The exploration of potato-associated bacteria in the Central Andean Highlands and their application in integrated crop management systems

Potato is the most important food crop after wheat and rice. A changing climate, coupled with a heightened consumer awareness of how food is produced and legislative changes governing the usage of agrochemicals, means that alternative more integrated and sustainable approaches are needed for crop management practices. Bioprospecting in the Central Andean Highlands resulted in the isolation and in vitro screening of 600 bacterial isolates. The best performing isolates, under in vitro conditions, were field trialled in their home countries. Six of the isolates, Pseudomonas sp. R41805 (Bolivia), Pseudomonas palleroniana R43631 (Peru), Bacillus sp. R47065, R47131, Paenibacillus sp. B3a R49541, and Bacillus simplex M3-4 R49538 (Ecuador), showed significant increase in the yield of potato. Using – omic technologies (i.e. volatilomic, transcriptomic, proteomic and metabolomic), the influence of microbial isolates on plant defence responses was determined. Volatile organic compounds of bacterial isolates were identified using GC/MS. RT-qPCR analysis revealed the significant expression of Ethylene Response Factor 3 (ERF3) and the results of this study suggest that the dual inoculation of potato with Pseudomonas sp. R41805 and Rhizophagus irregularis MUCL 41833 may play a part in the activation of plant defence system via ERF3. The proteomic analysis by 2-DE study has shown that priming by Pseudomonas sp. R41805 can induce the expression of proteins related to photosynthesis and protein folding in in vitro potato plantlets. The metabolomics study has shown that the total glycoalkaloid (TGA) content of greenhouse-grown potato tubers following inoculation with Pseudomonas sp. R41805 did not exceed the acceptable safety limit (200 mg kg-1 FW). As a result of this study, a number of bacteria have been identified with commercial potential that may offer sustainable alternatives in both Andean and European agricultural settings.

Differences between root traits of early- and late-successional trees influence below-ground competition and seedling establishment.

The competitive influence of the root system of the exotic grass Urochloa brizantha and the widespread forb Leonotis nepetifolia on the emergence, survival and early growth of the seedlings of eight tropical heliophilous herbaceous species, six early-successional woody species and five late-successional woody species from Brazil, grown in 3500-cm3 pots and in greenhouse without light restriction were assessed. The density of fine-root systems produced by the forb and the grass in pots were 6.8 cm cm-3 soil and 48.1 cm cm-3 soil, respectively. Seedlings survival of the heliophilous herbaceous, early- and late-successional woody species were 86%, 70% and 100% in presence of the forb root system and 12%, 14% and 100% in competition with grass root system, respectively. The competitive pressure applied by the grass root system on seedling growth of the heliophilous herbaceous, early- and late-successional woody species were 2.4, 1.9 and 1.4 times greater than the forb root system. Total root length of the heliophilous herbaceous, early- and late-successional woody species grown without competitors were 13, 33 and 5 times greater than in competition with forb, and were 66, 54 and 6 times greater than in competition with grass root system, respectively. The averages of fine-root diameter of plants grown without competitors were 209 microm for the heliophilous herbaceous, 281 microm for early-successional trees and 382 microm for late-successional trees. The root system of the forb did not avoid seedling establishment of most plant species, but the grass root system hampered more the establishment of heliophilous herbaceous and early-successional woody species than the seedling establishment of late-successional woody species. The different density of root systems produced in soil by the forb and the grass, and the distinct root traits (e.g. root diameter and root tissue density) of the early- and late-successional plant species can explain the differences in the establishment of seedlings of plant species belonging to different groups of tropical succession when exposed to below-ground competition.

Inoculação de bactérias diazotroficas e fungos micorrizico-arbusculares na cultura da mandioca

A versatilidade da mandioca (Manihot esculenta Crantz) em adaptar-se a solos de baixa fertilidade, apesar de possuir alto requerimento de nutrientes, tem sido relacionada à ocorrência de associações com os fungos micorrízico-arbusculares (FMA) e a bactérias diazotróficas. Visando avaliar o efeito da inoculação dos FMA e das bactérias diazotróficas, foi conduzido um experimento com plântulas micropropagadas em vasos de 3,5 litros de volume, com solo arenoso desinfestado como substrato. A inoculação das bactérias diazotróficas não apresentou efeito estimulatório, ao passo que inoculações dos FMA isoladamente e em conjunto com bactéria incrementaram todos os parâmetros de crescimento e nutricionais. A inoculação dos FMA, com a Bactéria E, aumentou a parte aérea e as raízes em até 50% e 105%, respectivamente, em relação à inoculação exclusiva com FMA. Efeitos sinergísticos também foram observados no acúmulo de N da parte aérea e das raízes com aumento de até 88% e 173% e no de fósforo em até 83% e 158%, respectivamente. A co-inoculação da Bactéria E com Glomus clarum também aumentou a colonização micorrízica em 40% e a esporulação em 168%, comparada à inoculação do fungo isolado. Estes efeitos benéficos podem ocorrer tanto pela maior absorção de nutrientes pela planta, como pelo estímulo na colonização dos fungos MA.

Ocorrência de micorrizas arbusculares e da bactéria diazotrofica Acetobacter diazotrophicus em cana-de-açúcar.

Foi avaliada a ocorrência e a distribuição de espécies de fungos micorrízicos arbusculares e A. diazotrophicus em plantios de cana-de-açúcar em diferentes tipos de manejo nos Estados do Rio de Janeiro e Pernambuco. Foram feitas 35 coletas de amostras de solo da rizosfera e de raízes de 14 variedades de cana-de-açúcar para extração de esporos e isolamento da bactéria. O numero de esporos variou de 18 a 2.070/ 100 mL de solo, e os maiores numero e diversidade de espécies foram verificados nos canaviais de Campos, RJ, especialmente naqueles que não adotam a queima de palhico. As espécies predominantes nas três localidades amostradas foram: Acaulospora sp., Scutellospora heterogama, Glomus etunicatum, Glomus occultum e Gigaspora margarita. A. diazotrophicus estava presente nas amostras de raízes colhidas em canaviais de Campos, com exceção de uma coleta de cana-de-açúcar plantada num solo usado como bacia de sedimentação de vinhaça. Não foi possível isolar essa bactéria a partir de esporos desinfestados dos FMAs nativos, apenas dos esporos lavados com agua estéril The occurrence and distribution of species of arbuscular mycorrhizae fungi and Acetobacter diazotrophicus in sugar cane (Saccharum officinarum) grown in different regimes of crop management in the States of Rio de Janeiro and Pernambuco were studied. Thirty five samples of the rhizosphere soil and roots were collected from 14 varieties of sugar cane for the extraction of spores and isolation of the bacterium. The number of spores varied from 18 to 2.070 per 100 mL of soil, and the greatest diversity of fungal species was found in the sugarcane fields of Campos (Rio de Janeiro State), especially in those where the sugarcane trash was not burned at harvest. The predominant species found in the three localities sampled were: Scutellospora heterogama, Glomus etunicatum, Glomus occultum, Glomus macrocarpum, Acaulospora sp. and Gigaspora margarita. A. diazotrophicus was present in almost all samples of root with the exception of one harvest of sugar cane taken from an area used for the sedimentation of vinasse (distillery waste). It was not possible to detect the bacterium from surface sterilised spores of native arbuscular mycorrhizal fungi (AMF), only from washed ones using sterile water.

Ocorrência de bactérias diazotroficas e fungos micorrizicos arbusculares na cultura da mandioca.

Este trabalho teve como objetivo avaliar a ocorrencia, isolar e identificar fungos micorrízicos carbunculares associados a cultura da mandioca (Manihot esculenta). Amostras de solo rizosférico e de varias partes da planta (raízes, tubérculos, manivas e folhas) de locais nos Estados do Rio de Janeiro, São Paulo e Paraná, foram inoculadas nos meios LGI-P, NFb-malato e NFb-GOC, avaliando-se o numero mais provável de células e a atividade de redução de acetileno. Bactérias diazotróficas foram isoladas de todas as partes da planta, com exceção das folhas, sendo identificadas como Klebsiella sp., Azospirillum lipoferum e uma bactéria denominada "E", provavelmente pertencente ao gênero Burkholderia. A bactéria E acumulou de 7,63 mg a 14,84 mg de N/g de C em meio semi-solido, isento de N, e conseguiu manter a capacidade de fixação biológica de N, mesmo apos uma dezena de repicagens consecutivas. A colonização micorrízica variou de 31% a 69%, e a densidade de esporos de 10 a 384 esporos/100 mL de solo, predominando as espécies Entrophospora colombiana e Acaulospora scrobiculata no Rio de Janeiro, A. scrobiculata e Scutellospora heterogama no Paraná e em Piracicaba (São Paulo) e A. appendicula e S. pellucida em Campinas (São Paulo). This study was performed to evaluate the occurrence and to isolate and identify diazotrophic bacteria and arbuscular mycorrhizal fungi associated with the cassava (Manihot esculenta) crop. Samples from rhizosperical soil, roots, tubers, stems and leaves from several localities of the States of Rio de Janeiro, Sao Paulo and Parana, in Brazil, were inoculated in three media specific for diazotrophic associative bacteria, LG1-P, NFb-malate and NFb-GOC, evaluating the most probable number of cells and the acetylene-reducing activity. Diazotrophic bacteria were detected in all plant parts except for the leaves, and were identified as Klebsiella sp., Azospirillum lipoferum and a bacterium called "E", probably belonging to the Burkholderia genus. Bacterium E was able to accumulate, in the N-free semi-solid media, from 7.63 to 14.84 mg of N/g of C and to maintain N fixation capacity after ten consecutive transferences. Mycorrhizal root colonization varied from 31% to 69% and spore density from 10 to 384 spores/100 mL of soil, predominanting the species Entrophospora colombiana and Acaulospora scrobiculata in Rio de Janeiro, A. scrobiculata and Scutellospora heterogama in Parana and in Piracicaba (Sao Paulo), and A. appendicula and S. pellucida in Campinas (Sao Paulo).

Interactions between transgenic trees and mycorrhizal and pathogenic fungi

The development of biotechnology techniques in plant breeding and the new commercial applications have raised public and scientific concerns about the safety of genetically modified (GM) crops and trees. To find out the feasibility of these new technologies in the breeding of commercially important Finnish hardwood species and to estimate the ecological risks of the produced transgenic plants, the experiments of this study have been conducted as a part of a larger project focusing on the risk assessment of GM-trees. Transgenic Betula pendula and Populus trees were produced via Agrobacterium mediated transformation. Stilbene synthase (STS) gene from pine (Pinus sylvestris) and chitinase gene from sugar beet (Beta vulgaris) were transferred to (hybrid) aspen and birch, respectively, to improve disease resistance against fungal pathogens. To modify lignin biosynthesis, a 4-coumarate:coenzyme A ligase (4CL) gene fragment in antisense orientation was introduced into two birch clones. In in vitro test, one transgenic aspen line expressing pine STS gene showed increased resistance to decay fungus Phellinus tremulae. In the field, chitinase transgenic birch lines were more susceptible to leaf spot (Pyrenopeziza betulicola) than the non-transgenic control clone while the resistance against birch rust (Melampsoridium betulinum) was improved. No changes in the content or composition of lignin were detected in the 4CL antisense birch lines. In order to evaluate the ecological effects of the produced GM trees on non-target organisms, an in vitro mycorrhiza experiment with Paxillus involutus and a decomposition experiment in the field were performed. The expression of a transgenic chitinase did not disturb the establishment of mycorrhizal symbiosis between birch and P. involutus in vitro. 4CL antisense transformed birch lines showed retarded root growth but were able to form normal ectomycorrhizal associations with the mycorrhizal fungus in vitro. 4CL lines also showed normal litter decomposition. Unexpected growth reductions resulting from the gene transformation were observed in chitinase transgenic and 4CL antisense birch lines. These results indicate that genetic engineering can provide a tool in increasing disease resistance in Finnish tree species. More extensive data with several ectomycorrhizal species is needed to evaluate the consequences of transgene expression on beneficial plant-fungus symbioses. The potential pleiotropic effects of the transgene should also be taken into account when considering the safety of transgenic trees.

Indigenous Arbuscular Mycorrhizal Fungal Assemblages Protect Grassland Host Plants from Pathogens

Plant roots can establish associations with neutral, beneficial and pathogenic groups of soil organisms. Although it has been recognized from the study of individual isolates that these associations are individually important for plant growth, little is known about interactions of whole assemblages of beneficial and pathogenic microorganisms associating with plants. We investigated the influence of an interaction between local arbuscular mycorrhizal (AM) fungal and pathogenic/saprobic microbial assemblages on the growth of two different plant species from semi-arid grasslands in NE Germany (Mallnow near Berlin). In a greenhouse experiment each plant species was grown for six months in either sterile soil or in sterile soil with one of three different treatments: 1) an AM fungal spore fraction isolated from field soil from Mallnow; 2) a soil pathogen/saprobe fraction consisting of a microbial community prepared with field soil from Mallnow and; 3) the combined AM fungal and pathogen/saprobe fractions. While both plant species grew significantly larger in the presence of AM fungi, they responded negatively to the pathogen/saprobe treatment. For both plant species, we found evidence of pathogen protection effects provided by the AM fungal assemblages. These results indicate that interactions between assemblages of beneficial and pathogenic microorganisms can influence the growth of host plants, but that the magnitude of these effects is plant species-specific.

Compositional divergence and convergence in arbuscular mycorrhizal fungal communities

In spite of the controversy that they have generated, neutral models provide ecologists with powerful tools for creating dynamic predictions about beta-diversity in ecological communities. Ecologists can achieve an understanding of the assembly rules operating in nature by noting when and how these predictions are met or not met. This is particularly valuable for those groups of organisms that are challenging to study under natural conditions (e.g., bacteria and fungi). Here, we focused on arbuscular mycorrhizal fungal (AMF) communities and performed an extensive literature search that allowed us to synthesize the information in 19 data sets with the minimal requisites for creating a null hypothesis in terms of community dissimilarity expected under neutral dynamics. In order to achieve this task, we calculated the first estimates of neutral parameters for several AMF communities from different ecosystems. Communities were shown either to be consistent with neutrality or to diverge or converge with respect to the levels of compositional dissimilarity expected under neutrality. These data support the hypothesis that divergence occurs in systems where the effect of limited dispersal is overwhelmed by anthropogenic disturbance or extreme biological and environmental heterogeneity, whereas communities converge when systems have the potential for niche divergence within a relatively homogeneous set of environmental conditions. Regarding the study cases that were consistent with neutrality, the sampling designs employed may have covered relatively homogeneous environments in which the effects of dispersal limitation overwhelmed minor differences among AMF taxa that would lead to environmental filtering. Using neutral models we showed for the first time for a soil microbial group the conditions under which different assembly processes may determine different patterns of beta-diversity. Our synthesis is an important step showing how the application of general ecological theories to a model microbial taxon has the potential to shed light on the assembly and ecological dynamics of communities.

Studies on arbuscular mycorrhiza (AM) in the Alentejo (Portugal) using pea mutants resistant to AM fungi as a control tool for field conditions

The utilization and management of arbuscular mycorrhiza (AM) symbiosis may improve production and sustainability of the cropping system. For this purpose, native AM fungi (AMF) were sought and tested for their efficiency to increase plant growth by enhanced P uptake and by alleviation of drought stress. Pot experiments with safflower (Carthamus tinctorius) and pea (Pisum sativum) in five soils (mostly sandy loamy Luvisols) and field experiments with peas were carried out during three years at four different sites. Host plants were grown in heated soils inoculated with AMF or the respective heat sterilized inoculum. In the case of peas, mutants resistant to AMF colonization were used as non-mycorrhizal controls. The mycorrhizal impact on yields and its components, transpiration, and P and N uptake was studied in several experiments, partly under varying P and N levels and water supply. Screening of native AMF by most probable number bioassays was not very meaningful. Soil monoliths were placed in the open to simulate field conditions. Inoculation with a native AMF mix improved grain yield, shoot and leaf growth variables as compared to control. Exposed to drought, higher soil water depletion of mycorrhizal plants resulted in a haying-off effect. The growth response to this inoculum could not be significantly reproduced in a subsequent open air pot experiment at two levels of irrigation and P fertilization, however, safflower grew better at higher P and water supply by multiples. The water use efficiency concerning biomass was improved by the AMF inoculum in the two experiments. Transpiration rates were not significantly affected by AM but as a tendency were higher in non-mycorrhizal safflower. A fundamental methodological problem in mycorrhiza field research is providing an appropriate (negative) control for the experimental factor arbuscular mycorrhiza. Soil sterilization or fungicide treatment have undesirable side effects in field and greenhouse settings. Furthermore, artificial rooting, temperature and light conditions in pot experiments may interfere with the interpretation of mycorrhiza effects. Therefore, the myc- pea mutant P2 was tested as a non-mycorrhizal control in a bioassay to evaluate AMF under field conditions in comparison to the symbiotic isogenetic wild type of var. FRISSON as a new integrative approach. However, mutant P2 is also of nod- phenotype and therefore unable to fix N2. A 3-factorial experiment was carried out in a climate chamber at high NPK fertilization to examine the two isolines under non-symbiotic and symbiotic conditions. P2 achieved the same (or higher) biomass as wild type both under good and poor water supply. However, inoculation with the AMF Glomus manihot did not improve plant growth. Differences of grain and straw yields in field trials were large (up to 80 per cent) between those isogenetic pea lines mainly due to higher P uptake under P and water limited conditions. The lacking N2 fixation in mutants was compensated for by high mineral N supply as indicated by the high N status of the pea mutant plants. This finding was corroborated by the results of a major field experiment at three sites with two levels of N fertilization. The higher N rate did not affect grain or straw yields of the non-fixing mutants. Very efficient AMF were detected in a Ferric Luvisol on pasture land as revealed by yield levels of the evaluation crop and by functional vital staining of highly colonized roots. Generally, levels of grain yield were low, at between 40 and 980 kg ha-1. An additional pot trial was carried out to elucidate the strong mycorrhizal effect in the Ferric Luvisol. A triplication of the plant equivalent field P fertilization was necessary to compensate for the mycorrhizal benefit which was with five times higher grain yield very similar to that found in the field experiment. However, the yield differences between the two isolines were not always plausible as the evaluation variable because they were also found in (small) field test trials with apparently sufficient P and N supply and in a soil of almost no AMF potential. This similarly occurred for pea lines of var. SPARKLE and its non-fixing mycorrhizal (E135) and non-symbiotic (R25) isomutants, which were tested in order to exclude experimentally undesirable benefits by N2 fixation. In contrast to var. FRISSON, SPARKLE was not a suitable variety for Mediterranean field conditions. This raises suspicion putative genetic defects other than symbiotic ones may be effective under field conditions, which would conflict with the concept of an appropriate control. It was concluded that AMF resistant plants may help to overcome fundamental problems of present research on arbuscular mycorrhiza, but may create new ones.

Glomus intraradices and Gigaspora margarita arbuscular mycorrhizal associations differentially affect nitrogen and potassium nutrition of Plantago lanceolata in a low fertility dune soil

Two controlled microcosm experiments aimed at a critical re-assessment of the contributions of divergent arbuscular mycorrhizal (AM) fungi to plant mineral nutrition were established that specifically targeted Plantago lanceolata–Glomus intraradices (B.B/E) and –Gigaspora margarita (BEG 34) symbioses developed in a native, nutrient limited, coastal dune soil. Plant tissue nitrogen (N), phosphorus (P) and potassium (K) status as well as plant growth parameters and levels of mycorrhizal colonization were assessed at harvest. In addition to the general well-established mycorrhizal facilitation of P uptake, the study was able to demonstrate a G. intraradices-specific contribution to improved plant nitrogen and potassium nutrition. In the two respective experiments, G. intraradices-inoculated plants had 27.8% and 40.8% more total N and 55.8% and 23.3% more total K when compared to Gi. margarita inoculated counterparts. Dissimilar overall contribution of the two isolates to plant nutrition was identified in AM-genus specific differences in plant tissue N:P:K ratios. G. intraradices inoculated and non-mycorrhizal plants generally exhibited N:P:K ratios indicative of P limitation whereas for Gi.margarita mycorrhizal plants, corresponding ratios strongly implied either N or K limitation. The study provides further evidence highlighting AM functional biodiversity in respect to plant nutrient limitation experienced by mycorrhizal P. lanceolata in an ecologically relevant soil system.

Arbuscular mycorrhizal modulation of diazotrophic and denitrifying microbial communities in the (mycor)rhizosphere of Plantago lanceolata

Impacts of divergent arbuscular mycorrhizal (AM) fungi, Glomus intraradices and Gigaspora margarita, on denitrifying and diazotrophic bacterial communities of Plantago lanceolata in nutrient-limited dune soil were assessed. We hypothesized AM species-related modifications that were confirmed in respective bacterial nirK and nifH sequence polymorphism -based community clustering and community variance allocation. The denitrifying community appeared more responsive to AM fungi than the nitrogen-fixing community. Nevertheless, the main explanatory variable, in both cases, was plant age. We conclude that AM fungi can modify N-cycling microbial rhizosphere communities and future work should aim to verify the functional significance and mechanistic basis.

Contrasting arbuscular mycorrhizal communities colonizing different host plants show a similar response to a soil phosphorus concentration gradient

High soil phosphorus (P) concentration is frequently shown to reduce root colonization by arbuscular mycorrhizal (AM) fungi, but the influence of P on the diversity of colonizing AM fungi is uncertain. We used terminal restriction fragment length polymorphism (T-RFLP) of 18S rDNA and cloning to assess diversity of AM fungi colonizing maize (Zea mays), soybean (Glycene max) and field violet (Viola arvensis) at three time points in one season along a P gradient of 10280mgl1 in the field. Percentage AM colonization changed between sampling time points but was not reduced by high soil P except in maize. There was no significant difference in AM diversity between sampling time points. Diversity was reduced at concentrations of P > 25mgl1, particularly in maize and soybean. Both cloning and T-RFLP indicated differences between AM communities in the different host species. Host species was more important than soil P in determining the AM community, except at the highest P concentration. Our results show that the impact of soil P on the diversity of AM fungi colonizing plants was broadly similar, despite the fact that different plants contained different communities. However, subtle differences in the response of the AM community in each host were evident.

Ecto- and arbuscular mycorrhizal symbiosis can induce tolerance to toxic pulses of phosphorus in jarrah (Eucalyptus marginata) seedlings

In common with many plants native to low P soils, jarrah (Eucalyptus marginata) develops toxicity symptoms upon exposure to elevated phosphorus (P). Jarrah plants can establish arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) associations, along with a non-colonizing symbiosis described recently. AM colonization is known to influence the pattern of expression of genes required for P uptake of host plants and our aim was to investigate this phenomenon in relation to P sensitivity. Therefore, we examined the effect on hosts of the presence of AM and ECM fungi in combination with toxic pulses of P and assessed possible correlations between the induced tolerance and the shoot P concentration. The P transport dynamics of AM (Rhizophagus irregularis and Scutellospora calospora), ECM (Scleroderma sp.), non-colonizing symbiosis (Austroboletus occidentalis), dual mycorrhizal (R. irregularis and Scleroderma sp.), and non-mycorrhizal (NM) seedlings were monitored following two pulses of P. The ECM and A. occidentalis associations significantly enhanced the shoot P content of jarrah plants growing under P-deficient conditions. In addition, S. calospora, A. occidentalis, and Scleroderma sp. all stimulated plant growth significantly. All inoculated plants had significantly lower phytotoxicity symptoms compared to NM controls 7 days after addition of an elevated P dose (30 mg P kg−1 soil). Following exposure to toxicity-inducing levels of P, the shoot P concentration was significantly lower in R. irregularis-inoculated and dually inoculated plants compared to NM controls. Although all inoculated plants had reduced toxicity symptoms and there was a positive linear relationship between rank and shoot P concentration, the protective effect was not necessarily explained by the type of fungal association or the extent of mycorrhizal colonization.

A novel plant-fungus symbiosis benefits the host without forming mycorrhizal structures

Most terrestrial plants form mutually beneficial symbioses with specific soil-borne fungi known as mycorrhiza. In a typical mycorrhizal association, fungal hyphae colonize plant roots, explore the soil beyond the rhizosphere and provide host plants with nutrients that might be chemically or physically inaccessible to root systems. Here, we combined nutritional, radioisotopic (33P) and genetic approaches to describe a plant growth promoting symbiosis between the basidiomycete fungus Austroboletus occidentalis and jarrah (Eucalyptus marginata), which has quite different characteristics. We show that the fungal partner does not colonize plant roots; hyphae are localized to the rhizosphere soil and vicinity and consequently do not transfer nutrients located beyond the rhizosphere. Transcript profiling of two high-affinity phosphate (Pi) transporter genes (EmPHT1;1 and EmPHT1;2) and hyphal-mediated 33Pi uptake suggest that the Pi uptake shifts from an epidermal to a hyphal pathway in ectomycorrhizal plants (Scleroderma sp.), similar to arbuscular mycorrhizal symbioses, whereas A. occidentalis benefits its host indirectly. The enhanced rhizosphere carboxylates are linked to growth and nutritional benefits in the novel symbiosis. This work is a starting point for detailed mechanistic studies on other basidiomycete–woody plant relationships, where a continuum between heterotrophic rhizosphere fungi and plant beneficial symbioses is likely to exist.