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  • 1
    Online Resource
    Online Resource
    Scientific Societies ; 2005
    In:  Molecular Plant-Microbe Interactions® Vol. 18, No. 5 ( 2005-05), p. 499-507
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 18, No. 5 ( 2005-05), p. 499-507
    Abstract: Rhizobium sp. strain NGR234, which is capable of interacting with a large number of legumes, utilizes a variety of signaling molecules to establish nitrogen-fixing symbioses. Among these are nodulation outer proteins (Nops) that transit through a type III secretion system (TTSS). Abolition of Nop secretion affects nodulation of certain legumes. Under free-living conditions, the secretion of Nops can be induced by the addition of flavonoids. Here, we show that an in-frame deletion of nopA abolishes secretion of all other Nops and has the same impact on nodule formation as mutations that lead to a nonfunctional TTSS. This secretionminus phenotype of the nopA mutant, as well as bioinformatics analysis of NopA itself, suggests that NopA could be an external component of the TTSS. Electron microscopy showed that NGR234 synthesizes fibrillar structures on the cell surface in a flavonoid-inducible and NopA-dependent manner. Purification of the macromolecular surface appendages revealed that NopA is a major component of these structures.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2005
    detail.hit.zdb_id: 2037108-1
    SSG: 12
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  • 2
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 16, No. 9 ( 2003-09), p. 743-751
    Abstract: The nitrogen-fixing symbiotic bacterium Rhizobium species NGR234 secretes, via a type III secretion system (TTSS), proteins called Nops (nodulation outer proteins). Abolition of TTSS-dependent protein secretion has either no effect or leads to a change in the number of nodules on selected plants. More dramatically, Nops impair nodule development on Crotalaria juncea roots, resulting in the formation of nonfixing pseudonodules. A double mutation of nopX and nopL, which code for two previously identified secreted proteins, leads to a phenotype on Pachyrhizus tuberosus differing from that of a mutant in which the TTSS is not functional. Use of antibodies and a modification of the purification protocol revealed that NGR234 secretes additional proteins in a TTSS-dependent manner. One of them was identified as NopA, a small 7-kDa protein. Single mutations in nopX and nopL were also generated to assess the involvement of each Nop in protein secretion and nodule formation. Mutation of nopX had little effect on NopL and NopA secretion but greatly affected the interaction of NGR234 with many plant hosts tested. NopL was not necessary for the secretion of any Nops but was required for efficient nodulation of some plant species. NopL may thus act as an effector protein whose recognition is dependent upon the hosts' genetic background.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2003
    detail.hit.zdb_id: 2037108-1
    SSG: 12
    Location Call Number Limitation Availability
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  • 3
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 24, No. 12 ( 2011-12), p. 1513-1521
    Abstract: In the presence of flavonoids, Rhizobium sp. strain NGR234 synthesizes a new lipopolysaccharide (LPS), characterized by a rhamnan O-antigen. The presence of this rhamnose-rich LPS is important for the establishment of competent symbiotic interactions between NGR234 and many species of leguminous plants. Two putative rhamnosyl transferases are encoded in a cluster of genes previously shown to be necessary for the synthesis of the rhamnose-rich LPS. These two genes, wbgA and rgpF, were mutated. The resulting mutant strains synthesized truncated rough LPS species rather than the wild-type rhamnose-rich LPS when grown with flavonoids. Based on the compositions of these purified mutant LPS species, we inferred that RgpF is responsible for adding the first one to three rhamnose residues to the flavonoid-induced LPS, whereas WbgA is necessary for the synthesis of the rest of the rhamnan O-antigen. The NGR234 homologue of lpsB, which, in other bacteria, encodes a glycosyl transferase acting early in synthesis of the core portion of LPS, was identified and also mutated. LpsB was required for all the LPS species produced by NGR234, in the presence or absence of flavonoids. Mutants (i.e., of lpsB and rgpF) that lacked any portion of the rhamnan O-antigen of the induced LPS were severely affected in their symbiotic interaction with Vigna unguiculata, whereas the NGRΩwbgA mutant, although having very few rhamnose residues in its LPS, was able to elicit functional nodules.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2011
    detail.hit.zdb_id: 2037108-1
    SSG: 12
    Location Call Number Limitation Availability
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  • 4
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 17, No. 9 ( 2004-09), p. 958-966
    Abstract: Formation of nitrogen-fixing nodules on legume roots by Rhizobium sp. NGR234 requires an array of bacterial factors, including nodulation outer proteins (Nops) secreted through a type III secretion system (TTSS). Secretion of Nops is abolished upon inactivation of ttsI (formerly y4xI), a protein with characteristics of two-component response regulators that was predicted to activate transcription of TTSS-related genes. During the symbiotic interaction, the phenotype of NGRΩttsI differs from that of a mutant with a nonfunctional secretion machine, however. This indicated that TtsI regulates the synthesis of other symbiotic factors as well. Conserved sequences, called tts boxes, proposed to act as binding sites for TtsI, were identified not only within the TTSS cluster but also in the promoter regions of i) genes predicted to encode homologs of virulence factors secreted by pathogenic bacteria, ii) loci involved in the synthesis of a rhamnose-rich component (rhamnan) of the lipopolysaccharides (LPS), and iii) open reading frames that play roles in plasmid partitioning. Transcription studies showed that TtsI and tts boxes are required for the activation of TTSS-related genes and those involved in rhamnose synthesis. Furthermore, extraction of polysaccharides revealed that inactivation of ttsI abolishes the synthesis of the rhamnan component of the LPS. The phenotypes of mutants impaired in TTSS-dependent protein secretion, rhamnan synthesis, or in both functions were compared to assess the roles of some of the TtsI-controlled factors during symbiosis.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2004
    detail.hit.zdb_id: 2037108-1
    SSG: 12
    Location Call Number Limitation Availability
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  • 5
    Online Resource
    Online Resource
    Scientific Societies ; 2009
    In:  Molecular Plant-Microbe Interactions® Vol. 22, No. 5 ( 2009-05), p. 519-528
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 22, No. 5 ( 2009-05), p. 519-528
    Abstract: Type III secretion systems (T3SS) have been found in several species of rhizobia. Proteins (termed effectors) secreted by this system are involved in host-range determination and influence nodulation efficiency. Mesorhizobium loti MAFF303099 possesses a functional T3SS in its symbiotic island whose expression is induced by flavonoids. As in other rhizobia, conserved cis-elements (tts box) were found in the promoter regions of genes or operons encoding T3SS components. Using a bioinformatics approach, we searched for other tts-box-controlled genes, and confirmed this transcriptional regulation for some of them using lacZ fusions to the predicted promoter regions. Translational fusions to a reporter peptide were created to demonstrate T3SS-mediated secretion of two new MAFF303099 effectors. Finally, we showed that mutation of the M. loti MAFF303099 T3SS affects its competitiveness on Lotus glaber and investigated, at the molecular level, responses of the model legume L. japonicus to the T3SS.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2009
    detail.hit.zdb_id: 2037108-1
    SSG: 12
    Location Call Number Limitation Availability
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