4.

3 AM
Siddhi K Lad
Introduction
 The mycorrhizal mutualistic association between fungi and the roots of plants has
contributed, from the earliest times, to the evolution of the Earth’s terrestrial
ecosystem.
 In this relationship the roots of the plant are infected by a fungus, but the rest of
the fungal mycelium continues to grow through the soil, digesting and absorbing
nutrients and water and sharing these with its plant host.
 This was discovered by a German botanist called Albert Bernhard Frank in 1885.
He began a study of the possibility of cultivating truffle groves in Prussia but his
study developed into a revolutionary theory of tree nutrition via symbiosis between
fungi and tree roots in a compound structure he called a Wurzelpilze, or root-
fungus.
 In fact, Frank used the words Wurzelsymbiose, Wurzelpilze and Mycorhiza in the
titles of three successive papers in 1885 (and mykorhiza and mycorrhiza in titles of
two later papers).
 The spelling has now been standardised to ‘mycorrhiza’, but it still means fungus-
root! Later studies have shown that just about all of Frank’s interpretations were
correct.
Arbuscular (AM) endomycorrhizas
 Arbuscular (AM) endomycorrhizas are the most common type
of mycorrhizal association, and were probably the first to
evolve; the fungi are members of the Glomeromycotina .
 AM fungi are obligate biotrophs, and they are associated
with roots of about 80% of plant species (that’s equivalent
to about two-thirds of all land plants, or around 90% of all
vascular plants), including many crop plants.
 The AM association is endotrophic, and has previously been
referred to as a Vesicular-Arbuscular Mycorrhiza (VAM).
 This name has since been dropped in favour of AM.
 There is a wide-ranging fungal mycelium within the
host root, and AM fungi explore the soil or other
substrata with an extensive extraradical mycelium.
 Externally to the host the fungal hyphae produce
the very large spores (often called chlamydospores).
 Formation of the mycorrhizal association is an
infection process .
 Spores germinate near a plant root and the germinating
hyphae penetrate the root in response to root exudates.
 Hyphae grow through the root tissues and in the root
cortex hyphal branches form appressoria that penetrate
the plant cells.
 The host plasmalemma invaginates and proliferates
around the fungal intrusion.
 Repeated dichotomous branching of the fungal ‘hypha’
produces the arbuscule inside the cortical cell.
 Arbuscules have a lifespan of 4-15 days, after which
they break down, and the plant cell returns to normal.
 Many AM fungi (except the family Gigasporaceae) also
produce vesicles within the roots (Fig. 6), either between
or within the cortical cells.
 These are swollen spherical or oval structures containing
lipids, which are thought to be used for storage.
 Vesicles are usually over 100 µm in diameter and are
swollen hyphal tips.
 All AM fungi are obligately biotrophic; that is they are
completely dependent on plants for their survival.
 This does not present a problem for AM fungi, as they
show little or no host specificity.
 Unlike ericoid and orchid mycorrhizas, AM fungi are not
restricted to any particular taxonomic group of plants, and
are found extensively in pteridophytes, gymnosperms and
angiosperms of all habitats and even ‘primitive’ plants such
as bryophytes and liverworts form AM-like symbioses.
 There are between 150 and 200 species currently known,
although the taxonomy is still largely based on
characteristics of the relatively large (40 to 800 µm
diameter) multinucleate spores that are formed on the
mycelium external to the host root.
 Studies using molecular markers have revealed a great deal
of diversity suggesting that the number of 200 or so
described ‘morphospecies’ might be an underestimate of
the true diversity of the phylum
 There is no evidence that the Glomeromycota
reproduce sexually and it is assumed that the spores are
formed asexually.
 The spores have a layered wall and features of this can
be used to describe species.
 Similarly, spores may be formed singly, in clusters or
aggregated in so-called sporocarps and the mode of
spore formation has been important in describing
genera and families.
 Ten genera are recognised in the Glomeromycotina.
 Glomus, is the largest genus in the subphylum, with more
than 70 morphospecies (spores, typically with layered
wall structure, are formed by budding from a hyphal
tip), placed in the Family Glomeraceae, Order
Glomerales.
 Gigaspora and Scutellospora are closely related genera in the family
Gigasporaceae; their spores form on a bulbous sporogenous cell and
germinate through a newly formed opening in the spore wall; the
genus Pacispora is related but separated from these.
 The family Acaulosporaceae includes the
genera Acaulospora and Entrophospora; which are related
to Diversispora.
 Paraglomus (in the Paraglomerales) is ancient, but forms a sister
group to the Archaeosporales, which includes the
genera Geosiphon and Archaeospora.
 Geosiphon pyriformis is interesting, as it forms an apparently unique
symbiosis with the cyanobacterium Nostoc punctiforme.
 This is structurally the reverse of the situation found in other AM fungi
because the photobionts is harboured in fungal bladders up to 2 mm
in size; so in this case the fungus is the macrosymbiont (or exhabitant)
and the prokaryotic photosynthetic partner is the microsymbiont
or inhabitant.
 Because AM fungi are obligately biotrophic, attempts to
grow them in axenic culture have so far failed; though
the spores will germinate in axenic culture, they
subsequently die.
 However, since the majority of commercially grown
plants form AM associations, interest in developing ways
of inoculating crops with mycorrhizal fungus continues to
increase (see below, section on the commercial
applications of mycorrhizas).
 The highly branched arbuscule produces a large surface
area which enhances exchange of nutrients between the
partners.
 Bidirectional nutrient transfer is a key feature of the
mycorrhizal symbiosis. In arbuscular mycorrhizas the
major nutrients exchanged are:
 reduced carbon, created through the photosynthetic
activity of the plant partner; and
 phosphate, mobilised and taken up by the fungal
hyphae through their exploration of soil microhabitats.
 however, although this is a neglected area of research,
these mycorrhizas probably also contribute to nitrogen-
cycling in the soil.
 This two-way exchange of nutrients takes place between
the plant root cortical cell and the fungal arbuscule that
has penetrated it;
 this is the symbiotic interface and on the one side it is
bordered by the plant plasma membrane and its
extracellular matrix, and on the other side by the fungal
plasma membrane and its extracellular matrix.
 Nutrient transfer between fungus and plant
is indirectly linked; that is, there is no ‘one-for-one’ linkage
between, say, glucose and phosphate exchange.
 This is indicated by the fact that the same host plant can be
in symbiosis with different AM fungi and, when this happens,
there can be very different carbohydrate-phosphate
exchange ratios between the different plant-fungus pair
wise combinations.
 In addition, carbohydrate transfer can be decoupled from
phosphate transfer if the soil has high phosphate availability.
 The process of transfer of nutrients between the plant and
the fungus is believed to involve passive efflux of solutes from
each donor organism into the interfacial apoplast
(the apoplast is the free diffusional space between the two,
plant and fungal, plasma membranes), followed by active
uptake by the receiver organism.
 All the active transporters that have been implicated so
far in this mechanism are proton-pumping ATPase
symporters.
 Specifically: on the fungal side there is an ATPase that
pumps protons into the arbuscule at the expense of ATP
and simultaneously takes in glucose, and on the plant
side there are mycorrhiza-inducible plant phosphate
and ammonium transporters in mycorrhizal roots.
 There is a tripartite symbiosis between leguminous plants,
their arbuscular mycorrhizal fungi and their rhizobial
bacteria that form nitrogen-fixing nodules on the legume
roots.
 This is an association at the molecular level that is crucial
for the nitrogen uptake by legumes.
 No eukaryotic enzymes can break the triple bond
between nitrogen atoms in atmospheric gaseous
nitrogen, N2.
 The reduction of nitrogen to ammonia (nitrogen fixation)
is limited to prokaryotes and is catalysed by the enzyme
nitrogenase.
 Most of the nitrogen entering the biosphere (around
100 million metric tonnes of N2 each year) does so
through the activity of nitrogenase (the other significant
source, lightning, contributes about 10%).
 Consequently, plants that can form a mutualism with
nitrogen-fixing bacteria not only have a significant
selective advantage under conditions of limiting nitrogen
but also make a major contribution to the availability of
reduced-nitrogen in the biosphere.
 By horizontal gene transfer, the bacteria in the Rhizobium-
legume symbiosis have inherited the ability to infect legume
roots and form nitrogen-fixing nodules from the tissues of the
root.
 These provide a habitat for the bacteria that is perfectly
suited for N2 reduction by providing to the bacteria a supply
of carbon (as malate from the plant) and an environment
with a low O2 tension (required for nitrogenase activity).
 Bacteria are housed in specialised intracellular membrane
compartments, formed by the host plant, which control the
bidirectional exchange of nutrients and, therefore, forms the
essential core of the endosymbiosis.
 Recent studies have demonstrated that the Rhizobium-
legume symbiosis makes use of a signalling pathway, receptor,
and regulators of exocytotic vesicle trafficking derived from the
more ancient arbuscular mycorrhizal symbiosis to form this
bacterium-plant symbiotic interface.
 Furthermore, the beneficial effects of the nitrogen-
fixing nodules can be communicated by the
mycorrhiza to non-leguminous plants with which it
also forms a mycorrhizal symbiosis.
 For example, inoculating with both arbuscular
mycorrhizal fungus and Rhizobium in a
soybean/maize intercropping system improved the
nitrogen fixation efficiency of soybean and
promoted nitrogen transfer from soybean to maize,
resulting in yield improvement in this legume/non-
legume intercropping system.