Specificity of fungal associations of Pyroleae and Monotropa hypopitys during germination and seedling development

Mycoheterotrophic plants obtain organic carbon from associated mycorrhizal fungi, fully or partially. Angiosperms with this form of nutrition possess exceptionally small ‘dust seeds’ which after germination develop ‘seedlings’ that remain subterranean for several years, fully dependent on fungi for supply of carbon. Mycoheterotrophs which as adults have photosynthesis thus develop from full to partial mycoheterotrophy, or autotrophy, during ontogeny. Mycoheterotrophic plants may represent a gradient of variation in a parasitism–mutualism continuum, both among and within species. Previous studies on plant–fungal associations in mycoheterotrophs have focused on either germination or the adult life stages of the plant. Much less is known about the fungal associations during development of the subterranean seedlings. We investigated germination and seedling development and the diversity of fungi associated with germinating seeds and subterranean seedlings (juveniles) in five Monotropoideae (Ericaceae) species, the full mycoheterotroph Monotropa hypopitys and the putatively partial mycoheterotrophs Pyrola chlorantha, P. rotundifolia, Moneses uniflora and Chimaphila umbellata. Seedlings retrieved from seed sowing experiments in the field were used to examine diversity of fungal associates, using pyrosequencing analysis of ITS2 region for fungal identification. The investigated species varied with regard to germination, seedling development and diversity of associated fungi during juvenile ontogeny. Results suggest that fungal host specificity increases during juvenile ontogeny, most pronounced in the fully mycoheterotrophic species, but a narrowing of fungal associates was found also in two partially mycoheterotrophic species. We suggest that variation in specificity of associated fungi during seedling ontogeny in mycoheterotrophs represents ongoing evolution along a parasitism–mutualism continuum.     

Fungal host specificity is not a bottleneck for the germination of Pyroleae species (Ericaceae) in a Bavarian forest

Plants that produce dust seeds can recruit fungi to meet their earliest requirements for carbon and other nutrients. This germination strategy, termed initial mycoheterotrophy, has been well investigated among the orchid family, but there are numerous other plant lineages that have independently evolved mycoheterotrophic germination strategies. One of these lineages is the tribe Pyroleae (Ericaceae). While the fungi associated with mature plants in Pyroleae have been fairly well documented, their mycobionts at the germination and seedling stages are largely unknown. Here, we use an in situ seed baiting experiment along with molecular fingerprinting techniques and phylogenetic tests to identify the fungi associated with seedlings of two Pyroleae species, Pyrola chlorantha and Orthilia secunda. Our results indicate that similar to adult plants, Pyroleae seedlings can associate with a suite of ectomycorrhizal fungi. Some seedlings harboured single mycobionts, while others may have been inhabited by multiple fungi. The dominant seedling mycobiont of both Pyroleae species was a fungus of unknown trophic status in the order Sebacinales. This taxon was also the only one shared among seedlings of both investigated Pyroleae species. We discuss these results juxtaposed to orchids and one additional Pyrola species in the context of ontogenetic shifts in fungal host specificity for mycoheterotrophic nutrition.     

Evidence for specificity to Glomus group Ab in two Asian mycoheterotrophic Burmannia species

Nonphotosynthetic mycorrhizal plants, so‐called mycoheterotrophic plants, have long attracted the curiosity of botanists and mycologists. Recent advances in molecular methods based on fungal‐specific PCR amplification have dramatically enhanced the identification of their host mycorrhizal fungi. However, studies investigating the fungal hosts of arbuscular mycorrhizae‐forming mycoheterotrophs are still limited in Asia, which is known as one of the diversity hot spots of mycoheterotrophs that parasitize arbuscular mycorrhizae (AM). Therefore, we aimed to reveal the mycorrhizal associations of two Asian, fully mycoheterotrophic Burmannia species by molecular identification. Sequences of the small subunit ribosomal DNA showed that both Burmannia species are associated with several distinct lineages of Glomus group Ab. Because Glomus group Ab fungi have been confirmed as fungal hosts of various mycoheterotrophic plants in Africa and South America, we suggest they are widely exploited by AM‐forming mycoheterotrophs globally.     

Isotopic evidence of partial mycoheterotrophy in Burmannia coelestis (Burmanniaceae)

The Burmanniaceae contain several lineages of achlorophyllous mycoheterotrophic plants that associate with arbuscular mycorrhizal fungi (AMF). Here we investigate the isotopic profile of a green and potentially mycoheterotrophic plant in situ, Burmannia coelestis, and associated reference plants. We generated δ ¹³C and δ ¹⁵N stable isotope profiles for five populations of B. coelestis. Burmannia coelestis was significantly enriched in ¹³C relative to surrounding C₃ reference plants and significantly depleted in ¹³C relative to C₄ reference plants. No significant differences were detected in ¹⁵N enrichment between B. coelestis and reference plants. The isotopic profiles measured are suggestive of partial mycoheterotrophy in B. coelestis. Within the genus Burmannia transitions to full mycoheterotrophy have occurred numerous times, suggesting that some green Burmannia species are likely to be partially mycoheterotrophic but in many conditions this mode of nutrition may only be detectable using natural abundance stable isotopic methods, such as when associated with C₄ mycorrhizal plants.     

Partial mycoheterotrophy in rhizoctonia-associated orchid Cheirostylis liukiuensis

Partially mycoheterotrophic plant species obtain organic carbon, via both photosynthesis and mycorrhizal symbiosis. In this study, we investigated the mycorrhizal fungi association and nutritional mode of Cheirostylis liukiuensis, which is suspected to be a partial mycoheterotrophic plant, due to its characteristic reduced underground organs, low-light growth environment, and some fully mycoheterotrophic species in the phylogenetically related genera. Molecular analysis of the dominant mycobiont and stable isotope analysis suggested that C. liukiuensis is a partial mycoheterotrophic plant predominantly associate with non-ectomycorrhizal Ceratobasidiaceae fungi. As examples of partial mycoheterotrophic orchids exploiting non-ectomycorrhizal rhizoctonia are still limited, this study provides valuable information on the nutritional modes of green orchids.     

带叶兜兰种子原地共生萌发及有效菌根真菌的分离与鉴定

为获得带叶兜兰(Paphiopedilum hirsutissimum)种子萌发的共生真菌,采用原地共生萌发技术获得了2株自然萌发的小幼苗,并分离和筛选出了有效的种子萌发共生菌——瘤菌根菌(Epulorhiza sp.)。为验证分离菌株对带叶兜兰种子萌发的有效性,将Phs34号菌株与带叶兜兰种子在灭菌后的原生境基质上进行室内共生萌发试验,结果表明,经过6周的培养,对照组没有观察到种子的萌发;接菌的种子胚明显膨大,突破种皮,形成原球茎,平均萌发率为(58.35±3.41)%。这表明分离得到的瘤菌根菌能促进带叶兜兰的种子萌发。     

Comparison of green and albino individuals of the partially mycoheterotrophic orchid Epipactis helleborine on molecular identities of mycorrhizal fungi,nutritional modes and gene expression in mycorrhizal roots

Some green orchids obtain carbon from their mycorrhizal fungi, as well as from photosynthesis. These partially mycoheterotrophic orchids sometimes produce fully achlorophyllous, leaf‐bearing (albino) variants. Comparing green and albino individuals of these orchids will help to uncover the molecular mechanisms associated with mycoheterotrophy. We compared green and albino Epipactis helleborine by molecular barcoding of mycorrhizal fungi, nutrient sources based on ¹⁵N and ¹³C abundances and gene expression in their mycorrhizae by RNA‐seq and cDNA de novo assembly. Molecular identification of mycorrhizal fungi showed that green and albino E. helleborine harboured similar mycobionts, mainly Wilcoxina. Stable isotope analyses indicated that albino E. helleborine plants were fully mycoheterotrophic, whereas green individuals were partially mycoheterotrophic. Gene expression analyses showed that genes involved in antioxidant metabolism were upregulated in the albino variants, which indicates that these plants experience greater oxidative stress than the green variants, possibly due to a more frequent lysis of intracellular pelotons. It was also found that some genes involved in the transport of some metabolites, including carbon sources from plant to fungus, are higher in albino than in green variants. This result may indicate a bidirectional carbon flow even in the mycoheterotrophic symbiosis. The genes related to mycorrhizal symbiosis in autotrophic orchids and arbuscular mycorrhizal plants were also upregulated in the albino variants, indicating the existence of common molecular mechanisms among the different mycorrhizal types.     

Arbuscular mycorrhizal fungal community structures differ between co-occurring tree species of dry Afromontane tropical forest, and their seedlings exhibit potential to trap isolates suited for reforestation

The diversity of arbuscular mycorrhizal (AM) fungi and their broad or narrow association with distinct plant species in natural environments are crucial information in the understanding of the ecological role of AM fungi on plant co-existence. This knowledge is also needed for appropriate mycorrhization of nursery-grown seedlings for forestation efforts. Here, we report results from comparative studies on three co-occurring indigenous tree species of the dry Afromontane forests of Ethiopia and their seedlings grown under controlled conditions in soil collected from the sites. AM fungal SSU rDNA fragment was amplified and sequenced from mycorrhizas of adult plants and seedlings of Olea europaea subsp. cuspidata and Prunus africana, and from Podocarpus falcatus seedlings. AM fungal identity, diversity and community structure were analyzed based on sequence types defined by the NS31-AM1 SSU rDNA fragment similarity in order to compare with data from other habitats. A total of 409 sequences, grouped in 32 sequence types, belonging to Glomeraceae, Diversisporaceae and Gigasporaceae were found. Some sequence types are close to the widespread Glomus intraradices, G. hoi, G. etunicatum, G. cf. etunicatum and Gigaspora margarita. However, the majority (59%) of sequence types are so far specific for the sites including 11 new types when compared with previous data from the same area. The AM fungal community associated with adult plants, including data previously obtained from adult Podocarpus falcatus seedlings, and seedlings of a host species differed significantly, where seedlings trapping a surprising large number of native fungi. AM fungal community structure also differed significantly between host species and sites, respectively. The results confirm previous results from the same area indicating distinct fungal communities associated with the diverse tree species and suggests the potential of these indigenous tree seedlings to trap a wide range of AM fungi appropriate for successful afforestation.     

Mycoheterotrophic germination of Pyrola asarifolia dust seeds reveals convergences with germination in orchids

Dust seeds that germinate by obtaining nutrients from symbiotic fungi have evolved independently in orchids and 11 other plant lineages. The fungi involved in this 'mycoheterotrophic' germination have been identified in some orchids and non-photosynthetic Ericaceae, and proved identical to mycorrhizal fungi of adult plants. We investigated a third lineage, the Pyroleae, chlorophyllous Ericaceae species whose partial mycoheterotrophy at adulthood has recently attracted much attention. We observed experimental Pyrola asarifolia germination at four Japanese sites and investigated the germination pattern and symbiotic fungi, which we compared to mycorrhizal fungi of adult plants. Adult P. asarifolia, like other Pyroleae, associated with diverse fungal species that were a subset of those mycorrhizal on surrounding trees. Conversely, seedlings specifically associated with a lineage of Sebacinales clade B (endophytic Basidiomycetes) revealed an intriguing evolutionary convergence with orchids, some of which also germinate with Sebacinales clade B. Congruently, seedlings clustered spatially together, but not with adults. This unexpected transition in specificity and ecology of partners could support the developmental transition from full to partial mycoheterotrophy, but probably challenges survival and distribution during development. We discuss the physiological and ecological traits that predisposed to the repeated recruitment of Sebacinales clade B for dust seed germination.     

Arbuscular mycorrhizal soil infectivity and spores distribution across plantations of tropical,subtropical and exotic tree species: a case study from the forest reserve of Bandia,Senegal

Several fast‐growing and multipurpose trees such as exotic and valuable native species have been widely used in West Africa to reverse the tendency of massive degradation of plant cover and restore soil productivity. Although benefic effects have been reported on soil stabilization, a lack of information about their impact on soil symbiotic microorganisms still remains. This investigation has been carried out in field trees of 28 years old in a forest reserve at Bandia. To determine the mycorrhizal inoculum potential (MIP) of soils, a mycorrhizal bioassay was conducted using seedlings of Zea mays L. Spores concentration, arbuscular mycorrhizal (AM) fungi morphotypes and mycorrhizal colonization of field plants were examined. Results showed that fungal communities were dominated in all samples by the genus Glomus. Nevertheless, the others genera Gigaspora and Scutellospora occurred preferentially out of the plantations. The number and richness of spores as well as the MIP of soils were decreased in the tree plantations. Accordingly, the amount of annual herbaceous plants kept out of the tree plantations was much greater than those under the tree plantations. The colonization was higher in field root systems of herb plants in comparison with that of the tree plants. Comparisons allowed us to conclude that vegetation type modifies the AM fungal communities, and the results suggest further adoption of management practices that could improve or sustain the development of herbaceous layers and thus promote the AM fungal communities.     

Effects of arbuscular mycorrhiza on community composition and seedling recruitment in temperate forest understory

Arbuscular mycorrhizal (AM) fungal communities can influence the species composition of plant communities. This influence may result from effects of AM on seedling recruitment, although the existing evidence is limited to experimental systems. We addressed the impact of AM fungi on the plant community composition and seedling recruitment of two species – Oxalis acetosella and Prunella vulgaris – in a temperate forest understory. We established a field experiment over two years in which soil fertility (using fertilizer to enhance and sucrose to decrease fertility) and the activity of AM fungi (using fungicide) was manipulated in a factorial design. Species richness, diversity and community composition of understory plants were not influenced by soil fertility or AM fungal activity treatments. However, plant community composition was marginally significantly affected by the interaction of these treatments as the effect of AM fungal activity became evident under enhanced soil fertility. Suppression of AM fungal activity combined with decreased soil fertility increased the number of shoots of herbaceous plants. Unchanged activity of AM fungi enhanced the growth of O. acetosella seedlings under decreased soil fertility, but did not influence the growth of P. vulgaris seedlings. We conclude that the role of AM fungi in structuring plant communities depends on soil fertility. AM fungi can have a strong influence on seedling recruitment, especially for those plants that are characteristic of the habitat.     

Specificity of assemblage,not fungal partner species,explains mycorrhizal partnerships of mycoheterotrophic Burmannia plants

Mycoheterotrophic plants (MHPs) growing on arbuscular mycorrhizal fungi (AMF) usually maintain specialized mycorrhizal associations. The level of specificity varies between MHPs, although it remains largely unknown whether interactions with mycorrhizal fungi differ by plant lineage, species, and/or by population. Here, we investigate the mycorrhizal interactions among Burmannia species (Burmanniaceae) with different trophic modes using high-throughput DNA sequencing. We characterized the inter- and intraspecific dynamics of the fungal communities by assessing the composition and diversity of fungi among sites. We found that fully mycoheterotrophic species are more specialized in their fungal associations than chlorophyllous species, and that this specialization possibly results from the gradual loss of some fungal groups. In particular, although many fungal species were shared by different Burmannia species, fully MHP species typically host species-specific fungal assemblages, suggesting that they have a preference for the selected fungi. Although no apparent cophylogenetic relationship was detected between fungi and plants, we observe that evolutionarily closely related plants tend to have a greater proportion of shared or closely related fungal partners. Our findings suggest a host preference and specialization toward fungal assemblages in Burmannia, improving understanding of interactions between MHPs and fungi.Subject terms: Fungi, Plant sciences, Evolution     

Mycorrhizal diversity,seed germination and long‐term changes in population size across nine populations of the terrestrial orchid Neottia ovata

In plant species that rely on mycorrhizal symbioses for germination and seedling establishment, seedling recruitment and temporal changes in abundance can be expected to depend on fungal community composition and local environmental conditions. However, disentangling the precise factors that determine recruitment success in species that critically rely on mycorrhizal fungi represents a major challenge. In this study, we used seed germination experiments, 454 amplicon pyrosequencing and assessment of soil conditions to investigate the factors driving changes in local abundance in 28 populations of the orchid Neottia ovata. Comparison of population sizes measured in 2003 and 2013 showed that nearly 60% of the studied populations had declined in size (average growth rate across all populations: ?0.01). Investigation of the mycorrhizal fungi in both the roots and soil revealed a total of 68 species of putatively mycorrhizal fungi, 21 of which occurred exclusively in roots, 25 that occurred solely in soil and 22 that were observed in both the soil and roots. Seed germination was limited and significantly and positively related to soil moisture content and soil pH, but not to fungal community composition. Large populations or populations with high population growth rates showed significantly higher germination than small populations or populations declining in size, but no significant relationships were found between population size or growth and mycorrhizal diversity. Overall, these results indicate that temporal changes in abundance were related to the ability of seeds to germinate, but at the same time they provided limited evidence that variation in fungal communities played an important role in determining population dynamics.     

Limitations on orchid recruitment: not a simple picture

Mycorrhizal fungi have substantial potential to influence plant distribution, especially in specialized orchids and mycoheterotrophic plants. However, little is known about environmental factors that influence the distribution of mycorrhizal fungi. Previous studies using seed packets have been unable to distinguish whether germination patterns resulted from the distribution of appropriate edaphic conditions or the distribution of host fungi, as these cannot be separated using seed packets alone. We used a combination of organic amendments, seed packets and molecular assessment of soil fungi required by three terrestrial orchid species to separate direct and indirect effects of fungi and environmental conditions on both seed germination and subsequent protocorm development. We found that locations with abundant mycorrhizal fungi were most likely to support seed germination and greater growth for all three orchids. Organic amendments affected germination primarily by affecting the abundance of appropriate mycorrhizal fungi. However, fungi associated with the three orchid species were affected differently by the organic amendments and by forest successional stage. The results of this study help contextualize the importance of fungal distribution and abundance to the population dynamics of plants with specific mycorrhizal requirements. Such phenomena may also be important for plants with more general mycorrhizal associations.     

Two widespread green Neottia species (Orchidaceae) show mycorrhizal preference for Sebacinales in various habitats and ontogenetic stages

Plant dependence on fungal carbon (mycoheterotrophy) evolved repeatedly. In orchids, it is connected with a mycorrhizal shift from rhizoctonia to ectomycorrhizal fungi and a high natural ¹³C and ¹⁵N abundance. Some green relatives of mycoheterotrophic species show identical trends, but most of these remain unstudied, blurring our understanding of evolution to mycoheterotrophy. We analysed mycorrhizal associations and ¹³C and ¹⁵N biomass content in two green species, Neottia ovata and N. cordata (tribe Neottieae), from a genus comprising green and nongreen (mycoheterotrophic) species. Our study covered 41 European sites, including different meadow and forest habitats and orchid developmental stages. Fungal ITS barcoding and electron microscopy showed that both Neottia species associated mainly with nonectomycorrhizal Sebacinales Clade B, a group of rhizoctonia symbionts of green orchids, regardless of the habitat or growth stage. Few additional rhizoctonias from Ceratobasidiaceae and Tulasnellaceae, and ectomycorrhizal fungi were detected. Isotope abundances did not detect carbon gain from the ectomycorrhizal fungi, suggesting a usual nutrition of rhizoctonia‐associated green orchids. Considering associations of related partially or fully mycoheterotrophic species such as Neottia camtschatea or N. nidus‐avis with ectomycorrhizal Sebacinales Clade A, we propose that the genus Neottia displays a mycorrhizal preference for Sebacinales and that the association with nonectomycorrhizal Sebacinales Clade B is likely ancestral. Such a change in preference for mycorrhizal associates differing in ecology within the same fungal taxon is rare among orchids. Moreover, the existence of rhizoctonia‐associated Neottia spp. challenges the shift to ectomycorrhizal fungi as an ancestral pre‐adaptation to mycoheterotrophy in the whole Neottieae.     

Fungal‐host diversity among mycoheterotrophic plants increases proportionally to their fungal‐host overlap

The vast majority of plants obtain an important proportion of vital resources from soil through mycorrhizal fungi. Generally, this happens in exchange of photosynthetically fixed carbon, but occasionally the interaction is mycoheterotrophic, and plants obtain carbon from mycorrhizal fungi. This process results in an antagonistic interaction between mycoheterotrophic plants and their fungal hosts. Importantly, the fungal‐host diversity available for plants is restricted as mycoheterotrophic interactions often involve narrow lineages of fungal hosts. Unfortunately, little is known whether fungal‐host diversity may be additionally modulated by plant–plant interactions through shared hosts. Yet, this may have important implications for plant competition and coexistence. Here, we use DNA sequencing data to investigate the interaction patterns between mycoheterotrophic plants and arbuscular mycorrhizal fungi. We find no phylogenetic signal on the number of fungal hosts nor on the fungal hosts shared among mycoheterotrophic plants. However, we observe a potential trend toward increased phylogenetic diversity of fungal hosts among mycoheterotrophic plants with increasing overlap in their fungal hosts. While these patterns remain for groups of plants regardless of location, we do find higher levels of overlap and diversity among plants from the same location. These findings suggest that species coexistence cannot be fully understood without attention to the two sides of ecological interactions.     

On the origins of extreme mycorrhizal specificity in the Monotropoideae (Ericaceae): performance trade-offs during seed germination and seedling development

Fungal-induced seed germination is a phenomenon characteristic of mycorrhizal plants that produce dust-like seeds with only minimal nutritional reserves. In such systems, fungi trigger germination and/or subsidize development. We studied mycorrhizal germination in relation to mycorrhizal specificity in the Monotropoideae, a lineage of dust-seeded non-photosynthetic plants that are dependent upon ectomycorrhizal fungi of forest trees. A total of 1695 seed packets, each containing two to five compartments with seeds from different sources, were buried for up to 2 years near known ectomycorrhizal fungi in six different native forest locations. Upon harvest, seedlings were analysed by cultivation-independent molecular methods to identify their mycorrhizal fungi. We report that (i) germination is only induced by the same fungus that associates with mature plants or by closely related congeners; (ii) seedlings associated with the latter fungi develop less than those associated with maternal fungal species in most settings; and (iii) exceptions to this pattern occur in allopatric settings, where novel plant-fungal associations can result in the greatest seedling development. We interpret these results as evidence of performance trade-offs between breadth of host range and rate of development. We propose that in conjunction with host-derived germination cues, performance trade-offs can explain the extreme mycorrhizal specificity observed at maturity. The allopatric exceptions support the idea that performance trade-offs may be based on a coevolutionary arms race and that host range can be broadened most readily when naive fungal hosts are encountered in novel settings.     

Symbiotic germination capability of four Epipactis species (Orchidaceae) is broader than expected from adult ecology

? Premise of the study: Both abiotic and biotic factors shape species distributions. Orchids produce minute seeds with few nutrient reserves, thus requiring mycorrhizal fungi for germination. Therefore, both environmental conditions and mycorrhizal fungi distribution affect their germination success, but these ecological requirements and their congruence with habitat preferences of adults remain poorly understood. We investigated the importance of these factors during germination in four forest orchid species of the genus Epipactis. ? Methods: We sowed seeds of three habitat specialists and one generalist in different forest types at sites harboring adults of at least one of these ecologically diverging species. We analyzed germination pattern and identified mycorrhizal fungi of both seedlings and adults. ? Key results: Habitat conditions had little influence on germination pattern as seedlings grew in more habitats than expected from the adults' ecology. Ectomycorrhizal fungi availability did not limit germination. Suitable mycorrhizal fungi, mostly pezizalean ascomycetes, were recruited in various forest types, though the fungal communities differed according to habitat type. Finally, orchids with divergent ecological preferences shared similar mycorrhizal fungi. ? Conclusions: Limited adult distribution contrasted with successful seed germination at diverse sites and indicates existence of niche differentiation between adults and seedlings. Ecological specialization may thus be determined by factors other than mycorrhizal fungi that act later in the ontogeny, perhaps during the transition to above-ground development.     

Availability and function of arbuscular mycorrhizal and ectomycorrhizal fungi during revegetation of dewatered reservoirs left after dam removal

Revegetation following dam removal projects may depend on recovery of arbuscular mycorrhizal (AM) and ectomycorrhizal (EM) fungal communities, which perform valuable ecosystem functions. This study assessed the availability and function of AM and EM fungi for plants colonizing dewatered reservoirs following a dam removal project on the Elwha River, Olympic Peninsula, Washington, United States. Availability was assessed via AM fungal spore density in soils and EM root tip colonization of Salix sitchensis (Sitka willow) in an observational field study. The effect of mycorrhizal fungi from 4 sources (reservoir soils, commercial inoculum, and 2 mature plant community soils) on growth and nutrient status of S. sitchensis was quantified in a greenhouse study. AM fungal spores and EM root tips were present in all field samples. In the greenhouse, plants receiving reservoir soil inoculum had only incipient mantle formation, while plants receiving inoculum from mature plant communities had fully formed EM root tips. EM formation corresponded with alleviation of phosphorus stress in plants (lower shoot nitrogen:phosphorus). Thus, revegetating plants have access to AM and EM fungi following dam removal, and EM formation may be especially important for plant P uptake in reservoir soils. However, availability of mycorrhizal fungi declines with distance from established plant communities. Furthermore, EM fungal communities in recently dewatered reservoirs may not be as effective at forming beneficial mycorrhizae as those from mature plant communities. Whole soil inoculum from mature plant communities may be important for the success of revegetating plants and recovery of mycorrhizal fungal communities.     

Do novel weapons that degrade mycorrhizal mutualisms promote species invasion?

Non-native plants often dominate novel habitats where they did not co-evolve with the local species. The novel weapons hypothesis suggests that non-native plants bring competitive traits against which native species have not adapted defenses. Novel weapons may directly affect plant competitors by inhibiting germination or growth, or indirectly by attacking competitor plant mutualists (degraded mutualisms hypothesis). Japanese knotweed (Fallopia japonica) and European buckthorn (Rhamnus cathartica) are widespread plant invaders that produce potent secondary compounds that negatively impact plant competitors. We tested whether their impacts were consistent with a direct effect on the tree seedlings (novel weapons) or an indirect attack via degradation of seedling mutualists (degraded mutualism). We compared recruitment and performance using three Ulmus congeners and three Betula congeners treated with allelopathic root macerations from allopatric and sympatric ranges. Moreover, given that the allelopathic species would be less likely to degrade their own fungal symbiont types, we used arbuscular mycorrhizal (AMF) and ectomycorrhizal (ECM) tree species to investigate the effects of F. japonica (no mycorrhizal association) and Rhamnus cathartica (ECM association) on the different fungal types. We also investigated the effects of F. japonica and R. cathartica exudates on AMF root colonization. Our results suggest that the allelopathic plant exudates impact seedlings directly by inhibiting germination and indirectly by degrading fungal mutualists. Novel weapons inhibited allopatric seedling germination but sympatric species were unaffected. However, seedling survivorship and growth appeared more dependent on mycorrhizal fungi, and mycorrhizal fungi were inhibited by allopatric species. These results suggest that novel weapons promote plant invasion by directly inhibiting allopatric competitor germination and indirectly by inhibiting mutualist fungi necessary for growth and survival.