Mycorrhiza

Landrace maize varieties differ from conventional and genetically modified hybrid maize in response to inoculation with arbuscular mycorrhizal fungi Abstract Land area planted with genetically modified (GM) crops has grown rapidly, and Brazil has the second largest area with those plants. There is, however, limited information on the possible effects of that technology on non-target organisms, especially root symbionts, such as arbuscular mycorrhizal fungi (AMF). We evaluated AMF symbiosis development in five maize genotypes: one landrace, two conventional hybrids (DKB 240 and Formula), and two GM hybrids (DKB 240-VT Pro and Formula TL). We evaluated symbiosis response in two separate experiments: one in autumn and the other in summer. Plants were inoculated with Rhizophagus clarus(Rc) and Gigaspora margarita (Gm) and compared to plants without inoculation. We evaluated root colonization, spore number, and plant biomass and phosphorous accumulation 30 and 60 days after inoculation. There were no consistent effects of GM crops, but AMF species and maize genotype affected symbiosis development. Formula genotype (isoline and GM) had a negative response to inoculation, with a decrease of around 30% in biomass and P concentration in Rc-inoculated plants. The maize landrace had a positive response, with increases of 17% and 14% in the same variables. DKB genotype (isoline and GM) showed negative, positive, and neutral effects. The results show that plant genetic identity is a determinant factor in symbiosis performance, suggesting that plants selected in low P availability can make better use of mycorrhizal symbiosis. Given the role that AMF play in different ecosystem processes, use of landrace maize may contribute to agrobiodiversity conservation.

The effects of co-colonising ectomycorrhizal fungi on mycorrhizal colonisation and sporocarp formation in Laccaria japonica colonising seedlings of Pinus densiflora Abstract Forest trees are colonised by different species of ectomycorrhizal (ECM) fungi that interact competitively or mutualistically with one another. Most ECM fungi can produce sporocarps. To date, the effects of co-colonising fungal species on sporocarp formation in ECM fungi remain unknown. In this study, we examined host plant growth, mycorrhizal colonisation, and sporocarp formation when roots of Pinus densiflora are colonised by Laccaria japonica and three other ECM fungal species (Cenococcum geophilum, Pisolithus sp., and Suillus luteus). Sporocarp numbers were recorded throughout the experimental period. The biomass, photosynthetic rate, and mycorrhizal colonisation rate of the seedlings were also measured at 45 days, 62 days, and 1 year after seedlings were transplanted. Results indicated that C. geophilum and S. luteus may negatively impact mycorrhizal colonisation and sporocarp formation in L. japonica. Sporocarp formation in L. japonica was positively correlated with conspecific mycorrhizal colonisation but negatively correlated with the biomass of seedlings of P. densiflora. The co-occurring ECM fungi largely competed with L. japonica, resulting in various effects on mycorrhizal colonisation and sporocarp formation in L. japonica. A variety of mechanisms may be involved in the competitive interactions among the different ECM fungal species, including abilities to more rapidly colonise root tips, acquire soil nutrients, or produce antibiotics. These mechanisms need to be confirmed in further studies.

Ectomycorrhizal fungal communities are dominated by mammalian dispersed truffle-like taxa in north-east Australian woodlands Abstract Mycorrhizal fungi are very diverse, including those that produce truffle-like fruiting bodies. Truffle-like fungi are hypogeous and sequestrate (produced below-ground, with an enclosed hymenophore) and rely on animal consumption, mainly by mammals, for spore dispersal. This dependence links mycophagous mammals to mycorrhizal diversity and, assuming truffle-like fungi are important components of mycorrhizal communities, to plant nutrient cycling and ecosystem health. These links are largely untested as currently little is known about mycorrhizal fungal community structure and its dependence on mycophagous mammals. We quantified the mycorrhizal fungal community in the north-east Australian woodland, including the portion interacting with ten species of mycophagous mammals. The study area is core habitat of an endangered fungal specialist marsupial, Bettongia tropica, and as such provides baseline data on mycorrhizal fungi-mammal interactions in an area with no known mammal declines. We examined the mycorrhizal fungi in root and soil samples via high-throughput sequencing and compared the observed taxa to those dispersed by mycophagous mammals at the same locations. We found that the dominant root-associating ectomycorrhizal fungal taxa (> 90% sequence abundance) included the truffle-like taxa Mesophellia, Hysterangium and Chondrogaster. These same taxa were also present in mycophagous mammalian diets, with Mesophellia often dominating. Altogether, 88% of truffle-like taxa from root samples were shared with the fungal specialist diet and 52% with diets from generalist mammals. Our data suggest that changes in mammal communities, particularly the loss of fungal specialists, could, over time, induce reductions to truffle-like fungal diversity, causing ectomycorrhizal fungal communities to shift with unknown impacts on plant and ecosystem health.

Phosphorus forms affect the hyphosphere bacterial community involved in soil organic phosphorus turnover Abstract Interactions between bacteria and arbuscular mycorrhizal (AM) fungi play a significant role in mediating organic phosphorus (P) transformations and turnover in soil. The bacterial community in soil is largely responsible for mobilization of the soil organic P pool, and the released P is taken up by extraradical AM hyphae, which mediate its use for plant growth. However, the functional microbiome involved in organic P mineralization in the hyphosphere remains poorly understood. The aim of this study was to determine how AM hyphae-associated bacterial communities related to P turnover in the hyphosphere of leek (Allium porrum) respond to different forms of soil P. Using a compartmented microcosm, leek was grown with the AM fungus Funneliformis mosseae, and the extraradical mycelium of F. mosseae was allowed to grow into a separate hyphal compartment containing either no added P, or P as KH2PO4 or phytin. High-throughput sequencing showed that the alkaline phosphatase (ALP)-harboring bacterial community associated with the AM hyphae was dominated by Sinorhizobium, Bradyrhizobium, Pseudomonas, and Ralstonia and was significantly changed in response to different P treatments, with Pseudomonas showing higher relative abundance in organic P treatments than in control and inorganic P treatments. Pseudomonas was also the major genus harboring the β-propeller phytase (BPP) gene in the hyphosphere, but the BPP-harboring community structure was not affected by the presence of different P forms. These results demonstrate the profound differences in ALP- and BPP-harboring bacterial communities in the hyphosphere at bacterial genus level, providing new insights to link bacteria and biogeochemical P cycling driven in association with mycorrhizal hyphae.

Comparative measurements of arbuscular mycorrhizal fungal responses to agricultural management practices Abstract Arbuscular mycorrhizal (AM) fungi are considered to be a key group of soil organisms for assessments of soil biological properties and developing relationships among crop production management practices, soil properties, crop performance, and ecosystem services. In a field study of cover crop treatments established during the transition from small grains to corn (Zea mays L), we assessed multiple measures of AM fungal responses to the management treatments: soil propagule numbers, biomass via lipid biomarkers, and root colonization extent. Our objectives were to determine response variables that reliably distinguished cover crop treatments and formed consistent relationships with grain yield, plant biomass, and mineral nutrient concentrations of the following corn crop. The number of soil AM fungal propagules and amount of the NLFA biomarker C16:1cis11 measured on fall-collected soils most consistently and significantly responded to fall cover crop treatments. Neither of these measures of soil inoculum potential was strongly related to measures of crop performance. The PLFA biomarker C16:1cis11 was marginally responsive to cover crop but did not strongly relate to crop performance parameters. Corn root colonization by AM fungi was not significantly affected by cover crop treatment, but significant negative relationships were found between root colonization and grain N concentration and plant biomass at maturity. In contrast, a significant positive relationship between root colonization and plant N concentration at the 6-leaf stage was found. Understanding the relative effectiveness and limitations of AM fungal response variables will inform their application in field studies of agricultural management practices.

A single ectomycorrhizal plant root system includes a diverse and spatially structured fungal community Abstract Although only a relatively small proportion of plant species form ectomycorrhizae with fungi, it is crucial for growth and survival for a number of widespread woody plant species. Few studies have attempted to investigate the fine scale spatial structure of entire root systems of adult ectomycorrhizal (EcM) plants. Here, we use the herbaceous perennial Bistorta vivipara to map the entire root system of an adult EcM plant and investigate the spatial structure of its root-associated fungi. All EcM root tips were sampled, mapped and identified using a direct PCR approach and Sanger sequencing of the internal transcribed spacer region. A total of 32.1% of all sampled root tips (739 of 2302) were successfully sequenced and clustered into 41 operational taxonomic units (OTUs). We observed a clear spatial structuring of the root-associated fungi within the root system. Clusters of individual OTUs were observed in the younger parts of the root system, consistent with observations of priority effects in previous studies, but were absent from the older parts of the root system. This may suggest a succession and fragmentation of the root-associated fungi even at a very fine scale, where competition likely comes into play at different successional stages within the root system.

Distinct arbuscular mycorrhizal fungal communities associate with different manioc landraces and Amazonian soils Abstract Manioc (Manihot esculenta Crantz) is an important tropical crop that depends on arbuscular mycorrhizal (AM) association for its nutrition. However, little is known about the richness and species composition of AM fungal communities associating with manioc and possible differences across soils and manioc landraces. We studied the diversity and composition of AM fungal communities present in the roots of different manioc landraces and surrounding soils in indigenous shifting cultivation fields on different Amazonian soil types. A total of 126 AM fungal virtual taxa (VT; phylogenetically defined taxonomic units) were recovered from soil and root samples using 454 sequencing of AM fungal SSU rRNA gene amplicons. Different AM fungal communities occurred in different soil types. Minor differences occurred in the composition of AM fungal community associating with different manioc landraces, but AM fungal richness was not different among them. There was a low similarity between the AM fungal communities colonizing manioc roots and those recorded in the soil, independently of differences in soil properties or the manioc landrace evaluated. Rhizophagus manihotis and Glomus VT126 were the most abundant AM fungal species colonizing manioc roots. Contrasting with the results of earlier spore-based investigations, all the AM fungi identified as indicator species of particular manioc landraces were morphologically unknown Glomus species. In conclusion, different manioc landraces growing in common conditions associated with distinct AM fungal communities, whereby AM fungal communities in soils did not necessarily reflect the AM fungal communities colonizing manioc roots.

Towards the conservation of ectomycorrhizal fungi on endangered trees: native fungal species on Pinus amamiana are rarely conserved in trees planted ex situ Abstract Ectomycorrhizal (ECM) symbiosis is essential for the survival of both host trees and associated ECM fungi. However, during conservation activities of endangered tree species, their ECM symbionts are largely ignored. Here, we investigated ECM fungi in ex situ populations established for the conservation of Pinus amamiana, an endangered species distributed on Yakushima Island, Japan. Our objective was to determine whether ECM fungi in natural forests are conserved in ex situ populations on the same island. In particular, we focused on the existence of Rhizopogon yakushimensis, which is specific to P. amamiana and the most dominant in natural P. amamiana forests. Molecular identification of ECM fungi in resident tree roots and soil propagule banks indicated that ECM fungal species native to natural forests were rarely conserved in ex situ populations. Furthermore, R. yakushimensis was not confirmed in any of the resident root or spore bioassay samples from the ex situ populations. Thus, ECM fungal spores may not be effectively dispersed from natural forests located on the same island. Instead, ECM fungi distributed in other geographical regions occurred more frequently in the ex situ populations, indicating unintentional introductions of non-native ECM fungi from the nurseries where seedlings were raised before transplanting. These findings imply that the current ex situ conservation practices of endangered tree do not work for the conservation of native ECM fungi, and instead may need modification to avoid the risk of introducing non-native ECM fungi near the endangered forest sites.

Maize varieties can strengthen positive plant-soil feedback through beneficial arbuscular mycorrhizal fungal mutualists Abstract Plant-soil feedback (PSF) describes the process whereby plant species modify the soil environment, which subsequently impacts the growth of the same or another plant species. Our aim was to explore PSF by two maize varieties (a landrace and a hybrid variety) and three arbuscular mycorrhizal fungi (AMF) species (Funneliformis mosseae, Claroideoglomus etunicatum, Gigaspora margarita, and the mixture). We carried out a pot experiment with a conditioning and a feedback phase to determine PSF with different species of AMF and with a non-mycorrhizal control. Sterilized soil was conditioned separately by each variety, with or without AMF; in the feedback phase, each soil community was used to grow each in its "home" soil and in the "away" soil. Plant performance was assessed as shoot biomass, phosphorus (P) concentration and P content, and fungal performance was assessed as mycorrhizal colonization and hyphal length density. Both maize varieties were differentially influenced by AMF in the conditioning phase. In the feedback phase, PSF was generally negative for non-mycorrhizal plants or when plants were colonized by G. margarita, whereas PSF was positive in the other three AMF treatments. When plants were grown on home soil, hyphal length density was larger than on away soil. We conclude that different maize varieties can strengthen positive plant-soil feedback for themselves through beneficial mutualists for themselves, but not across the maize varieties.

New insights into black truffle biology: discovery of the potential connecting structure between a Tuber aestivum ascocarp and its host root Abstract According to isotopic labeling experiments, most of the carbon used by truffle (Tuber sp.) fruiting bodies to develop underground is provided by host trees, suggesting that trees and truffles are physically connected. However, such physical link between trees and truffle fruiting bodies has never been observed. We discovered fruiting bodies of Tuber aestivum adhering to the walls of a belowground quarry and we took advantage of this unique situation to analyze the physical structure that supported these fruiting bodies in the open air. Observation of transversal sections of the attachment structure indicated that it was organized in ducts made of gleba-like tissue and connected to a network of hyphae traveling across soil particles. Only one mating type was detected by PCR in the gleba and in the attachment structure, suggesting that these two organs are from maternal origin, leaving open the question of the location of the opposite paternal mating type.