Patterns of diversity and adaptation in Glomeromycota from three prairie grasslands

Arbuscular mycorrhizal (AM) fungi are widespread root symbionts that often improve the fitness of their plant hosts. We tested whether local adaptation in mycorrhizal symbioses would shape the community structure of these root symbionts in a way that maximizes their symbiotic functioning. We grew a native prairie grass (Andropogon gerardii) with all possible combinations of soils and AM fungal inocula from three different prairies that varied in soil characteristics and disturbance history (two native prairie remnants and one recently restored). We identified the AM fungi colonizing A. gerardii roots using PCR amplification and cloning of the small subunit rRNA gene. We observed 13 operational taxonomic units (OTUs) belonging to six genera in three families. Taxonomic richness was higher in the restored than the native prairies with one member of the Gigaspora dominating the roots of plants grown with inocula from native prairies. Inoculum source and the soil environment influenced the composition of AM fungi that colonized plant roots. Correspondingly, host plants and AM fungi responded significantly to the soil–inoculum combinations such that home fungi often had the highest fitness and provided the greatest benefit to A. gerardii. Similar patterns were observed within the soil–inoculum combinations originating from two native prairies, where five sequence types of a single Gigaspora OTU were virtually the only root colonizers. Our results indicate that indigenous assemblages of AM fungi were adapted to the local soil environment and that this process occurred both at a community scale and at the scale of fungal sequence types within a dominant OTU.     

Fungal response from oat (Avena sativa) plants and surface residue in relation to soil aggregation and organic carbon

Abstract

The influence of soil fungi on soil organic carbon (OC) from surface residue was tested in outdoor plots in southern Ontario, Canada, 2004. Fungal hyphal length, soil aggregation, OC and light and heavy fractions of organic matter were compared with factors of plant growth (with or without oat [Avena sativa]) and surface residue (no residue, oat straw (low C:N) or corn (Zea mays) stalks (high C:N)) in a factorial arrangement. Significant increases were observed in soil OC from the oat plants, and from corn stalks compared to straw residue, in the growing season with very moist, high OC, sandy soil. In treatments with corn stalk residue, fungal hyphal length was increased with interaction from the oat plants and residue and was positively correlated with the heavy fraction organic matter along with soil OC. Fungal hyphae, plant roots and high C:N residue were all factors in soil OC increases.     

Glomalean mycorrhizal fungi from tropical Australia

 A comparison of different methods for isolation of vesicular-arbuscular mycorrhizal (VAM) fungi into open-pot cultures was undertaken as part of a study of the diversity of these fungi. Four different isolation techniques using spores separated from soil, soil trap cultures, root samples, or transplanted seedlings grown in intact soil cores were used to obtain as many fungi as possible from each site. Isolation methods were compared using paired samples from the same locations within natural (savanna, rocky hill, wetland, rainforest) and disturbed (minesite) habitats in a seasonally dry tropical region in the Northern Territory of Australia. There were large differences in (i) the efficiency (rate of increase in mycorrhizal colonisation), (ii) the proportion of successful cultures, (iii) fungal diversity (number of fungal species in each culture) and (iv) specificity (identity of species isolated) between these four procedures. However, the less-efficient procedures generally resulted in a higher proportion of cultures of one fungus, which could be used without further isolation steps. Most species of Scutellospora, Acaulospora and Gigaspora were obtained primarily from field-collected spores, but only 50% of these culture attempts were successful. Spores from these initial cultures produced mycorrhizas much more rapidly and successfully when used to start second-generation cultures. Several species of fungi, rarely recovered as living spores from field soils, were dominant in many trap cultures started from soil or roots. Most of these fungi were Glomus species, that were first distinguished by colonisation patterns in roots and eventually identified after sporulation in second- or third-generation trap cultures. These experiments demonstrated that glomalean fungi in the habitats sampled belonged to two functional categories, based on whether or not spores were important propagules. The "non-sporulating" fungi were dominant in many trap cultures, which suggests that these fungi had higher total inoculum levels in soils than other fungi. Pot-culturing methods provided additional information on fungal diversity which complemented spore occurrence data obtained using the same soil samples and provided valuable new information about the biology of these fungi. Accepted: 26 December 1998     

Mycorrhizal colonization mediated by species interactions in arctic tundra

The Alaskan tussock tundra is a strongly nutrient-limited ecosystem, where almost all vascular plant species are mycorrhizal. We established a long-term removal experiment to document effects of arctic plant species on ecto- and ericoid mycorrhizal fungi and to investigate whether species interactions and/or nutrient availability affect mycorrhizal colonization. The treatments applied were removal of Betula nana (Betulaceae, dominant deciduous shrub species), removal of Ledum palustre (Ericaceae, dominant evergreen shrub species), control (no removal), and each of these three treatments with the addition of fertilizer. After 3 years of Ledum removal and fertilization, we found that overall ectomycorrhizal colonization in Betula was significantly reduced. Changes in ectomycorrhizal morphotype composition in removal and fertilized treatments were also observed. These results suggest that the effect of Ledum on Betula 's mycorrhizal roots is due to sequestration of nutrients by Ledum, leading to reduced nutrient availability in the soil. In contrast, ericoid mycorrhizal colonization was not affected by fertilization, but the removal of Betula and to a lower degree of Ledum resulted in a reduction of ericoid mycorrhizal colonization suggesting a direct effect of these species on ericoid mycorrhizal colonization. Nutrient availability was only higher in fertilized treatments, but caution should be taken with the interpretation of these data as soil microbes may effectively compete with the ion exchange resins for the nutrients released by plant removal in these nutrient-limited soils.     

<Emphasis Type="Italic">Pterostylis nutans</Emphasis> (Orchidaceae) has a specific association with two <Emphasis Type="Italic">Ceratobasidium</Emphasis> root-associated fungi across its range in eastern Australia

In this study, we have identified the root-associated fungi of a common species of terrestrial orchid across its range in eastern Australia. We have amplified and cloned fungal ITS DNA extracted from roots of 15 Pterostylis nutans R. Br. plants from six separate geographic localities. Sequencing and GenBank comparison demonstrated two species of Ceratobasidium fungi as the main fungal partners of the orchid. Uncommon fungal associates included homobasidiomycete species such as a Gymnomyces sp. and a Tricholoma sp., Leptodontidium orchidicola, and an unidentified soil fungus. These results demonstrate that specificity for fungal partners occurs in P. nutans and reinforces the idea that conservation measures for endangered Australian orchids must include ex situ perpetuation of fungal symbionts as well as plant material.     

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.     

Mixotrophy in orchids: insights from a comparative study of green individuals and nonphotosynthetic individuals of Cephalanthera damasonium

Some green orchids obtain carbon (C) from their mycorrhizal fungi and photosynthesis. This mixotrophy may represent an evolutionary step towards mycoheterotrophic plants fully feeding on fungal C. Here, we report on nonphotosynthetic individuals (albinos) of the green Cephalanthera damasonium that likely represent another evolutionary step. Albino and green individuals from a French population were compared for morphology and fertility, photosynthetic abilities, fungal partners (using microscopy and molecular tools), and nutrient sources (as characterized by 15N and 13C abundances). Albinos did not differ significantly from green individuals in morphology and fertility, but tended to be smaller. They harboured similar fungi, with Thelephoraceae and Cortinariaceae as mycorrhizal partners and few rhizoctonias. Albinos were nonphotosynthetic, fully mycoheterotrophic. Green individuals carried out photosynthesis at compensation point and received almost 50% of their C from fungi. Orchid fungi also colonized surrounding tree roots, likely to be the ultimate C source. Transition to mycoheterotrophy may require several simultaneous adaptations; albinos, by lacking some of them, may have reduced ecological success. This may limit the appearance of cheaters in mycorrhizal networks.     

Impact of soil warming and shading on colonization and community structure of arbuscular mycorrhizal fungi in roots of a native grassland community

Arbuscular mycorrhizal (AM) fungi have a major influence on the structure, responses and below‐ground C allocation of plant communities. Our lack of understanding of the response of AM fungi to factors such as light and temperature is an obstacle to accurate prediction of the impact of global climate change on ecosystem functioning. In order to investigate this response, we divided a grassland site into 24 plots, each either unshaded or partly shaded with soil either unheated or heated by 3°C at 2 cm depth. In both short‐term studies in spring and autumn, and in a 1‐year‐long study, we measured root length colonization (L_RC) by AM and non‐AM fungi. For selected root samples, DNA sequences were amplified by PCR with fungal‐specific primers for part of the small sub‐unit (SSU) rRNA gene. In spring, the total L_RC increased over 6 weeks from 12% to 25%. Shading significantly reduced AM but increased non‐AM fungal colonization, while soil warming had no effect. In the year‐long study, colonization by AM fungi peaked in summer, whereas non‐AM colonization peaked in autumn, when there was an additive effect of shading and soil warming that reduced AM but increased non‐AM fungi. Stepwise regression revealed that light received within the 7 days prior to sampling was the most significant factor in determining AM L_RC and that mean temperature was the most important influence on non‐AM L_RC. Loglinear analysis confirmed that there were no seasonal or treatment effects on the host plant community. Ten AM fungal sequence types were identified that clustered into two families of the Glomales, Glomaceae and Gigasporaceae. Three other sequence types were of non‐AM fungi, all Ascomycotina. AM sequence types showed seasonal variation and shading impacts: loglinear regression analysis revealed changes in the AM fungal community with time, and a reduction of one Glomus sp. under shade, which corresponded to a decrease in the abundance of Trifolium repens. We suggest that further research investigating any impacts of climate change on ecosystem functioning must not only incorporate their natural AM fungal communities but should also focus on niche separation and community dynamics of AM fungi.