Review

Blackwell Publishing Ltd

Tansley review
A journey through signaling in
arbuscular mycorrhizal symbioses 2006

Author for correspondence:

Uta Paszkowski
Tel: +41 22 379 3022
Fax: +41 22 379 3107
Email: uta.paskowski@bioveg.unige.ch

Uta Paszkowski
University of Geneva, Department of Plant Biology, 1211 Geneva, Switzerland

Received: 5 April 2006

Accepted: 6 June 2006

Contents
Summary

35

IV. Mature symbiotic phase haustoria and mineral nutrition

41

I.

Introduction

35

V.

Concluding remarks

43

II.

Presymbiotic dialogue recognition and anticipation

36

Acknowledgements

43

39

References

43

III. Early symbiotic phase contact and penetration

Summary
Key words: arbuscular mycorrhizal
symbiosis, Glomus, nitrogen, phosphate,
signaling.

Recent years have seen fascinating contributions to our understanding of the

molecular dialogue between fungi and plants entering into arbuscular mycorrhizal
(AM) symbioses. Attention has shifted from descriptions of physiological and cellular
events to molecular genetics and modern chemical diagnostics. Genes, signal
transduction pathways and the chemical structures of components relevant to the
symbiosis have been defined. This review examines our current knowledge of signals
and mechanisms involved in the establishment of AM symbioses.
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The Authors (2006). Journal compilation New Phytologist (2006)
doi: 10.1111/j.1469-8137.2006.01840.x

I. Introduction
The arbuscular mycorrhizal (AM) symbiosis occurs between
fungi of the Glomeromycota (Schssler et al., 2001) and the
majority of terrestrial plants. It is the most prevalent mutualistic
association between plants and microbial organisms and involves
an intimate relationship between plant roots and fungal hyphae.
The mutualism of the AM symbiosis is manifested in bidirectional nutrient exchange: the fungus is nourished by plant
photosynthates, and plant mineral nutrition particularly

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phosphate is enhanced by the fungus (reviewed in Smith &

Read, 1997). AM fungi are obligate biotrophs, depending
on living root tissue for carbohydrate supply to complete their
asexual life cycle. Although typically the outcome of the AM
association is beneficial for both partners, the effectiveness
varies in individual plantfungus combinations (Ravenskov
& Jakobsen, 1995; Burleigh et al., 2002; Smith et al., 2003).
In natural habitats, this influences ecosystem variability and
productivity (van der Heijden et al., 1998). The Glomeromycota
consists of approximately 150 isolates, which colonize a wide

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range of both mono- and dicotyledonous plant species. Under

laboratory conditions, single fungal isolates do not exhibit host
specificity and will associate with taxonomically distant plants.
The widespread distribution of this symbiosis within the plant
kingdom has been attributed to its antiquity; fossil records of
mycorrhizal land plants place the origin of the AM symbiosis at
least 400 million years ago (mya) (Remy et al., 1994), predating
the divergence of plant lineages. Nonmycorrhizal plant species
arose later in evolution, dating back to 100 mya (Brundrett,
2002). Their occurrence within different plant clades suggests
loss of compatibility with AM fungi to be of polyphyletic
origin (Brundrett, 2002).
The sequence of steps leading to an AM symbiosis is largely
conserved among different combinations of fungal and plant
species and appears to be independent of the effectiveness of
the symbiosis. The process can be divided into major developmental stages: (i) the presymbiotic phase; (ii) contact and
entrance of the fungus into root tissue; (iii) intraradical fungal
proliferation; and (iv) cell invagination and nutrient transfer.
These stages describe a chronological series of events at an
individual infection site. However, establishment of the AM
symbiosis in the whole root is highly asynchronous, with the
described stages all present simultaneously once the fungus has
commenced root penetration.
Recognition in concert with complex morphological and
physiological alterations of both symbiotic partners suggests
that the AM symbiosis is the result of multifaceted, fine-tuned
signaling events. Forward genetic screens have allowed estimation of the scope of processes under plant control, and genes
vital to the establishment of the symbiosis have been isolated.
Application of genomic tools to molecular studies of AM
symbioses has revealed a multitude of potentially relevant
plant genes that respond to the development of the symbiosis.
The absence of efficient transformation protocols and a lack
of genetic tools for asexual fungi have so far hindered the
description of signaling events in the fungus. This article
summarizes the progress made in understanding AM symbiosis
signaling since the reviews by Harrison (2005) and Hause &
Fester (2005).

II. Presymbiotic dialogue recognition and

anticipation
1. Fungal responses to plant-derived signals
For both symbionts, the period before physical contact (appressoria
formation) involves recognition and attraction of appropriate
partners and other events promoting an alliance. The survival
of the biotrophic fungus is enhanced by economic germination
and rapid colonization of host plants. Spores of AM fungi
persist in the soil and germinate spontaneously, independent
of plant-derived signals (Mosse, 1959). However, root exudates
and volatiles may promote or suppress spore germination,
indicating the existence of spore receptors responsive to altera-

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tions in the chemical composition of the environment (reviewed

in Giovanetti & Sbrana, 1998; Bcard et al., 2004; Harrison,
2005). After germination, the hyphal germ tube grows through
the soil. In the absence of a potential host (asymbiotic phase),
hyphal growth is limited by the utilization of low amounts of
stored carbon (Bcard & Pich, 1989; Bago et al., 1999; Bago
et al., 2000) and eventually ceases; however, the spore retains
sufficient carbon to allow repeated germination and further
possibilities to encounter an appropriate host. The particularly
large spores of Gigaspora gigantea can germinate up to 10 times
(Koske, 1981).
In the vicinity of a host root, fungal morphology shifts towards
enhanced hyphal growth and extensive hyphal branching
(Fig. 1a) (Giovanetti et al., 1993b; Buee et al., 2000). Such a
response can be triggered by host root exudates or volatiles
but not by exposure to nonhost roots or their exudates. These
observations suggest that the fungus senses a host-derived
signal (branching factor), leading to intensified hyphal ramification that is likely to increase the probability of contact
with a host root. Hence, distinction between host and nonhost
occurs to a certain degree at this early point in the interaction.
A recent major breakthrough was the identification of the host
branching factor 5-deoxy-strigol, belonging to the strigolactones
(Akiyama et al., 2005). Strigolactones have been isolated from
a wide range of mono- and dicotyledonous plants and were
previously found to stimulate seed germination of parasitic
weeds such as Striga and Orobanche (reviewed in Bouwmeester
et al., 2003). The concentration of strigolactones in root
exudates coincides with the host specificity of AM fungi
(Akiyama et al., 2005). The nonmycotrophic Arabidopsis
thaliana, for example, produces very low amounts of strigolactones compared with hosts of AM fungi such as carrot and
tobacco (Westwood, 2000). The biosynthetic pathway for
strigolactones is not well understood. They were previously
described as sesquiterpenes (Yokota et al., 1998; Akiyama et al.,
2005); however, the use of carotenoid mutants of maize and
inhibitors of isoprenoid pathways in maize, sorghum and
cowpea showed that strigolactones are derived from the
carotenoid biosynthetic pathway, via the plastid, nonmevalonate, methylerythritol phosphate (MEP) pathway (Matusova
et al., 2005). Interestingly, the carotenoid cleavage products
mycorradicin (yellow pigment) and cyclohexenones specifically accumulate in mycorrhizal roots (Klingner et al., 1995;
Maier et al., 1997; Walter et al., 2000). Use of the same maize
carotenoid mutations revealed the lack of mycorradicin production and a reduction in mycorrhizal colonization (Fester
et al., 2002). These authors examined later stages of the AM
interaction. It will now be interesting to analyze the early
symbiotic properties of these maize mutants in relation to
strigolactone as a stimulant of presymbiotic fungal branching.
Nevertheless, the results do indicate that processed carotenoid
derivatives are involved at multiple stages in the development
of the AM symbiosis, possibly by stimulating intraradical fungal
branching.

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Review

Fig. 1 Stages of root colonization by an arbuscular mycorrhizal (AM) fungus. (a) Hyphal branching occurs upon perceiving plant-released
strigolactone; (b) pENOD11::GUS expression upon perceiving Myc-factor(s); (c) appressoria formation and passage through outer root cell
layers; (d) longitudinal apoplastic fungal spreading; (e) arbuscule formation in the inner cortex. Microphotographs display chlorazole black E
stained rice roots colonized by Glomus intraradices. Bars, 25 mm.

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Tomato mutants impaired in the development of AM

symbiosis with Gigaspora and Glomus isolates were shown to
be affected in the presymbiotic stimulation of spore germination and hyphal growth (David-Schwartz et al., 2001, 2003).
The conclusion could be drawn that the lack of a stimulant
caused the mutant phenotype. However, the same group
observed that exudates of mutant root suppressed spore
germination and hyphal tip growth, which they attributed to
the presence of an active inhibitory compound (Gadkar et al.,
2003).
Although the fungal strigolactone receptor has not yet been
isolated, the consequences of strigolactone detection were
investigated in Gigaspora rosea and Glomus intraradices exposed
to exudates from carrot roots (Tamasloukht et al., 2003). Hyphal
branching commenced shortly after initial rapid changes
in transcript abundance of putative mitochondrial genes,
followed by enhanced oxygen consumption and reducing
activity. Hence, induction of a subset of genes appeared to lead
to elevated respiratory activity and an energy status necessary
for extensive hyphal branching (Tamasloukht et al., 2003).
Thus, upon detection of a host root there is vigorous investment in the production of fungal hyphae, which can then
rapidly make contact with essential carbon sources (Bcard
et al., 2004).
2. Plant responses to fungus-derived signals
The plant responds to the microbial profile of the rhizosphere
in different ways, depending upon the type of organism present.
Detection of pathogen-derived elicitors triggers plant signaling
cascades that lead to a defense response, a phenomenon that
has been widely studied (for a recent review, see Glazebrook,
2005). On the other hand, plant defense responses are either
not mounted at all or mounted only transiently before being
suppressed during AM symbiosis (reviewed in Harrison, 2005;
Hause & Fester, 2005). Little is known about AM fungal
signal(s), their perception or the consequences triggered in
the plant. One study used an inducible transgenic reporter line
(Journet et al., 2001) to document the detection of an as yet
unknown fungal factor (Kosuta et al., 2003). GUS reporter
expression before contact was monitored in root sections
adjacent to intensely branching hyphae (Kosuta et al., 2003).
A cellophane membrane was placed between the two organisms
to prevent physical contact while allowing exchange of signals.
The intensity and distribution of GUS expression indicated the
detection by the plant of a compound released by the fungus
(Fig. 1b). This putative Myc factor (in analogy to Nod factor)
was produced by the AM fungi tested but was absent from three
pathogenic fungi (Kosuta et al., 2003). Interestingly, when
contact and penetration were permitted, GUS expression was
restricted to infected and associated cells (Chabaud et al., 2002;
Genre et al., 2005), indicating the induction of a suppressor
activity in noncolonized neighboring cells (Parniske, 2004).
Thus, it is possible that a larger region of the plant root

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becomes primed for an encounter with the fungus by the

turning on of a symbiont anticipation program that includes
MtENOD11 expression. When contact is made and a
precise area of penetration and colonization is established,
gene expression is restricted to cells in direct contact with the
penetrating fungus. Similarly, during preinfection and infection
stages in the interaction of Medicago truncatula with nitrogenfixing Rhizobia, GUS expression was found in the rhizodermis
of larger root section before contact and upon subsequent
bacterial entrance into the root tissue becomes confined to
the area of infection, namely the infection site and the invaded
nodule (Journet et al., 2001). This resemblance in expression
patterns between the two types of root symbioses suggests that
the responses are part of a rather general symbiont anticipation
program. The M. truncatula mutant Does not make infections 2
(dmi2 ) does not support fungal penetration (Catoira et al., 2000).
However, hyphal branching and MtENOD11 expression before
contact is induced to levels comparable to that of wild-type
plants, indicating that the presymbiotic properties of dmi2
are not affected (Chabaud et al., 2002). However, no GUS
expression was monitored in dmi2 upon contact with the fungus.
Therefore while presymbiotic anticipation signaling operates
independently of MtDMI2 the signaling pathway involved in
contact-associated MtENOD11 expression requires functional
MtDMI2.
Interestingly, an independent report showed induction
in the number of lateral roots of M. truncatula in response to
perception of a diffusible factor from AM fungi before contact
(Olah et al., 2005). Taking under consideration that lateral roots
are the preferred site for fungal colonization, it is tempting
to speculate that the plant prepares for the association by
producing an enhanced amount of favored tissue (Harrison,
2005). How far the fungal factors inducing MtENOD11
expression or lateral root formation relate to the same molecule
is not clear. However, in contrast to MtENOD11 expression,
lateral root formation was dependent on MtDMI2 (and
MtDMI1), which suggests that the two processes are triggered
either by different input molecules or by employing different
signaling pathways. Evidence for an early and rather broad
transcriptional response of the plant to perception of the
fungus in its vicinity comes from the recent identification of
11 M. truncatula genes induced by inoculation with Glomus
mosseae before contact (Weidmann et al., 2004). Up-regulation
of these genes was shown to require DMI3 and gene induction was abolished in dmi3 mutants that do not permit fungal
penetration (Catoira et al., 2000). These data indicate that
presymbiotic signaling partially depends on the functioning
of DMI3, which is further required for penetration of the
rhizodermal cell layer.
A different kind of supporting evidence for the concept
of a symbiont anticipation program comes from the recent
finding that higher amounts of starch accumulate in roots of
Lotus japonicus following the detection of approaching
fungi during the presymbiotic phase (Gutjahr et al., 2005).

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Such starch accumulation might promote and sustain the

symbiosis.
Plant receptors for presymbiotic Myc factor-like signals have
not yet been described. However, the absence of appressoria
formation on roots of the recently identified maize mutant
nope1 indicated a defect in early presymbiotic signaling
before physical contact (Paszkowski et al., 2006). The
transposon-associated sectorial restoration of wild-type
function indicated cell autonomous functioning of ZmNOPE1,
arguing against a defect in a plant signal essential for recognition, but pointing towards its involvement in the perception
of a fungal component.

III. Early symbiotic phase contact and

penetration
1. Appressoria development
The onset of the symbiosis is marked morphologically by the
formation of appressoria, the first cell-to-cell contact between
fungus and plant and the site of fungal ingress into the host root
(Fig. 1c). The development of appressoria can be considered to
be the result of successful presymbiotic recognition events when
fungal and plant partners are committed to an interaction
(Giovanetti et al., 1993a). Structurally, appressoria differ from
hyphae by being flattened, elliptical hyphal tips that adhere
by unknown means to the surface of host rhizodermal cells
(Garriock et al., 1989). This morphological switch is reflected
by changes in fungal gene transcription (Breuninger & Requena,
2004). Appressoria do not occur on the rhizodermal cells of
nonhosts (Garriock et al., 1989; Giovanetti et al., 1993a) or
on artificial surfaces such as cellulose or nylon (Giovanetti et al.,
1993a). Interestingly, in a further study, isolated rhizodermal
cell walls of a host but not vascular or cortical cell walls of
the same origin triggered appressoria development without
inducing penetration (Nagahashi & Douds, 1997). Thus,
while physical and/or chemical rhizodermal cell wall features
are required and sufficient to elicit appressoria formation,
penetration needs the support of intact cells, whose coordinated,
matching response accommodates the fungus. The phenotypes
of numerous plant mutants affected at the stage of penetration
following appressoria development fosters this view (reviewed
in Marsh & Schultze, 2001). Although penetration is compromised in all of these mutants, specific plant-regulated
processes associated with penetration have been delineated.
In L. japonicus, for example, an epidermal cleft opens between
two adjacent rhizodermal cells through which the fungus enters.
From here, invasion of the rhizodermal cell(s) is initiated
and the fungus trespasses through the underlying exodermal
cell (reviewed in Parniske, 2004). Mutations in LjSYM2
( LjSYMRK ), LjSYM4 (LjCASTOR) and LjSYM15 affect
these processes, specifically the epidermal opening (LjSYM15;
Demchenko et al., 2004), the intracellular accommodation
of the fungus within rhizo- and exodermal cells (LjSYM2,

Review

LjSYM4; Bonfante et al., 2000) and the penetration of

cortical cells (LjCASTOR, LjSYM15; Novero et al., 2002).
Discovery of the corresponding mutants assisted the
definition of these plant-regulated steps (reviewed in Parniske,
2004).
In addition to entering through a rhizodermal cleft, the
fungus can also traverse rhizodermal cells directly, as shown
in detail for G. gigantea penetrating roots of M. truncatula
(Genre et al., 2005). Interestingly, the green fluorescent
labeling of cytoskeleton and endoplasmatic reticulum proteins
revealed that M. truncatula rhizodermal cells form a prepenetration apparatus after appressoria development (but before
fungal invagination of the plant cell) that guides the invading
hypha through the cell lumen (Genre et al., 2005). This
process commences with nuclear repositioning adjacent to
the appressorium. Subsequently, transcellular migration of the
nucleus is associated with the formation of a hollow column
composed of microtubules, microfilaments and endoplasmic
reticulum cisternae connecting the moving nucleus to its
previous position below the appressorium. Finally, the fungal
hypha enters the preformed channel and follows the route
earlier outlined by the plant cell nucleus. Hence, infection only
occurs after preparatory activities in the plant cell. These cytological studies illustrate nicely the extensive complementary
contribution made by host cell activity to fungal penetration.
Moreover, the prepenetration apparatus is not induced in
dmi2 and dmi3 in which fungal infection is blocked following
appressoria formation. This emphasizes the significance and
indispensable participation of plant processes in the coordinated invasion (Genre et al., 2005). It will be important to
investigate how far equivalent rearrangements are involved
in the accommodation of other endosymbionts. As Genre
et al. (2005) show, elaboration of a tunnel for fungal passage
through the plant cell is accompanied by the synthesis of a
perifungal membrane that encloses the intracellular fungal hypha
and confines the apoplastic interface compartment. The formation of a symbiosome membrane and an interface matrix have
been recognized as features common to biotrophic plant
microbial interactions involving intracellular growth of the
microsymbiont (Parniske, 2000). Therefore, it will be an exciting
task in the future to elucidate mechanistic conservation among
phylogenetically distant but strategically analogous infection
forms.
2. The common SYM pathway
All of the legume mutants described so far are common sym
mutants impaired in root endosymbioses, mycorrhiza and
nodulation. These mutants indicate the existence in legumes
of partially shared genetic programs required for successful
interaction with fungal and bacterial symbionts. A large number
of legume mutants, including dmi1, dmi2 and dmi3 mutants
(Catoira et al., 2000), and one tomato mutant (Barker et al., 1998),
have been identified, which support appressoria formation

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Fig. 2 A mycocentric view of the common SYM/DMI signaling pathway. The signaling pathway as it appears in this figure has emerged from
genetic investigations on Medicago truncatula, Lotus japonicus and Medicago sativa. Abbreviations in squares refer to identified genes. The
Myc factor(s) released by arbuscular mycorrhizal (AM) fungi has not been identified and also the link (possibly a transcription factor, TF) between
DMI3 and mycorrhiza-induced alterations in gene expression is unknown. (Modified after Oldroyd et al. (2005).)

but are compromised in intraradical invasion (reviewed

in Marsh & Schultze, 2001). While in M. truncatula, three
classes of mutants (dmi1, dmi2 and dmi3) have been reported,
seven genes (LjSYMRK, LjCASTOR, LjPOLLUX, LjSYM3,
LjSYM6, LjSYM15, LjSYM24) were found to be required for
fungal penetration in L. japonicus (reviewed in Harrison, 2005;
Kistner et al., 2005). The use of model legumes paved the way
for isolating corresponding genes from mutant backgrounds
and defining a signal transduction pathway essential for the
two root symbioses (Fig. 2). As several authors have discussed
this topic in detail (Parniske, 2004; Harrison, 2005; Hause &
Fester, 2005; Oldroyd et al., 2005; Oldroyd & Downie, 2006),
this article will only briefly summarize the current knowledge
and describe recent progress taking a mycocentric view. The
genes revealed by these mutations all control fungal passage
through the outer cell layers. The corresponding proteins,
namely the Leu-rich repeat receptor-like kinase LjSYMRK/
MsNORK/MtDMI2 (Endre et al., 2002; Stracke et al., 2002)
as well as the plastid ion channels LjCASTOR/LjPOLLUX/
MtDMI1 (Ane et al., 2004; Imaizumi-Anraku et al., 2004)
and the nuclear localized calcium- and calmodulin-dependent
protein kinase MtCCamK/MtDMI3 (Levy et al., 2004; Mitra
et al., 2004), are needed for the early steps in Nod factor
signaling and are probably also involved in signaling during
the early stages of mycorrhizal colonization. The first genes
isolated encoded the LjSYMRK/MsNORK/MtDMI2 receptorlike kinases (Endre et al., 2002; Stracke et al., 2002). The
structure of the protein with an extensive extracellular domain
suggests it is involved in the extracellular binding of a ligand
and the intracellular transduction of the signaling event via
its protein kinase domain. At present, however, it is not
clear if these proteins in fact operate as primary receptors
for a compound released by AM fungi. It is equally plausible

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that additional receptors are involved in the perception of a Myc

factor as was shown for the Nod factor receptors LjNFR1/
MtLYK3 and LjNFR5 (Limpens et al., 2003; Madsen et al.,
2003; Radutoiu et al., 2003). One of the early responses of
legume root hair cells to the perception of Rhizobia or purified
Nod factors is intracellular calcium oscillation (for a recent
review, see Oldroyd & Downie, 2006). During Nod factor
signaling, DMI1 (LjCASTOR and LjPOLLUX) and MtDMI2
(LjSYMRK, MsNORK) type proteins are necessary for the
induction of calcium spiking, while MtDMI3 (MtCCamK)
type proteins are assumed to be involved in deciphering the
calcium signal (Fig. 2) (Levy et al., 2004; Mitra et al., 2004).
Although calcium oscillation has not been shown for mycorrhizal
signaling, the participation of proteins associated with calcium
signaling suggests calcium also functions as a second messenger
within the fungus-related signaling pathway. Recently, another
member of the SYM pathway required for calcium spiking
during nodulation has been discovered: the gene affected in
the L. japonicus sym3 mutant encodes a homologue of the
mammalian nucleoporin NUP133 and was therefore termed
Lotus NUP133 (Kanamori et al., 2006). Consistent with a
function as a nucleoporin, an eYFP-NUP133 fusion protein
localized to the nuclear rim assembly with the nuclear pore
complex. The absence of pleiotropic phenotypes of Lotus nup133
mutant plants indicates the specific involvement of the
LjNUP133 protein in root symbioses. Although its function
during symbiosis is not clear, it is intriguing that the early
symbiotic signal transduction pathway includes ion fluxes,
plastid ion channels and nuclear pore complexes. Cloning of
the mutated genes from the remaining Lotus sym mutants is
currently in progress in different laboratories. The establishment
of a comprehensive picture of how the individual components
of the SYM pathway tie up should be possible in the near

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future. Furthermore, because of the ancient nature of the AM

symbiosis, a high degree of functional conservation of these
SYM factors among members of plant families other than
legumes can be anticipated. Noticeably, a first supporting
example showed that the nodulation phenotype of the M.
truncatula dmi3 mutant can be rescued by introducing a
rice gene putatively orthologous to the leguminous CCamK
(Godfroy et al., 2006). It will be important to verify complementation of the mycorrhizal mutant phenotype of the transformed dmi3 line and also to address the function of this
gene for the interaction of rice with AM fungi. Similarly, for
other putative SYM factor orthologs from non-leguminous
dicotyledonous or from monocotyledonous plants (Zhu et al.,
2006), the relevance for the AM symbiosis can be addressed
via reverse genetics.
The identification of additional components within the
symbiotic nitrogen fixation signaling cascade indicates that
proteins with related functions might be involved in mycorrhizal
signaling. For example, a functional equivalent of the putative
Nod factor receptors LjNFR1 and LjNFR5 (reviewed in
Oldroyd et al., 2005; Oldroyd & Downie, 2006) could exist for
Myc factor perception (Fig. 2). It is also likely that components
corresponding to the GRAS transcription factors MtNSP1
and MtNSP2 (Kalo et al., 2005; Smit et al., 2005) are present
to modulate gene expression in response to calcium oscillation during mycorrhizal signaling (Fig. 2). In summary,
genetically dissecting the common SYM signal transduction pathway required for bacterial and fungal root
endosymbiosis not only unraveled the players involved but also
provided a first glimpse at conservation and specialization
of signaling cascades essential for nodulation and mycorrhiza
development.
3. Apoplastic fungal spread
Transcellular passage of the outer root cell layers appears to
be a bottleneck in the development of the AM symbiosis,
while entrance to the cortex apoplast permits rapid spread of
the fungus along the axes of the root (Fig. 1d, Parniske, 2004).
In the maize mutant taci1, the fungus enters the apoplast
and initiates hyphal growth along the longitudinal axes of
the root. However, these hyphae become septate and die, and
horizontal spreading is not accomplished (Paszkowski et al.,
2006). Furthermore, silencing of the M. truncatula CDPK1
gene, which encodes a calcium-dependent protein kinase
required for root development, led to significantly reduced
longitudinal spread of the fungus and similarly decreased progression of nodulation-associated infection threads through
cortical cells (Ivashuta et al., 2005). Reduced microbial growth
was suggested to occur as a result of altered cell wall composition and cytoskeleton organization of silenced CDPKi lines.
The phenotypes of taci1 and CDPKi provide evidence that
plant-encoded factors are required for apoplastic proliferation
of the fungus.

Review

IV. Mature symbiotic phase haustoria and

mineral nutrition
1. Arbuscule development
Arbuscules are the key feature of the AM symbiosis as they
represent an extreme form of intimacy and compatibility.
Formation of arbuscules within host cells is associated with
dramatic morphological and physiological changes in both
symbiotic partners. The fungus invaginates inner cortex cells
where it undergoes extensive dichotomous branching into
a tree-like fungal structure that may entirely fill the living
cortical cell (Fig. 1e). Consequently the architecture of the host
cell undergoes remarkable changes: for example, the nucleus
moves from a peripheral to a central position, the vacuole becomes
fragmented and an extensive periarbuscular membrane is
synthesized that is in continuum with the plant plasma
membrane (reviewed in Harrison, 1999). Despite this intense
activity of both partners leading towards arbusculated cells,
arbuscules collapse after several days, leaving an intact cortical
cell that is then able to host another arbuscule. It is not known
what triggers fungal entrance into the cell but the perception
of a radial sugar gradient between the vascular tissue and the
outer cell layers may be involved in induction of arbuscule
formation (Blee & Anderson, 1998). The signals that induce
intracellular fungal branching or arbuscule decay have also
not been identified. However, as mentioned in Section II.1,
the recent identification of strigolactone as a stimulant for
presymbiotic fungal branching (Akiyama et al., 2005), combined
with the finding that part of its putative biosynthetic pathway
is activated in colonized roots (Maier et al., 1997; Walter et al.,
2000; Matusova et al., 2005), indicates that related molecules
may elicit intracellular hyphal branching. It has been suggested
that arbuscule senescence is a result of plant responses countering intracellular colonization (Salzer et al., 1999; Walter et al.,
2000). Alternatively, the initiation of arbuscule collapse
may be caused by endogenous fungal signaling or coordinated
signaling cross-talk.
Development of arbuscules is at least partially under the
control of the host genetic program. So far, one legume mutant
has been described (pea sym36 ) in which reduced arbuscules
with fewer and shorter branches are formed (Engvild, 1987;
Gianinazzi-Pearson et al., 1991). In addition, proteins of the
SYM pathway are either required for arbuscule development,
such as LjCASTOR, LjSYM15 and likely LjSYM6, or contribute
to arbuscule formation, such as LjPOLLUX, LjNup133
and LjSYM24 (Kistner et al., 2005). The recovery of further
mutants in forward genetic screens is an urgent task in order
to unravel plant factors essential to the formation of this key
structure of the AM symbiosis. In recent years, several laboratories
have undertaken global gene expression profiling to identify AM
regulated genes (Martin-Laurent et al., 1997; Lapopin et al.,
1999; van Buuren et al., 1999; Journet et al., 2002; Liu et al.,
2003; Taylor & Harrier, 2003; Wulf et al., 2003; Brechenmacher

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et al., 2004; Grunwald et al., 2004; Weidmann et al., 2004;

Guimil et al., 2005; Hohnjec et al., 2005). In many cases gene
expression was monitored at the stage of a mature symbiosis
and long lists of induced and suppressed genes have been
created. Reporter lines using the promoters of up-regulated
genes in conjunction with GUS or GFP revealed either
exclusive expression in arbusculated cells or expression in
tissue patches including cells with arbuscules and cells in
the vicinity of arbuscules (reviewed in Harrison, 2005;
Hohnjec et al., 2005). The list of candidate genes probably
includes those essential for signaling during the initiation and
formation of arbuscules.
With the help of reverse genetics it is possible to determine
the relevance of differentially regulated genes for the development of an AM symbiosis, especially where extensive resources
are available, such as L. japonicus, M. truncatula and rice (Perry
et al., 2003; Hirochika et al., 2004; Tadege et al., 2005).
Alternatively, efficient transformation protocols can be applied
to RNAi (or antisense)-mediated gene silencing in a variety of
AM host plants. Knock-down of the expression of a M. truncatula
gene encoding the jasmonic acid-biosynthetic enzyme allene
oxide cyclase by transformation with antisense cDNA resulted
in lower amounts of jasmonic acid (Isayenkov et al., 2005). A
delay in colonization, accompanied by reduced arbuscle
formation was observed indicating that jasmonic acid plays
a role during the development of an AM symbiosis. Further
exciting discoveries facilitated by the availability of these
resources can be predicted in the near future in various AM
host plants.
2. Symbiotic phosphate and nitrogen acquisition
Arbuscular mycorrhizal fungi obtain carbohydrates from
their hosts and simultaneously improve plant acquisition of
mineral nutrients, in particular phosphate (Pi). It was recently
shown that, depending on the particular plantfungus combination, symbiotic phosphate uptake may partially participate
in, or even dominate, overall plant Pi acquisition (Smith et al.,
2003). The proposed metabolic route of symbiotic Pi acquisition starts with the assimilation of inorganic Pi at the hyphal
soil interface by fungal high-affinity transporters (Harrison
& van Buuren, 1995; Maldonado-Mendoza et al., 2001;
Benedetto et al., 2005). Inside the fungus, inorganic Pi is
translocated in the form of polyphosphate from fungal structures
outside of the root to those inside (Solaiman et al., 1999;
Ohtomo & Saito, 2005). Before release into the periarbuscular
interface, phosphate becomes depolymerized to inorganic Pi
(Ohtomo & Saito, 2005). Pi is acquired from the interface by
plant-encoded phosphate transporters. Such transporters have
been identified from several plant backgrounds and were shown
to be transcriptionally induced during the development of
the AM symbiosis (Rausch et al., 2001; Harrison et al., 2002;
Paszkowski et al., 2002; Glassop et al., 2005; Nagy et al., 2005).
While only one mycorrhiza-induced Pi transporter coding gene

New Phytologist (2006) 172: 3546

Table 1 Mycorrhiza-specific plant phosphate transporters

Gene name

Plant species

Reference

MtPT4
LePT4
LePT5
StPT4
StPT5
OsPT11
TaPT1

Medicago truncatula
Tomato
Tomato
Potato
Potato
Rice
Wheat

Harrison et al. (2002)

Karandashov et al. (2004)
Nagy et al. (2005)
Karandashov & Bucher (2005)
Nagy et al. (2005)
Paszkowski et al. (2002)
Glassop et al. (2005)

has currently been identified from M. truncatula (MtPT4;

Harrison et al., 2002), three have been found in solanaceous
species (StPT3/LePT3, StPT4/LePT4, and StPT5/LePT5;
Karandashov & Bucher, 2005; Nagy et al., 2005), two in rice
(OsPT11 and OsPT13; Paszkowski et al., 2002; Guimil et al.,
2005) and one in maize, barley and wheat (Glassop et al.,
2005). The mycorrhiza-specific subfamily comprises the
orthologous monocot proteins OsPT11 and TaPT1, and the
dicot proteins MtPT4, StPT4/LePT4 and StPT5/LePT5
(Table 1), indicating a gene duplication event in the case of
the solanaceous transporters (Nagy et al., 2005). The StPT3
protein is nonorthologous to the MtPT4 type proteins and to
OsPT13. While OsPT13 is weakly expressed in mycorrhizal
roots (Guimil et al., 2005), StPT3 exhibits a strong up-regulation
(Rausch et al., 2001). One of these mycorrhiza-induced Pi
transporters has been localized to the periarbuscular membrane,
consistent with a function in plant uptake of Pi from the
interface between fungus and plant (Harrison et al., 2002). A
pressing project is to confirm the function of these transporter
proteins in symbiotic Pi acquisition and to examine their
influence on the establishment of the symbiosis. The fact that
mycorrhiza-induced Pi transporters in some plant species are
present in small gene families has so far hampered approaches to
this question because of functional redundancy (Nagy et al.,
2005). Moreover, it is of great interest to elucidate the nature
of the signal specifically inducing symbiotic Pi transporter
genes and also to determine the mechanism leading to the
periarbuscular membrane localization of the corresponding Pi
transporter proteins.
It has been long recognized that in addition to Pi, AM fungi
also promote nitrogen nutrition of their hosts (Raven et al.,
1978). Until recently, very little was known about how AM
fungi take up nitrogen from the environment and enable
transfer of a portion to the plant. Stable isotope labeling has now
suggested that inorganic nitrogen is taken up by the extraradical
mycelium, incorporated into amino acids, translocated from
extra- to intraradical fungal structures as arginine and then
released as ammonium to the plant (Govindarajulu et al.,
2005; Jin et al., 2005). Further support for this uptake route
comes from the finding that transcript abundance of key
enzymes of nitrogen assimilation and of arginine breakdown
preferentially accumulate in extra- or intraradical mycelium,

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Tansley review

respectively (Govindarajulu et al., 2005). In analogy to the path

of symbiotic Pi uptake, the arbuscule may be the site of
symbiotic nitrogen uptake involving plant-encoded nitrogen
transporters located within the periarbuscular plant membrane.
Promising candidates such as mycorrhiza-induced nitrate and
ammonium transporters have been identified in transcriptome
analyses of mycorrhizal M. truncatula and rice (Frenzel et al.,
2005; Guimil et al., 2005; Hohnjec et al., 2005). It will be
interesting to investigate their role in symbiotic nitrogen
uptake to determine the contribution of AM symbioses to
nitrogen acquisition of plants.

V. Concluding remarks
Although our understanding of molecular mechanism and
signaling pathways coupled to AM symbioses need further
refinement, the past few years have brought exciting discoveries
in this area. Elucidation of molecular events associated with
signaling and nutrient acquisition processes have moved rapidly
forward. It is foreseeable that the availability of forward and
reverse genetics resources as well as established transformation
protocols will stimulate further work towards identification
and characterization of plant (and fungal) factors relevant to
AM symbioses. Furthermore, central questions concerning their
conservation among mono- and dicotyledonous plants, and
also their relevance in other plantmicrobial interactions, can
be addressed in the near future.

Acknowledgements
I apologize to all those researchers whose work I have overlooked
or could not include because of space considerations. I am
grateful to Patrick King, Caroline Gutjahr and Ruairidh Sawers
for critical reading of the manuscript and to the University of
Geneva and Swiss National Science Foundation for funding
(grant 3100A0-104132).

References
Akiyama K, Matsuzaki K, Hayashi H. 2005. Plant sesquiterpenes
induce hyphal branching in arbuscular mycorrhizal fungi. Nature
435: 824827.
Ane JM, Kiss GB, Riely BK, Penmetsa RV, Oldroyd GE, Ayax C, Levy J,
Debelle F, Baek JM, Kalo P, Rosenberg C, Roe BA, Long SR,
Denarie J, Cook DR. 2004. Medicago truncatula DMI1 required
for bacterial and fungal symbioses in legumes. Science 303:
13641367.
Bago B, Pfeffer PE, Douds DD Jr, Brouillette J, Becard G, Shachar-Hill Y.
1999. Carbon metabolism in spores of the arbuscular mycorrhizal fungus
Glomus intraradices as revealed by nuclear magnetic resonance
spectroscopy. Plant Physiology 121: 263 272.
Bago B, Pfeffer PE, Shachar-Hill Y. 2000. Carbon metabolism and
transport in arbuscular mycorrhizas. Plant Physiology 124: 949 958.
Barker S, Stummer B, Gao L, Dispain I, OConnor P, Smith S. 1998.
A mutant in Lycopersicon esculentum Mill. with highly reduced VA
mycorrhizal colonization: isolation and preliminary characterization. Plant
Journal 15: 791797.

Review

Bcard G, Kosuta S, Tamasloukht M, Sjalon-Delmas N, Roux C. 2004.

Partner communication in the arbuscular mycorrhizal interaction.
Canadian Journal of Botany 82: 11861197.
Bcard G, Pich Y. 1989. New aspects on the acquisition of biotrophic status
by a vesicular-arbuscular mycorrhizal fungus, Gigaspora margarita. New
Phytologist 112: 7783.
Benedetto A, Magurno F, Bonfante P, Lanfranco L. 2005. Expression
profiles of a phosphate transporter gene (GmosPT) from the
endomycorrhizal fungus Glomus mosseae. Mycorrhiza 15: 620627.
Blee K, Anderson A. 1998. Regulation of arbuscule formation by carbon in
the plant. The Plant Journal 16: 523530.
Bonfante P, Genre A, Faccio A, Martini I, Schauser L, Stougaard J, Webb
J, Parniske M. 2000. The Lotus japonicus LjSym4 gene is required for the
successful symbiotic infection of root epidermal cells. Molecular Plant
Microbe Interactions 13: 11091120.
Bouwmeester HJ, Matusova R, Zhongkui S, Beale MH. 2003. Secondary
metabolite signalling in hostparasitic plant interactions. Current Opinions
in Plant Biology 6: 358364.
Brechenmacher L, Weidmann S, Van Tuinen D, Chatagnier O, Gianinazzi
S, Franken P, Gianinazzi-Pearson V. 2004. Expression profiling of
up-regulated plant and fungal genes in early and late stages of Medicago
truncatulaGlomus mosseae interactions. Mycorrhiza 14: 253262.
Breuninger M, Requena N. 2004. Recognition events in AM symbiosis:
analysis of fungal gene expression at the early appressorium stage. Fungal
Genetics Biology 41: 794804.
Brundrett M. 2002. Coevolution of roots and mycorrhizas of land plants.
New Phytologist 154: 275304.
Buee M, Rossignol M, Jauneau A, Ranjeva R, Becard G. 2000. The
pre-symbiotic growth of arbuscular mycorrhizal fungi is induced by
a branching factor partially purified from plant root exudates.
Molecular PlantMicrobe Interactions 13: 693698.
Burleigh SH, Cavagnaro T, Jakobsen I. 2002. Functional diversity of
arbuscular mycorrhizas extends to the expression of plant genes involved
in P nutrition. Journal of Experimental Botany 53: 15931601.
van Buuren ML, Maldonado-Mendoza IE, Trieu AT, Blaylock LA,
Harrison MJ. 1999. Novel genes induced during an arbuscular
mycorrhizal (AM) symbiosis formed between Medicago truncatula and
Glomus versiforme. Molecular PlantMicrobe Interactions 12: 171181.
Catoira R, Galera C, de Billy F, Penmetsa RV, Journet EP, Maillet F,
Rosenberg C, Cook D, Gough C, Denarie J. 2000. Four genes of
Medicago truncatula controlling components of a nod factor transduction
pathway. Plant Cell 12: 16471666.
Chabaud M, Venard C, Defaux-Petras A, Becard G, Barker D. 2002.
Targeted inoculation of Medicago truncatula in vitro root cultures reveals
MtENOD11 expression during early stages of infection by arbucular
mycorrhizal fungi. New Phytologist 156: 265273.
David-Schwartz R, Badani H, Smadar W, Levy AA, Galili G, Kapulnik Y.
2001. Identification of a novel genetically controlled step in mycorrhizal
colonization: plant resistance to infection by fungal spores but not
extra-radical hyphae. Plant Journal 27: 561569.
David-Schwartz R, Gadkar V, Wininger S, Bendov R, Galili G, Levy AA,
Kapulnik Y. 2003. Isolation of a premycorrhizal infection (pmi2) mutant
of tomato, resistant to arbuscular mycorrhizal fungal colonization.
Molecular PlantMicrobe Interactions 16: 382388.
Demchenko K, Winzer T, Stougaard J, Parniske M, Pawlowska K. 2004.
Distinct roles of Lotus japonicus SYMRK and SYM15 in root colonization
and arbuscule formation. New Phytologist 163: 381392.
Endre G, Kereszt A, Kevei Z, Mihacea S, Kalo P, Kiss GB. 2002. A receptor
kinase gene regulating symbiotic nodule development. Nature 417: 962
966.
Engvild K. 1987. Nodulation and nitrogen fixation mutants of pea, Pisum
sativum L. Theoretical and Applied Genetics 74: 711713.
Fester T, Schmidt D, Lohse S, Walter MH, Giuliano G, Bramley PM,
Fraser PD, Hause B, Strack D. 2002. Stimulation of carotenoid
metabolism in arbuscular mycorrhizal roots. Planta 216: 148154.

The Authors (2006). Journal compilation New Phytologist (2006) www.newphytologist.org

New Phytologist (2006) 172: 3546

43

44 Review

Tansley review

Frenzel A, Manthey K, Perlick AM, Meyer F, Puhler A, Kuster H,

Krajinski F. 2005. Combined transcriptome profiling reveals a novel
family of arbuscular mycorrhizal-specific Medicago truncatula lectin
genes. Molecular Plant MicrobeInteractions 18: 771782.
Gadkar V, David-Schwartz R, Nagahashi G, Douds DD Jr, Wininger S,
Kapulnik Y. 2003. Root exudate of pmi tomato mutant M161 reduces
AM fungal proliferation in vitro. FEMS Microbiological Letters 223:
193198.
Garriock M, Peterson R, Ackerley C. 1989. Early stages in colonization of
Allium porrum (leek) roots by the vesicular-arbucular mycorrhizal fungus,
Glomus versiforme. New Phytologist 112: 85 92.
Genre A, Chabaud M, Timmers T, Bonfante P, Barker DG. 2005.
Arbuscular mycorrhizal fungi elicit a novel intracellular apparatus in
Medicago truncatula root epidermal cells before infection. Plant Cell 17:
34893499.
Gianinazzi-Pearson V, Gianinazzi S, Guillemin J, Trouvelot A, Duc G.
1991. Genetic and cellular analysis of resistance to vesicular arbuscular
(VA) mycorrhizal fungi in pea mutants. In: Verma HHAD, ed. Advances
in molecular genetics of plantmicrobe interactions. Dordrecht, the
Netherlands: Kluwer Academic Publishers, 336 342.
Giovanetti M, Avio L, Sbrana C, Citernesi A. 1993a. Factors affecting
appressoria development in the vesicular-arbuscular mycorrhizal fungus
Glomus mosseae (Nicol. & Gerd.) Gerd. and Trappe. New Phytologist 123:
114122.
Giovanetti M, Sbrana C. 1998. Meeting a non-host: the behaviour of AM
fungi. Mycorrhiza 8: 123 130.
Giovanetti M, Sbrana C, Avio L, Citernesi A, Logi C. 1993b.
Differential hyphal morphogenesis in arbuscular mycorrhizal fungi
during pre-infection stages. New Phytologist 125: 587 593.
Glassop D, Smith SE, Smith FW. 2005. Cereal phosphate transporters
associated with the mycorrhizal pathway of phosphate uptake into roots.
Planta 222: 688698.
Glazebrook J. 2005. Contrasting mechanisms of defense against biotrophic
and necrotrophic pathogens. Annual Review of Phytopathology 43: 205
227.
Godfroy O, Debelle F, Timmers T, Rosenberg C. 2006. A rice calcium- and
calmodulin-dependent protein kinase restores nodulation to a legume
mutant. Molecular PlantMicrobe Interactions 19: 495 501.
Govindarajulu M, Pfeffer PE, Jin H, Abubaker J, Douds DD, Allen JW,
Bucking H, Lammers PJ, Shachar-Hill Y. 2005. Nitrogen transfer in
the arbuscular mycorrhizal symbiosis. Nature 435: 819 823.
Grunwald U, Nyamsuren O, Tamasloukht M, Lapopin L, Becker A,
Mann P, Gianinazzi-Pearson V, Krajinski F, Franken P. 2004.
Identification of mycorrhiza-regulated genes with arbuscule
development-related expression profile. Plant Molecular Biology 55:
553566.
Guimil S, Chang H-S, Zhu T, Sesma A, Osbourn A, Roux C, Ionnidis V,
Oakeley E, Docquier M, Descombes P, Briggs S, Paszkowski U. 2005.
Comparative transcriptomics of rice reveals an ancient pattern of response
to microbial colonization. Proceedings of the National Academy of Sciences,
USA 102(22): 80668070.
Gutjahr C, Novero M, Genre A, Welham T, Wang T, Bonfante P. 2005.
Prior to colonization Gigaspora margarita induces starch accumulation in
Lotus japonicus roots. In: 12th International Conference on Molecular
PlantMicrobe Interactions, Merida, Mexico. (In press.)
Harrison MJ. 1999. Molecular and cellular aspects of the arbuscular
mycorrhizal symbiosis. Annual Review of Plant Physiological Plant
Molecular Biology 50: 361 389.
Harrison M. 2005. Signaling in the arbuscular mycorrhizal symbiosis.
Annual Review of Microbiology 59: 19 42.
Harrison MJ, Dewbre GR, Liu J. 2002. A phosphate transporter from
Medicago truncatula involved in the acquisition of phosphate released by
arbuscular mycorrhizal fungi. Plant Cell 14: 2413 2429.
Harrison MJ, van Buuren ML. 1995. A phosphate transporter from the
mycorrhizal fungus Glomus versiforme. Nature 378: 626 629.

New Phytologist (2006) 172: 3546

Hause B, Fester T. 2005. Molecular and cell biology of arbuscular

mycorrhizal symbiosis. Planta 221: 184196.
Heijden MVD, Klironomos J, Ursic M, Moutoglis P, Streitwolf-Engel R,
Boller T, Wiemken A, Sanders I. 1998. Mycorrhizal fungal diversity
determines plant biodiversity, ecosystem variability and productivity.
Nature 396: 6972.
Hirochika H, Guiderdoni E, An G, Hsing YI, Eun MY, Han CD,
Upadhyaya N, Ramachandran S, Zhang Q, Pereira A, Sundaresan V,
Leung H. 2004. Rice mutant resources for gene discovery. Plant Molecular
Biology 54: 325334.
Hohnjec N, Vieweg MF, Puhler A, Becker A, Kuster H. 2005. Overlaps in
the transcriptional profiles of Medicago truncatula roots inoculated with
two different Glomus fungi provide insights into the genetic program
activated during arbuscular mycorrhiza. Plant Physiology 137: 1283 1301.
Imaizumi-Anraku H, Takeda N, Charpentier M, Perry J, Miwa H,
Umehara Y, Kouchi H, Murakami Y, Mulder L, Vickers K, Pike J,
Allan Downie J, Wang T, Sato S, Asamizu E, Tabata S, Yoshikawa M,
Murooka Y, Wu GJ, Kawaguchi M, Kawasaki S, Parniske M,
Hayashi M. 2004. Plastid proteins crucial for symbiotic fungal
and bacterial entry into plant roots. Nature 433(7025): 527531.
Isayenkov S, Mrosk C, Stenzel I, Strack D, Hause B. 2005. Suppression
of allene oxide cyclase in hairy roots of Medicago truncatula reduces
jasmonate levels and the degree of mycorrhization with Glomus
intraradices. Plant Physiology 139: 14011410.
Ivashuta S, Liu J, Lohar DP, Haridas S, Bucciarelli B, VandenBosch KA,
Vance CP, Harrison MJ, Gantt JS. 2005. RNA interference identifies
a calcium-dependent protein kinase involved in Medicago truncatula
root development. Plant Cell 17: 29112921.
Jin H, Pfeffer PE, Douds DD, Piotrowski E, Lammers PJ, Shachar-Hill Y.
2005. The uptake, metabolism, transport and transfer of nitrogen in an
arbuscular mycorrhizal symbiosis. New Phytology 168: 687696.
Journet EP, El-Gachtouli N, Vernoud V, de Billy F, Pichon M, Dedieu A,
Arnould C, Morandi D, Barker DG, Gianinazzi-Pearson V. 2001.
Medicago truncatula ENOD11: a novel RPRP-encoding early nodulin
gene expressed during mycorrhization in arbuscule-containing cells.
Molecular PlantMicrobe Interactions 14: 737748.
Journet EP, van Tuinen D, Gouzy J, Crespeau H, Carreau V, Farmer MJ,
Niebel A, Schiex T, Jaillon O, Chatagnier O, Godiard L, Micheli F,
Kahn D, Gianinazzi-Pearson V, Gamas P. 2002. Exploring root
symbiotic programs in the model legume Medicago truncatula using
EST analysis. Nucleic Acids Research 30: 55795592.
Kalo P, Gleason C, Edwards A, Marsh J, Mitra RM, Hirsch S, Jakab J,
Sims S, Long SR, Rogers J, Kiss GB, Downie JA, Oldroyd GE. 2005.
Nodulation signaling in legumes requires NSP2, a member of the GRAS
family of transcriptional regulators. Science 308: 17861789.
Kanamori N, Madsen LH, Radutoiu S, Frantescu M, Quistgaard EM,
Miwa H, Downie JA, James EK, Felle HH, Haaning LL, Jensen TH,
Sato S, Nakamura Y, Tabata S, Sandal N, Stougaard J. 2006. A
nucleoporin is required for induction of Ca2+ spiking in legume nodule
development and essential for rhizobial and fungal symbiosis. Proceedings
of the National Academy of Sciences, USA 103: 359364.
Karandashov V, Bucher M. 2005. Symbiotic phosphate transport
in arbuscular mycorrhizas. Trends in Plant Science 10: 2229.
Karandashov V, Nagy R, Wegmuller S, Amrhein N, Bucher M. 2004.
Evolutionary conservation of a phosphate transporter in the arbuscular
mycorrhizal symbiosis. Proceedings of the National Academy of Sciences,
USA 101: 62856290.
Kistner C, Winzer T, Pitzschke A, Mulder L, Sato S, Kaneko T, Tabata S,
Sandal N, Stougaard J, Webb KJ, Szczyglowski K, Parniske M. 2005.
Seven Lotus japonicus genes required for transcriptional reprogramming
of the root during fungal and bacterial symbiosis. Plant Cell 17: 2217
2229.
Klingner A, Bothe H, Wray V, Marner FJ. 1995. Identification of a yellow
pigment formed in maize roots upon mycorrhizal colonization.
Phytochemistry 38: 5355.

www.newphytologist.org The Authors (2006). Journal compilation New Phytologist (2006)

Tansley review
Koske R. 1981. Multiple germination by spores of Gigaspora gigantea.
Transactions of the British Mycological Society 76: 328 330.
Kosuta S, Chabaud M, Lougnon G, Gough C, Denarie J, Barker DG,
Becard G. 2003. A diffusible factor from arbuscular mycorrhizal fungi
induces symbiosis-specific MtENOD11 expression in roots of Medicago
truncatula. Plant Physiology 131: 952 962.
Lapopin L, Gianinazzi-Pearson V, Franken P. 1999. Comparative
differential RNA display analysis of arbuscular mycorrhiza in Pisum
sativum wild type and a mutant defective in late stage development. Plant
Molecular Biology 41: 669 677.
Levy J, Bres C, Geurts R, Chalhoub B, Kulikova O, Duc G, Journet EP,
Ane JM, Lauber E, Bisseling T, Denarie J, Rosenberg C, Debelle F.
2004. A putative Ca2+ and calmodulin-dependent protein kinase required
for bacterial and fungal symbioses. Science 303: 13611364.
Limpens E, Franken C, Smit P, Willemse J, Bisseling T, Geurts R. 2003.
LysM domain receptor kinases regulating rhizobial Nod factor-induced
infection. Science 302: 630 633.
Liu J, Blaylock LA, Endre G, Cho J, Town CD, VandenBosch KA, Harrison
MJ. 2003. Transcript profiling coupled with spatial expression analyses
reveals genes involved in distinct developmental stages of an arbuscular
mycorrhizal symbiosis. Plant Cell 15: 2106 2123.
Madsen EB, Madsen LH, Radutoiu S, Olbryt M, Rakwalska M,
Szczyglowski K, Sato S, Kaneko T, Tabata S, Sandal N, Stougaard J.
2003. A receptor kinase gene of the LysM type is involved in legume
perception of rhizobial signals. Nature 425: 637 640.
Maier W, Hammer K, Dammann U, Schulz B, Strack D. 1997.
Accumulation of sesquiterpenoid cyclohexonone derivatives induced by
an arbuscular mycorrhizal fungus in members of the Poaceae. Planta 202:
3642.
Maldonado-Mendoza IE, Dewbre GR, Harrison MJ. 2001. A phosphate
transporter gene from the extra-radical mycelium of an arbuscular
mycorrhizal fungus Glomus intraradices is regulated in response to
phosphate in the environment. Molecular PlantMicrobe Interactions
14: 11401148.
Marsh J, Schultze M. 2001. Analysis of arbuscular mycorrhizas using
symbiosis-defective plant mutants. New Phytologist 150: 525 532.
Martin-Laurent F, van Tuinen D, Dumas-Gaudot E, Gianinazzi-Pearson
V, Gianinazzi S, Franken P. 1997. Differential display analysis of RNA
accumulation in arbuscular mycorrhiza of pea and isolation of a novel
symbiosis-regulated plant gene. Molecular General Genetics 256: 3744.
Matusova R, Rani K, Verstappen FW, Franssen MC, Beale MH,
Bouwmeester HJ. 2005. The strigolactone germination stimulants of the
plant-parasitic Striga and Orobanche spp. are derived from the carotenoid
pathway. Plant Physiology 139: 920 934.
Mitra RM, Gleason CA, Edwards A, Hadfield J, Downie JA, Oldroyd GE,
Long SR. 2004. A Ca2+/calmodulin-dependent protein kinase required
for symbiotic nodule development: Gene identification by transcriptbased cloning. Proceedings of the National Academy of Sciences, USA 101:
47014705.
Mosse B. 1959. The regular germination of resting spores and some
observations on the growth requirements of an Endogone sp. causing
vesicular-arbuscular mycorrhiza. Transactions of the British Mycological
Society 42: 273286.
Nagahashi G, Douds D. 1997. Appressorium formation by AM fungi on
isolated cell walls of carrot roots. New Phytologist 136: 299 304.
Nagy R, Karandashov V, Chague V, Kalinkevich K, Tamasloukht M, Xu G,
Jakobsen I, Levy AA, Amrhein N, Bucher M. 2005. The characterization
of novel mycorrhiza-specific phosphate transporters from Lycopersicon
esculentum and Solanum tuberosum uncovers functional redundancy in
symbiotic phosphate transport in solanaceous species. Plant Journal 42:
236250.
Novero M, Faccio A, Genre A, Stougaard J, Webb K, Mulder L,
Parniske M, Bonfante P. 2002. Dual requierment of the LjSYM4 gene
for mycorrhizal development in epidermal and cortical cells of Lotus
japonicus roots. New Phytologist 154: 741749.

Review

Ohtomo R, Saito M. 2005. Polyphosphate dynamics in mycorrhizal roots

during colonization of an arbuscular mycorrhizal fungus. New Phytology
167: 571578.
Olah B, Briere C, Becard G, Denarie J, Gough C. 2005. Nod factors and
a diffusible factor from arbuscular mycorrhizal fungi stimulate lateral
root formation in Medicago truncatula via the DMI1/DMI2 signalling
pathway. Plant Journal 44: 195207.
Oldroyd GE, Downie JA. 2006. Nuclear calcium changes at the core of
symbiosis signalling. Current Opinion in Plant Biology 9(4): 351357.
Oldroyd GE, Harrison MJ, Udvardi M. 2005. Peace talks and trade deals.
Keys to long-term harmony in legume-microbe symbioses. Plant
Physiology 137: 12051210.
Parniske M. 2000. Intracellular accommodation of microbes by plants:
a common developmental program for symbiosis and disease? Current
Opinion in Plant Biology 3: 320328.
Parniske M. 2004. Molecular genetics of the arbuscular mycorrhizal
symbiosis. Current Opinion in Plant Biology 7: 414421.
Paszkowski U, Jakovleva L, Boller T. 2006. Maize mutants affected at
distinct stages of the arbuscular mycorrhizal symbiosis. The Plant Journal
47(2): 165173.
Paszkowski U, Kroken S, Roux C, Briggs SP. 2002. Rice phosphate
transporters include an evolutionarily divergent gene specifically activated
in arbuscular mycorrhizal symbiosis. Proceedings of the National Academy of
Sciences, USA 99: 1332413329.
Perry JA, Wang TL, Welham TJ, Gardner S, Pike JM, Yoshida S,
Parniske M. 2003. A TILLING reverse genetics tool and a web-accessible
collection of mutants of the legume Lotus japonicus. Plant Physiology 131:
866871.
Radutoiu S, Madsen LH, Madsen EB, Felle HH, Umehara Y, Gronlund M,
Sato S, Nakamura Y, Tabata S, Sandal N, Stougaard J. 2003. Plant
recognition of symbiotic bacteria requires two LysM receptor-like kinases.
Nature 425: 585592.
Rausch C, Daram P, Brunner S, Jansa J, Laloi M, Leggewie G, Amrhein N,
Bucher M. 2001. A phosphate transporter expressed in arbusculecontaining cells in potato. Nature 414: 462470.
Raven J, Smith S, Smith F. 1978. Ammonium assimilation and the role
of mycorrhizas in climax communities in Scotland. Botanical Society
of Edinburgh 43: 2735.
Ravenskov S, Jakobsen I. 1995. Functional compatibility in arbuscular
mycorrhizas measured as hyphal P transport to the plant. New Phytologist
129: 611618.
Remy W, Taylor TN, Hass H, Kerp H. 1994. Four hundred-millionyear-old vesicular arbuscular mycorrhizae. Proceedings of the National
Academy of Sciences, USA 91: 1184111843.
Salzer P, Corbiere H, Boller T. 1999. Hydrogen peroxide accumulation in
Medicago truncatula root colonized by the arbuscular mycorrhiza-forming
fungus. Planta 208: 319325.
Schssler A, Schwarzott D, Walker C. 2001. A new fungal phylum, the
Glomeromycota: phylogeny and evolution. Mycological Research 105:
14131421.
Smit P, Raedts J, Portyanko V, Debelle F, Gough C, Bisseling T,
Geurts R. 2005. NSP1 of the GRAS protein family is essential for
rhizobial Nod factor-induced transcription. Science 308: 17891791.
Smith SE, Read DJ. 1997. Mycorrhizal symbiosis. San Diego: Academic Press.
Smith SE, Smith FA, Jakobsen I. 2003. Mycorrhizal fungi can dominate
phosphate supply to plants irrespective of growth responses. Plant
Physiology 133: 1620.
Solaiman MZ, Ezawa T, Kojima T, Saito M. 1999. Polyphosphates in
intraradical and extraradical hyphae of an arbuscular mycorrhizal
fungus, Gigaspora margarita. Applied Environmental Microbiology 65:
56045606.
Stracke S, Kistner C, Yoshida S, Mulder L, Sato S, Kaneko T, Tabata S,
Sandal N, Stougaard J, Szczyglowski K, Parniske M. 2002. A plant
receptor-like kinase required for both bacterial and fungal symbiosis.
Nature 417: 959962.

The Authors (2006). Journal compilation New Phytologist (2006) www.newphytologist.org

New Phytologist (2006) 172: 3546

45

46 Review

Tansley review

Tadege M, Ratet P, Mysore KS. 2005. Insertional mutagenesis: a Swiss

Army knife for functional genomics of Medicago truncatula. Trends in
Plant Science 10: 229 235.
Tamasloukht M, Sejalon-Delmas N, Kluever A, Jauneau A, Roux C, Becard
G, Franken P. 2003. Root factors induce mitochondrial-related gene
expression and fungal respiration during the developmental switch from
asymbiosis to presymbiosis in the arbuscular mycorrhizal fungus Gigaspora
rosea. Plant Physiology 131: 1468 1478.
Taylor J, Harrier LA. 2003. Expression studies of plant genes differentially
expressed in leaf and root tissues of tomato colonised by the arbuscular
mycorrhizal fungus Glomus mosseae. Plant Molecular Biology 51: 619
629.
Walter MH, Fester T, Strack D. 2000. Arbuscular mycorrhizal fungi induce
the non-mevalonate methylerythritol phosphate pathway of isoprenoid
biosynthesis correlated with accumulation of the yellow pigment and
other apocarotenoids. Plant Journal 21: 571 578.
Weidmann S, Sanchez L, Descombin J, Chatagnier O, Gianinazzi S,

Gianinazzi-Pearson V. 2004. Fungal elicitation of signal transductionrelated plant genes precedes mycorrhiza establishment and requires the
dmi3 gene in Medicago truncatula. Molecular PlantMicrobe Interactions
17: 13851393.
Westwood J. 2000. Characterization of the Orobanche-Arabidopsis
system for studying parasitehost interactions. Weed Science 48: 742
748.
Wulf A, Manthey K, Doll J, Perlick AM, Linke B, Bekel T, Meyer F,
Franken P, Kuster H, Krajinski F. 2003. Transcriptional changes in
response to arbuscular mycorrhiza development in the model plant
Medicago truncatula. Molecular PlantMicrobe Interactions 16: 306 314.
Yokota T, Sakal H, Okuno K, Yoneyama K, Takeucji Y. 1998. Alectrol and
Orobanchol, germination stimulants for Orobanche minor, from its host
red clover. Phytochemistry 49: 19671973.
Zhu H, Riely BK, Burns NJ, Ane JM. 2006. Tracing non-legume orthologs
of legume genes required for nodulation and arbuscular mycorrhizal
symbioses. Genetics 172(4): 24912499.

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