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J. Bacteriol. doi:10.1128/JB.01160-06
Copyright (c) 2006, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

The algT gene of Pseudomonas syringae pv. glycinea and new insights into the transcriptional organization of the algT-muc gene cluster

Alexander Schenk, Michael Berger, Lisa M. Keith, Carol L. Bender, Georgi Muskhelishvili, and Matthias S. Ullrich*

International University Bremen, School of Engineering and Sciences, Campus Ring 1, D-28759 Bremen, Germany. Department of Entomology and Plant Pathology, Oklahoma State University, 127 Noble Research Center, Stillwater, OK 74078, USA

* To whom correspondence should be addressed. Email: m.ullrich{at}iu-bremen.de.


   Abstract

The phytopathogenic bacterium, Pseudomonas syringae pv. glycinea, infects soybean plants and causes bacterial blight. In addition to P. syringae, the human pathogen Pseudomonas aeruginosa and the soil bacterium Azotobacter vinelandii produce the exopolysaccharide alginate, a copolymer of D-mannuronic and L-guluronic acids. Alginate production in P. syringae has been associated with increased fitness and virulence in planta. Alginate biosynthesis is tightly controlled by proteins encoded by the algT-muc regulatory gene cluster in P. aeruginosa and A. vinelandii. These genes encode the alternative sigma factor AlgT ({sigma}22), its anti-sigma factors MucA and MucB, MucC, a protein with a controversial function that is absent in P. syringae, and MucD, a periplasmic serine protease and homolog of HtrA in Escherichia coli. We compared an alginate-deficient algT mutant of P. syringae pv. glycinea with an alginate-producing derivative in which algT is intact. The alginate-producing derivative grew significantly slower in vitro growth but showed increased epiphytic fitness and better symptom development in planta. Evaluation of expression levels for algT, mucA, mucB, mucD and algD, which encodes an alginate biosynthesis gene, showed that mucD transcription is not dependent on AlgT in P. syringae in vitro. Promoter mapping using primer extension experiments confirmed this finding. Results of RT-PCR demonstrated that algT, mucA and mucB are co-transcribed as an operon in P. syringae. Northern blot analysis revealed that mucD was expressed as a 1.75 kb monocistronic mRNA in P. syringae.




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