Oomycetes and Fungi: Similar Weaponry To Attack Plants

462
Review
TRENDS in Microbiology
Vol.11 No.10 October 2003
Oomycetes and fungi: similar

weaponry to attack plants
Maita Latijnhouwers1,2, Pierre J.G.M. de Wit1 and Francine Govers1
1
2
Laboratory of Phytopathology, Wageningen University, Binnenhaven 5, 6709 PD Wageningen, Netherlands

Current address: Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK
Fungi and Oomycetes are the two most important

groups of eukaryotic plant pathogens. Fungi form a
separate kingdom and are evolutionarily related to
animals. Oomycetes are classified in the kingdom
Protoctista and are related to heterokont, biflagellate,
golden-brown algae. Fundamental differences in physiology, biochemistry and genetics between fungi and
Oomycetes have been described previously. These
differences are also reflected in the large variations
observed in sensitivity to conventional fungicides.
Recently, more pronounced differences have been
revealed by genomics approaches. However, in this
review we compare the mode of colonization of the two
taxonomically distinct groups and show that their strategies have much in common.
Together, fungi and Oomycetes cover the majority of
eukaryotic plant pathogens. The modes of colonization
observed among fungal and Oomycete species are
described in Box 1. At first glance the growth patterns of
these two groups of pathogens are similar. For this reason
Oomycetes were long considered a class within the
kingdom Fungi. Both fungi and Oomycetes show filamentous growth in their vegetative stage, produce mycelia and
form spores for asexual and sexual reproduction. However,
taxonomic analyses of phenotypic characteristics and
sequence comparisons have unambiguously shown that
the two groups are placed on different eukaryotic branches
of phylogenetic trees. Based on comparisons of genes
encoding, among others, the small ribosomal subunit,
actin and tubulin, it was concluded that fungi share a
common ancestor with animals. By contrast, the Oomycetes closest relatives are the heterokont golden-brown
algae [1]. Although the reclassification of the Oomycetes is
still under construction, it is no longer disputed that they
are taxonomically unrelated to fungi (Figure 1).
Several important differences between fungi and
Oomycetes have previously been described (Box 2).
Among these are the differences observed in sensitivities
to conventional fungicides. Azole fungicides, for example,
act by inhibition of the ergosterol biosynthesis pathway.
However, Oomycetes do not synthesize ergosterol and are
therefore insensitive to this important group of fungicides
[2]. Because chitin is only a minor component of Oomycete
cell walls [3,4], chitin synthase inhibitors, such as
Nikkomycin and Polyoxin D, also have no inhibitory

effects. Conversely, phenylamides that interfere with RNA
polymerases, such as metalaxyl, are highly selective
against Oomycetes [5].
As their vegetative stage is diploid, and homologous
recombination has not been found to occur, Oomycetes are
far less tractable to genetic manipulation than many fungi.
Of all Oomycetes, Phytophthora is the best-studied genus,
causing diseases not only of the economically important
staple crops such as potato, cocoa and soybean, but also of
valuable forest trees in California and Australia. Useful
resources including genetic linkage maps, bacterial
artificial chromosome (BAC) libraries and expressed
sequence tags (ESTs) of numerous different developmental
stages have been generated in the last few years [6]. In
addition to the conventional polyethyleneglycol (PEG)mediated protoplast transformation, three new DNA
transformation methods have recently been developed.
These are based on zoospore electroporation (B. Tyler and
Box 1. Different lifestyles among fungi and Oomycetes [12]

Biotrophs
Biotrophs grow and reproduce in living plant tissue. They obtain
nutrients through intimate interactions with living plant cells.
Examples of plant-pathogenic fungi with a biotrophic lifestyle are
Cladosporium fulvum, the causal agent of tomato leaf mold, and the
species causing smut diseases, such as Ustilago maydis. Some
biotrophs, for example, the species causing rusts and powdery
mildews, cannot be cultured on artificial media and are therefore
called obligate biotrophs. Obligate biotrophic Oomycetes are found
within the families Peronosporaceae and Albuginaceae, and cause
diseases called downy mildews and white rusts. Obligate biotrophs
usually form haustoria for retrieval of nutrients from plants, whereas
biotrophs such as C. fulvum and U. maydis behave like endophytes
and do not produce haustoria.
Necrotrophs
Necrotrophic species feed on dead plant cells. They kill host tissue
before colonizing it. Cochliobolus and Botrytis species are examples
of fungal necrotrophs, whereas Oomycete necrotrophs are found
among the genera Pythium and Aphanomyces.
Hemibiotrophs
Hemibiotrophs, such as the rice blast fungus Magnaporthe grisea
and several fungal species belonging to the genera Colletotrichum
and Venturia, have intermediate lifestyles. They initially establish a
biotrophic relationship with their host but subsequently, the host
cells die as the infection proceeds. A similar lifestyle is observed in
species of the Oomycete genera Phytophthora and Pythium.
Corresponding author: Francine Govers (Francine.Govers@wur.nl).

http://www.trends.com 0966-842X/$ - see front matter q 2003 Elsevier Ltd. All rights reserved. doi:10.1016/j.tim.2003.08.002
Review
green algae
land plants
heterokont algae
oomycetes
ciliates
dinoflagellates
red algae
animals
basidiomycete fungi
ascomycete fungi
Figure 1. Phylogenetic tree showing the evolutionary relationships between the

major eukaryotic groups. The Oomycetes and the (ascomycetous and basidiomycetous) fungi are highlighted in green. Note the evolutionary distance between the
Oomycetes and the fungi. Reproduced from [71] and adapted from [70].
F. Govers, unpublished), microprojectile bombardment [7]

and Agrobacterium tumefaciens-mediated transformation
[8]. To circumvent the need for homologous recombination
to obtain gene-knockout strains, gene silencing has been
successfully used as a method to accomplish targeted
knockdowns of some genes [9 11], but this still requires
further optimization.
Because of the development of these new methods and
resources, significant progress has been made in elucidating infection strategies of Oomycetes. In this review,
important aspects of fungal and Oomycete pathology are
compared, including histopathology of infection structures, the production of pathogenicity factors, and the role
of conserved signaling pathways in development and
pathogenicity.
Spores and infection structures
Efficient spore production and dispersal is a prerequisite
for successful infection. Both Oomycete and fungal asexual
spores can be dispersed by wind or rain. The asexual
spores of most Oomycetes (sporangia) can undergo
cytoplasmic cleavage, resulting in the formation of
Box 2. Important morphological and physiological

differences between Oomycetes and fungi [13]
Fungi are haploid or dikaryotic during the major part of their
lifecycle, whereas Oomycetes are diploid.
Fungal hyphae are septate, whereas Oomycete hyphae are nonseptate.
Many Oomycetes are (partial) sterol auxotrophs. Their membranes contain lipids with unusual structures and long-chain fatty
acids that presumably replace sterols in mycelial membranes.
Fungi and Oomycetes synthesize lysine by different pathways. The
Oomycetes use the a,1-diaminopimelic acid pathway, whereas
fungi synthesize this amino acid by the so-called a-aminoadipic
acid pathway.
Cell walls of most Oomycetes consist mainly of 1,3-b-glucans,
some 1,6-b-glucans and 1,4-b glucans (cellulose). Chitin, which is a
major constituent of fungal cell walls, has been detected in small
amounts in only a few Oomycetes.
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zoospores: wall-less, uninucleate cells possessing two

flagella of unequal length. Zoospores are dispersed in
water drops, water films or through water-rich soils. The
mechanism by which these spores are attracted to the host
is not fully understood, but there are indications that
chemotaxis, electrotaxis and autoaggregation play a role
[14,15]. Once zoospores have reached the host, they retract
or shed their flagella and become immobile cysts. Fungal
and Oomycete sexual spores can usually survive under
extreme environmental conditions (cold, heat, or drought)
encountered during overwintering or oversummering on
leaf debris or in the soil.
Fungal spores generally use a combination of waterinsoluble glycoproteins, lipids and polysaccharides for host
surface-attachment [16,17]. Magnaporthe grisea secretes
pre-formed adhesive material from the spore tip, whereas
in other fungi, spore attachment requires de novo protein
synthesis. Adhesive substances of the fungi Blumeria
graminis and Uromyces viciae-fabae include enzymes that
can modify the surface of the host plant to improve spore
attachment [16]. Germinating sporangia of the Oomycete
Peronospora parasitica have been shown to secrete an
adhesive layer containing glycoproteins and b-1,3-glucans
that might contribute to germling attachment [18]. The
Oomycetes Pythium and Phytophthora store adhesive
material in small vesicles inside zoospores, which is
released upon encystment [19]. In addition, germinating
cysts of Phytophthora infestans produce proteins that
contain many repeats that have homology with human
mucins, and are thought to play a role in the protection of
germlings from desiccation or physical damage [20].
Some fungi, such as the tomato leaf mould fungus
Cladosporium fulvum, avoid breaching the cell wall by
entering the plant tissue exclusively through stomata and
remain confined to the apoplastic space. Other fungi, and
many Oomycetes, form specialized infection structures
named appressoria to enter the plant tissue (Figure 2a c).
An appressorium is a swelling of the tip of a germ tube
from which a penetration peg emerges that serves to pierce
the epidermal cell wall or to enter the epidermis through
stomatal apertures [16,17]. Many fungi require a hard,
hydrophobic surface for the induction of appressorium
formation and the topology of the surface, such as the
presence of ridges, can be an important trigger (thigmotropism) [21]. Fungal appressoria are usually separated
from the spore by a septum. Appressoria of M. grisea
accumulate high concentrations of glycerol causing a
turgor pressure of up to 8 MPa [16]. The presence of
melanin in appressorial cell walls is also required for
turgor pressure to buildup [16]. During the past five years,
many genes have been identified that are required for
appressorium formation, turgor generation or for appressorial penetration in M. grisea and an overall picture of
appressorium function is beginning to emerge [16,22 24].
Although many Oomycetes produce appressoria, those
of Phytophthora, Pythium and Peronospora species are
generally small compared with those of the fungi [25,26].
They are not melanized or pigmented, and the appressoria
are only separated from germ tubes by so-called false
septa, which do not actually divide the two cells [18,25,27].
As in fungi, topological features on the surface of host
464
(a)
Review
the case of infection by the rust fungus Puccinia

hemerocallidis, the extrahaustorial membrane was
recently reported to form tubular extensions into the
plant cytoplasm, possibly to enlarge the interacting
surface [31]. Genes that encode hexose and amino acid
transporters and thiamine biosynthesis genes are highly
expressed in haustoria of the rust fungus U. viciae-fabae,
adding credence to the idea that haustoria play a central
role in primary metabolism and the importing of nutrients
[32,33]. Uptake of nutrients was proposed to be driven by a
proton gradient generated by H-ATPases [34]. Haustoria
of biotrophic Oomycetes such as Pe. parasitica are similar
in size to those of rust fungi, but the organization,
structure and composition of the electron-dense neckband
are different [35]. By contrast, hemibiotrophic Oomycete
species belonging to the genera Phytophthora and
Pythium form small finger- or digit-like haustoria during
the biotrophic phase [27,36] (Figure 2g).
Cytological examinations suggest that many of the
events following spore and cyst germination are highly
similar in fungi and Oomycetes. However, more in-depth
molecular and biochemical analyses will be required,
especially in Oomycetes, to enable detailed comparisons of
the composition of appressoria and extrahaustorial
matrices and the identification of transport proteins and
mechanisms in haustoria.
(b)
(c)
(d)
(f)
(e)
(g)
(f)
Figure 2. Infection structures of plant-pathogenic fungi (a,d,f) and Oomycetes

(b,c,e,g). (a) Cryoscanning electron micrograph of mature appressorium (indicated
by an asterisk) of Magnaporthe grisea (micrograph kindly provided by Richard
J. Howard, DuPont Crop Genetics). (b) Light microscopy showing a cyst of Phytophthora infestans germinated on an artificial surface. At the tip of the germ tube
an appressorium is formed (indicated by an asterisk). (c) Cryoscanning electron
micrograph of P. infestans cyst (left) with the germ tube forming an appressorium
(indicated by an asterisk). Site of penetration is adjacent to a stoma [72]. (d) Haustorium of Erisyphe graminis f. sp. hordei with lobed structure in epidermal host
cell of Hordeum vulgare, stained with DiOC7, as viewed by differential interference
microscopy [73]. (e) Micrograph showing fluorescent trypan blue-stained tissue
containing branching hyphae of P. infestans with haustoria in the mesophyll cells
of the potato cultivar Bintje [74]. (f) Transmission electron micrograph showing
haustorium of E. graminis f. sp. hordei in H. vulgare [75]. (g) Transmission electron
micrograph showing haustorium of P. infestans in the potato cultivar Majestic
[36]. Note digit-like shape of P. infestans haustoria in (e) and (g). Scale bars (a)
5 mm, (c) 8 mm, (d) 20 mm, (e) 35 mm, (f) 1 mm, (g) 2 mm.
tissues, such as ridges and irregularities, can induce

formation of appressoria in Pe. parasitica, P. infestans,
Phytophthora palmivora and Phytophthora sojae
[14,25,28,29]. In contrast to fungi, turgor pressure in
Oomycete appressoria has not been quantified, nor has the
chemical composition of the low molecular weight solutes
in appressoria been analyzed.
Intercellular hyphae of biotrophic fungi, as well as
Oomycetes, form spherical or lobed structures named
haustoria, which penetrate the adjacent plant cells
(Figure 2d g). Haustoria are structures specialized in
retrieving nutrients from the host plant [30]. At the site of
haustorium formation, the plant plasma membrane
invaginates and surrounds the pathogens cell wall. The
electron-dense material between the pathogen cell wall
and the plant plasma membrane is known as the
extrahaustorial matrix. A so-called neckband separates
the extrahaustorial matrix from the plants intercellular
space to prevent leakage of the material in the matrix. In
Pathogenicity factors
Compounds synthesized by pathogens are designated
pathogenicity factors when they are essential for the
successful infection and colonization, and are called
aggressiveness factors when they contribute to the
efficiency of the infection process. Whether or not a
compound is a pathogenicity or aggressiveness factor can
be established by gene disruption or gene silencing, which
should result in non-pathogenic mutants or less pathogenic mutants, respectively. Cell-wall-degrading enzymes
(CWDEs), proteins involved in protection against plant
defense compounds (counter-defense), and toxins are
examples of pathogenicity or aggressiveness factors.
Pathogen-derived compounds can also induce the arrest
of pathogen infection. Many of these so-called elicitors, or
avirulence factors, were discovered owing to their ability to
elicit numerous defense responses that culminate in host
plant resistance to pathogen attack [37,38]. Some of these
compounds might in fact have intrinsic functions as
pathogenicity or aggressiveness factors, but, as plants
have evolved the ability to recognize them, they have
become the telltale signals of the pathogens presence
(Box 3) [39]. For this reason, they were initially called
avirulence factors but gradually the terminology is
changing; the word effector is taking root.
Cell-wall-degrading enzymes
CWDEs are often expressed in germ tubes and infection
hyphae, which suggests that they are used to loosen the
plant cell wall, facilitating successful penetration [40]. By
contrast, necrotrophic fungi are thought to use CWDEs to
break down the plant cell wall and feed on the released
nutrients. A large number of fungal CWDE genes that
encode polygalacturonases, pectin-lyases, pectinases,
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465
Box 3. Elicitors
Fungal and Oomycete elicitors were thoroughly reviewed recently
[38,55]. Elicitors are pathogen-derived molecules, mostly secreted
proteins, that are specifically recognized by plants. The recognition
event results in the triggering of efficient defense responses, often
including a hypersensitive response (HR). It is assumed at present that
many elicitors are pathogenicity or aggressiveness factors, and that
plants have evolved the ability to recognize such factors at a later stage
of (co)evolution with their pathogens [39]. This would explain why
elicitors form such a heterogeneous group of molecules. However, as a
consequence of this heterogeneity, searching for Oomycete elicitors
based on homology with those of fungi, or vice versa, is not expected to
be a successful approach. To date there is only one example of a fungal
elicitor that has counterparts in Oomycetes. However, it also has a
counterpart in bacteria, which suggests a horizontal gene transfer event
across kingdoms. This elicitor was initially identified as a necrosisinducing protein in Fusarium oxysporum [56]. The Oomycete counterpart was discovered when a secreted Phytophthora protein with elicitor
activity was sequenced [57]. The gene encoding this protein showed up
independently in a computational analysis that selected for those genes
that encode proteins secreted by Phytophthora species. This also
demonstrates that such an approach can be used successfully to identify
putative elicitors [58 60]. Unraveling the elicitors intrinsic functions
and their roles in pathogenicity can be very informative to understand
the infection strategies of pathogens. Table 1 gives an overview of: (i)
putative functions of the few elicitors known to date that are
homologous to known proteins in the databases; and (ii) the elicitors
that have a role in pathogenicity or aggressiveness as shown by the
disruption or silencing of the encoding gene.
Table 1. Putative functions of elicitors with homology to known proteins and elicitors of which disruption or silencing of the
encoding gene revealed a role in pathogenicity or aggressivenessa
Elicitor
Organism
Putative function
Gene disrupted or silenced
Role in pathogenicity or
aggressiveness?b
Fungi
AvrPita
ECP1
ECP2
AVR4
Magnaporthe grisea
Cladosporium fulvum
Cladosporium fulvum
Cladosporium fulvum
No
Yes
Yes
Yes
NIP1
ACE1c
Rhynchosporium secalis
Magnaporthe grisea
zinc metallo protease

Unknown
Unknown
chitin binding or protection
against chitinases
unknown
polyketide synthese or nonribosomal
peptide synthetase
Yes
No
Yes
Oomycetes
INF1d
GPE1
Phytophthora infestans
Phytophthora sojae
Yes
No
No
CBEL
Phytophthora parasitica
sterol binding
transglutaminase; crosslinking of cell
wall proteins
cellulose binding glycoprotein
Yes
No
Yes
Yes
No
Refs
[48]
[10]
references [38,55] unless otherwise indicated.

only relevant for the genes of which the function in pathogenicity was investigated by gene disruption or silencing.
it is hypothesized that ACE1 is an enzyme involved in the synthesis of a so far unknown secondary metabolite elicitor (I. Fudal et al. XXII Fungal Genetics Conference,
Asilomar, CA, 2003).
d
INF1 belongs to a class of proteins named elicitins. The sterol binding property of this class of proteins was initially shown for elicitins from other Phytophthora species [61].
b
c
cellulases, xylanases and glucanases have been cloned

[41]. Targeted disruption of many of these genes clearly
shows their role in fungal pathogenicity [41]. However,
there are several examples where the disruption or
replacement of genes encoding CWDEs does not alter
pathogenicity or aggressiveness, possibly because these
genes are redundant and other isozymes encoded by
paralogous genes are able to take over the function of the
defective enzyme [41].
Oomycete species of the genera Phytophthora and
Pythium are known to produce and secrete CWDEs in
liquid cultures and on solid media [26,42]. Recently, a gene
that encodes an extracellular endopolygalacturonase was
identified in P. infestans. Polygalacturonases (PGs) are
pectinases that hydrolyze the homogalacturonan backbone of pectin [41]. The P. infestans PG gene Pipg1 is
expressed in germinating cysts, suggesting that this PG
plays a role in the initial stage of infection. However, the
gene is also expressed during all stages of growth in
tomato. Therefore, the same enzyme could be involved in
inducing or maintaining necrotrophic growth [43]. A
remarkably large PG gene family that has 19 members
was identified in Phytophthora cinnamomi [44]. In
addition, five genes that encode exo- and endo-1,3(1,4)-bglucanases were cloned from P. infestans, and lowstringency hybridization studies indicated the presence
of additional glucanase genes in the P. infestans genome
[45]. Phytophthora PGs and glucanases are more similar to
the corresponding fungal and insect enzymes than to their
plant counterparts, which seems to contradict the previously described evolutionary distance between Oomycetes and fungi. However, the enzymes probably perform
similar functions and encounter a similar selection
pressure. It has therefore been suggested that sequence
convergence could explain the similarities between the
Phytophthora and fungal CWDEs [43 45].
Plants produce CWDEs directed against fungal and
Oomycete cell walls. These CWDEs presumably function
by affecting the invaders cell wall integrity [41]. In
addition, CWDE-mediated release of oligosaccharides
from the pathogens cell wall is known to elicit defense
responses in plants [46]. To escape from damage by plant
glucanases, the Oomycete P. sojae secretes proteins that
can inhibit those CWDEs. One of the glucanase inhibiting
proteins, GIP1, specifically inhibits EgaseA, the soybean
endo-b-1,3-glucanase that is responsible for the release of
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Review
glucan oligosaccharide elicitors from the pathogens cell

wall. By secreting this protein into the apoplast, P. sojae
prevents the release of oligo-b-glucans and therefore
avoids or slows down early recognition by the host [47].
Similarly, P. infestans has evolved mechanisms to inhibit
the activity of plant extracellular proteases. This pathogen
secretes protease-inhibiting proteins, one of which, EPI1,
interacts with and inhibits P69, which is a tomato serine
protease involved in plant defense. Homologues of this
protein have not been found in fungi (M. Tian and
S. Kamoun, XXII Fungal Genetics Conference, Asilomar,
CA, 2003). Fungi have also evolved strategies to escape
from the action of plant CWDEs, in particular plant
chitinases. AVR4 is an extracellular protein produced by
C. fulvum while growing in planta. It contains a domain
that, by binding to chitin, can protect the chitin in the cell
wall of C. fulvum against the deleterious effects of basic
tomato chitinases. AVR4 is also an avirulence factor, but
its primary function probably involves a passive defense
against plant chitinases [48]. AVR4 can also protect other
fungi such as Trichoderma viride and Fusarium solani
against the lytic effects of basic plant chitinases. Results
show that Oomycetes and fungi have both found ways to
defend themselves against plant CWDEs and other plant
extracellular enzymes involved in defense, some by
binding to the enzyme itself, but in the case of AVR4, by
binding to chitin and preventing the enzyme binding to
and acting on its substrate.
Inactivators of plant defense compounds and toxins
Plants not only produce CWDEs to inhibit pathogen
proliferation, they also synthesize antimicrobial secondary
metabolites such as phytoanticipins (pre-existing) and
phytoalexins (induced by pathogens) for the same purpose.
However, fungi have developed ways to prevent damage by
these compounds. Some fungi produce enzymes that
detoxify antimicrobial plant secondary metabolites, some
of which have been shown to contribute to pathogenicity
[49]. Another mechanism of tolerance to antimicrobial
plant compounds in plant-pathogenic fungi seems to be
based on the active secretion of these compounds. ABCtransporters are membrane pumps that are involved in
transporting a wide variety of compounds over the plasma
membrane: ABC-transporter knockout strains of Botrytis
cinerea and Gibberella pulicaris are more sensitive to
secondary metabolites of their respective hosts, which
suggests that ABC-transporters actively secrete these
compounds. The aggressiveness of these knockout strains,
as well as the ABC-transporter disruptants of Mycosphaerella graminicola and M. grisea, is thereby reduced
[50]. Often, Oomycete plant pathogens have to cope with
the same antimicrobial compounds in their hosts as these
fungi, which suggests that they too have evolved mechanisms to withstand them. To discover counter-defensive
mechanisms that occur in pathogenic Oomycetes, the
mechanisms described for fungi should be fully explored.
Toxin biosynthesis also remains unexplored in Oomycetes. Some necrotrophic fungi produce toxins that kill
host cells, and the genes involved in their biosynthesis
have been studied in detail. Two important groups of
fungal phytotoxins are non-ribosomally synthesized linear
or cyclic peptides and polyketide derivatives [51]. Peptide

and polyketide synthetases involved in the synthesis of the
respective groups of toxins contain domains that are
conserved among different fungal species. A large number
of Oomycete pathogens are necrotrophic or have a
necrotrophic growth phase. Therefore, the possibility
that they too use toxic compounds to kill their host cells
should be considered.
Plants produce reactive oxygen species (ROS) as an
early response to infection. These are frequently associated with the hypersensitive response (HR) [52]: a defense
response that involves plant cell death around the site of
infection. ROS are thought to be involved in limiting
pathogen growth. Enzymes produced by pathogens, such
as superoxide dismutases, catalases and peroxidases, are
thought to be involved in the breakdown or inactivation of
ROS. Several genes encoding ROS-inactivating enzymes
have been cloned from fungi. However, their role in
pathogenicity or aggressiveness remains uncertain [53].
Sequences derived from ROS-inactivating enzymes were
identified in Phytophthora EST databases, indicating that
Oomycetes can also defend themselves against damage by
host-generated ROS [54].
Signal transduction involved in pathogenesis
Recently, signaling pathways that have been conserved
throughout evolution and underlie fungal development
and pathogenicity have received much attention. These
pathways transmit extracellular signals across the plasma
membrane to the interior of the cell, allowing the pathogen
to respond adequately and timely to changes in its
environment [62]. The plant-pathogenic fungi Cryphonectria parasitica, M. grisea and Ustilago maydis have served
as models for the dissection of the G-protein, cAMP and
MAPK (mitogen-activated protein kinase) signaling pathways. Signaling through heterotrimeric G-proteins controls a wide range of developmental processes, including
conidiation, vegetative growth, fertility, dimorphic growth
and appressorium development, and plays a role in
infectious, invasive growth [62,63]. In addition, a
B. cinerea Ga subunit (BCG1) is involved in the secretion
of proteases [64].
There is ample evidence that G-proteins feed into the
cAMP pathway in several different fungal plant-pathogens; the addition of pathway constituents was able to
restore normal development in fungal strains that
contained mutations in the G-protein subunits. Also,
strains with mutations in the catalytic subunits of
cAMP-dependent protein kinase (PKA) or adenylyl cyclase
displayed phenotypes similar to those of G-protein deletion
mutants [62].
The role of MAP kinases in the mating pathway and
pathogenicity of U. maydis has been extensively studied
[62,65]. In addition, the MAPK PMK1 of M. grisea plays a
key role in the onset of appressorium formation, in the
transfer of storage carbohydrate and lipid reserves to the
appressorium, and is also required for pathogenicity ([66]
and reviewed in [62,67]). Another MAPK of M. grisea,
MPS1, is thought to be involved in cell-wall integrity and is
indispensable for appressorial penetration. The role of
Review
orthologues of PMK1 in pathogenicity has been studied in

a wide range of other fungal species [67].
P. infestans is the only Oomycete in which the functions
of G-protein subunits have been analyzed. Silencing of the
Gb subunit gene Pigpb1 resulted in a defect in sporangia
formation [11], reminiscent of the sporulation defect
observed in the C. parasitica Gb disruptant strain [68].
Mutants deficient in the Ga subunit PiGPA1 show a
reduction in zoospore release and their aggressiveness is
severely impaired. The zoospores of the PiGPA1-deficient
mutants change direction frequently and fail to aggregate
or be attracted to chemotactic compounds. This suggests
that PiGPA1 plays a role in the sensing capabilities
of zoospores that are required to control motility
(M. Latijnhouwers et al., XXII Fungal Genetics Conference, Asilomar, CA, 2003). As the genes that encode the
respective kinases are well represented in the Phytophthora EST databases, a role for the cAMP and the
MAPK pathway downstream of G-proteins is conceivable,
but this has not yet been investigated.
It can be concluded that both Oomycetes and fungi rely
on a functional G-protein pathway for normal development and pathogenicity, whereas downstream pathways
and targets of G-proteins have been studied only in fungi.
To better understand how environmental cues affect
development in these pathogens, the identification of
receptors and ligands deserves more attention in future
research.
Concluding remarks
In spite of the distinct evolutionary origin of the two
groups, fungi and Oomycetes use infection strategies that
have much in common. Their specialized infection structures (appressoria, infection hyphae and haustoria) share
many features. A signaling pathway (the heterotrimeric
G-protein pathway) that is a key-regulator in development
and pathogenicity of fungi also seems to govern crucial
processes in Oomycetes. Moreover, they both use an
extensive toolbox of CWDEs to penetrate into and to
degrade plant cell walls. This in itself may not be
surprising, but the protein sequences of CWDEs of
fungi and Oomycetes are more conserved than would
be expected from their evolutionary relationship alone.
This high degree of similarity in attacking and
defensive mechanisms suggests that the in planta
conditions are very similar for plant-pathogenic fungi
and Oomycetes where they encounter a wide range of
preformed and induced antimicrobial compounds. Thus,
convergent evolution seems to have forced the development of similar infection strategies in these two
types of plant pathogens.
Horizontal gene transfer might also have occurred,
either between the fungi and Oomycetes, or between these
organisms and other species. It is hypothesized that fungi
have acquired gene clusters from other organisms [69] and
that the occurrence of homologues of the Fusarium
oxysporum f.sp. erythroxyli necrosis-inducing protein in
many Oomycete species might have been the result of
horizontal gene transfer [59]. However, the frequency of
occurrence of this phenomenon and its impact on evolution
is not clear.
467
Important differences between the infection strategies of fungi and Oomycetes involve classes of
pathogenicity or aggressiveness factors that occur
only in one of the two groups of plant pathogens. For
example, antimicrobial plant secondary metabolitedetoxifying enzymes and fungal phytotoxins have not
yet been detected in Oomycetes. Nearly complete genomic
sequences of plant-pathogenic fungi have recently been
released (http://www-genome.wi.mit.edu/seq/fgi/). In
addition, funds have been allocated to produce genomic
sequences of P. sojae and P. ramorum and recently random
shotgun reads of the P. sojae genome corresponding to a
depth of 8X have been released (http://www.jgi.doe.org).
Complete sequence information for these Oomycetes will
be very useful in aiding the discovery of novel pathogenicity and aggressiveness factors, based on either homology
with fungi, or in high-throughput functional genomics
approaches that employ gene silencing.
Acknowledgements
We thank the Dutch Science Foundation, Council Earth and Life Sciences
(NWO-ALW) for financial support. Thanks to Andre van t Slot and Arjen
ten Have for critically reading the manuscript.
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Vol.11 No.10 October 2003