Bacillus brevis DX01 Labeling by Green Fluorescent Protein Gene and Its Colonization in Rice Seedling Roots

Since the gfp cDNA was cloned from Aequoreavictoria1], the green fluorescence protein (GFP)has attracted considerable attention as a marker/reporter system2, 3]. In recent years, the greenfluorescent protein gene has been used as a markersystem for detection of marked cells in soil.Tombolini et al.4]used a red-shift mutant to de-tect individual bacterial cells on the surface ofroots. They also investigated the stability of theprotein under nutrient starvation conditions. Theprotein was s…

Bacillus brevis DX01 Labeling by Green Fluorescent Protein Gene and Its Colonization in Rice Seedling Roots

A constitutive expression vector pHY300-F1gfp was constructed to test the function of a promoter F1 subcloned from a rice epiphyte Bacillus brevis strain DX01. The DX01 cells harboring plasmid pHY300-F1gfp were detected to produce bright green fluorescence. Subsequently, the gfp-tagged B. brevis strain was released into the soil and its survival was investigated by PCR and the detection of green fluorescence. The spatial location of in situ gfp-tagged bacterial cells on the root surface of rice seedlings was visualized. All these results indicated that green fluorescent protein is an ideal molecular marker for detection of the activities of promoter F1, and it is also a reliable probe to monitor specific B. brevis bacteria in the environment.