula and Medicago sativa ssp. falcata lines improved in somatic embryogenesis. Plant Cell Reports 17, 345–355.

    Article  CAS  Google Scholar 

  • Van Spronsen PC, Grønlund M, Pacios Bras C, Spaink HP, and Kijne JW. (2001) Cell biological changes of outer cortical root cells in early determinate nodulation. Molecular Plant-Microbe Interactions 14, 839–847.

    PubMed  Google Scholar 

  • Weijers D, Franke-van Dijk M, Vencken R-J, Quint A, Hooykaas, P, and Offringa R. (2001) An Arabidopsis Minute-like phenotype caused by a semi-dominant mutation in a ribosomal protein S5 gene. Development 128, 4289–4299.

    CAS  PubMed  Google Scholar 

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Correspondence to Herman P Spaink .

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Díaz, C.L., Schlaman, H.R.M., Spaink, H.P. (2005). Concurrent visualization of gusA and lacZ reporter gene expression. In: Márquez, A.J. (eds) Lotus japonicus Handbook. Springer, Dordrecht. https://doi.org/10.1007/1-4020-3735-X_7

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