Abstract
An ATP-binding cassette (ABC) transporter, called the PseEF
efflux system, was identified at the left border of the syr-syp
genomic island of Pseudomonas syringae pv. syringae strain
B301D. The PseEF efflux system was located within a 3.3-kb
operon that encodes a periplasmic membrane fusion protein
(PseE), and an ABC-type cytoplasmic membrane protein
(PseF). The PseEF efflux system exhibited amino acid homology
to a putative ABC efflux system (MacAB) of E. coli
W3104 with identities of 47.2% (i.e., PseE to MacA) and
57.6% (i.e., PseF to MacB). A nonpolar mutation within the
pseF gene was generated by nptII insertional mutagenesis.
The resultant mutant strain showed significant reduction in
secretion of syringomycin (74%) and syringopeptin (71%),
as compared to parental strain B301D. Quantitative real-time
RT-PCR was used to determine transcript levels of the syringomycin
(syrB1) and syringopeptin (sypA) synthetase
genes in strain B301D-HK7 (a pseF mutant). Expression of
the sypA gene by mutant strain B301D-HK7 was approximately
6.9% as compared to that of parental strain B301D,
while the syrB1 gene expression by mutant strain B301D-HK7
was nearly 14.6%. In addition, mutant strain B301D-HK7
was less virulent by approximately 67% than parental strain
B301D in immature cherry fruits. Mutant strain B301D-HK7
was not reduced in resistance to any antibiotics used in this
study as compared to parental strain B301D. Expression
(transcript levels) of the pseF gene was induced approximately
six times by strain B301D grown on syringomycin
minimum medium (SRM) supplemented with the plant signal
molecules arbutin and D-fructose (SRMAF), as compared to
that of strain B301D grown on SRM (in the absence of plant
signal molecules). In addition, during infection of bean
plants by P. syringae pv. syringae strain B728a, expression
of the pseF gene increased at 3 days after inoculation (dai).
More than 180-fold induction was observed in transcript
levels of the pseF gene by parental strain B728a as compared
to strain B728a-SL7 (a salA mutant). Thus, the PseEF
efflux system, an ABC-type efflux system, has an important
role in secretion of syringomycin and syringopeptin, and is
required for full virulence in P. syringae pv. syringae.
Citations
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