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Appl Environ Microbiol ; 73(16): 5075-82, 2007 Aug.
Article in English | MEDLINE | ID: mdl-17557843

ABSTRACT

Application of glyphosate (N-[phosphonomethyl] glycine) to Bradyrhizobium sp. (Lupinus)-nodulated lupin plants caused modifications in the protein pattern of bacteroids. The most significant change was the presence of a 44-kDa polypeptide in bacteroids from plants treated with the higher doses of glyphosate employed (5 and 10 mM). The polypeptide has been characterized by the amino acid sequencing of its N terminus and the isolation and nucleic acid sequencing of its encoding gene. It is putatively encoded by a single gene, and the protein has been identified as a putative porin. Protein modeling revealed the existence of several domains sharing similarity to different porins, such as a transmembrane beta-barrel. The protein has been designated BLpp, for Bradyrhizobium sp. (Lupinus) putative porin, and would be the first porin described in Bradyrhizobium sp. (Lupinus). In addition, a putative conserved domain of porins has been identified which consists of 87 amino acids, located in the BLpp sequence 30 amino acids downstream of the N-terminal region. In bacteroids, mRNA of the BLpp gene shows a basal constitutive expression that increases under glyphosate treatment, and the expression of the gene is seemingly regulated at the transcriptional level. By contrast, in free-living bacteria glyphosate treatment leads to an inhibition of BLpp mRNA accumulation, indicating a different effect of glyphosate on BLpp gene expression in bacteroids and free-living bacteria. The possible role of BLpp in a metabolite interchange between Bradyrhizobium and lupin is discussed.


Subject(s)
Bacterial Proteins/metabolism , Bradyrhizobium/drug effects , Glycine/analogs & derivatives , Porins/metabolism , Amino Acid Sequence , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Blotting, Northern , Blotting, Southern , Bradyrhizobium/genetics , Bradyrhizobium/metabolism , Electrophoresis, Polyacrylamide Gel , Fabaceae/microbiology , Gene Expression Regulation, Bacterial/drug effects , Glycine/pharmacology , Models, Molecular , Molecular Sequence Data , Molecular Weight , Phylogeny , Porins/chemistry , Porins/genetics , Protein Structure, Secondary , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Analysis, Protein , Glyphosate
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