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Nodule-specific expression of Rhizobium meliloti symbiotic promoters P1 and P2 in chickpea-Rhizobium sp. symbiosis
Authors:S. P. S. Khanuja  A. Suman  G. Singh  V. Kumar
Affiliation:(1) Biotechnology Centre, Indian Agricultural Research Institute, 110 012 New Delhi, India;(2) Division of Microbiology, Indian Agricultural Research Institute, 110 012 New Delhi, India
Abstract:Developmentally specific expression of Rhizobium spp. genes involved in symbiotic N2 fixation is known to operate through cascade regulation of various nif and fix operons. Fusion constructs of lacZ under symbiotic promoters P1 (for nifHDK operon) and P2 (for fixABCX operon) of Rhizobium meliloti were mobilized into Rhizobium spp. (Cicer) strains Rcd301 and RCR13. The assays for beta-galactosidase activity to monitor the expression of lacZ under these promoters was performed in host backgrounds of Escherichia coli, R. meliloti, and Rhizobium spp. (Cicer). The enzyme assays indicated significant levels of expression from P1 and P2 promoters in chickpea rhizobia, specifically in symbiotic cells from nodules. However, as in R. meliloti, these promoters did not induce strong expression in free-living cells of Rhizobium spp. (Cicer). This indicates functional homology of R. meliloti promoters in rhizobium spp. (Cicer). Functional cross-reactivity of trans regulatory factors like NtrA, NtrC, and NifA between these rhizobia seems evident from the nodule-specific expression of P1 and P2 cis elements.
Keywords:Symbiotic promoters  Interspecific expression  Rhizobium meliloti  Rhizobium spp.  Cicer  lacZ fusions  Nif and Fix operons  Plasmids
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