Quantification of plasticity of plant traits in response to light intensity: comparing phenotypes at a common weight

Summary Plasticity of plant traits is commonly quantified by comparing different phenotypes at the same age. In this paper, we present a method in which the effect of resource conditions on plant weight is used as a basis for quantifying the plasticity of individual plant traits. Abutilon theophrasti individuals were grown in, and some transferred between, high and low intensity light conditions, resulting in four phenotypes. Plant traits were found to exhibit different degrees of plasticity, decreasing in this order: height; specific leaf area; allocation to branch roots; allocation to leaf area; number of nodes; allocation to tap roots; allocation to stem; allocation to leaf weight. Under these conditions, individuals of the four phenotypes had very similar heights when compared at the same age, but very different heights when compared at the same plant weight. The latter comparison indicates that light intensity influences height independently of its influence on plant weight. Individuals that were transferred from high to low light had greater allocation that had not been transferred, but individuals of all phenotypes had nearly the same leaf weight allocation when compared at the same plant weight. The latter comparison indicates that light intensity influeces leaf weight allocation mostly by influencing plant weight. In the phenotype resulting from the transfer of plants from low to high light, reproduction was stimulated much less than plant weight and axillary leaf growth, and reproductive allocation was delayed relative to the other three phenotypes. We conclude that when plasticity is measured by comparing phenotypes at the same plant weight, the effects of resources on plant size can be excluded from the quantification.     

Effect of arbuscular mycorrhizal fungi inoculation on root traits and root volatile organic compound emissions of Sorghum bicolor

Volatile organic compounds (VOCs) emitted by plant roots have important functions that can influence the rhizospheric environment. The aim of this study was to examine the effects of arbuscular mycorrhizal (AM) fungi on the profile of root VOCs. Sorghum (Sorghum bicolor) plants were grown in pots inoculated with either Glomus mosseae or Glomus intraradices, which formed mycorrhiza with the roots. Control plants were grown in pots inoculated with sterile inoculum and did not form mycorrhiza. Forty-four VOCs were determined using headspace solid-phase microextraction (HS-SPME) and gas chromatography–mass spectrometry (GC-MS). Alkanes were the most abundant type of VOCs emitted by both mycorrhizal and non-mycorrhizal plants. Both the quantity and type of volatiles were dramatically altered by the presence of AM fungi, and these changes had species specificity. Compared with non-mycorrhizal plants, mycorrhizal plants emitted more alcohols, alkenes, ethers and acids but fewer linear-alkanes. The AM fungi also influenced the morphological traits of the host roots. The total root length and specific root length of mycorrhizal plants were significantly greater than those of non-mycorrhizal plants; however, both the incidence and length of root-hair were dramatically decreased. Our findings confirm that AM fungi can alter the profile of VOCs emitted by roots as well as the root morphology of sorghum plants, indicating that AM fungi have the potential to help plants adapt to and alter soil environments.     

Exotic cheatgrass and loss of soil biota decrease the performance of a native grass

Soil disturbances can alter microbial communities including arbuscular mycorrhizal (AM) fungi, which may in turn, affect plant community structure and the abundance of exotic species. We hypothesized that altered soil microbial populations owing to disturbance would contribute to invasion by cheatgrass (Bromus tectorum), an exotic annual grass, at the expense of the native perennial grass, squirreltail (Elymus elymoides). Using a greenhouse experiment, we compared the responses of conspecific and heterospecific pairs of cheatgrass and squirreltail inoculated with soil (including live AM spores and other organisms) collected from fuel treatments with high, intermediate and no disturbance (pile burns, mastication, and intact woodlands) and a sterile control. Cheatgrass growth was unaffected by type of soil inoculum, whereas squirreltail growth, reproduction and nutrient uptake were higher in plants inoculated with soil from mastication and undisturbed treatments compared to pile burns and sterile controls. Squirreltail shoot biomass was positively correlated with AM colonization when inoculated with mastication and undisturbed soils, but not when inoculated with pile burn soils. In contrast, cheatgrass shoot biomass was negatively correlated with AM colonization, but this effect was less pronounced with pile burn inoculum. Cheatgrass had higher foliar N and P when grown with squirreltail compared to a conspecific, while squirreltail had lower foliar P, AM colonization and flower production when grown with cheatgrass. These results indicate that changes in AM communities resulting from high disturbance may favor exotic plant species that do not depend on mycorrhizal fungi, over native species that depend on particular taxa of AM fungi for growth and reproduction.     

Specificity in Arabidopsis thaliana recruitment of root fungal communities from soil and rhizosphere

Biotic and abiotic conditions in soil pose major constraints on growth and reproductive success of plants. Fungi are important agents in plant soil interactions but the belowground mycobiota associated with plants remains poorly understood. We grew one genotype each from Sweden and Italy of the widely-studied plant model Arabidopsis thaliana. Plants were grown under controlled conditions in organic topsoil local to the Swedish genotype, and harvested after ten weeks. Total DNA was extracted from three belowground compartments: endosphere (sonicated roots), rhizosphere and bulk soil, and fungal communities were characterized from each by amplification and sequencing of the fungal barcode region ITS2. Fungal species diversity was found to decrease from bulk soil to rhizosphere to endosphere. A significant effect of plant genotype on fungal community composition was detected only in the endosphere compartment. Despite A. thaliana being a non-mycorrhizal plant, it hosts a number of known mycorrhiza fungi in its endosphere compartment, which is also colonized by endophytic, pathogenic and saprotrophic fungi. Species in the Archaeorhizomycetes were most abundant in rhizosphere samples suggesting an adaptation to environments with high nutrient turnover for some of these species. We conclude that A. thaliana endosphere fungal communities represent a selected subset of fungi recruited from soil and that plant genotype has small but significant quantitative and qualitative effects on these communities.     

A suite of rare microbes interacts with a dominant,heritable, fungal endophyte to influence plant trait expression

Endophytes are microbes that live, for at least a portion of their life history, within plant tissues. Endophyte assemblages are often composed of a few abundant taxa and many infrequently observed, low-biomass taxa that are, in a word, rare. The ways in which most endophytes affect host phenotype are unknown; however, certain dominant endophytes can influence plants in ecologically meaningful ways—including by affecting growth and immune system functioning. In contrast, the effects of rare endophytes on their hosts have been unexplored, including how rare endophytes might interact with abundant endophytes to shape plant phenotype. Here, we manipulate both the suite of rare foliar endophytes (including both fungi and bacteria) and Alternaria fulva–a vertically transmitted and usually abundant fungus–within the fabaceous forb Astragalus lentiginosus. We report that rare, low-biomass endophytes affected host size and foliar %N, but only when the heritable fungal endophyte (A. fulva) was not present. A. fulva also reduced plant size and %N, but these deleterious effects on the host could be offset by a negative association we observed between this heritable fungus and a foliar pathogen. These results demonstrate how interactions among endophytic taxa determine the net effects on host plants and suggest that the myriad rare endophytes within plant leaves may be more than a collection of uninfluential, commensal organisms, but instead have meaningful ecological roles.Subject terms: Microbial ecology, Community ecology, Microbial ecology     

Restoration of the mycobiome of the endangered Hawaiian mint Phyllostegia kaalaensis increases its resistance to a common powdery mildew

Beneficial microbes such as plant mutualistic fungi, hold the promise of ameliorating challenges faced in native plant conservation such as disease management. As an alternative to costly chemical pest control, conservation efforts could potentially harness the benefits of plant mutualistic fungi to aid in defense and disease resistance, but there are few tests of this notion. We set out to test the efficacy of controlling a common foliar pathogen, the powdery mildew Neoerysiphe galeopsidis, by inoculating the endangered Hawaiian plant species Phyllostegia kaalaensis with potentially beneficial members of its wild-type mycobiome. We tested whether inoculating plants with above or belowground fungal mutualists, or both, led to increased disease resistance in the host. We found that while all treatments reduced average disease incidence, colonization by the foliar yeast Moesziomyces aphidis was the only treatment to do so significantly. These results provide an exciting new strategy for plant conservation practices.     

丛枝菌根真菌对植物繁殖的影响研究进展

丛枝菌根真菌(arbuscular mycorrhizal fungi, AMF)与宿主植物所形成的互惠共生体系是生态系统中广泛分布的共生体系代表类型之一。AMF除能够促进宿主植物生长发育外,也可以对宿主植物的繁殖过程产生多方面影响。研究宿主植物在AMF共生状态下繁殖策略的变化规律,对于深入理解植物繁殖适合度的变化具有重要理论意义。该文综述了AMF对宿主植物繁殖影响的相关研究,包括AMF的侵染对宿主植物繁殖分配、花部特征、虫媒传粉和花期的影响。目前已有研究发现某些AMF能够促进宿主植物增加繁殖资源投入,提高花朵产生的数量或花冠直径,同时增加花粉数量和花蜜量来影响访花昆虫的行为,以及造成开花提前及花期延长,但其作用的具体机制尚不明确,且因宿主植物的差异,并未有完全统一的结论。然而,由于AMF与植物共生的普遍性,其在植物繁殖过程中发挥的重要作用不可忽略。今后除了在以上各方面开展更深入的研究外,还需在AMF对宿主植物繁殖性状的影响机制、AMF共生条件下植物有性繁殖过程中雌雄功能的资源分配,以及对无性繁殖和后代适应性的影响等方面进行更深入的研究。     

Root endophyte symbiosis in vitro between the ectomycorrhizal basidiomycete Tricholoma matsutake and the arbuscular mycorrhizal plant Prunus speciosa

We previously reported that Tricholoma matsutake and Tricholoma fulvocastaneum, ectomycorrhizal basidiomycetes that associate with Pinaceae and Fagaceae, respectively, in the Northern Hemisphere, could interact in vitro as a root endophyte of somatic plants of Cedrela odorata (Meliaceae), which naturally harbors arbuscular mycorrhizal fungi in South America, to form a characteristic rhizospheric colony or “shiro”. We questioned whether this phenomenon could have occurred because of plant–microbe interactions between geographically separated species that never encounter one another in nature. In the present study, we document that these fungi formed root endophyte interactions and shiro within 140 days of inoculation with somatic plants of Prunus speciosa (=Cerasus speciosa, Rosaceae), a wild cherry tree that naturally harbors arbuscular mycorrhizal fungi in Japan. Compared with C. odorata, infected P. speciosa plants had less mycelial sheath surrounding the exodermis, and the older the roots, especially main roots, the more hyphae penetrated. In addition, a large number of juvenile roots were not associated with hyphae. We concluded that such root endophyte interactions were not events isolated to the interactions between exotic plants and microbes but could occur generally in vitro. Our pure culture system with a somatic plant allowed these fungi to express symbiosis-related phenotypes that varied with the plant host; these traits are innately programmed but suppressed in nature and could be useful in genetic analyses of plant–fungal symbiosis.     

The role of in vitro cultivation on symbiotic trait and function variation in a single species of arbuscular mycorrhizal fungus

In vitro propagation of AM fungi using transformed root cultures (TRC) is commonly used to obtain pure AM fungal propagules for use in research and industry. Early observations indicate that such an artificial environment can alter traits and function of AM fungi over time. We hypothesized that increased in vitro cultivation may promote ruderal strategies in fungi by enhancing propagule production and reducing mutualistic quality. To examine the effect of in vitro cultivation on the trait and function of AM fungi, we inoculated plants with 11 Rhizoglomus irregulare isolates which fell along a cultivation gradient spanning 80 generations. We harvested plants at 10, 20 and 30 d post inoculation to observe differences in fungal and plant traits post infection. In vitro cultivation led to increased spore production but reduced plant shoot phosphorus. Our results indicate that in vitro propagation may indirectly select for traits that affect symbiotic quality.     

The interactions between plant life form and fungal traits of arbuscular mycorrhizal fungi determine the symbiotic community

Arbuscular mycorrhizal (AM) fungi have traditionally been considered generalist symbionts. However, an increasing number of studies are pointing out the selectivity potential of plant hosts. Plant life form, determined by plant life history traits, seems to drive the AM fungal community composition. The AM fungi also exhibit a wide diversity of functional traits known to be responsible for their distribution in natural ecosystems. However, little is known about the role of plant and fungal traits driving the resultant symbiotic assemblages. With the aim of testing the feedback relationship between plant and fungal traits on the resulting AM fungal community, we inoculated three different plant life forms, i.e. annual herbs, perennial herbs and perennial semi-woody plants, with AM fungal communities sampled in different seasons. We hypothesized that the annual climate variation will induce changes in the mean traits of the AM fungal communities present in the soil throughout the year. Furthermore, the association of plants with different life forms with AM fungi with contrasting life history traits will show certain preferences according to reciprocal traits of the plants and fungi. We found changes in the AM fungal community throughout the year, which were differentially disrupted by disturbance and altered by plant growth form and plant biomass. Both plant and fungal traits clearly contributed to the resultant AM fungal communities. The revealed process can have implications for the functioning of ecosystems since changes in dominant plant life forms or climatic variables could influence the traits of AM fungal communities in soil and hence ecosystem processes.     

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.     

Linking flowering and reproductive allocation in response to nitrogen addition in an alpine meadow

Aims Plants can change in phenology and biomass allocation in response to environmental change. It has been demonstrated that nitrogen is the most limiting resource for plants in many terrestrial ecosystems. Previous studies have usually focused on either flowering phenology or biomass allocation of plants in response to nitrogen addition; however, attempts to link flowering phenology and biomass allocation are still rare. In this study, we tested the effects of nitrogen addition on both flowering phenology and reproductive allocation in 34 common species. We also examined the potential linkage between flowering time and reproductive allocation in response to nitrogen addition.Methods We conducted a 3-year nitrogen addition experiment in Tibetan alpine meadow. We measured first flowering date and the reproductive allocation for 34 common plant species in control, low and high nitrogen added plots, respectively. One-way analysis of variance was used to examine differences of first flowering date and reproductive allocation among treatments. The relationships between the change in species first flowering date and change in reproductive allocation in response to nitrogen addition were examined by calculating Pearson correlation coefficients.Important findings For most species, both first flowering date and reproductive allocation significantly responded to nitrogen addition. Nitrogen addition significantly delayed the first flowering date and reduced the reproductive allocation for all graminoid species, but accelerated flowering and increased reproductive allocation for most forb species. We found that changes in first flowering date significantly negatively correlated with the changes in reproductive allocation over species in response to nitrogen, which indicated a positive relationship between flowering response and plant performance in reproductive allocation. Species that advanced their flowering time with nitrogen addition increased their reproductive allocation, whereas those that delayed flowering time tended to decline in reproductive allocation with nitrogen addition. Our results suggest that species-specific switch from vegetative growth to reproductive growth could influence species performance.     

Foliar damage modifies floral attractiveness to pollinators in Alstroemeria exerens

Pollination is a requisite for sexual reproduction in plants and its success may determine the reproductive output of individuals. Pollinator preference for some floral designs or displays that are lacking or poorly developed in focal plants may constrain the pollination process. Foliar herbivory may affect the expression of floral traits, thus reducing pollinator attraction. Natural populations of the Andean species Alstroemeria exerens (Alstromeriaceae) in central Chile show high levels of foliar herbivory, and floral traits show phenotypic variation. In the present field study, we addressed the attractive role of floral traits in A. exerens and the effect of foliar damage on them. Particularly, we posed the following questions: (1) Is there an association between floral display and design traits and the number and duration of pollinator visits? and (2) Does foliar damage affect the floral traits associated with pollinator visitation? To assess the attractiveness of floral traits for pollinators, we recorded the number and duration of visits in 101 focal plants. To evaluate the effects of foliar damage on floral traits, 100 plants of similar size were randomly assigned to control or damage groups during early bud development. Damaged plants were clipped using scissors (50% of leaf area) and control plants were manually excluded from natural herbivores (<5% of leaf area eaten). During the peak of flowering, we recorded the number of open flowers, and estimated corolla and nectar guide areas. The number and duration of pollinator visits was statistically associated with floral design and display traits. Plants with larger displays, corollas and nectar guide areas received more visits. Visits lasted longer as display increases. In addition, foliar damage affected attractive traits. Damaged plants had fewer open flowers and smaller nectar guide areas. We conclude that foliar damage affects plant attractiveness for pollinators and hence may indirectly affect plant fitness.     

Complex Genetic Effects on Early Vegetative Development Shape Resource Allocation Differences Between Arabidopsis lyrata Populations

Costs of reproduction due to resource allocation trade-offs have long been recognized as key forces in life history evolution, but little is known about their functional or genetic basis. Arabidopsis lyrata, a perennial relative of the annual model plant A. thaliana with a wide climatic distribution, has populations that are strongly diverged in resource allocation. In this study, we evaluated the genetic and functional basis for variation in resource allocation in a reciprocal transplant experiment, using four A. lyrata populations and F₂ progeny from a cross between North Carolina (NC) and Norway parents, which had the most divergent resource allocation patterns. Local alleles at quantitative trait loci (QTL) at a North Carolina field site increased reproductive output while reducing vegetative growth. These QTL had little overlap with flowering date QTL. Structural equation models incorporating QTL genotypes and traits indicated that resource allocation differences result primarily from QTL effects on early vegetative growth patterns, with cascading effects on later vegetative and reproductive development. At a Norway field site, North Carolina alleles at some of the same QTL regions reduced survival and reproductive output components, but these effects were not associated with resource allocation trade-offs in the Norway environment. Our results indicate that resource allocation in perennial plants may involve important adaptive mechanisms largely independent of flowering time. Moreover, the contributions of resource allocation QTL to local adaptation appear to result from their effects on developmental timing and its interaction with environmental constraints, and not from simple models of reproductive costs.     

Trade-off between seed number and weight: Influence of a grass–endophyte symbiosis

Plant fitness is enhanced by resource allocation to seed number (offspring number) or weight (offspring survival). Besides, there is a well known trade-off in resource allocation between both traits. Symbiotic interactions can influence plant resource allocation to reproduction, yet little research has been performed in this direction. We studied the consequences of a grass–fungus symbiosis on the trade-off between seed number and weight, using Lolium multiflorum and the endophyte Neotyphodium occultans as our study system. In ecological terms, we experimentally removed N. occultans from L. multiflorum plants, and compared reproductive allocation to seed number and weight in endophyte-symbiotic vs. non-symbiotic plants at different levels of nutrient availability (small pots vs. large pots). In evolutionary terms, we compared reproductive allocation between symbiotic vs. non-symbiotic plants for different host genotypes. All plants showed a negative association between seed number and weight, once standardized for total reproductive biomass. Under high nutrient availability, endophyte-symbiotic plants showed higher seed weight than non-symbiotic plants for any seed number. However, no differences were observed under low nutrient availability. Endophyte influence also varied according to L. multiflorum genotype; specifically, endophyte-symbiotic plants showed a lower slope in the relationship between seed number and weight than non-symbiotic plants for the ‘Marshall’ genotype but no endophyte influence was found for the “Pampean” genotype. The results implied a higher plasticity in seed weight and lower plasticity in seed number for symbiotic plants. Indeed, endophyte-symbiotic plants showed an overall lower slope in the association between seed number and total reproductive biomass than non-symbiotic plants. Our results suggest that N. occultans induces heavier seeds in L. multiflorum plants under environmental conditions favorable to plant growth or for certain plant genotypes. We propose that symbiotic interactions may influence the evolution of seed number and weight trade-off.     

Mutually beneficial legume symbioses with soil microbes and their potential for plant production

Legumes develop different mutually beneficial symbioses with soil microbes, such as arbuscular mycorrhizal (AM) fungi, nodule bacteria and plant growth promoting bacteria. Symbioses supply the plants with nutrients (predominantly with nitrogen and phosphorus), protect them from pathogens and abiotic stresses and improve soil microbial biodiversity and fertility. The synergistic activity of beneficial soil microbes (BSM) on the plants has great importance for the use of multi-component symbiotic systems in low-input sustainable environmentally-friendly agrotechnologies. However, the complex nature of the AM symbiosis when in a multi-component symbiosis (plant-fungus-bacteria) creates complications for the fungus to produce AM fungal propagules and poses questions (a) about the effectiveness of the fungus per se in interactions with the plants, without associates, and (b) about the necessity of using sterile/axenic conditions for the production of the AM fungi based inoculants because of any mixing and competition by microbes from the inoculants with the local soil microbial consortia. The legume genes controlling interactions with BSM (including genes responsible for effectiveness of such interactions) should be considered as a united genetic system. The plant genome is more stable than that of microbes and therefore crop plants should select beneficial microbes and control the effectiveness of the whole plant-microbe system in the field for the benefit of the crop and therefore of human beings. There is clearly a need to breed legume crops with improved performance under sustainable conditions involving interactions with BSM and optimising the use of agrochemicals.     

Effects of the endophyte Epichloë coenophiala on the root microbial community and growth performance of tall fescue in different saline-alkali soils

Soil salinization is detrimental to plant growth and yield in agroecosystems worldwide. Epichloë endophytes, a class of clavicipitaceous fungi, enhance the resistance of host plants to saline-alkali stress. This study explored the effects of the systemic fungal endophyte Epichloë coenophiala on the root microbial community and growth performance of tall fescue (Lolium arundinaceum) growing under different saline-alkali stress conditions. Structural equation modeling (SEM) was conducted to analyze the direct and indirect effects (mediated by root microbial community diversity and soil properties) of the endophyte on the growth of tall fescue under saline-alkali stress. The endophyte-infected plants produced higher shoot and root biomass compared to endophyte-free plants under saline-alkali stress (200 and 400 mM). Endophyte infection increased the fungal community diversity and altered its composition in the roots, decreasing the relative abundance of Ascomycota and increasing that of Glomeromycota. Furthermore, endophyte infection decreased the bacterial community diversity and the relative abundance of dominant Proteobacteria. SEM showed that endophyte infection increased the shoot and root biomass under saline-alkali stress (200 and 400 mM) by increasing the arbuscular mycorrhizal fungal diversity in the roots, and soil total nitrogen and phosphorus concentrations. Therefore, it is important to examine aboveground microbes as factors influencing plant growth in saline-alkali stress by affecting belowground microbes and soil chemical properties.     

The significance of host-fungus combinations in ectomycorrhizal symbioses for the chemical quality of willow foliage

The majority of plants interacts with mycorrhizal fungi, which predominantly provide mutual benefits, but also costs. We tested the hypothesis that specific combinations of host plants (four commercial varieties of Salix spp.) and ectomycorrhizal partners (species of Laccaria, Paxillus, Tricholoma and Hebeloma) differ in their effects on the host foliar chemistry. Twenty specific host - mycorrhiza combinations were pot-grown outdoors under low-N conditions. Foliar concentrations of total phenolics and salicylic acid were decreased by mycorrhizas in S. schwerinii x S. viminalis and S. x dasyclados genotypes, but increased in S. viminalis. Mycorrhiza effects on host biomass production were positive, indifferent or negative, depending on the specific host—mycorrhiza combination. The host plant genotype influenced mainly the direction of mycorrhizal effects on foliar chemistry, whereas the magnitude of mycorrhizal effects varied mostly among the fungal genotypes. The results are relevant with respect to the possible interactions between mycorrhizas, plants and leaf herbivores.     

Consequences of simultaneous interactions of fungal endophytes and arbuscular mycorrhizal fungi with a shared host grass

Plants simultaneously associate with multiple microbial symbionts throughout their lifetimes. To address the question of whether the effects of simultaneous symbionts are contingent on the specific identities, we conducted a greenhouse experiment manipulating the presence and identities of arbuscular mycorrhizal fungi (AMF) and fungal endophytes on the shared host grass Elymus hystrix. Each plant host was inoculated with one of two AMF species having varying effects on host growth, or a sterile soil control. Further, we used naturally occurring endophyte‐infected (E+) and uninfected (E–) individuals from two populations of the endophyte Epichloë elymi that varied in their interaction with E. hystrix. We then measured responses of plants, AMF, and fungal endophytes. Overall, we found that the combined effects of AMF and fungal endophytes on plant growth were additive, reflecting the mutualistic quality of each symbiont independently interacting with host plants. However, fungal endophyte infection differentially altered hyphal colonization of the two AMF species and the identity of the coinfecting AMF species affected fungal endophyte fitness traits. The results of this study demonstrate that the outcome of interspecific symbiotic interactions varies with partner identity such that the effects of simultaneous symbioses can not be generalized.     

Nitrogen form, availability, and mycorrhizal colonization affect biomass and nitrogen isotope patterns in Pinus sylvestris

Nitrogen (N) isotope patterns are useful for understanding carbon and nitrogen dynamics in mycorrhizal systems but questions remain about how different N forms, fungal symbionts, and N availabilities influence δ¹⁵N signatures. Here, we studied how biomass allocation and δ¹⁵N patterns in Pinus sylvestris L. cultures were affected by nitrogen supply rate (3% per day or 4% per day relative to the nitrogen already present), nitrogen form (ammonium versus nitrate), and mycorrhizal colonization by fungi with a greater (Laccaria laccata) or lesser (Suillus bovinus) ability to assimilate nitrate. Mycorrhizal (fungal) biomass was greater with ammonium than with nitrate nutrition for Suillus cultures but similar for Laccaria cultures. Total biomass was less with nitrate nutrition than with ammonium nutrition for nonmycorrhizal cultures and was less in mycorrhizal cultures than in nonmycorrhizal cultures. The sequestration of available N by mycorrhizal fungi limited plant N supply. This limitation and the higher energetic cost of nitrate reduction than ammonium assimilation appeared to control plant biomass accumulation. Colonization decreased foliar δ¹⁵N by 0.5 to 2.2‰ (nitrate) or 1.7 to 3.5‰ (ammonium) and increased root tip δ¹⁵N by 0 to 1‰ (nitrate) or 0.6 to 2.3‰ (ammonium). Root tip δ¹⁵N and fungal biomass on root tips were positively correlated in ammonium treatments (r ²?=?0.52) but not in nitrate treatments (r ²?=?0.00). Fungal biomass on root tips was enriched in ¹⁵N an estimated 6–8‰ relative to plant biomass in ammonium treatments. At high nitrate availability, Suillus colonization did not reduce plant δ¹⁵N. We conclude that: (1) transfer of ¹⁵N-depleted N from mycorrhizal fungi to plants produces low plant δ¹⁵N signatures and high root tip and fungal δ¹⁵N signatures; (2) limited nitrate reduction in fungi restricted transfer of ¹⁵N-depleted N to plants when nitrate is supplied and may account for many field observations of high plant δ¹⁵N under such conditions; (3) plants could transfer assimilated nitrogen to fungi at high nitrate supply but such transfer was without ¹⁵N fractionation. These factors probably control plant δ¹⁵N patterns across N availability gradients and were here incorporated into analytical equations for interpreting nitrogen isotope patterns in mycorrhizal fungi and plants.