320

Establishing compatibility between plants and

obligate biotrophic pathogens
Ralph Panstruga

The apparent under-representation of the term ‘plant disease sively cultured in vitro and that form specialized infection
susceptibility’ as opposed to ‘plant disease resistance’ in the structures, haustoria, within infected host cells [1]. Con-
current scientific literature might indicate that ‘compatibility’ has vergent evolution of haustoria has apparently occurred in
not gained the same appreciation as ‘resistance’ in the past. oomycetes (e.g. Peronospora parasitica), ascomycetes (e.g.
However, these seemingly contrary phenomena are intimately powdery mildews, such as Blumeria and Erysiphe ssp.), and
linked, and progress in understanding one process inherently basidiomycetes (e.g. rusts, such as Puccinia ssp.). This
contributes to our comprehension of the other. Recent progress suggests that the ability to form haustoria is either indis-
in analyzing plant–biotroph compatibility includes the molecular pensable for obligate biotrophs or creates a selective
isolation and functional characterization of haustorium-specific advantage to these pathogens (e.g. facilitating access to
cDNAs that encode presumptive hexose- and amino-acid- intracellular pools of solutes that are not available in the
transporter proteins for proton-driven nutrient uptake. apoplast [1]). In contrast to necrotrophic and hemibio-
Accumulating evidence from cytological, pharmacological, trophic fungal and oomycete pathogens, which have no or
phytopathological and molecular studies indicates that only a limited biotrophic phase, obligate biotrophs are
pathogens mediate the suppression of host defenses in a range entirely dependent on living plant tissue for their growth
of plant–biotroph interactions. Arabidopsis thaliana mutants that and propagation. The initial phases of pathogenesis do
are resistant to powdery or downy mildew but that do not not differ fundamentally among obligate biotrophs, hemi-
exhibit constitutively activated defense could be affected in biotrophs and necrotrophs, and commonly include spore
host-compatibility factors. adhesion, appressorium formation and penetration [2]. At
later stages, differences become evident as biotrophs
Addresses establish haustoria within living plant cells and redirect
Max-Planck-Institut für Züchtungsforschung, Department of the host’s metabolism to meet their own needs without
Plant–Microbe Interactions, Carl-von-Linné-Weg 10,
causing the death of host cells. By contrast, necrotrophic
50829 Köln, Germany
e-mail: panstrug@mpiz-koeln.mpg.de and hemibiotrophic fungi trigger host cell death either
immediately (e.g. by secretion of toxins) or during the
course of infection. Lack of host cell death in plant–
Current Opinion in Plant Biology 2003, 6:320–326 biotroph interactions might be accomplished by the
This review comes from a themed issue on
pathogen’s ability to avoid or durably suppress pre-
Biotic interactions formed and induced host defenses [2]. It is plausible that
Edited by Barbara Baker and Jane Parker specific host genes and/or proteins are targeted by bio-
trophs to achieve these goals, and as such may be con-
1369-5266/03/$ – see front matter
ß 2003 Elsevier Science Ltd. All rights reserved. sidered to be compatibility factors that are essential for
successful pathogenesis.
DOI 10.1016/S1369-5266(03)00043-8
In this review, I summarize recent findings on the poten-
Abbreviations tial role of haustoria in the biotrophic lifestyle, on defense
AAT1 AMINO-ACID TRANSPORTER1 suppression in various plant–biotroph interactions, and
Bgh Blumeria graminis forma specialis hordei on host mutants in which potential compatibility factors
CaM calmodulin
are affected.
DMR DOWNY MILDEW RESISTANT
EHM extrahaustorial membrane
fis1 flax inducible sequence1 Haustoria: efficient nutrient robbers and
GFP green fluorescent protein productive metabolite factories
HXT1 HEXOSE TRANSPORTER1 Haustoria can be envisaged as determinate branches of
mlo powdery mildew resistance gene o
PM plasma membrane
intracellular, intercellular, or epicuticular hyphae that are
PMR POWDERY MILDEW RESISTANT formed upon host-cell penetration and terminate within
THI1 THIAMINE BIOSYNTHESIS1 the penetrated cell [1]. However, haustoria are not truly
intracellular: they are separated from the host cytoplasm
by a specific derivative of the host plasma membrane
Introduction (PM), the extrahaustorial membrane (EHM), which is
Obligate biotrophs, such as powdery mildews, downy formed upon haustorium formation and tightly surrounds
mildews and rust fungi, constitute a phylogenetically this fungal organ ([1,3,4]; Figure 1). The amorphous, gel-
unrelated group of phytopathogens that cannot be exten- like layer between the invaginated EHM and the

Current Opinion in Plant Biology 2003, 6:320–326 www.current-opinion.com

 Plant–biotroph compatibility Panstruga 321

Figure 1

Haustorial neck band

Plant cell wall Hechtian strands
Cell wall apposition
PMR4?

Plant plasma
membrane Pathogen plasma
membrane

Extrahaustorial ATPase PMR6?

membrane (EHM) H+ Pathogen cell
wall
AAT1/2
H+
Amino acids

Vitamin B synthesis
HXT1
H+
Hexoses
Extrahaustorial
matrix (EHMAT)

Cell death suppression MLO? Defense suppression

bax inhibitor?
Current Opinion in Plant Biology

A central role for haustoria in compatible plant–biotroph interactions. The scheme represents a virtual haustorium of a fictitious phytopathogenic
biotroph within a host cell. The cartoon collates findings from various studies of plant–pathogen interactions (see text). The host PM and the
extrahaustorial membrane (which are separated by the haustorial neck band) are shown in distinct colors to highlight their differing qualities. The
haustorium absorbs nutrients (e.g. hexoses and amino acids) from the extrahaustorial matrix via proton-symport transporters. The necessary proton
gradient is established by a PM-localized pathogen Hþ-ATPase. The haustorium is a location of extensive vitamin B synthesis and mediates the
suppression of host defense and cell death (the latter possibly by targeting the host bax inhibitor). A reduction of Hechtian strands around the
penetration site might contribute to reduced cell-wall-associated defense responses. The subcellular localization and sites of action of the host
proteins MLO, PMR6 (a potential pectin lyase) and PMR4 (a putative callose synthase) might be the extrahaustorial membrane, the extrahaustorial
matrix and the plant cell wall at sites of infection, respectively. It should be stressed that the projection of findings from studies of a number of
plant–microbe interactions in a single scheme is artificial and does not reflect the realities of any one host–biotroph interaction.

haustorial cell wall is enriched in carbohydrates and is
termed the extrahaustorial matrix ([1,4]; Figure 1).
Although the EHM appears to be contiguous with the
host PM, cytological and biochemical studies (reviewed
in [1,4]) suggest that it differs structurally from the PM.
These earlier studies were extended by a combined
molecular/cytological analysis which demonstrated that
the EHM lacks several proteins that are usually found in
the plant PM. Transgenic Arabidopsis thaliana lines were
identified that expressed translational fusions of Arabi-
dopsis cDNAs with the green fluorescent protein (GFP)
coding sequence that were targeted to the PM [5]. Sub-
cellular changes in GFP fluorescence in eight of these
PM-tagged marker lines were investigated upon powdery
mildew (Erysiphe cichoracearum) inoculation [6]. In each
case, GFP fluorescence was not seen in the EHM when
mature haustoria were formed. A sharp boundary was
visible between the non-fluorescent EHM and the fluor-
escently labeled host PM in the haustorial neckband area,
a collar-like region that represents the junction between
PM and EHM and is thought to seal the extrahaustorial
matrix from the host cytoplasm ([1,3,4]; Figure 1).
There has been a lengthy debate on the potential role(s)
of haustoria. It was suggested that they might serve as
feeding organs or contribute to the establishment or
maintenance of compatibility [4,7,8]. Recent molecular
studies provide accumulating evidence for a pivotal role
of haustoria in nutrient uptake in the broad bean (Vicia
faba)–rust (Uromyces fabae) interaction. A library of rust
genes that are induced in planta has been isolated from a
haustorium-specific cDNA library [9], and includes genes
with high similarity to hexose (HXT1) and amino acid
(AAT1, AAT2, AAT3) transporters from other fungi
[9,10,11

,12]. HXT1 and AAT2 are expressed preferen-
tially, if not exclusively, in haustoria; whereas the expres-
sion of AAT1 is less confined [10,11

,12]. Functional
characterization revealed that HXT1p has a transport
preference for D-glucose and D-fructose, suggesting that
these might be the main carbohydrates assimilated by rust

www.current-opinion.com Current Opinion in Plant Biology 2003, 6:320–326

322 Biotic interactions
haustoria [11

]. Similarly, AAT1p functions as a high-
affinity amino-acid permease that has preference for
histidine and lysine [12]. Indirect data from inhibitor
experiments in yeast and electrophysiological studies in
transfected Xenopus oocytes indicate that the uptake of
hexoses and amino acids involves a proton-symport
mechanism [11

,12]. The required proton gradient might
be generated by a fungal plasma membrane Hþ-ATPase
[13]. The frequencies of cDNAs in the non-amplified
haustorium-specific cDNA library suggest that HXT1,
AAT1, AAT2, and AAT3 transcripts represent about 3%
of total haustorial mRNAs [9]. Collectively, these data
strongly support the idea that the haustoria of U. fabae are
indeed the primary fungal structures for the uptake of
host-derived sugars and amino acids (and can thus be
considered to be ‘hidden robbers’ [3]). Furthermore,
nutrient retrieval is likely to be accomplished by a pro-
ton-symport mechanism.
What amount of nutrients may be absorbed by a sole
haustorium? Transient expression experiments in a pow-
dery mildew (Blumeria graminis forma specialis hordei
[Bgh]) resistant barley mutant have shown the potential
of a single haustorium to funnel sufficient nutrients for a
mildew colony to produce abundant aerial mycelium and
complete its asexual life cycle. Upon inoculation with
powdery mildew spores, only single barley epidermal
cells that had been biolistically transformed with a plas-
mid that conferred susceptibility supported a compatible
interaction, which finally gave rise to a sporulating fungal
colony [14]. The haustorium of primarily epiphytically
growing powdery mildew is supposed to represent the
sole organ of nutrient uptake [3,7]. Furthermore, the
resistant nature of (non-transformed) cells adjacent to
those that supported a compatible interaction did not
permit the formation of secondary haustoria. Hence, the
entire mildew colony seems to gain its nutrients from a
solitary haustorium.
Putative gene products of two genes that are highly
expressed in haustoria (THI1 and THI2), which together
account for about 5% of haustorial mRNAs, are very
similar to proteins that are involved in thiamine (vitamin
B1) biosynthesis [15]. Vitamin B1 is an essential cofactor
for several enzymes of central carbon metabolism. This
suggests an additional role for haustoria in the biosynthesis
of fundamental metabolites that might not be available in
sufficient amounts from the host. The isolation of genes of
a further ascomycete, the wheat leaf rust fungus (Puccinia
triticina), that are induced in planta has been reported
recently [16
]. Homologues of genes that are also found in
the U. fabae library (e.g. THI1, THI2 and cyclophilin) as
well as eight novel genes were identified among 21 rust
transcripts that were induced in planta [16
]. The identi-
fication of homologous genes that are induced in planta
in different biotrophs may indicate a vital and conserved
role in pathogenesis for their encoded polypeptides.
Current Opinion in Plant Biology 2003, 6:320–326
Switching off the alarm bells: defense
suppression for compatibility
It has long been reasoned that pathogens, particularly
those that form long-term biotrophic relationships with
their hosts, may suppress host defense reactions [17].
However, experimental substantiation of this assumption
has been scarce. Recent studies provide accumulating
cytological, pharmacological, phytopathological, and mol-
ecular evidence that obligate biotrophs indeed have the
capacity to disable host defense. Alternatively, it is con-
ceivable that the apparent defense suppression is an
indirect consequence of biotroph-mediated re-program-
ming of host cells so that they enter a developmental
stage or physiological state that does not allow the execu-
tion of defense programs.
One cytological study suggests that biotrophs might
employ distinct strategies for defense suppression.
Cell-wall-associated defense responses, such as the gen-
eration of reactive oxygen intermediates or the formation
of cell-wall appositions, are common non-specialized
reactions to fungal attack that may serve as a first line
of defense against various intruders [18]. Mellersh and
Heath [19

] demonstrated that these general responses
require adhesion between the cell wall and the PM via
thin connections that are known as Hechtian strands. The
disruption of Hechtian strands by specific peptides
reduced cell-wall-associated defense responses and led
to increased frequencies of fungal penetration on host and
non-host plants, suggesting that cytosolic components are
required for the expression of cell-wall-associated defense.
Both compatible host (Erysiphe polygoni) and incompatible
non-host (E. cichoracearum) powdery mildew fungi trig-
gered an increase in the number of Hechtian strands during
the penetration phase on cowpea plants. The cowpea rust
fungus Uromyces vignae, however, caused a transient loca-
lized reduction of cell-wall–PM adhesion sites, which
coincided with a reduction of cell-wall-associated defenses,
on host but not on non-host plants [19

]. These findings
suggest that the rust fungus, in contrast to the powdery
mildews, might have evolved means to suppress the cell-
wall-associated defense responses of its host through a local
reduction of Hechtian strands.
Pharmacological evidence for defense suppression comes
from an analysis in wheat. Application of syringolin A
(a small elicitor-active peptide secreted by the bacterial
pathogen Pseudomonas syringae pv. syringae) to wheat plants
that were infected with powdery mildew (B. graminis f. sp.
tritici) eradicated the fungal infection [20

]. This eradica-
tion coincided with a reactivation of pathogenesis-related
wheat genes. The transcript levels of these genes initially
increase during compatible and incompatible interactions
but then decline during the course of successful infection,
possibly because of defense suppression by the pathogen.
As syringolin A has no apparent fungicidal activity, it was
hypothesized that it revived host defense responses that
www.current-opinion.com

had been suppressed by the powdery mildew fungus
[20

]. The re-activation of effective defense upon appli-
cation of syringolin A argues against defense suppression
being an indirect result of host cells being re-programmed
so as to enter a developmental state that is incapable of
activating defense programs.
Indirect evidence for the suppression of host defense
by fungi in compatible barley–powdery mildew (Bgh)
interactions is provided by the phenomena of ‘induced
accessibility’ and ‘induced inaccessibility’. Single host
cells that were successfully penetrated by a compatible
‘inducer’ isolate were rendered more susceptible to other-
wise weakly virulent or avirulent ‘challenger’ isolates
(‘induced accessibility’). Basal [21], broad spectrum (mlo;
[22]), race-specific (Mla1; [23]) and non-host [24] resis-
tance were shown to be partially compromised in this
manner, suggesting that defense suppression can effi-
ciently disable genetically distinct defense pathways.
Conversely, failure of the ‘inducer’ isolate to penetrate
the host led to enhanced host resistance to a virulent
‘challenger’ isolate (‘induced inaccessibility’) [21,22].
Micro-extraction of RNA from single epidermal cells of
barley plants that were infected with powdery mildew
and subsequent reverse northern blot analysis provided
molecular support for defense suppression in compatible
interactions. Pathogenesis-related genes (e.g. the perox-
idase-encoding gene Prx8, the oxalate oxidase-encoding
gene GRP94, and BH6-12, a gene that encodes a protein
of unknown function) were transcriptionally upregulated
in blots probed with RNA that was extracted from cells on
which fungal sporelings had failed to penetrate the host
cell wall [25]. By contrast, blots probed with RNA
extracted from haustorium-containing cells showed basal
transcript levels of these genes, suggesting that the upre-
gulation is repressed upon successful fungal colonization
(M Lyngkjaer, T Gjetting, unpublished results). Taken
together, these findings indicate that the deactivation of
host defenses appears to correlate with haustorium for-
mation, suggesting that haustoria might play a key role
not only in nutrient absorption but also in mediating
defense suppression (Figure 1).
The host gene flax inducible sequence1 (fis1) is induced in
the course of a compatible flax (Linum utissativum)–flax
rust (Melampsora lini) interaction [26
]. This gene
encodes a putative D1-pyrroline-5-carboxylate dehydro-
genase (P5CDH) that catalyzes a step in the degradation
of proline, specifically the conversion of D1-pyrroline-5-
carboxylate (P5C) to glutamate. Proline is known to
accumulate upon exposure to abiotic and biotic stresses
[27], and elevated levels of proline may serve as an
endogenous stress signal in plants. It has been suggested
that activation of fis1 could provide protection from pro-
line toxicity [26
]. Alternatively, expression of fis1 might
be induced by the biotroph to control proline levels in
www.current-opinion.com
Plant–biotroph compatibility Panstruga 323
(and thus indirectly the stress status of) host tissue. In this
scenario, activation of fis1 would indirectly serve the
purpose of defense suppression.
Keeping the host alive: suppression of cell
death for compatibility
A localized programmed cell death at infection sites (fre-
quently termed the ‘hypersensitive reaction’) is a common
host response to pathogen attack that generally coincides
with cessation of the attempted colonization. As host cell
death is evidently detrimental to the success of the infec-
tion, biotrophs must avoid the triggering of this process by,
for example, elicitors released during cell-wall penetration.
Evidence that host cell death is indeed suppressed during
compatible biotrophic interactions is provided by the
‘green island’ effect: leaf areas around successful infection
sites display delayed senescence in comparison with the
rest of the leaf tissue [28]. Little is known, however, about
how the suppression of cell death might be mediated. It has
been reported that the Pseudomonas type-III effector AvrP-
toB antagonizes programmed host cell death [29]. On the
plant side, bax inhibitor, a protein that is conserved in
eukaryotes and that acts as an anti-apoptotic factor in
animal cells, is one of the few proteins that have been
implicated in the regulation of cell death in plants. Over-
expression of bax inhibitor was shown to suppress the
induction of cell death by fungal elicitors in rice suspen-
sion-cultured cells [30]. Although conceivable, it remains
to be shown whether effector proteins from obligate bio-
trophs are also capable of mediating the suppression of cell
death, and whether bax inhibitor is one of the plant
proteins targeted to achieve this goal.
‘Susceptibility genes’: host genes required
for compatibility
Loss of function of host genes that are required for a
compatible plant–pathogen interaction is generally pre-
dicted to result in incompatibility. Thus, recessively inher-
ited resistance to single or closely related pathogen species
without constitutive activation of defense responses indi-
cate the lack of a pathogen-specific host compatibility
factor. Genetic analysis of resistant lines in natural plant
populations and of induced mutations have revealed single
recessive resistance loci that confer resistance to a wide
range of pathogens in a range of plant species. They
include loci that confer resistance to fungi (mlo in barley
[31], er-1 conferring powdery mildew [Erysiphe pisi] resis-
tance in pea [32], ol-2 conferring resistance to Oidium
lycopersici in tomato [33] and POWDERY MILDEW RESIS-
TANT [PMR] genes in Arabidopsis [34,35

]); oomycetes
(DOWNY MILDEW RESISTANT [DMR] genes in Arabi-
dopsis [M Van Damme, G Van den Ackerveken, personal
communication]); bacteria (xa13 conferring resistance to
Xanthomonas oryzae in rice [36] and RRS1-R in Arabidopsis
[37]); viruses (bc-12 conferring resistance to bean common
mosaic potyvirus in bean [38]) and nematodes (rk3 con-
ferring resistance to root-knot nematode in cowpea [39]).
Current Opinion in Plant Biology 2003, 6:320–326
324 Biotic interactions
For some of these mutants (mlo, pmr, dmr) at least, it has
been demonstrated that resistance is not associated with
the constitutive expression of known defense marker
genes. It remains a formal possibility, however, that the
loss of gene function in these mutants triggers novel and as
yet unidentified defense pathways.
On the basis of the criteria outlined above, barley MLO is
a candidate compatibility factor. A functional Mlo copy is
a prerequisite for basal compatibility of barley and Bgh. In
mlo mutant plants, fungal penetration attempts terminate
within the host cell wall. Barley Mlo encodes the proto-
type of a novel plant-specific family of integral PM
proteins that have seven transmembrane helices
[31,40]. The wildtype MLO protein is predicted to act
as a control element of cell death/senescence and as
modulator of mutually inhibitory defense pathways,
and is possibly exploited by Bgh for ‘molecular docking’
and/or defense suppression (reviewed in [41,42]). Recently,
it has been demonstrated that barley MLO interacts with
calmodulin (CaM), a protein that acts as a cytoplasmic
calcium (Ca2þ) sensor. CaM functions as an in vivo
activator of MLO, and its Ca2þ-dependent binding to
MLO contributes half of the MLO activity in transient
expression-based complementation assays [43
]. A rapid
and transient increase of cytosolic Ca2þ concentration has
been reported as an immediate early host response upon
pathogen attack, and is thought to be required to trigger
various defense responses [44–46]. The decreased com-
plementation efficiency of MLO variants that are defec-
tive in CaM binding suggests that increased Ca2þ
concentration not only might activate defense responses
but also may promote compatibility, at least in the barley–
Bgh interaction. It is possible that the powdery mildew
fungus has evolved means to exploit host defense signal-
ling to its own advantage.
A genetic screen in Arabidopsis for genes that are required
for a compatible powdery mildew (E. cichoracearum) inter-
action revealed 26 mutant plants that exhibited enhanced
resistance to the fungal pathogen without constitutive
expression or infection-induced hyper-activation of known
defense marker genes [34,35

]. These mutants consti-
tute six recessive loci, pmr1–pmr6. The efficiency of
primary fungal penetration was unaffected but mycelial
growth was greatly reduced and sporulation abolished in
all of the analyzed mutants. Resistance was not associated
with host cell death and, with the exception of pmr4
plants that are also resistant to the oomycete P. parasitica,
confined to powdery mildews. Map-based cloning of
PMR6 revealed a gene that encodes a putative pectate
lyase [35

]. This protein has a unique carboxy-terminal
extension of about 90 amino acids that possibly mediates
apoplastic localization [35

]. Vogel et al. [35

] hypothe-
size that the loss of PMR6 might result in pectin accu-
mulation in the EHM, subsequently leading to decreased
nutrient accumulation.
Current Opinion in Plant Biology 2003, 6:320–326
The lack of callose accumulation upon powdery mildew
infection in pmr4 plants gave rise to the idea that these
mutants might be defective in the synthesis of this b-1,3
glucan polymer. Indeed, analysis of the DNA sequence of
a candidate callose synthase gene that is located at the
same relative map position on chromosome IV as pmr4
revealed mutations in the coding region of pmr4 mutants,
strongly suggesting that PMR4 encodes a callose synthase
(M Nishimura, M Stein, S Somerville, personal commu-
nication). In an independent approach, the effect of
double-stranded RNA interference (dsRNAi)-mediated
silencing of callose synthase genes in transgenic Arabi-
dopsis was investigated. Transgenic plants silenced for
one particular callose synthase gene (one of the three
genes selected on the basis of their transcriptional upre-
gulation upon pathogen challenge) showed no detectable
callose deposition at wound and pathogen infection sites.
This callose synthase gene turns out to be identical to the
presumptive PMR4 gene (A Jacobs et al., unpublished
results). Callose deposition at sites of attempted penetra-
tion is a common host response to pathogen attack and was
suggested to serve as a structural barrier to arrest invading
microbes [47]. Counterintuitively, the absence of callose
deposition in the papilla of pmr4 mutants results in
enhanced resistance to powdery mildew disease [34]. This
surprising result might indicate that, contrary to previous
supposition, the presence of callose promotes powdery
mildew pathogenesis in Arabidopsis, possibly by acting
as an inductive cue or by forming a structural scaffold
for the fungus. Alternatively, breakdown products of cal-
lose (e.g. those released during enzymatic degradation by
the fungus) may serve as suppressors of host defenses.
In an approach analogous to that used to identify the pmr
mutants [34], a genetic screen for downy mildew (P.
parasitica)-resistant Arabidopsis plants revealed three dmr
mutants that did not support growth of the oomycete
pathogen and did not exhibit enhanced induced defense
responses (M Van Damme, G Van den Ackerveken, per-
sonal communication). Thus, it is possible that these
mutants are also affected in host genes that are required
to establish a compatible interaction. The map positions of
the three dmr mutants deviate from those described for pmr
mutants [34]. The prevalence of mutants with selective
resistance to either powdery mildews or P. parasitica sug-
gests predominantly distinct host gene requirements for
these biotrophs, possibly reflecting their diverse modes of
infection. However, the fact that the pmr4 mutant is
resistant to both powdery mildews and Peronospora demon-
strates that the requirements also partially overlap.
Conclusions
Despite considerable recent progress, our understanding
of many aspects of plant–biotroph compatibility is still in
its infancy. An exponentially increasing number of patho-
genicity genes have been isolated from phytopathogenic
hemibiotrophic and necrotrophic fungi in recent years
www.current-opinion.com

(reviewed in [48,49]). Nevertheless, the lack of suitable
methods for the molecular analysis of obligate biotrophic
pathogens (e.g. in vitro cultivation, transformation and
gene disruption) has so far hindered the identification of
corresponding genes in these species. Although the first
attempts to establish such procedures have been made [50–
53], current techniques are not robust and there remains an
urgent need to extend the molecular tool kit for biotrophs.
Likewise, knowledge of the mechanisms that govern the
suppression of defense and cell death, as well as those
controlling the redirection of host nutrients [54], is still
sparse. These topics represent additional bottlenecks in
our comprehension of plant–biotroph compatibility. How-
ever, novel techniques that are suitable for the micro-
extraction and subsequent diagnostic analysis of various
molecules (e.g. nucleic acids, metabolites, and proteins)
from single host cells [55,56
] promise to be of great value
in tackling these questions in the future.
Acknowledgements
I am grateful for helpful suggestions on the manuscript proposed by Volker
Lipka and Paul Schulze-Lefert.
References and recommended reading
Papers of particular interest, published within the annual period of
review, have been highlighted as:

of special interest


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For the first time, this study provides convincing evidence that the uptake
of host sugars in plant–biotrophic interactions is likely to occur via fungal
haustoria. The authors describe the molecular analysis of HXT1, which
encodes a bean rust fungus hexose transporter. They demonstrate the
haustorium-specific expression of the HXT1 protein by northern blot
analysis and immunocytology. Transport specificity of HXT1p upon
heterologous expression of HXT1 was convincingly demonstrated in
two different organisms (Saccharomyces cerevisiae and Xenopus
oocytes) by two distinct methods (competition experiments and electro-
physiological studies).
12. Struck C, Ernst M, Hahn M: Characterization of a
developmentally regulated amino acid transporter (AAT1p) of
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13. Struck C, Siebels C, Rommel O, Wernitz M, Hahn M: The plasma
membrane Hþ-ATPase from the biotrophic rust fungus
Uromyces fabae: molecular characterization of the gene
(PMA1) and functional expression of the enzyme in yeast.
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14. Shirasu K, Nielsen K, Piffanelli P, Oliver R, Schulze-Lefert P:
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16. Thara VK, Fellers JP, Zhou JM: In planta induced genes of

Puccinia triticina. Mol Plant Pathol 2003, 4:51-56.
This study provides the second example (following that in U. fabae [9]) of
the analysis of genes of an obligate biotrophic pathogen that are induced
in planta. The availability of a second dataset (although small at present)
will allow the first comparative analysis of genes that are induced in planta
in different biotrophic phytopathogens. It will be interesting to broaden
this comparison and to include non-ascomycetes, such as powdery or
downy mildews, for which equivalent data are currently lacking.
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19. Mellersh DG, Heath MC: Plasma membrane–cell wall adhesion is


required for expression of plant defense responses during
fungal penetration. Plant Cell 2001, 13:413-424.
The authors describe their beautiful cytological study, which provided two
major novel findings. First, they provide evidence that cell-wall-asso-
ciated host defense responses (e.g. the generation of reactive oxygen
species) depend on adhesion between the plant cell wall and the PM via
cytoplasmic strands (known as Hechtian strands). Second, analysis by
sucrose-induced plasmolysis of the state of Hechtian strands during
fungal penetration revealed that rust fungi (in contrast to powdery mil-
dews) might have evolved a means to decrease PM–cell-wall adhesion
during compatible interactions.
20. Wäspi U, Schweizer P, Dudler R: Syringolin reprograms wheat to


undergo hypersensitive cell death in a compatible interaction
with powdery mildew. Plant Cell 2001, 13:153-161.
This paper has two aspects of outstanding interest. First, the authors
describe a study that provides direct evidence for reversible pathogen-
mediated defense suppression in the course of a compatible wheat–
powdery mildew interaction. Second, it demonstrates the potency of a
bacterial elicitor to override installed fungal defense suppression effec-
tively, possibly by synergism with mildew-derived elicitors.
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Current Opinion in Plant Biology 2003, 6:320–326
326 Biotic interactions
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26. Ayliffe MA, Roberts JK, Mitchell HJ, Zhang R, Lawrence GJ,

Ellis JG, Pryor TJ: A plant gene up-regulated at rust infection
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To my knowledge, this is the first report to describe a host gene that is
locally upregulated at compatible biotroph infection sites. Upregulation of
fis1 homologues upon rust infection was found to be conserved in maize
and barley. The protein encoded by this gene is involved in proline
metabolism and might play an indirect role in the suppression of defense
and/or cell death.
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This study provides the first example of the molecular isolation of a gene
that encodes a presumptive host compatibility factor. The gene was
isolated by a targeted search for host mutants that had enhanced pathogen
(in this case, powdery mildew) resistance that was not associated with a
constitutive activation of host defense responses [34]. The exact role of the
encoded protein, PMR6, during a compatible interaction remains spec-
ulative, but map-based cloning of further candidate compatibility genes
promises to shed more light on the basis of biotrophic interactions.
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Current Opinion in Plant Biology 2003, 6:320–326
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This study demonstrates that the same intracellular signal — in this case,
an increase in cytosolic Ca2þ concentrations — is able to serve seemingly
antagonistic purposes. Increases in cytosolic Ca2þ concentrations upon
biotic stress are currently believed to contribute to the activation of host
defenses, but this study shows that Ca2þ-triggered binding of CaM to
MLO enhances the penetration success of the powdery mildew fungus.
44. Mithöfer A, Ebel J, Bhagwat AA, Boller T, Neuhaus-Url G:
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45. Blume B, Nürnberger T, Nass N, Scheel D: Receptor-mediated
increase in cytoplasmic free calcium required for activation of
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46. Romeis T, Ludwig AA, Martin R, Jones JDG: Calcium-dependent
protein kinases play an essential role in a plant defence
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47. Skou JP: Callose formation responsible for the powdery
mildew resistance in barley with genes in the ml-o locus.
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fungi. Mol Plant Pathol 2001, 2:241-255.
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50. Chaure P, Gurr SJ, Spanu P: Stable transformation of Erysiphe
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56. Nakazono M, Qiu F, Borsuk LA, Schnable PS: Laser-capture

microdissection, a tool for the global analysis of gene
expression in specific plant cell types: identification of genes
differently expressed in epidermal cells or vascular tissues of
maize. Plant Cell 2003, 15:583-596.
Laser-capture microdissection is a novel and powerful technique that
enables high-throughput single-cell sampling of particular plant cell types
(e.g. pathogen-infected epidermal and/or mesophyll cells). Single cells
are captured from frozen tissue sections by means of a laser beam. This
technique might pave the way for a thorough molecular analysis of
changes that take place in individual pathogen-infected cells. In this
study, laser-capture microdissection was used to study the expression
profile of particular maize cell types.
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