Fusions between green fluorescent protein and beta-glucuronidase as sensitive and vital bifunctional reporters in plants

• Published in: Plant molecular biology 1998-11, Vol.38 (5), p.861-873
• Main Authors: Quaedvlieg, N.E.M. (Leiden Univ. (Netherlands). Inst. of Molecular Plant Sciences), Schlaman, H.R.M, Admiraal, P.C, Wijting, S.E, Stougaard, J, Spaink, H.P
• Format: Article
• Language: English
• Subjects: Arabidopsis - chemistry; Arabidopsis - genetics; ARABIDOPSIS THALIANA; Cloning, Molecular; EXPRESION GENICA; EXPRESSION DES GENES; Fluorescence; FORMATION DE NODOSITES; GENE EXPRESSION; Gene Expression Regulation, Plant; Genes, Reporter - genetics; Genes, Reporter - physiology; GENETIC TRANSFORMATION; Glucuronidase - genetics; Green Fluorescent Proteins; Immunoblotting; LOTUS; Lotus japonicus; Luminescent Proteins - genetics; MICROSCOPIA; MICROSCOPIE; MICROSCOPY; Microscopy, Confocal; Nicotiana tabacum; NODULACION; Plants - genetics; Plants, Genetically Modified; Plasmids - genetics; Potato virus X; Recombinant Fusion Proteins - analysis; Recombinant Fusion Proteins - genetics; ROOT NODULATION; Sensitivity and Specificity; TRANSFORMACION GENETICA; TRANSFORMATION GENETIQUE; Transformation, Genetic
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1023/A:1006182623165

By fusing the genes encoding green fluorescent protein (GFP) and beta-glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its high sensitivity in histochemical staining, with the advantages of GFP as a vital marker. The fusion proteins were functional in transient expression studies in tobacco using either DNA bombardment or potato virus X as a vector, and in stably transformed Arabidopsis thaliana and Lotus japonicus plants. The results show that high level of expression does not interfere with efficient stable transformation in A. thaliana and L. japonicus. Using confocal laser scanning microscopy we show that the fusion constructs are very suitable for promoter expression studies in all organs of living plants, including root nodules. The use of these reporter constructs in the model legume L. japonicus offers exciting new possibilities for the study of the root nodulation process.