Bacterias Fitopatógenas

1.- INTRODUCCIÓN

- Procariotas que causan enfermedad

- Interacciones planta bacteria. Tipos

2.- Enzimas, Hormonas, LPS y EPS, toxinas.

3.- Los efectores y la manipulación del huésped

 Effector: pathogen delivered molecules into host cells to
suppress immune responses and promote pathogen
proliferation.

Effector molecules are important for pathogen virulence and

promote penetration into host tissues, persistence inside the host,
suppression of immune responses, access to nutrients,
proliferation, and growth. Pathogens secrete effectors into
intercellular spaces (apoplast) and into host cells. Bacteria are
known to deliver effectors via their type-2- or type-3-secretion
systems (T3SS); fungi and oomycetes release effectors via
exocytosis. Also, effectors can be subject to horizontal gene
transfer and adaptive evolution. (Göhre and Robatzek 2008)
 In plant-bacterial interactions –
At least 2 ways to trigger the innate immunity system
Flagella
TTSS effector

TTSS

FLS2

effector-triggered PAMP-triggered
Innate immunity R Innate immunity
protein
Avr proteins are Pathogen-associated molecular
unique to pathogens patterns are conserved
generally delivered molecules in nonpathogenic
by type III systems and pathogenic bacteria

HR and other
Plant Defenses Much overlap between these Plant Defenses
pathways
 Concepts in plant immunity

Comparing and contrasting

PAMP/MAMP Effector

PRR R Protein

PTI ETI

Non-host resistance Basal resistance

Preinvasive immunity postinvasive immunity

Methods for identifying effectors suppressing plant defense responses

•Supresion of gene expression induced by PAMP:

NHO1, FRK1/SIRK and RAP2.6…..
Mesophyll protoplast system carrying a promoter-luciferase fusion
promoter luciferase fusion in planta

•Monitoring immune responses:

ROS production
Callose deposition.
Vascular flow in minor veins

•Monitoring bacterial growth rates + ectopic espression of effectors.

•Delivery of effectors via model pathogens.

•Suppression of PCD (HR). Overlapping PTI/ETI.

 Genetic evidence that specific TTSS effectors acted
as HR suppressors

?
R
Virulent pathogen Susceptible plant
effector
TTSS ?
Disease

R
Avirulent pathogen effector Resistant plant
TTSS

HR

Bacterial cell Plant cell

Experiments did not determine
whether the “masking” occurred
Jackson et al. 1999. PNAS 96:10875 in the bacterial or plant cell???
 Where are Avr/effector proteins
located in the cell?
• P. syringae effectors:
– Several contain potential amino-terminal
myristoylation and palmitoylation sites
(normally in eukaryotes)
– These groups ‘tether’ the fatty-acid-modified
proteins to membranes

• Are the myristoylation sites required for

location and function of effector proteins?
Putative myristolyation sites

White et al. 2000. Curr. Opin. Plant Biol. 3: 291.

Robert H. Dowen et al. 2009 JBC VOL. 284, NO. 23, pp. 15867–15879
AvrRpm1 and AvrB green fluorescence fusion proteins localize to
the protoplast plasma membrane

•G2A: site directed mutation changing WT G2A

G at the 2 position of the myristoylation
site to an A in each avr gene
AvrRpm1
•Stacked laser confocal micrographs
demonstrate wt Avr proteins enriched
in the PM, whereas mutant localized AvrB
like non-targeted GFP control

GFP

Nimchuk et al. 2000. Cell 101: 353

 AvrPto also localizes to the plant plasma membrane

• A G2A mutation in
AvrPto disrupts
membrane
localization (A) and
the avirulence
function of avrPto on
tomato (B) and
tobacco (C )

Shan et al. 2000. Plant Cell 12: 2323.

 AvrBs3 family members localize to
the plant nucleus
• Contain three NLS sequences
• NLS sequences required for avirulence
function of avrBs3, avrXa10, avrXa7, etc.

Van den Ackerveken et al., 1996

Other effectors ????:

Coronatine: JA mimetic. COR inhibits MAMP and bacterium-triggered stomatal

closure, which is dependent on SA signaling [14,19], and COR and JA suppress
MAMP-induced callose deposition in leaf mesophyll cells and root cells [26,27] in
Arabidopsis. Taken together, these results collectively indicate that a major function
of COR is to suppress interconnected MAMP- and SA-activated defense responses
during bacterial infection.

Melotto M, Underwood W, Koczan J, Nomura K, He SY (2006) Plant stomata function in innate immunity
against bacterial invasion. Cell 126: 969–980.
Zeng W, He SY (2010) A prominent role of the flagellin receptor FLAGELLINSENSING2 in mediating stomatal
response to Pseudomonas syringae pv tomato DC3000 in Arabidopsis. Plant Physiol 153: 1188–1198.

EPS: Ca2+ chelator. Inhibit Ca2+ entry and downstream responses such as the
oxidative burst induced by MAMPs.

Aslam SN, Newman M-A, Erbs G, Morrissey KL, Chinchilla D, et al. 2008. Bacterial polysaccharides suppress
induced innate immunity by calcium chelation. Curr. Biol. 18:1078–83
 Interference of type III effector proteins with plant cellular pathways.

Daniela Büttner FEMS Microbiol Rev 2016;femsre.fuw026

© FEMS 2016.
Cytoplasmic effectors
In Pseudomonas syringae:
 The Zigzag model showing evolution of the plant immune system

Jones and Dangl. 2006. Nature 444: 323.

More in the Resistance Mechanisms Classes
Minimal functional set of
eight effectors (AvrPtoB,
HopE1, HopG1, HopAM1,
AvrE, HopM1, HopAA1 and
HopN1), which enhanced
the growth of C3000D28E
in N.benthamiana by more
than 3 logs to near wild type
levels and enabled
production of
necrotic/chlorotic disease
lesions.
 Erwinia chrysanthemi hrp genes and their involvement in soft rot pathogenesis and elicitation of the
hypersensitive response.
Bauer DW, Bogdanove AJ, Beer SV, Collmer A.
Mol Plant Microbe Interact. 1994 Sep-Oct;7(5):573-81.

hrp genes of Erwinia chrysanthemi 3937 are important virulence factors.

Yang CH, Gavilanes-Ruiz M, Okinaka Y, Vedel R, Berthuy I, Boccara M, Chen JW, Perna NT, Keen NT.
Mol Plant Microbe Interact. 2002 May;15(5):472-80.

Use of a pooled transposon mutation grid to demonstrate roles in disease development for Erwinia
carotovora subsp. atroseptica putative type III secreted effector (DspE/A) and helper (HrpN) proteins.
Holeva MC, Bell KS, Hyman LJ, Avrova AO, Whisson SC, Birch PR, Toth IK.
Mol Plant Microbe Interact. 2004 Sep;17(9):943-50.

Pectobacterium carotovorum elicits plant cell death with DspE/F but the P. carotovorum
DspE does not suppress callose or induce expression of plant genes early in plant-microbe
interactions.
Kim HS, Thammarat P, Lommel SA, Hogan CS, Charkowski AO.
Mol Plant Microbe Interact. 2011 Jul;24(7):773-86.