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Alteration of the exopolysaccharide production and the transcriptional profile of free-living Frankia strain CcI3 under nitrogen-fixing conditions

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Abstract

We investigated the effect of different nitrogen (N) sources on exopolysaccharide (EPS) production and composition by Frankia strain CcI3, a N2-fixing actinomycete that forms root nodules with Casuarina species. Frankia cells grown in the absence of NH4Cl (i.e., under N2-fixing conditions) produced 1.7-fold more EPS, with lower galactose (45.1 vs. 54.7 mol%) and higher mannose (17.3 vs. 9.7 mol%) contents than those grown in the presence of NH4Cl as a combined N-source. In the absence of the combined N-source, terminally linked and branched residue contents were nearly twice as high with 32.8 vs. 15.1 mol% and 15.1 vs. 8.7 mol%, respectively, than in its presence, while the content of linearly linked residues was lower with 52.1 mol% compared to 76.2 mol%. To find out clues for the altered EPS production at the transcriptional level, we performed whole-gene expression profiling using quantitative reverse transcription PCR and microarray technology. The transcription profiles of Frankia strain CcI3 grown in the absence of NH4Cl revealed up to 2 orders of magnitude higher transcription of nitrogen fixation-related genes compared to those of CcI3 cells grown in the presence of NH4Cl. Unexpectedly, microarray data did not provide evidence for transcriptional regulation as a mechanism for differences in EPS production. These findings indicate effects of nitrogen fixation on the production and composition of EPS in Frankia strain CcI3 and suggest posttranscriptional regulation of enhanced EPS production in the absence of the combined N-source.

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Acknowledgments

We thank Dr. Raymond Jones in the Genomics Core Facility at the University at Texas-Arlington for providing the technical support for the qRT-PCR analysis and Dr. Schwartz, senior scientist from Mycroarray.com, for his assistance with the microarray data analysis. This research was supported by a Research Enhancement Program (REP) grant from the University of Texas-Arlington (WSC) and in part by Hatch grant NH530 (LST).

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Correspondence to Woo-Suk Chang.

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Hae-In Lee and Andrew J. Donati contributed equally.

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Lee, HI., Donati, A.J., Hahn, D. et al. Alteration of the exopolysaccharide production and the transcriptional profile of free-living Frankia strain CcI3 under nitrogen-fixing conditions. Appl Microbiol Biotechnol 97, 10499–10509 (2013). https://doi.org/10.1007/s00253-013-5277-z

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