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Construction of green fluorescent protein (GFP)-marked strains of Bradyrhizobium for ecological studies.

Abstract：

AIM:To introduce the gfp gene encoding green fluorescent protein (GFP) into bradyrhizobia for their identification in nodules, soil and carrier-based inoculants. METHODS AND RESULTS:Bradyrhizobium sp. strains M29 and GN7, which nodulate mungbean (Vigna radiata), were conjugated with Escherichia coli S17-1 carrying plasmid EDS 15 (a suicide plasmid carrying a promoterless gfp gene fused with Tn5). The GFP-marked strain expressed the gfp gene from a Bradyrhizobium promoter and gave green fluorescence when observed under an epifluorescent microscope or u.v. transilluminater. All the GFP-marked strains were able to nodulate mungbean and fix nitrogen. The GFP-marked bradyrhizobia were recovered at a frequency of 90-100% and 16-63% from nodules formed under sterilized and unsterilized conditions, respectively. The GFP-marked bradyrhizobia were identified from soil and from charcoal-based inoculants on the basis of green fluorescence. CONCLUSIONS:The GFP-marked Bradyrhizobium was successfully identified on the basis of green fluorescence to study its competition and survival in the soil and in charcoal-based inoculants. SIGNIFICANCE AND IMPACT OF THE STUDY:Introduction of the gfp gene into Bradyrhizobium provides a simple, specific and cost-effective method of strain identification for ecological studies.

journal_name

J Appl Microbiol

journal_title

Journal of applied microbiology

authors

Bhatia R,Dogra RC,Sharma PK

doi

10.1046/j.1365-2672.2002.01768.x

subject

Has Abstract

pub_date

2002-01-01 00:00:00

pages

835-9

issue

5

eissn

1364-5072

issn

1365-2672

pii

1768

journal_volume

93

pub_type

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