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Research article

J. Microbiol. Biotechnol. 2010; 20(11): 1491-1499

Published online November 28, 2010 https://doi.org/10.4014/jmb.1006.06016

Copyright © The Korean Society for Microbiology and Biotechnology.

Effect of transgenic rhizobacteria over-expressing Citrobacter braakii appA on phytate–P availability to mung bean plants.

Kuldeep J. Patel 1, Saurabh Vig 1, G. Naresh Kumar 2 and G. Archana 1*

1Department of Microbiology and Biotechnology Centre, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara -390002, Gujarat, India, 2Department of Biochemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara - 390002, Gujarat, India.

Received: June 10, 2010; Accepted: August 12, 2010

Abstract

Rhizosphere microorganisms possessing phytase activity are considered important for rendering phytate-P available to plants. In the present study, Citrobacter braakii phytase gene (appA) was over-expressed in rhizobacteria possessing plant growth promoting (PGP) traits for increasing their potential as bioinoculants. AppA was cloned under the lac promoter in the broad host-range expression vector pBBR1MCS2. Transformation of the recombinant construct pCBappA resulted in high constitutive phytase activity in all of the eight rhizobacterial strains belonging to genera Pantoea, Citrobacter, Enterobacter, Pseudomonas (two strains), Rhizobium (two strains) and Ensifer that were studied. Transgenic rhizobacterial strains were found to display varying level of phytase activity, ranging from 10 folds to 538 folds higher than the corresponding control strains. Transgenic derivative of Pseudomonas fluorescens CHA0, a well-characterized plant growth promoting rhizobacterium, showed highest expression of phytase (~8 U/ mg) activity in crude extracts. Although all transformants showed high phytase activity, rhizobacteria having ability to secrete organic acid, showed significantly higher release of P from Ca-phytate in buffered minimal media. AppA over-expressing rhizobacteria showed increased P content, dry weight (shoot) or shoot/ root ratio of mung bean (Vigna radiata) plants, to different extents, when grown in semi solid agar (SSA) medium containing Na-phytate or Ca-phytate as the P sources. This is the first report of over-expression of phytase in rhizobacterial strains and its exploitation for plant growth enhancement.

Keywords: appA over-expression, phytate P-availability, Phytase, Plant growth promotion, Transgenic rhizobacteria