Volume No. 8 Issue No.: 2 Page No.: 254-260 Oct - Dec 2013

 

TRANSFORMED FLUORESCENT Rhizobium AS REPORTER BIOINOCULANT IN SOIL ENVIRONMENT

 

Tile Vaishali*, Bholay A.D. and Wagh Pratima

Department of Microbiology, KTHM College Nashik, University of Pune (INDIA)

 

Received on : June 05, 2013

 

ABSTRACT

 

Rhizobium spp isolated from root nodules of Cicer arietinum plant was transformed using GFP (Green Fluorescent Protein) isolated from the aquatic jelly fish, fluoresces green in the UV range. The aim of this work is to simplify the studies based on Rhizobium spps. GFP gene obtained from plasmid of GFP cloned E. coli DH5αused as reporter gene to transform isolated Rhizobium spp to study its life cycle, mode of nutrition and survival rate without the use of the conventional radioactive reporters. The transformed strain was ampicillin resistant, the character which is by virtue of the plasmid pUC18 that it possessed. This eliminated the competitors when that antibiotic is applied at a certain level. Survival studies of the transformed Rhizobium spp in the soil environments showed that the organism survived well at 4°C as well as at 25°C, but not at 40°C. The root and shoot lengths of plants which were inoculated with transformed Rhizobium spp were more than that of uninoculated control plants. Nodule formation was observed only in the plants inoculated with transformed Rhizobium spp with mean number of nodules being 13.8. The organism reisolated from the nodules of the test plant showed characteristics similar to that isolated from Cicer arietinum plant from the field. The transformed Rhizobium spp was effective in nodule formation in Cicer arietinum plant and thus can act as a bioinoculant reporter in soil environment.

 

Keywords : Rhizobium, Cicer arietinum, GFP, Plasmid pUC18, E. coli DH5α

 

 

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