Diversity and classification of mycorrhizal associations

Most mycorrhizas are 'balanced' mutualistic associations in which the fungus and plant exchange commodities required for their growth and survival. Myco-heterotrophic plants have 'exploitative' mycorrhizas where transfer processes apparently benefit only plants. Exploitative associations are symbiotic (in the broad sense), but are not mutualistic. A new definition of mycorrhizas that encompasses all types of these associations while excluding other plant-fungus interactions is provided. This definition recognises the importance of nutrient transfer at an interface resulting from synchronised plant-fungus development. The diversity of interactions between mycorrhizal fungi and plants is considered. Mycorrhizal fungi also function as endophytes, necrotrophs and antagonists of host or non-host plants, with roles that vary during the lifespan of their associations. It is recommended that mycorrhizal associations are defined and classified primarily by anatomical criteria regulated by the host plant. A revised classification scheme for types and categories of mycorrhizal associations defined by these criteria is proposed. The main categories of vesicular-arbuscular mycorrhizal associations (VAM) are 'linear' or 'coiling', and of ectomycorrhizal associations (ECM) are 'epidermal' or 'cortical'. Subcategories of coiling VAM and epidermal ECM occur in certain host plants. Fungus-controlled features result in 'morphotypes' within categories of VAM and ECM. Arbutoid and monotropoid associations should be considered subcategories of epidermal ECM and ectendomycorrhizas should be relegated to an ECM morphotype. Both arbuscules and vesicles define mycorrhizas formed by glomeromycotan fungi. A new classification scheme for categories, subcategories and morphotypes of mycorrhizal associations is provided.     

Specificity in host-fungus associations: Do mutualists differ from antagonists?

Summary Physically intimate interactions between organisms are assumed to be highly specific, yet intimate mutualisms exhibiting little specificity are common and important in many communities. We compare host records for ectomycorrhizal fungi (mutualists) to those for biotrophic shoot fungi and necrotrophic root fungi (both antagonists) in order to test two alternative predictions: (1) intimate physical associations (biotrophy) are more specific than less intimate ones (necrotrophy); (2) antagonisms are more specific than mutualisms. Specificity of fungi for hosts supports prediction (1): ectomycorrhizal fungi and shoot biotrophs are more host specific than root necrotrophs. Fungal symbiont ranges of hosts supports prediction (2): woody hosts are associated with a greater number of mutualistic fungi than antagonistic fungi. The numbers of fungi in the three groups infecting hosts are all significantly positively correlated. This result suggests that some hosts are resistant to fungal invasion and others are quite susceptible. Thus, plants may not be able to erect selective barriers to only antagonistic fungi. The marked asymmetry of specificity from the perspectives of hosts vs fungi suggests that evolutionary and ecological processes act differently on partners in symbioses.     

Dark septate endophytes – are they mycorrhizal?

Dark septate endophytes (DSE) are a miscellaneous group of ascomycetous anamorphic fungi that colonize root tissues intracellularly and intercellularly. The limited selection of studies quoted here exemplifies the range of host responses to symbiotic DSE fungi. Like mycorrhizal associations, DSE associations vary from negative to neutral and positive when measured by host performance or host tissue nutrient concentrations. This range of host responses is partially attributable to variation between different fungus taxa and strains. Similarly, hosts differ in their responses to a single DSE strain. Experimental conditions may also govern the nature of the symbiotic association. It is concluded that DSE are capable of forming mutualistic associations functionally similar to mycorrhizas. If the variation in host response to mycorrhizal fungi is considered to represent a continuum ranging from parasitism to mutualism, DSE symbiosis must be considered mycorrhizal, at least under some conditions.     

Further advances in orchid mycorrhizal research

Orchid mycorrhizas are mutualistic interactions between fungi and members of the Orchidaceae, the world’s largest plant family. The majority of the world’s orchids are photosynthetic, a small number of species are myco-heterotrophic throughout their lifetime, and recent research indicates a third mode (mixotrophy) whereby green orchids supplement their photosynthetically fixed carbon with carbon derived from their mycorrhizal fungus. Molecular identification studies of orchid-associated fungi indicate a wide range of fungi might be orchid mycobionts, show common fungal taxa across the globe and support the view that some orchids have specific fungal interactions. Confirmation of mycorrhizal status requires isolation of the fungi and restoration of functional mycorrhizas. New methods may now be used to store orchid-associated fungi and store and germinate seed, leading to more efficient culture of orchid species. However, many orchid mycorrhizas must be synthesised before conservation of these associations can be attempted in the field. Further gene expression studies of orchid mycorrhizas are needed to better understand the establishment and maintenance of the interaction. These data will add to efforts to conserve this diverse and valuable association.     

A leaf-rolling weevil benefits from general saprophytic fungi in polysaccharide degradation

Insects, especially those feeding on leaf litter, widely form symbiosis with fungi. As dead plant tissues provide insects with poor-quality diets, which contain relatively high levels of indigestible lignin and cellulose, some saprophytic fungi may increase nutrient availability by polysaccharide degradation. Although the inherited, obligate bacterial symbionts are well documented, the non-inherited, facultative fungal symbionts are relatively overlooked. Females of the leaf-rolling weevil Heterapoderopsis bicallosicollis, a specialist of Triadica sebifera, construct leaf-rolls that serve as retreats from which larvae feed internally. We found that fungi associated with leaf-rolls were not transported by the female, but likely originated from the soil. To determine the effects of fungi on H. bicallosicollis development, fungal growth was reduced by a dry treatment. This treatment decreased adult weight and survival, and prolonged larval duration significantly. We further tested the hypothesis that fungi degrade leaf-roll polysaccharides, by a fungus inoculation experiment. Three dominant fungi (Penicillium sp., Aspergillus sp. and Cladosporium sp.) decreased the levels of soluble carbohydrate, cellulose, and lignin in inoculation experiments. Soluble carbohydrate, cellulose, and lignin of leaf-rolls all were found to decrease gradually during insect development. We conclude that these saprophytic fungi form facultative associations with H. bicallosicollis and benefit weevil nutrition by polysaccharide decomposition. Our study highlights the significance of fungal symbionts in insect nutritional ecology.     

A novel ascomycetous endophytic association in the rhizoids of the leafy liverwort family, Schistochilaceae (Jungermanniidae, Hepaticopsida)

Liverworts form diverse associations with endophytic fungi similar to mycorrhizas in vascular plants. Whereas the widespread occurrence of glomeromycotes in the basal liverwort lineages is well documented, knowledge of the distribution of ascomycetes and basidiomycetes in derived thalloid and leafy clades is more fragmented. Our discovery that the ramified and septate rhizoids of the Schistochilaceae, the sister group to all other ascomycete-containing liverworts, are packed with fungal hyphae prompted this study on the effects of the fungi on rhizoid morphology, host specificity, the cytology of the association, and a molecular analysis of the endophytes. Two species of Pachyschistochila and their fungi were grown axenically. Axenic rhizoids were unbranched and nonseptate. Reinfected with their own fungus and that from the other species, both Pachyschistochila species produced branched and septate rhizoids identical to those in nature. Woronin bodies and simple septa identified the fungus as an ascomycete referable, according to phylogenetic analyses of ITS sequences, to the Rhizoscyphus (Hymenoscyphus) ericae aggregate, also found in other liverwort-ascomycete associations and in mycorrhizas in the Ericales. Healthy hyphae and host cytoplasm suggest that the Schistochila-fungus association reflects a balanced mutualistic relationship. The recent dating of the divergence of the Jungermanniales from the fungus-free Porellales in the Permian and the origins of the Schistochilaceae in the Triassic indicate that these associations in liverworts predate the appearance of the Ericales.     

Infection structures of biotrophic and hemibiotrophic fungal plant pathogens

Biotrophic plant pathogenic fungi are one of the major causes of crop losses. The infection processes they exhibit are typified by infected host plant cells remaining alive for several days. This requires the development of specialized infection structures such as haustoria which are produced by obligate biotrophs, and intracellular hyphae which are produced by many hemibiotrophs. These infection hyphae are surrounded by the host plant plasma membrane, and in the case of haustoria the extrahaustorial membrane differs biochemically and structurally from the normal membrane. An interfacial matrix separates haustoria and intracellular hyphae from the invaginated membrane and this seems to be characteristic of biotrophic interactions. There is clear evidence for molecular differentiation of the haustorial plasma membrane in powdery mildews and rusts in comparison with the other fungal membranes. Relatively few pathogenicity genes related to biotrophy, and the switch from biotrophy to necrotrophy in hemibiotrophs, have been identified.     

Plants,mycorrhizal fungi and endobacteria: a dialog among cells and genomes

This review focuses on mycorrhizas, which are associations between fungi and the roots of 90% of terrestrial plants. These are the most common symbioses in the world; they involve about 6000 species of fungi distributed through all the fungal phyla and about 240000 species of plants, including forest and crop plants. Thanks to mycorrhizal symbiosis and nutrient exchanges, regulated by complex molecular signals, the plant improves its vegetative growth, while the fungus accomplishes its life cycle. Molecular and cellular analyses demonstrate that during colonization the cellular organization of the two eukaryotes is completely remodeled. For example, in cortical cells, structural modifications involve both the host and the microbiont. Recent studies revealed that in arbuscular mycorrhizas (AM), system complexity is increased by the presence of a third symbiont: a bacterium living inside the fungus. The presence of this resident genome makes the investigation of the molecular dialogues among the symbiotic partners even more complex. Molecular analysis showed that the bacterium has genes involved in the acquisition of mineral nutrients. The experimental data support the current view that mycorrhizal symbioses are often tripartite associations.     

Asymmetric interaction specificity between two sympatric termites and their fungal symbionts

Abstract.  1. Fungus-growing termites live in an obligate mutualistic symbiosis with Termitomyces fungi. The functions of the fungal symbiont have been hypothesised to differ between species and to range from highly specific roles of providing plant-degrading enzymes complementary to termite gut enzymes, to non-specific roles of providing protein-rich food to the termites.
2. Termite species with unspecialised fungal symbionts are predicted to be associated with a wider range of symbionts than species with specialised symbionts. Recent DNA data have confirmed this prediction, but evidence for differences in functional specificity has been sparse and indirect.
3. Here the consequences of symbiont interaction specificity are experimentally tested by reciprocally exchanging the fungal symbionts of sympatric colonies of Macrotermes natalensis and Odontotermes badius , which were inferred to have specialised and non-specialised symbionts respectively.
4. As expected, survival of O. badius termites on M. natalensis fungus was not significantly worse than on their own fungus, but survival of M. natalensis termites on O. badius fungus was significantly reduced.
5. This asymmetric result confirms that symbiont roles differ significantly between macrotermitine genera and indicates that symbiont transplantation experiments are a powerful tool for testing the functional details of mutualistic symbioses.     

Rhizosphere communication of plants, parasitic plants and AM fungi

Plants use an array of secondary metabolites to defend themselves against harmful organisms and to attract others that are beneficial. However, the attraction of beneficial organisms could also lead to abuse by malevolent organisms. An exciting example of such abuse is the relationship between plants, beneficial mutualistic arbuscular mycorrhizal fungi and harmful parasitic plants. Signalling molecules called strigolactones, which are secreted by plant roots in low concentrations, induce the growth of both obligate biotrophs. Here, we review the importance of strigolactones for these two interactions and discuss possible developments that should further clarify the role of these signalling molecules in rhizosphere processes.     

Challenges and progress towards understanding the role of effectors in plant-fungal interactions

Both mutualistic and biotrophic pathogenic fungi rely on living host plants for growth and reproduction and must modify host cell structure and function for successful infection. The deployment of a diverse set of secreted virulence determinants referred to as 'effectors', many of which are directly delivered into the host cell, is postulated to be the key to host infection. This review provides a snapshot of the current progress in fungal effector biology. Recent genome sequencing of rust and powdery mildew obligate biotrophs has provided insight into the repertoires of potential effectors of these highly specialised pathogens. Identification of the first host-translocated effectors from mutualistic fungi has revealed that these fungi also manipulate host cells through effectors. The biological activities of some fungal effectors are just beginning to be revealed, while much uncertainty still surrounds the mechanisms of transport into host cells.     

Changing partners in the dark: isotopic and molecular evidence of ectomycorrhizal liaisons between forest orchids and trees

In the mycorrhizal symbiosis, plants exchange photosynthates for mineral nutrients acquired by fungi from the soil. This mutualistic arrangement has been subverted by hundreds of mycorrhizal plant species that lack the ability to photosynthesize. The most numerous examples of this behaviour are found in the largest plant family, the Orchidaceae. Although these non-photosynthetic orchid species are known to be highly specialized exploiters of the ectomycorrhizal symbiosis, photosynthetic orchids are thought to use free-living saprophytic, or pathogenic, fungal lineages. However, we present evidence that putatively photosynthetic orchids from five species which grow in the understorey of forests: (i) form mycorrhizas with ectomycorrhizal fungi of forest trees; and (ii) have stable isotope signatures indicating distinctive pathways for nitrogen and carbon acquisition approaching those of non-photosynthetic orchids that associate with ectomycorrhizal fungi of forest trees. These findings represent a major shift in our understanding of both orchid ecology and evolution because they explain how orchids can thrive in low-irradiance niches and they show that a shift to exploiting ectomycorrhizal fungi precedes viable losses of photosynthetic ability in orchid lineages.     

GR24, a synthetic analog of strigolactones, stimulates the mitosis and growth of the arbuscular mycorrhizal fungus Gigaspora rosea by boosting its energy metabolism

Arbuscular mycorrhizal (AM) fungi are obligate biotrophs that participate in a highly beneficial root symbiosis with 80% of land plants. Strigolactones are trace molecules in plant root exudates that are perceived by AM fungi at subnanomolar concentrations. Within just a few hours, they were shown to stimulate fungal mitochondria, spore germination, and branching of germinating hyphae. In this study we show that treatment of Gigaspora rosea with a strigolactone analog (GR24) causes a rapid increase in the NADH concentration, the NADH dehydrogenase activity, and the ATP content of the fungal cell. This fully and rapidly (within minutes) activated oxidative metabolism does not require new gene expression. Up-regulation of the genes involved in mitochondrial metabolism and hyphal growth, and stimulation of the fungal mitotic activity, take place several days after this initial boost to the cellular energy of the fungus. Such a rapid and powerful action of GR24 on G. rosea cells suggests that strigolactones are important plant signals involved in switching AM fungi toward full germination and a presymbiotic state.     

Mycorrhizal status of epiphytes in Malaysian oil palm plantations

Epiphytic plants were collected from four oil palm plantations in Peninsular Malaysia and their mycorrhizal status determined. Conspecific plants with a terrestrial habit (16 species) and rhizosphere soils were also examined for mycorrhizal colonization and glomalean fungi, respectively. Twelve species of glomalean fungi were recovered from the four oil palm plantation soils. Of the 29 epiphytic species in 16 families belonging to the bryophytes, pteridophytes and angiosperms, only four species of angiosperms that were facultative epiphytes and a hemiepiphyte growing within 0.4 m of ground level had vesicular-arbuscular mycorrhizal (VAM) fungi. Bioassays of organic debris from oil palm trunks did not produce vesicular-arbuscular mycorrhizas on maize. Six epiphytic species grown in the greenhouse in pots containing oil palm rhizosphere soils rooted and had VAM fungi and thus may be facultative epiphytes. Five other epiphyte species failed to grow in pots and are probably obligate epiphytes. Seven epiphyte species that established themselves in pots failed to form vesicular-arbuscular mycorrhizas.     

Novel genes induced during an arbuscular mycorrhizal (AM) symbiosis formed between Medicago truncatula and Glomus versiforme

Many terrestrial plant species are able to form symbiotic associations with arbuscular mycorrhizal fungi. Here we have identified three cDNA clones representing genes whose expression is induced during the arbuscular mycorrhizal symbiosis formed between Medicago truncatula and an arbuscular mycorrhizal fungus, Glomus versiforme. The three clones represent M. truncatula genes and encode novel proteins: a xyloglucan endotransglycosylase-related protein, a putative arabinogalactan protein (AGP), and a putative homologue of the mammalian p110 subunit of initiation factor 3 (eIF3). These genes show little or no expression in M. truncatula roots prior to formation of the symbiosis and are significantly induced following colonization by G. versiforme. The genes are not induced in roots in response to increases in phosphate. This suggests that induction of expression during the symbiosis is due to the interaction with the fungus and is not a secondary effect of improved phosphate nutrition. In situ hybridization revealed that the putative AGP is expressed specifically in cortical cells containing arbuscules. The identification of two mycorrhiza-induced genes encoding proteins predicted to be involved in cell wall structure is consistent with previous electron microscopy data that indicated major alterations in the extracellular matrix of the cortical cells following colonization by mycorrhizal fungi.     

Evolutionary stability in a 400-million-year-old heritable facultative mutualism

Many eukaryotes interact with heritable endobacteria to satisfy diverse metabolic needs. Some of these interactions are facultative symbioses, in which one partner is not essential to the other. Facultative symbioses are expected to be transitional stages along an evolutionary trajectory toward obligate relationships. We tested this evolutionary theory prediction in Ca. Glomeribacter gigasporarum, nonessential endosymbionts of arbuscular mycorrhizal fungi (Glomeromycota). We found that heritable facultative mutualisms can be both ancient and evolutionarily stable. We detected significant patterns of codivergence between the partners that we would only expect in obligate associations. Using codiverging partner pairs and the fungal fossil record, we established that the Glomeromycota-Glomeribacter symbiosis is at least 400 million years old. Despite clear signs of codivergence, we determined that the Glomeribacter endobacteria engage in recombination and host switching, which display patterns indicating that the association is not evolving toward reciprocal dependence. We postulate that low frequency of recombination in heritable endosymbionts together with host switching stabilize facultative mutualisms over extended evolutionary times.     

Specific, non-nutritional association between an ascomycete fungus and Allomerus plant-ants

Ant-fungus associations are well known from attine ants, whose nutrition is based on a symbiosis with basidiomycete fungi. Otherwise, only a few non-nutritional ant-fungus associations have been recorded to date. Here we focus on one of these associations involving Allomerus plant-ants that build galleried structures on their myrmecophytic hosts in order to ambush prey. We show that this association is not opportunistic because the ants select from a monophyletic group of closely related fungal haplotypes of an ascomycete species from the order Chaetothyriales that consistently grows on and has been isolated from the galleries. Both the ants' behaviour and an analysis of the genetic population structure of the ants and the fungus argue for host specificity in this interaction. The ants' behaviour reveals a major investment in manipulating, growing and cleaning the fungus. A molecular analysis of the fungus demonstrates the widespread occurrence of one haplotype and many other haplotypes with a lower occurrence, as well as significant variation in the presence of these fungal haplotypes between areas and ant species. Altogether, these results suggest that such an interaction might represent an as-yet undescribed type of specific association between ants and fungus in which the ants cultivate fungal mycelia to strengthen their hunting galleries.     

Die arbuskuläre Mykorrhiza: Eine unterirdische Lebensgemeinschaft

During evolution, plants and some fungi have developped an intimate underground association which established a very successful symbiosis, the arbuscular mycorrhiza. Mycorrhizal fungi play a vital role in water and mineral nutrient supply for plant growth as well as in development of diversity and increased productivity in plant associations. In addition mycorrhizal plants exhibit increased resistance towards pathogens. Plants ‚pay’︁ for these benefits by supplying the fungus with carbohydrates (glucose, fructose). With some success in mycorrhiza research on the metabolic and genetic levels, we gradually touch complexity of its molecular interactions. We expect that the growing interest in mycorrhiza research will lead in the near future to new insights into the strategies of plants and fungi to develop mutualistic symbiotic associations.