Innate Immunity - Has Poplar PDF

Tansley review
Innate immunity: has poplar made its

BED?
Author for correspondence:
Hugo Germain
Tel: +1 418 648 4925
Email: hugo.germain@nrcan.gc.ca
Received: 15 June 2010
Accepted: 5 October 2010
Hugo Germain and Armand Seguin
Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, 1055 du PEPS, PO
Box 10380, Stn Sainte-Foy, Quebec, QC, G1V 4C7, Canada
New Phytologist (2011) 189: 678687
doi: 10.1111/j.1469-8137.2010.03544.x
Key words: BED-NB-LRR, poplar, qualitative
resistance, R-genes, salicylic acid.
Summary
The perennial plant model species Populus trichocarpa has received considerable
attention in the last 5 yr because of its potential use as a bioenergy crop. The
completion of its genome sequence revealed extensive homologies with the herba-
ceous annual species Arabidopsis thaliana. This review highlights the similarities
and differences at the qualitative defence response components level, notably in
putative NBS-LRR protein content and downstream defence regulators. With
almost a twofold NBS-LRR gene complement compared with A. thaliana,
P. trichocarpa also encodes some putative R-proteins with unusual architectures
and possible DNA-binding capacity. P. trichocarpa also possesses all the known
main components characteristic of TIR-NB-LRR and CC-NB-LRR signalling.
However, very little has been done with regard to the components involved in the
poplar qualitative response to pathogens. In addition, the relationship between
plant-biotroph perception signalling and the role of salicylic acid, an important
defence compound, remains uncertain. This review aims to identify the genomic
components present in poplar that could potentially participate in the qualitative
response and highlights where efforts should be devoted to obtain a better under-
standing of the poplar qualitative defence response.
Contents
Summary 678
I. Introduction 679
II. R-gene-mediated response in plants:
an introductory overview
679
III. R-protein pathways in poplar 680
IV. What is the role of BED-NB-LRR? 681
V. Downstream of R-proteins 683
VI. The role of salicylic acid in biotrophic interaction 683
VII. Concluding remarks 684
References 685
New
Phytologist
Review
678 New Phytologist (2011) 189: 678687
www.newphytologist.com
No claim to original US government works
New Phytologist 2010 New Phytologist Trust
I. Introduction
The never-ending battle between plants and their pathogens
is mediated by a two-layer immune system in which the
rst component, plasma membrane resident pathogen
recognition receptors (PRRs), perceives microbe-associated
molecular patterns (MAMPs). Upon MAMP recognition,
PRRs will trigger MAMP-triggered immunity (MTI,
formerly called basal resistance), a low-amplitude defence
response usually sufcient to thwart the pathogen attack.
However, pathogens use secreted MTI-inhibiting effectors
that may then be recognized (or their resulting effect) by
cognate resistance proteins (R-proteins), leading to a power-
ful defence response termed effector-triggered immunity
(ETI, or race-specic resistance). The ETI response is
characterized by the production of reactive oxygen species,
the accumulation of nitric oxide, the stomatal closure and
the activation of MAPK cascades, and it often culminates
in the hypersensitive response (Delledonne et al., 1998;
Bolwell, 1999; Zhang & Klessig, 2001; Jones & Dangl,
2006; Boller & Felix, 2009). Once an effector is recognized,
it reduces the tness of the carrier strain, and thus other
effectors that were not under selective pressure may arise
through population evolution as new virulence factors.
Conversely, to avoid plant population extinction, one of the
various polymorphic resistance loci must recognize these
new effectors, which will confer heritable resistance (Jones
& Dangl, 2006). This elegant model received broad support
by the community and was coined the zigzag model.
Perennial plants are different from annual herbaceous
plants at several levels and the question arises whether the
ndings obtained in Arabidopsis can translate to trees. For
example, the zigzag model entails that the plant population
can acquire genetic loci encoding R-proteins for every new
virulence factor that will arise in the pathogen population.
This model is conceivable for annual plants that have short
generation time, but is it conceptually applicable to trees
that have a life span of several decades or even centuries?
How could these long-lived plants keep up with the short-
generation, fast-evolving pathogens they face? Moreover,
for Arabidopsis and most dicotyledonous species, salicylic
acid (SA) is believed to be a very important component of
the defence response to biotrophic and systemic-acquired
resistance (SAR). It is also not clear whether the SA concen-
tration can be modulated in the various clones of hybrid
poplar, raising questions about the role of SA and SAR in
poplar, and perhaps in trees in general. SAR may seem like
a good strategy for plants of modest relative sizes but how
could this systemic process in plants operate in plants that
can be 5 m tall? In trees the poplar-rust pathogen has
recently emerged as a model pathosystem and has been
thoroughly studied at the transcriptome level thanks to the
availability of microarray chips, leading to a better under-
standing of the quantitative and qualitative defence
responses (Miranda et al., 2007; Rinaldi et al., 2007; Azaiez
et al., 2009; Duplessis et al., 2009; Hacquard et al., 2010).
This review will try to infer how poplar may respond to
pathogens based on the knowledge we have of poplar
defence and the knowledge gained from the poplar genome
analysis (Tuskan et al., 2006).
II. R-gene-mediated response in plants: an
introductory overview
Traditional plant breeders have relied on crosses between
cultivars for the introgression of R-gene into susceptible
crops. The response resulting from an incompatible inter-
action between a plant carrying a resistance protein and an
avirulence gene product carried by the pathogen was
described by Flor (1971) in his gene-for-gene model. In this
model, Flor hypothesized that the interaction between a
plant that possesses a resistance factor and a pathogen that
has a cognate avirulence (Avr) factor would result in a resist-
ance response. If the plant does not possess the R-gene or if
the pathogens do not possess the Avr factor, the infection
will prevail (Flor, 1971); this model ts with a receptor
ligand model. One important characteristic of poplar is that
different poplar species can also be crossed to generate
fertile F
1
hybrids. This feature greatly increases gene ow in
these obligate outcrossers, allowing new combinations of
R-gene alleles and enhancing the potential of breeding for
resistance (Bradshaw, 1996).
Although known Avrs are generally small molecules and
R-proteins have a LRR domain believed to be involved in
proteinprotein interactions, only a few direct R AVR
interactions have been reported (Jia et al., 2000; Deslandes
et al., 2003; Ueda et al., 2006), including Flors original
ax-rust (or Melampsora lini ax) model pathosystem
(Dodds et al., 2006). A renement to Flors hypothesis was
suggested in which the role of the NB-LRR protein would
be to guard or monitor the status of a host protein that is
the target of an AVR. This rened model was initially used
to describe the Prf Pto AvrPto interaction (Van der Biezen
& Jones, 1998) and was later coined the guard model
(Dangl & Jones, 2001). However, this model is also not
perfect and another variant has emerged: the decoy model.
In this new model, the plant protein targeted by the patho-
gen effector would have no function in host defence but
would mimic a plant defence component (van der Hoorn
& Kamoun, 2008). A good example of this would be Pto
mimicking a defence component (such as the kinase
domain of FLS2) to interact with AvrPto; and that this
interaction is monitored by Prf (a NB-LRR), which sub-
sequently triggers defence signalling (Xiang et al., 2008).
Regardless of these hypotheses, R-proteins remain at
the centre stage of how plants perceive the pathogen. Most
identied plant R-proteins belong to the large group of
NB-LRR proteins in which NB is a nucleotide-binding site
New
Phytologist Tansley review Review 679
that is required for ATP binding and hydrolysis (Tameling
et al., 2002) and LRR stands for leucine-rich repeat.
NB-LRR can be further separated into two distinct groups
based on their N-termini (Martin et al., 2003; Belkhadir
et al., 2004). Group one includes the toll-interleukin recep-
tor domain (TIR) and the second group has an N-terminal
coiled-coil domain (CC). The NB domain also contains
motifs that are specic to TNL and CNL (Meyers et al.,
2003). A total of 51 CC-NB-LRRs (CNL) and 93 TIR-
NB-LRRs (TNL) were found in the genome of A. thaliana
ecotype Columbia (Meyers et al., 2003) (Fig. 1). The TIR
resistance pathway is mediated by the EDS1 PAD4
SAG101 (enhanced disease susceptibility 1 phytoalexin-
decient 4 senescence-associated genes 101) complexes.
EDS1 forms distinct cytosolic and nuclear protein com-
plexes with PAD4 and SAG101 (Feys et al., 2001, 2005).
The CC pathway signals through NDR1 (nonrace-specic
disease resistance 1), which localizes to the plasma mem-
brane via a C-terminal glycosylphosphatidyl-inositol (GPI)
anchor (Century et al., 1997; Aarts et al., 1998; Coppinger
et al., 2004). The CC and TIR pathways converge at the
synthesis of the defence hormone SA (Fig. 1). Following
biotrophic pathogen detection by R-protein, SA accumu-
lates in the infected plants. This pathogen-triggered accumu-
lation is dependent on ISOCHORISMATE SYNTHASE 2
(Wildermuth et al., 2001). SA is a sufcient and necessary
signal for SAR (Vernooij et al., 1994), a broad-spectrum
and long-lasting systemic resistance (Durrant & Dong,
2004) mediated by the positive regulator NPR1 (nonexpres-
sor of PR-1 genes) (Dong, 2004). The SA-dependent
defence signalling pathway is associated with interactions
with biotrophic pathogens, while the ethylene and jasmonic
acid pathways (ET and JA), which are generally thought to
be antagonistic to the SA pathway, are associated with necro-
trophic pathogens (Glazebrook, 2005). Recent evidence
shows that this antagonistic effect would be mediated, at
least in part, by the transcription factor EIN3 (ethylene
insensitive 3), which can directly bind to the SID2 promoter
(SA synthesis) (Chen et al., 2009). Consistent with these
observations, the ein3eil1 double mutant accumulates very
high concentrations of SA and the double mutant displays
enhanced resistance to virulent and avirulent strains of
Pseudomonas syringae (Chen et al., 2009). Recent results also
show that the JA pathway can be made insensitive to SA sup-
pression if the ET pathway is induced (Leon-Reyes et al.,
2010). Despite a large and old consensus among the
community regarding the antagonistic relationship and the
selectivity of the JA and SA pathways for necrotrophic or
biotrophic pathogens, it was recently demonstrated that the
SA pathway can positively contribute to the response to nec-
rotrophic pathogens and that the ET and the JA pathways
can also positively contribute to the response to biotrophic
pathogens (Tsuda et al., 2009). Using infection and genetic
interaction, the Katagiri group made quantitative measure-
ments using combinatorial mutants of the ET, JA and SA
pathways to identify the role of the wild-type genes rather
than to analyse the effect of the mutant phenotypes (Tsuda
et al., 2009). There is nowaccumulating evidence that, upon
activation, R-proteins can go the nucleus themselves (Burch-
Smith et al., 2007; Shen et al., 2007; Wirthmueller et al.,
2007; Cheng et al., 2009) and the nucleocytoplasmic shut-
tling of the defence components is highly reminiscent of
NF-jB nuclear import (Wiermer et al., 2010).
III. R-protein pathways in poplar
Although there has not been any R-AVR combination iden-
tied in poplar, there is mounting evidence to suggest that
resistance to some pathogens would depend on R-proteins.
One of the major threats faced by poplar is the foliar rust
caused by the obligate biotrophic fungus Melampsora spp.
Responses to Melampsora can be categorized into qualitative
or quantitative defence responses. Different physiological
races of rust can elicit incompatible or compatible (qualita-
tive) reactions on a given clone of pure poplar species or on
hybrid clones. For example, the hybrid Populus deltoides
Populus nigra clone Ogy displays an incompatible reaction
with Melampsora larici-populina isolates belonging to race
Fig. 1 Main actors of the R-protein pathway
in Arabidopsis and their homologues in
poplar. NDR1, nonrace-specic disease
resistance 1; NPR1, nonexpressor of PR-1
genes; EDS1, enhanced disease susceptibility
1; SA, salicylic acid; PAD4, phytoalexin-
decient 4; BNL, BED-NB-LRR; CNL,
CC-NB-LRR; TNL, TIR-NB-LRR.
680 Review Tansley review
New
Phytologist
E1 but a compatible reaction with race E2 M. larici-
populina isolates. The incompatible reaction is characterized
by highly localized early collapse and disorganization of the
cytoplasm within 2 h after the appearance of the rst hausto-
ria (Laurans & Pilate, 1999). This reaction is indicative of
R-protein-mediated hypersensitive response. Both types of
response have been studied at the transcriptomics level. As in
A. thaliana, the compatible and incompatible responses were
found to trigger largely overlapping gene sets, including the
well-known genes PR1, PR5, PR10 and NPR1 (Rinaldi et al.,
2007). More recently, the quantitative defence response,
which compares the plant response with the growth of two
Melampsora species, was also analysed by microarray (Azaiez
et al., 2009). The studied plant responses to a fully compati-
ble Melampsora species with unrestricted growth, and another
compatible species that elicits plant defence resulting in
restricted growth, revealed gene sets partly overlapping but
also specic to both responses (Azaiez et al., 2009).
For a given R AVR combination leading to qualitative
resistance, crosses have shown that resistance was controlled
by one single dominant gene or closely located genes.
Mendelian segregation of qualitative resistance phenotypes
in interhybrid crosses also suggests that resistance is con-
trolled by one single dominant gene or closely located genes
(Cervera et al., 1996; Villar et al., 1996; Tabor et al., 2000;
Stirling et al., 2001; Zhang et al., 2001; Yin et al., 2004).
The rst putative R-gene-mediated resistance locus to be
ne mapped was MER (Zhang et al., 2001). MER confers
resistance to races E1, E2 and E3 of M. laricina-populina
and it segregated in a Mendelian fashion when the resistant
female parent P. deltoides was used in interhybrid crosses
with male P. nigra or P. trichocarpa to generate a segregating
mapping population (Zhang et al., 2001). The sequenced
amplied fragment length polymorphism (AFLP) markers
linked to the MER locus revealed the presence of three NB-
LRRs in the region associated with resistance (Zhang et al.,
2001); the locus was later mapped to chromosome XIX
(Yin et al., 2004). The MXC3 locus conferring resistance
Melampsora X columbiana pathotype 3 also segregated in a
Mendelian manner in an F
1
interspecic hybrid poplar ped-
igree (P. trichocarpa P. deltoides), indicating again that
the gene is a single dominant gene. Unfortunately, despite
the saturation of genetic markers around the MXC3 locus,
the gene responsible for resistance could not be identied
because of a lack of recombination close to the marker
(Stirling et al., 2001) as was previously observed for other
R-gene loci (Ganal & Tanksley, 1996; Wei et al., 1999;
Behura et al., 2004; Yang & Hua, 2004). MXC3 was later
mapped to chromosome IV (Yin et al., 2004). Intriguingly,
no NBS-LRR genes were found in the vicinity of MXC3.
Two thaumatin-like pathogenesis-related proteins (PR5s),
two receptor-like kinases (RLKs), one receptor-like protein
(lacking a kinase domain) and one TIR-RLK were found.
The four receptor-like genes (excluding the two thaumatins)
were located c. 20 cM from the marker linked to the MXC3
locus, making them unlikely candidates. Although thauma-
tin-like pathogenesis-related (PR5) proteins have been
shown to be involved in resistance, they are usually involved
in downstream secondary responses and do not act as bona
de resistance genes per se. Gaps still exist in the poplar gen-
ome and the MXC3 gene could still be a NB-LRR located in
one of those gaps (Yin et al., 2004).
IV. What is the role of BED-NB-LRR?
At the genomic level, poplar possesses a set of R-genes
nearly twofold that of Arabidopsis (Kohler et al., 2008;
Yang et al., 2008) (Fig. 1). Both grapevine and poplar dis-
play a higher degree of recent tandem duplication and gene
conversion than Arabidopsis and rice (Yang et al., 2008).
This feature could increase overall recombination events
and thus lead to a larger number of disease-resistant alleles
and curtail the longer generation time of perennial species.
Poplar contains a small subset of atypical putative
R-genes that seem to have arisen from domain fusion, such
as TIR-NBS-LRR-TIR, TIR-NBS-LRR-NBS and NBS-LRR-
TIR, and others also found in the CNL family or a mix of
the two families (TCNL) (Kohler et al., 2008; Yang et al.,
2008). Such chimeric putative R-proteins were also
reported in Arabidopsis (Meyers et al., 2003). The presence
of a putative R-gene family in poplar, the BED-NB-LRR
family (henceforth called BNL) comprising 32 members,
seems to be unique to poplar in the dicots family (Table 1).
Blast search does not reveal any hits with a BNL architec-
ture outside of the poplar species in the dicots except for a
TIR-NB-LRR-BED-TIR in Vitis vinifera. In monocots,
there are eight occurrences of BNL in the rice genome. One
of these genes, Xa1, was shown to confer resistance to bacte-
rial blight caused by Xanthomonas oryzae pv. oryzae
(Yoshimura et al., 1998). We inferred a phylogenetic tree of
all BNLs using the NB-ARC domain (Fig. 2). Poplar BNLs
are supported on a different clade from V. vinifera and rice
BNL. In addition, and as expected, BNLs on chromosome
XIX are more alike than those located on other chromo-
somes (see grey-shaded box in Fig. 2). Some BNLs, pres-
ently positioned on the scaffold, also fall within the
grey-shaded box. The discovery of the BED domain was
rst published in 2000 and was named BED nger after
two Drosophila known proteins named BEAF and DREF
containing this domain (Aravind, 2000). It is rather surpris-
ing that both rice and poplar seem to have independently
acquired this gene architecture. The BED domain is a ubiq-
uitous zinc nger DNA-binding domain and its DNA-
binding sequence was only recently identied using CHIP
sequencing in mouse cells (Markljung et al., 2009).
Unfortunately, the DNA-binding sequence of the BED
domain is relatively short (8 bp), making its random pres-
ence more frequent. We performed an in silico search of a
New
poplar promoter repository using the BED nucleotide-
binding sequence to assess whether its presence would be
enriched in promoters located upstream of defence-related
genes. Perhaps owing to the fact that the BED-binding
domain is very short, and therefore not very specic, we
observed no correlation between the occurrence of BED-
binding sites and a defence function for the genes associated
with promoters that contained the BED-binding domain
(H. Germain & A. Seguin, pers. obs.). Most BNLs in poplar
arose from common ancestors and 27 out of 32 have close
homology with At4g27220, four show close homology with
At4g10780 and one shows close homology with At4g26090;
all are closely located genes on A. thaliana chromosome IV
(Table 1 and Supporting Information Table S1) for
sequence and alternate gene model). Intriguingly, most
BNLs are found in the upper peritelomeric end of poplar
chromosome XIX (Fig. 3), a genomic region rich in putative
R-genes where MER is located and thought to be an incipient
sex chromosome where segregation distortion and sup-
pressed recombination are observed (Yin et al., 2004).
Zhang et al. (2001) mapped the MER locus several years
before the full genome sequence of poplar became available
and yet they found, through cloning and sequencing, that
some of their AFLP markers were NB-LRRs encoding genes.
We now know that the AFLP marker AF393739 used by
Zhang et al. (2001) is in fact Poptr0019s00510, a BNL
(Fig. 3). A total of 20 BNLs (out of 32) are associated with
the MER locus, making them good candidates for being the
MER gene, if they are indeed R-genes (for the precise chro-
mosomal location of these BNLs, see Table S2).
Froma signalling standpoint, the fusion of a DNA-binding
domain with a NB-LRRdomain is of great interest. Although
some R-proteins have recently been shown to localize to the
nucleus (Burch-Smith et al., 2007; Shen et al., 2007;
Wirthmueller et al., 2007; Cheng et al., 2009), none have
been shown to bind DNA or regulate transcription directly.
Another alternative is that the BED domain of the putative
resistance protein could act as a decoy (van der Hoorn &
Kamoun, 2008) for another BED-containing transcriptional
regulator, which would be the true target of the virulence
Table 1 List of all members of the BED-NB-LRR family (BNLs) and the presence of NLS or NES in their amino acid sequence
New annotation
Arabidopsis
thaliana
orthologues
NLS prediction
Psort NES (number and position of amino acid)
POPTR_0001s41540 At4g27220 Negative (0.00) 1 (686)
POPTR_0001s41570 At4g27220 Negative (0.00) 2 (569, 571)
POPTR_0011s12620 At4g27220 Negative (0.00) 5 (430, 433-6)
POPTR_0011s12500 At4g27220 notclr (0.40) 3 (430, 673, 675)
POPTR_0019s00410 At4g27220 notclr (0.30) 12 (670-8, 964, 969, 972)
POPTR_0019s00430 At4g27220 notclr (0.60) 5 (661-2, 664, 896, 898)
POPTR_0019s00510 At4g27220 Positive (0.70) 8 (578-584, 940)
POPTR_0019s00540 At4g10780 Positive (0.70) 1 (854)
POPTR_0019s00570 At4g27220 Negative (0.00) 6 (543, 937, 940, 942, 945, 948)
POPTR_0019s00620 At4g26090 notclr (0.30) 4 (498, 500, 599, 601)
POPTR_0019s01010 At4g27220 notclr (0.60) 3 (688, 1282, 1366)
POPTR_0019s01020 At4g27220 notclr (0.30) 4 (1028, 1385, 1387, 1470)
POPTR_0019s01080 At4g27220 Negative (0.00) 9 (488-96)
POPTR_0019s01670 At4g27220 Negative (0.00) 7 (234, 564, 566, 610, 892, 895, 897)
POPTR_0019s02150 At4g10780 Positive (0.70) 2 (918, 926)
POPTR_0019s02170 At4g27220 notclr (0.40) 2 (568, 840)
POPTR_0019s02200 At4g27220 notclr (0.60) 2 (676, 942)
POPTR_0019s03720 At4g27220 notclr (0.30) 7 (443, 445, 818, 1041, 1043-4, 1090)
POPTR_0031s00350 At4g10780 Negative (0.00) 9 (743-51)
POPTR_0031s00430 At4g27220 notclr (0.30) 1 (333)
POPTR_0060s00250 At4g27220 notclr (0.30) 3 (537-8, 540)
POPTR_0190s00200 At4g27220 notclr (0.30) 9 (603-11)
POPTR_0190s00220 At4g27220 Negative (0.15) 3 (567, 572, 1055)
POPTR_0287s00220 At4g27220 notclr (0.30) 2 (806,1049)
fgenesh4_pg.C_LG_XIX000056,
new annotation not found
At4g27220 Positive (0.80) 1 (631)
New
Phytologist
factor. Supporting their possible role in the nucleus is the
presence of a nuclear localization signal and a nuclear export
signal in some of the BNL proteins (Table 1, Fig. S1).
V. Downstream of R-proteins
R-proteins are quite numerous and they converge to major
regulatory nodes depending on their respective classications.
TNL signals through the EDS1 PAD4 complex while CNL
signals via the NDR1 pathway. According to the new Poplar
2.0 genome annotation on Phytozome (http://www.phyto
zome.net/), three copies of EDS1 would be found in poplar
with 42.0, 42.1 and 27.2% identity, along with two copies
of PAD4 having 48.4 and 46.4% identity with their
Arabidopsis orthologues. In the CNL pathway, two copies
of NDR1 are found, having 45.7 and 46.9% identity with
their Arabidopsis counterparts. Although poplar seems to
possess EDS1 PAD4 and NDR1, a role in defence has not
yet been demonstrated. Whether the BNLs are functional
R-proteins in poplar and whether they signal through the
EDS1 PAD4 node or via the NDR1 node has not yet been
demonstrated. Based on our observations, a hypothesis with
regard to BED-NB-LRR signalling can be made. First is the
presence and positive R-gene-like activity of a BNL in rice,
even though rice lacks TNL. Second, all BNLs relate
strongly to only three Arabidopsis NB-LRRs and all three
are CNLs. Additionally, previous work by Meyers et al.
(2003) identied different patterns in the NBS domain,
namely the RNBS-A, RNBS-C and RNBS-D motifs that
were different between the CNLs and TNLs (Meyers et al.,
2003). Kohler et al. (2008) did a similar analysis using the
poplar NB-LRR proteins and we nd there is slightly more
homology between BNL and CNL than with TNL, particu-
larly in the RNBS-D motif (see the CAI LFPxD section)
and the RNBS-A motif (in the WxxVSQDFxxxxxEEL sec-
tion) (Fig. S2). Based on these observations, BNL signalling
via the NDR1 node or via a novel pathway would be more
likely than via the EDS1 PAD4 node.
VI. The role of salicylic acid in biotrophic
interaction
In Arabidopsis, race-specic pathogen recognition usually
leads to localized HR and results in SAR in uninfected parts
Fig. 2 Phylogenetic tree of all BNLs (BED-NB-LRR) from poplar, rice and one from Vitis vinifera. The tree was inferred using the method of
neighbour-joining and used the full NB-ARC domain.
New
of the plants. The SAR response is dependent on the protein
NPR1 and on the accumulation of SA, and leads to an
increase in PR gene expression. In addition, NPR1 is also
involved in the negative regulation of the JA pathway (for a
review on NPR1, see Dong, 2004). In rice, which has a very
high endogenous SA concentration, initial work has shown
that SA-depleted plants were not more sensitive to pathogen
infection than wild-type plants (Yang et al., 2004). Recent
results showed that WRKY gene OsWRKY33 overexpression
can trigger PR gene expression and increase SA accumula-
tion, which leads to HR-like cell death (Koo et al., 2009),
indicating that SA may have a role in the rice defence mech-
anism. Additionally, following probenazole treatment (a
SAR inducer) in rice, free SA concentration increase is
observed as well as OsPR1 transcript abundance, resulting in
plants that are more resistant to infection by Magnaporthe
grisea (Iwai et al., 2007).
The role of SA in poplar, or more generally in tree defence,
has not been claried, and whether the concentration of SA
can be modulated in poplar remains controversial. Results by
Morse et al. (2007) demonstrate that transgenic poplars
overexpressing the widely used bacterial salicylate hydrolase
NahG had unchanged free SA and catechol concentrations.
However, glycosyl-conjugated SA was reduced by > 90% in
two transgenic lines and there was a vefold increase in glycosyl-
conjugated catechol (downstream metabolite of SA deg-
radation), indicating that free SA concentration is tightly
regulated (Morse et al., 2007). Diara et al. (2005) used
the ozone (O
3
)-induced hypersensitive response to assess
whether SA, 1-aminocylcyclopropane-1-carboxylic acid
(ACC, precursor of ethylene synthesis), ethylene and per-
oxide had similar kinetics and scale of production in the
ozone-sensitive clone Eridano (P. deltoides P. maximowizii)
and the O
3
-resistant clone I-214 (P. deltoides P. eur-
americana). In their treatment, a modest but signicant
increase in SA was observed in the sensitive clone (Eridano)
while the I-214 clone, which had a much higher basal SA
concentration, remained unchanged (Diara et al., 2005).
The varying kinetics and magnitudes observed in specic
hybrid to HR-inducing condition argues against a general
and conserved role of SA in poplar defence response. The
rapid up-regulation of poplar genes involved in JA and ET
biosynthesis, such as allene oxide synthase (AOS) and ACC
synthase, following Melampsora sp. infection, supports the
positive contribution of JA and ET in response to biotrophic
pathogens in trees (Azaiez et al., 2009). In glasshouse trials
we observed that the exogenous application of isonicotinic
acid (a SAR inducer) could trigger PR-gene expression and
restrict the growth of the fully compatible fungal pathogen
M. larici-populina in hybrid clones of P. tricocarpa P.
deltoides 3225 juveniles. These results suggest that young
hybrids rely on SA for pathogen resistance. Whether this role
is maintained as the tree ages remains to be claried.
VII. Concluding remarks
Poplar and Arabidopsis both belong to the Eurosid clade of
dicots and are thus relatively close cousins in comparison
with species such as cereal crops (monocotyledon) or conifer
trees (gymnosperm). This phylogenetic relationship should
sustain assumptions made from knowledge acquired in
Arabidopsis pathosystems and thus circumvent the lack of
data for poplar. Little is known about the active players
involved in the poplar qualitative defence response, but
solely based on genomic comparison with Arabidopsis, it
seems that poplar has all the proper tools to launch an
R-gene-mediated response. Poplar also has unique features,
such as the BNL family, for which a role in tree defence is
yet to be demonstrated. To account for the disadvantage of
its long juvenile stage, which gives poplar (and trees in
general) less generation per pathogen generation to evolve
new R alleles, poplar has a higher degree of tandem dupli-
cation and gene conversion than Arabidopsis, and is an
obligate outcrosser, which promotes genetic exchange and
heterozygosity (Ingvarson, 2010), in contrast to A. thaliana,
which is almost an obligate inbreeder and a highly homozy-
gous species. The divergence with Arabidopsis probably
occurs downstream of proteinaceous defence regulators at
the hormonal level. There is a well-established consensus
among plant pathologists that the plant response to bio-
trophic pathogens is mediated by SA, while the response to
necrotrophic pathogens is mediated by JA and that both
pathways are antagonistic (Koornneef & Pieterse, 2008).
Fig. 3 Location of BNL (BED-NB-LRR) on the peritelomeric region
of chromosome 19 where MER is located.
New
Phytologist
Only recently have we seen evidence that JA could have a
positive output on biotrophic response and SA on necro-
troph response (Tsuda et al., 2009). The extent of this
contribution may be different in trees and Arabidopsis. For
instance, auxin, gibberellic acid and abscisic acid also inu-
ence plantpathogen interactions (Grant & Jones, 2009),
but their contribution to the poplar defence response has
not been investigated. Nonetheless, the fact that the poplar
SA concentration appears to respond so differently to stim-
ulus between hybrid clones and the fact that some hybrid
clones appear insensitive to SA-stimulating conditions
points to the fact that SA may not be the only hormone
leading the response to biotrophic pathogens or that it
is mediated by SA derivatives to keep the free SA pool
constant for developmental purposes or other reasons.
The lack of a method to transiently assess gene functions
in poplar (technical limitation) seriously impedes the inves-
tigation of gene functions and defence regulators. Transient
gene silencing and gene overexpression technologies must
be adapted to poplar to quickly move forward the func-
tional analysis of genes in poplar. The successful complete
sequencing of the poplar mosaic virus genome (PopMV) by
Dr Malcolm Campbells group (Smith & Campbell, 2004)
could potentially lead to the use of virus-induced gene
silencing in poplar. However, since PopMV virulence varies
greatly between hybrids, other less species-specic viral vec-
tors should also be assessed. Because poplar is an obligate
outcrosser, the T-DNA insertion strategy used to generate
homozygous knock-out, such as the one so widely used in
Arabidopsis, is irrelevant in this species. However, the gain
of function could be investigated in activation-tagging pop-
lar lines such as the 1800 independent activation-tagged
lines produced and are available through collaboration with
the Regan laboratory at Queens University (Harrison et al.,
2007).
The availability of transcriptome analysis for poplar is of
limited help when it comes to nding defence regulators.
Unlike the downstream components of signalling cascades,
the regulation of R-genes at the transcriptional level is usu-
ally relatively limited. Despite time and labour require-
ments, generation of stable transgenic lines remains the best
approach to perform functional analyses in poplar.
Complementation of Arabidopsis knock-out mutants with
poplar genes is another strategy that needs to be imple-
mented to conrm poplar protein function. Although this
method has its limitations and does not allow the discovery
of new processes or poplar-specic processes, it could help
to conrm protein activity or function. Another aspect
essential to a better understanding of poplar defence would
be the availability of cloned Avr and a delivery system that
could be used to dissect the poplar ETI pathway and analyse
whether all the downstream regulators of R-gene signalling
are redundant. At the present stage, functional research on
poplar qualitative defence response still faces many hurdles.
Overcoming these technical limitations would greatly help
poplar to become a better model organism.
References
Aarts N, Metz M, Holub E, Staskawicz BJ, Daniels MJ, Parker JE. 1998.
Different requirements for EDS1 and NDR1 by disease resistance
genes dene at least two R gene-mediated signaling pathways in
Arabidopsis. Proceedings of the National Academy of Sciences, USA 95:
1030610311.
Aravind L. 2000. The BED nger, a novel DNA-binding domain in
chromatin-boundary-element-binding proteins and transposases. Trends
in Biochemical Sciences 25: 421423.
Azaiez A, Boyle B, Levee V, Seguin A. 2009. Transcriptome proling in
hybrid poplar following interactions with Melampsora rust fungi.
Molecular Plant-Microbe Interactions 22: 190200.
Behura SK, Valicente FH, Rider SDJ, Shun-Chen M, Jackson S, Stuart
JJ. 2004. A physically anchored genetic map and linkage to avirulence
reveals recombination suppression over the proximal region of Hessian
y chromosome A2. Genetics 167: 343355.
Belkhadir Y, Subramaniam R, Dangl JL. 2004. Plant disease resistance
protein signaling: NBS-LRR proteins and their partners. Current
Opinion in Plant Biology 7: 391399.
Boller T, Felix G. 2009. A renaissance of elicitors: perception of microbe-
associated molecular patterns and danger signals by pattern-recognition
receptors. Annual Review of Plant Biology 60: 379406.
Bolwell GP. 1999. Role of active oxygen species and NO in plant defence
responses. Current Opinion in Plant Biology 2: 287294.
Bradshaw HD Jr. 1996. Molecular genetics of Populus. In: Stettler RF,
Bradshaw HD Jr, Heilman PE, Hinckley TM, eds. Biology of Populus
and its implications for management and conservation. Ottawa, Canada:
NRC Research Press, 183199.
Burch-Smith TM, Schiff M, Caplan JL, Tsao J, Czymmek K, Dinesh-
Kumar SP. 2007. A novel role for the TIR domain in association with
pathogen-derived elicitors. PLoS Biology 5: e68.
Century KS, Shapiro AD, Repetti PP, Dahlbeck D, Holub E, Staskawicz
BJ. 1997. NDR1, a pathogen-induced component required for
Arabidopsis disease resistance. Science 278: 19631965.
Cervera MT, Gusmao J, Steenackers M, Peleman J, Storme V, Broeck A,
Montagu M, Boerjan W. 1996. Identication of AFLP molecular
markers for resistance against Melampsora larici-populina in Populus.
Theoretical and Applied Genetics 93: 733737.
Chen H, Xue L, Chintamanani S, Germain H, Lin H, Cui H, Cai R, Zuo
J, Tang X, Li X et al. 2009. ETHYLENE INSENSITIVE3 and
ETHYLENE INSENSITIVE3-LIKE1 repress SALICYLIC ACID
INDUCTION DEFICIENT2 expression to negatively regulate plant
innate immunity in Arabidopsis. Plant Cell 21: 25272540.
Cheng YT, Germain H, Wiermer M, Bi D, Xu F, Garc a AV,
Wirthmueller L, Despres C, Parker JE, Zhang Y et al. 2009. Nuclear
pore complex component MOS7 Nup88 is required for innate
immunity and nuclear accumulation of defense regulators in
Arabidopsis. Plant Cell 21: 25032516.
Coppinger P, Repetti PP, Day B, Dahlbeck D, Mehlert A, Staskawicz BJ.
2004. Overexpression of the plasma membrane-localized NDR1 protein
results in enhanced bacterial disease resistance in Arabidopsis thaliana.
Plant Journal 40: 225237.
Dangl JL, Jones JDG. 2001. Plant pathogens and integrated defence
responses to infection. Nature 411: 826833.
Delledonne M, Xia Y, Dixon RA, Lamb C. 1998. Nitric oxide functions
as a signal in plant disease resistance. Nature 394: 585588.
Deslandes L, Olivier J, Peeters N, Feng DX, Khounlotham M, Boucher
C, Somssich I, Genin S, Marco Y. 2003. Physical interaction between
RRS1-R, a protein conferring resistance to bacterial wilt, and PopP2, a
New
type III effector targeted to the plant nucleus. Proceedings of the National
Academy of Sciences, USA 100: 80248029.
Diara C, Castagna A, Baldan B, Sodi AM, Sahr T, Langebartels C,
Sebastiani L, Ranieri A. 2005. Differences in the kinetics and scale of
signalling molecule production modulate the ozone sensitivity of hybrid
poplar clones. New Phytologist 168: 351364.
Dodds PN, Lawrence GJ, Catanzariti A-M, Teh T, Wang C-IA, Ayliffe
MA, Kobe B, Ellis JG. 2006. Direct protein interaction underlies gene-
for-gene specicity and coevolution of the ax resistance genes and ax
rust avirulence genes. Proceedings of the National Academy of Sciences,
USA 103: 88888893.
Dong X. 2004. NPR1, all things considered. Current Opinion in Plant
Biology 7: 547552.
Duplessis S, Major I, Martin F, Seguin A. 2009. Poplar and pathogen
interactions: insights from Populus genome-wide analyses of resistance
and defense gene families and gene expression proling. Critical Reviews
in Plant Sciences 28: 309334.
Durrant WE, Dong X. 2004. Systemic acquired resistance. Annual Review
of Phytopathology 42: 185209.
Feys BJ, Moisan LJ, Newman M-A, Parker JE. 2001. Direct interaction
between the Arabidopsis disease resistance signaling proteins, EDS1 and
PAD4. EMBO Journal 20: 54005411.
Feys BJ, Wiermer M, Bhat RA, Moisan LJ, Medina-Escobar N, Neu C,
Cabral A, Parker JE. 2005. Arabidopsis SENESCENCE-ASSOCIATED
GENE101 stabilizes and signals within an ENHANCED DISEASE
SUSCEPTIBILITY1 complex in plant innate immunity. Plant Cell 17:
26012613.
Flor HH. 1971. Current status of the gene-for-gene concept. Annual
Review of Phytopathology 9: 275296.
Ganal MW, Tanksley SD. 1996. Recombination around the Tm2a and
Mi resistance genes in different crosses of Lycopersicon peruvianum.
Theoretical and Applied Genetics 92: 101108.
Glazebrook J. 2005. Contrasting mechanisms of defense against biotrophic
and necrotrophic pathogens. Annual Review of Phytopathology 43:
205227.
Grant MR, Jones JDG. 2009. Hormone (dis)harmony moulds plant
health and disease. Science 324: 750752.
Hacquard S, Delaruelle C, Legue V, Tisserant E, Kohler A, Frey P,
Martin F, Duplessis S. 2010. Laser capture microdissection of uredinia
formed by Melampsora larici-populina revealed a transcriptional switch
between biotrophy and sporulation. Molecular Plant-Microbe Interaction
23: 12751286.
Harrison EJ, Bush M, Plett JM, McPhee DP, Vitez R, OMalley B,
Sharma V, Bosnich W, Seguin A, MacKay J et al. 2007. Diverse
developmental mutants revealed in an activation-tagged population of
poplar. Canadian Journal of Botany 85: 10711081.
van der Hoorn RAL, Kamoun S. 2008. Fromguard to decoy: a newmodel
for perception of plant pathogen effectors. Plant Cell 20: 20092017.
Ingvarson PK. 2010. Nucleotide polymorphism, linkage disequilibrium
and complex trait dissection in Populus. In: Jansson S, Bhalerao RP,
Groover AT, eds. Genetics and genomics of Populus. New York, NY,
USA: Springer, 91112.
Iwai T, Seo S, Mitsuhara I, Ohashi Y. 2007. Probenazole-induced
accumulation of salicylic acid confers resistance to Magnaporthe grisea in
adult rice plants. Plant and Cell Physiology 48: 915924.
Jia Y, McAdams SA, Bryan GT, Hershey HP, Valent B. 2000. Direct
interaction of resistance gene and avirulence gene products confers rice
blast resistance. EMBO Journal 19: 40044014.
Jones JDG, Dangl JL. 2006. The plant immune system. Nature 444:
323329.
Kohler A, Rinaldi C, Duplessis S, Baucher M, Geelen D, Duchaussoy F,
Meyers BC, Boerjan W, Martin F. 2008. Genome-wide identication
of NBS resistance genes in Populus trichocarpa. Plant Molecular Biology
66: 619636.
Koo SC, Moon BC, Kim JK, Kim CY, Sung SJ, Kim MC, Cho MJ,
Cheong YH. 2009. OsBWMK1 mediates SA-dependent defense
responses by activating the transcription factor OsWRKY33.
Biochemical and Biophysical Research Communications 387: 365370.
Koornneef A, Pieterse CMJ. 2008. Cross talk in defense signaling. Plant
Physiology 146: 839844.
Laurans F, Pilate G. 1999. Histological aspects of a hypersensitive
response in poplar to Melampsora larici-populina. Phytopathology 89:
233238.
Leon-Reyes A, Du Y, Koornneef A, Proietti S, Korbes AP, Memelink J,
Pieterse CMJ, Ritsema T. 2010. Ethylene signaling renders the
jasmonate response of Arabidopsis insensitive to future suppression by
salicylic acid. Molecular Plant-Microbe Interactions 23: 187197.
Markljung E, Jiang L, Jaffe JD, Mikkelsen TS, Wallerman O,
Larhammar M, Zhang X, Wang L, Saenz-Vash V, Gnirke A et al.
2009. ZBED6, a novel transcription factor derived from a domesticated
DNA transposon regulates IGF2 expression and muscle growth. PLoS
Biology 7: e1000256.
Martin GB, Bogdanove AJ, Sessa G. 2003. Understanding the functions
of plant disease resistance proteins. Annual Review of Plant Biology 54:
2361.
Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW. 2003.
Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant
Cell 15: 809834.
Miranda M, Ralph SG, Mellway R, White R, Heath MC, Bohlmann J,
Constabel CP. 2007. The transcriptional response of hybrid poplar
(Populus trichocarpa P. deltoides) to infection by Melampsora medusae
leaf rust involves induction of avonoid pathway genes leading to the
accumulation of proanthocyanidins. Molecular Plant-Microbe
Interactions 20: 816831.
Morse AM, Tschaplinski TJ, Dervinis C, Pijut PM, Schmelz EA, Day W,
Davis JM. 2007. Salicylate and catechol levels are maintained in nahG
transgenic poplar. Phytochemistry 68: 20432052.
Rinaldi C, Kohler A, Frey P, Duchaussoy F, Ningre N, Couloux A,
Wincker P, Le Thiec D, Fluch S, Martin F et al. 2007. Transcript
proling of poplar leaves upon infection with compatible and
incompatible strains of the foliar rust Melampsora larici-populina. Plant
Physiology 144: 347366.
Shen Q-H, Saijo Y, Mauch S, Biskup C, Bieri S, Keller B, Seki H, U
lker
B, Somssich IE, Schulze-Lefert P. 2007. Nuclear activity of MLA
immune receptors links isolate-specic and basal disease-resistance
responses. Science 315: 10981103.
Smith CM, Campbell MM. 2004. Complete nucleotide genome sequence
of the genomic RNA of poplar mosaic virus (genus Carlavirus). Archives
of Virology 149: 18311841.
Stirling B, Newcombe G, Vrebalov J, Bosdet I, Bradshaw HDJ. 2001.
Suppressed recombination around the MXC3 locus, a major gene for
resistance to poplar leaf rust. Theoretical and Applied Genetics 103:
11291137.
Tabor GM, Kubisiak TL, Klopfenstein NB, Hall RB, McNabb HJ. 2000.
Bulked segregant analysis identies molecular markers linked to
Melampsora medusae resistance in Populus deltoides. Phytopathology 90:
10391042.
Tameling WIL, Elzinga SDJ, Darmin PS, Vossen JH, Takken FLW,
Haring MA, Cornelissen BJC. 2002. The tomato R gene products I-2
and Mi-1 are functional ATP binding proteins with ATPase activity.
Plant Cell 14: 29292939.
Tsuda K, Sato M, Stoddard T, Glazebrook J, Katagiri F. 2009.
Network properties of robust immunity in plants. PLoS Genetics 5:
e1000772.
Tuskan GA, DiFazio S, Jansson S, Bohlmann J, Grigoriev I, Hellsten U,
Putnam N, Ralph S, Rombauts S, Salamov A et al. 2006. The genome
of black cottonwood, Populus trichocarpa (Torr. & Gray). Science 313:
15961604.
New
Phytologist
Ueda H, Yamaguchi Y, Sano H. 2006. Direct interaction between the
tobacco mosaic virus helicase domain and the ATP-bound resistance
protein, N factor during the hypersensitive response in tobacco plants.
Plant Molecular Biology 61: 3145.
Van der Biezen EA, Jones JDG. 1998. Plant disease-resistance proteins
and the gene-for-gene concept. Trends in Biochemical Sciences 23:
454456.
Vernooij B, Friedrich L, Morse A, Reist R, Kolditz-Jawhar R, Ward E,
Uknes S, Kessmann H, Ryals J. 1994. Salicylic acid is not the
translocated signal responsible for inducing systemic acquired resistance
but is required in signal transduction. Plant Cell 6: 959965.
Villar M, Lefevre F, Bradshaw HD Jr, du-Cros ET. 1996. Molecular
genetics of rust resistance in poplars (Melampsora larici-populina
Kleb Populus sp.) by bulked segregant analysis in a 2 2 factorial
mating design. Genetics 143: 531536.
Wei F, Gobelman-Werner K, Morroll SM, Kurth J, Mao L, Wing R,
Leister D, Schulze-Lefert P, Wise RP. 1999. The Mla (powdery
mildew) resistance cluster is associated with three NBS-LRR gene
families and suppressed recombination within a 240-kb DNA interval
on chromosome 5S (1HS) of barley. Genetics 153: 19291948.
Wiermer M, Germain H, Cheng Y, Garc a AV, Parker JE, Li X. 2010.
Nucleoporin MOS7 Nup88 contributes to plant immunity and nuclear
accumulation of defense regulators. Nucleus 1: 332336.
Wildermuth MC, Dewdney J, Wu G, Ausubel FM. 2001. Isochorismate
synthase is required to synthesize salicylic acid for plant defence. Nature
414: 562565.
Wirthmueller L, Zhang Y, Jones JDG, Parker JE. 2007. Nuclear
accumulation of the Arabidopsis immune receptor RPS4 is necessary for
triggering EDS1-dependent defense. Current Biology 17: 20232029.
Xiang T, Zong N, Zou Y, Wu Y, Zhang J, Xing W, Li Y, Tang X, Zhu
LH, Chai J et al. 2008. Pseudomonas syringae effector AvrPto blocks
innate immunity by targeting receptor kinases. Current Biology 18:
7480.
Yang S, Hua J. 2004. A haplotype-specic Resistance gene regulated by
BONZAI1 mediates temperature-dependent growth control in
Arabidopsis. Plant Cell 16: 10601071.
Yang S, Zhang X, Yue J-X, Tian D, Chen J-Q. 2008. Recent duplications
dominate NBS-encoding gene expansion in two woody species.
Molecular Genetics and Genomics 280: 187198.
Yang Y, Qi M, Mei C. 2004. Endogenous salicylic acid protects rice plants
from oxidative damage caused by aging as well as biotic and abiotic
stress. Plant Journal 40: 909919.
Yin TM, DiFazio SP, Gunter LE, Jawdy SS, Boerjan W, Tuskan GA.
2004. Genetic and physical mapping of Melampsora rust resistance genes
in Populus and characterization of linkage disequilibrium and anking
genomic sequence. New Phytologist 164: 95105.
Yoshimura S, Yamanouchi U, Katayose Y, Toki S, Wang ZW, Kono I,
Kurata N, Yano M, Iwata N, Sasaki T. 1998. Expression of Xa1, a
bacterial blight-resistance gene in rice, is induced by bacterial
inoculation. Proceedings of the National Academy of Sciences, USA 95:
16631668.
Zhang J, Steenackers M, Storme V, Neyrinck S, Van Montagu M, Gerats
T, Boerjan W. 2001. Fine mapping and identication of nucleotide
binding site leucine-rich repeat sequences at the MER locus in Populus
deltoides S9-2. Phytopathology 91: 10691073.
Zhang S, Klessig DF. 2001. MAPK cascades in plant defense signaling.
Trends in Plant Science 6: 520527.
Supporting Information
Additional supporting information may be found in the
online version of this article.
Fig. S1 Output le from NetNES showing the precise loca-
tion of each NES in the amino acid sequence.
Fig. S2 MEME analysis of the NB-ARC domain of all
poplar BNL and comparison of their RNBS-A and D
motifs with TNL and CNL.
Table S1 List of all poplar BNL, their former gene model,
alternative transcript and full amino acid sequence
Table S2 BNLs associated with the MER locus and their
precise location on chromosome 19
Please note: Wiley-Blackwell are not responsible for the
content or functionality of any supporting information
supplied by the authors. Any queries (other than missing
material) should be directed to the New Phytologist Central
Ofce.
New

Innate Immunity - Has Poplar PDF

Cargado por

Información del documento

Descripción original:

Título original

Derechos de autor

Formatos disponibles

Compartir este documento

Compartir o incrustar documentos

Opciones para compartir

¿Le pareció útil este documento?

¿Este contenido es inapropiado?

Copyright:

Formatos disponibles

Innate Immunity - Has Poplar PDF

Cargado por

Copyright:

Formatos disponibles

Tansley review

Innate immunity: has poplar made its

También podría gustarte