Transferring of Exopolysaccharide (EPS) Gene Cluster from Rhizobium leguminosarum to Pseudomonas fluorescens by Horizontal Gene Transfer Techniques

Abstract

To increase drought resistance of efficient plant growth promoting Psuedomonas  fluorescens isolated from Erbil governorate soils, exopolysaccharide (EPS) gene cluster was transferred from Rhizobium leguminosarum to P. fluorescens. EPS gene cluster is responsible for generating of capsular polysaccharides which are tightly attached to the cell surface and extracellular polysaccharides that are directly secreted to the environment. For P. fluorescens and R. leguminosarum isolations, different soil samples were collected in Erbil governorate.  According to morphological, biochemical and molecular examinations 75 P. fluorescens isolates and 58 R. leguminosarum isolates were identified from 110 soil samples. Most P. fluorescens isolates showed efficiency in phosphate solubilization, hydrogen cyanide, siderophores and indol acidic acid production, the most efficient P. fluorescens (Mpf16) was selected for conjugation process.  Potential transferring of EPS genes, included EPS1, EPS2, and EPS3, was investigated in this study. Gene cluster was transferred successfully from R. leguminosarum into P. fluorescens by horizontal conjugation gene transfer technique after confirmation via molecular techniques using different primers and enzymes. The expression of transferred EPS gene cluster from transconjugant P. fluorescens was confirmed by testing capsule formations from viable cells. The new generated capsule former P. fluorescens can be used later on as an efficient bio-fertilizer agent for enhancing plant growth and crop production in drought cultivated soils.

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