Characterization of flagella genes of Agrobacterium tumefaciens, and the effect of a bald strain on virulence

Olga Chesnokova, John B. Coutinho, Imran Khan, Maurice S. Mikhail, Clarence I. Kado

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Abstract

Agrobacterium tumefaciens produces flagella that are arranged circumthecally near one end of the bacilliform cell. The flagella are required for motility to facilitate reaching the root surface, and possibly aid in orientating the bacterial cells at various sites for infection, We have identified three flagella genes designated flaA, flaB and flaC. Mutations in flaA, flaB and flaC result in abberant swimming behaviour. Electron microscopic examination of these mutants revealed the defective flagella. A non-motile, bald mutant strain was generated by deleting all three fla genes. Nucleotide sequencing of flaA, flaB, and flaC showed that they have a potential coding capacity for polypeptides of 307, 321, and 314 amino acid residues, respectively. The predicted amino acid sequences of the A. tumefaciens FlaA and FlaB proteins are similar (66% average identity) to the FlaA and FlaB proteins encoded by flaA and flaB genes, respectively, in Rhizobium meliloti. There was no counterpart FlaC protein reported in R. meliloti, but the A. tumefaciens FlaC is similar in amino acid sequence to the R. meliloti FlaA (59.8% identity) and FlaB (66.7% identity). Distinct from FlaA and FlaB of R. meliloti is the absence of histidine and cysteine residues and their shorter length (by 88 amino acid residues fewer than FlaA and FlaB of R. meliloti). The transcriptional start sites of each fla gene determined by primer extension revealed consensus-sequence boxes representing potential binding sites for σ28 RNA polymerase (RNAP) upstream of the transcriptional start of each fla gene. Besides the potential σ28-binding site upstream of flaC, also present are additional putative conserved sequences, GC at -11 and GG at -21 from the transcriptional start, that resemble potential binding motifs for σ54. Because the σ54 promoter is associated with genes regulated by physiological changes in various bacteria, the flaC gene might he similarly regulated in response to A. tumefaciens responding to host plant stimuli. Virulence studies showed that the bald strain was consistently reduced in virulence below that of the parental wild-type strain by at least 38%. The difference is statistically significant and suggests that the flagella may play a role in facilitating virulence.

Original languageEnglish (US)
Pages (from-to)579-590
Number of pages12
JournalMolecular Microbiology
Volume23
Issue number3
StatePublished - 1997

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ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

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