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Rhizobium leguminosarum bv. trifolii PssP Protein Is Required for Exopolysaccharide Biosynthesis and Polymerization

April 2002 , Volume 15 , Number  4
Pages  388 - 397

Andrzej Mazur , Jarosław E. Król , Jerzy Wielbo , Teresa Urbanik-Sypniewska , and Anna Skorupska

Department of General Microbiology, M. Curie-Skłodowska University, Akademicka 19, 20-033 Lublin, Poland

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Accepted 8 December 2001.

Rhizobium leguminosarum bv. trifolii produces an acidic exopolysaccharide (EPS) that is important for the induction of nitrogen-fixing nodules on clover. Recently, three genes, pssN, pssO, and pssP, possibly involved in EPS biosynthesis and polymerization were identified. The predicted protein product of the pssP gene shows a significant sequence similarity to other proteins belonging to the PCP2a family that are involved in the synthesis of high-molecular-weight EPS. An R. leguminosarum bv. trifolii TA1 mutant with the entire coding region of pssP deleted did not produce the EPS. A pssP mutant with the 5′ end of the gene disrupted produced exclusively low-molecular-weight EPS. A mutant that synthesized a functional N-terminal periplasmic domain but lacked the C-terminal part of PssP produced significantly reduced amounts of EPS with a slightly changed low to high molecular form ratio. Mutants affected in the PssP protein carrying a stable plasmid with a constitutively expressed gusA gene induced nodules on red clover that were not fully occupied by bacteria. A mutant with the entire pssP gene deleted infected only a few plant cells in the nodule. The pssP promoter-gusA reporter fusion was active in bacteroids during nodule development.

Additional keywords: exopolysaccharide-deficient mutant , nitrogen fixation .

© 2002 The American Phytopathological Society