Colonization of Green Fluorescent Protein Gene Tagged Bacillus subtilis R31 in Celery Rhizosphere

doi:10.11924/j.issn.1000-6850.2013-1713

Abstract

Abstract: In order to study the colonization of Bacillus subtilis R31 in celery rhizosphere, to assess its colonization ability and provide basis to celery wilt biocontrol. Plasmid pHT315-gfp was introduced into cells of Bacillus subtilis R31 by protoplasts conversion and gfp- tagged R31 was acquired, gfp- tagged R31 was irrigated around the celery root to study its colonization style and ability. Results showed the recombined plasmid was stable in gfp-tagged R31 cells. The R31-gfp cells could emit bright green fluorescence under blue light stably. Green fluorescent film on the surface of the celery root inoculated R31-gfp could be observed under fluorescent microscopy. It was found that R31-gfp cells could colonize in celery rhizosphere quickly. Three days after inoculation, cells on the surface of celery roots and in the soil around the roots were about 1.4×10⁴ cfu/g and 7.6×10⁴ cfu/g, respectively. The colonization was very stably. The number of cells on celery root surface and soil around the roots were still 9.1×10⁴ cfu/g and 3.17×10⁵ cfu/g at the 65 days after inoculation separately. Conclusion: Bacillus subtilis R31 can form biofilm on the surface of the celery roots, and can colonize stably for a long-term in celery rhizosphere.

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