Quorum Sensing Regulation

And Its Role In The Legume-

Rhizobia Symbiosis

PhD Seminar
By Abraham Mikru Teklemichael

Instructor: Fasil Assefa (PhD)

Dpt. Biology, AAU 2010

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 Contents
• Introduction
• General principles of quorum sensing
• The Lux paradigm
• QS in some rhizobia
• Response of host plant
• Concluding remarks

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 Introduction
• Microbes are not solitary
• Microbes interact with each other and
their environment
• Microbial interaction is widespread &
vast
• The focus here is on quorum sensing (QS)
via acylhomoserine lactones (AHLs)

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 What is QS?
• Cell Density-dependent regulation of gene
expression mediated by diffusible signal
molecules

• “Molecular dialogue”
• Bacterial ‘talk’ to each other & ‘ccrosstalk’
• Bacterial chemical communication or
signaling

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 Why do bacteria communicate?

• Regulate gene expression concertedly

• Orchestrate activities not achievable by a
single cell
• Coordinate population-wide behavioral
response
• Behave as a multicellular organism
• Rapid adaptation and survival

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 Behaviors controlled by QS
Among others:
• Bioluminescence
• Virulence
• Symbiosis
• Biofilm
• Horizontal gene transfer

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 QS molecules (autoinducer)
• N-Acyl homoserine lactones (AHLs) or AI-1
• In Gram negatives as well as rhizobia
• Exclusively intraspecies communication

Hydrophilic head
Hydrophobic tail
Acyl side chain Homoserine lactone
ring

Variability
•n= C4 -C18
•R= H, O, OH
•Unsaturation
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 Synthesis and accumulation

SAM= S-adenosyl methionine

ACP= acyl-carrier protein
2. Movement across membrane 3. Accumulation depends on:

•Diffusion •Cell density

•Active/carrier mediated •Biofilm
•Microcolony
•Microenvironment
•Degraders

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 QS regulator
(LuxR and homologues)
• AHL receptor and transcriptional regulator
• Two regions of sequence conservation

 Amino terminus binds to cognate AHL

 Carboxy terminus binds to target DNA
(Lux box)

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 AHL degraders
• Unstable under alkaline pH
• AHL degrading enzymes
• Produced by several rhizobacteria
• Potentially interfere with QS

 Lactonase (1) and

 Acylase (3)

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 When does QS occur?

At threshold [AHL] or when population is quorate

growth
Quorum
size

Quorum sensing-regulated gene expression

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time 11
How many cells in a Quorate population?
Depends…
Critical [AHL]
Specific Genes
ON

[AHLs]

Increasing Cell Density

Low cell density High cell density

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 General principle of QS and the
Lux paradigm
• Cells make a signalling molecule (AHL) constitutively
• The AHL diffuses freely out of the cell into the
surrounding medium
• As the population of the cells grow, so does the
concentration of AHL in the growth medium
• When the population (and hence AHL) concentration
reaches a threshold, the AHL moves back down its
concentration gradient into the cell.

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 Cont.
• The concentration of AHL inside the cell
now increases and AHL can bind to luxR –
the transcriptional activator protein
• The luxR-AHL conjugate activates gene
expression
• This system is often referred to as
“autoinduction” or
• Positive feedback loop control

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 luxI codes for AHL
Synthase

luxR codes for

Transcriptional
regulator

QS in Photobacterium fischeri (V.fischeri)

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Bioluminescence at high cell density
in culture and light organ

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 QS in Rhizobiacae
• QS now considered one of the requisite
symbiotic signals
• Reported for many rhizobia

• In all cases AHLs are the signaling

molecules
• The genI/genR (luxI/luxR homologues)
system still remains
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 QS in R. leguminosarum
Four QS systems known

• cinI/cinR
• raiI/raiR
• rhiI/rhiR
• traI/traR

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 cinI/cinR
• Located on chromosome
QS molecule is:
• 3-OH-C14:1-HSL
Functions
• Overall control of other QS systems
• Stationary phase adaptation and survival
• Growth inhibition (bacteriocin–like action)

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 raiI/raiR
• Located on non-symbiotic plasmid
QS molecules:
• C8-HSL, 3-OH-C8-HSL
• Function: unknown

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 rhiI/rhiR
• Located on symbiotic plasmid
• Close to nod box
QS molecules
• C6-HSL, C7-HSL and C8-HSL
Target genes
• rhiI, rhiABC
Function
• Influence nodulation

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 traI/traR
• Located on symbiotic plasmid
QS molecules
• 3-oxo-C8-HSL
• 3-OH-C14:1-HSL from recipient

Other regulatory genes

• bisR, traM

Target genes and functions

• Induce plasmid transfer operon

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QS network in R. leguminosarum

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 traI/traR

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 The R. etli Qs systems
• Possess orthologues of the cinI/cinR,
raiI/raiR and traI/traR genes
• Target genes and functions are different
• cinI/cinR: growth inhibition, symbiosome
development, nitrogen fixation and
swarming motility
• raiI/raiR : Mutants show higher nodulation
• traI/traR: conjugal plasmid (p42a)transfer
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Interaction of two QS in R. etli CNPAF 512

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 S. meliloti QS systems
• sinI/sinR: EPS production, swarming motility and
nodulation efficiency
• Involves several C12 to C18 AHLs
• Some symbiotic genes dependent on sinR but not on sinI.
• What could be the signal molecules other than SinI- made
AHLS controls these genes?

• expR: role in EPSII synthesis

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S. meliloti (Rm1021 and Rm41)

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 Mesorhizobium spp
Reported for several species

• M. huakuii: QS involved in biofilm formation

• M. loti: three luxI-like genes influence
nodulation
• M. tianshanense: QS involved in nodulation

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 Rhizobium sp.NGR234
• traI/traR genes on symbiotic plasmid
pNGR234a
• QS molecule is 3-oxo-C8-HSL
Functions
• Conjugal plasmid transfer
• Growth inhibition

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 Bradyrhizobium japonicum
• QS inhibit nod gene expression
• QS molecule is bradyoxetin not an AHL
• Exhibit siderophor-like property
• Maximal in iron depleted condition
• AHLs detected in B. japonicum and B. elkani
but the target genes not known

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Bradyrhizobium japonicum

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 Responses of host plants to QS
Plants ‘listen’ & respond to bacterial AHL QS
• Medicago truncatula root respond to two
different AHLs by up regulating 154 proteins
• The proteins function in defense,
transcriptional regulation, stress and plant
hormone response
• Host plants also produce AHL-mimics
potentially interfering with bacterial QS

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 Concluding remarks
AHL QS plays several role in rhizobia-legume
symbiosis
• EPS production,symbiosome development,
Nodulation efficiency,Nitrogen fixation
• Plasmid transfer
• Stationary phase adaptation and Growth inhibition
Host plants also “listen” and respond to bacterial AHL-
QS

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Thank You!

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Variations in structure of AHLs

A. tumefaciens, Rhizobium sp. NGR234, R. leguminosarum,

B. S. meliloti, R. etli CFN42

R. leguminosarum, S. meliloti, R. etli CFN42

R. leguminosarum, S. meliloti

R. leguminosarum

S. meliloti

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