Extreme drought impacts have been underestimated in grasslands and shrublands globally.

Climate change is increasing the frequency and severity of short-term (~1 y) drought events-the most common duration of drought-globally. Yet the impact of this intensification of drought on ecosystem functioning remains poorly resolved. This is due in part to the widely disparate approaches ecologists have employed to study drought, variation in the severity and duration of drought studied, and differences among ecosystems in vegetation, edaphic and climatic attributes that can mediate drought impacts. To overcome these problems and better identify the factors that modulate drought responses, we used a coordinated distributed experiment to quantify the impact of short-term drought on grassland and shrubland ecosystems. With a standardized approach, we imposed ~a single year of drought at 100 sites on six continents. Here we show that loss of a foundational ecosystem function-aboveground net primary production (ANPP)-was 60% greater at sites that experienced statistically extreme drought (1-in-100-y event) vs. those sites where drought was nominal (historically more common) in magnitude (35% vs. 21%, respectively). This reduction in a key carbon cycle process with a single year of extreme drought greatly exceeds previously reported losses for grasslands and shrublands. Our global experiment also revealed high variability in drought response but that relative reductions in ANPP were greater in drier ecosystems and those with fewer plant species. Overall, our results demonstrate with unprecedented rigor that the global impacts of projected increases in drought severity have been significantly underestimated and that drier and less diverse sites are likely to be most vulnerable to extreme drought.

Local Community Assembly Mechanisms and the Size of Species Pool Jointly Explain the Beta Diversity of Soil Fungi.

Fungi play vital regulatory roles in terrestrial ecosystems. Local community assembly mechanisms, including deterministic and stochastic processes, as well as the size of regional species pools (gamma diversity), typically influence overall soil microbial community beta diversity patterns. However, there is limited evidence supporting their direct and indirect effects on beta diversity of different soil fungal functional groups in forest ecosystems. To address this gap, we collected 1606 soil samples from a 25-ha subtropical forest plot in southern China. Our goal was to determine the direct effects and indirect effects of regional species pools on the beta diversity of soil fungi, specifically arbuscular mycorrhizal (AM), ectomycorrhizal (EcM), plant-pathogenic, and saprotrophic fungi. We quantified the effects of soil properties, mycorrhizal tree abundances, and topographical factors on soil fungal diversity. The beta diversity of plant-pathogenic fungi was predominantly influenced by the size of the species pool. In contrast, the beta diversity of EcM fungi was primarily driven indirectly through community assembly processes. Neither of them had significant effects on the beta diversity of AM and saprotrophic fungi. Our results highlight that the direct and indirect effects of species pools on the beta diversity of soil functional groups of fungi can significantly differ even within a relatively small area. They also demonstrate the independent and combined effects of various factors in regulating the diversities of soil functional groups of fungi. Consequently, it is crucial to study the fungal community not only as a whole but also by considering different functional groups within the community.

Host Defense Metabolites Alter the Interactions between a Bark Beetle and its Symbiotic Fungi.

Successful host plant colonization by tree-killing bark beetle-symbiotic fungal complexes depends on host suitability, which is largely determined by host defense metabolites such as monoterpenes. Studies have shown the ability of specific blends of host monoterpenes to influence bark beetles or their fungal symbionts, but how biologically relevant blends of host monoterpenes influence bark beetle-symbiotic fungal interaction is unknown. We tested how interactions between two host species (lodgepole pine or jack pine) and two fungal symbionts of mountain pine beetle (Grosmannia clavigera or Ophiostoma montium) affect the performance of adult female beetles in vitro. Beetles treated with the propagules of G. clavigera or O. montium or not treated (natural fungal load) were introduced into media amended with a blend of the entire monoterpene profile of either host species and beetle performance was compared. Overall, host blends altered beetle performance depending on the fungal species used in the beetle amendment. When beetles were amended with G. clavigera, their performance was superior over beetles amended with O. montium in either host blend. Furthermore, G. clavigera-amended beetles performed better in media amended with host blends than without a host blend; in contrast, O. montium-amended beetles performed better in media without a host blend than with a host blend. Overall, this study showed that host defense metabolites affect host suitability to bark beetles through influencing their fungal symbionts and that different species of fungal symbionts respond differentlly to host defense metabolites.

Global change effects on plant communities are magnified by time and the number of global change factors imposed.

Global change drivers (GCDs) are expected to alter community structure and consequently, the services that ecosystems provide. Yet, few experimental investigations have examined effects of GCDs on plant community structure across multiple ecosystem types, and those that do exist present conflicting patterns. In an unprecedented global synthesis of over 100 experiments that manipulated factors linked to GCDs, we show that herbaceous plant community responses depend on experimental manipulation length and number of factors manipulated. We found that plant communities are fairly resistant to experimentally manipulated GCDs in the short term (<10 y). In contrast, long-term (≥10 y) experiments show increasing community divergence of treatments from control conditions. Surprisingly, these community responses occurred with similar frequency across the GCD types manipulated in our database. However, community responses were more common when 3 or more GCDs were simultaneously manipulated, suggesting the emergence of additive or synergistic effects of multiple drivers, particularly over long time periods. In half of the cases, GCD manipulations caused a difference in community composition without a corresponding species richness difference, indicating that species reordering or replacement is an important mechanism of community responses to GCDs and should be given greater consideration when examining consequences of GCDs for the biodiversity-ecosystem function relationship. Human activities are currently driving unparalleled global changes worldwide. Our analyses provide the most comprehensive evidence to date that these human activities may have widespread impacts on plant community composition globally, which will increase in frequency over time and be greater in areas where communities face multiple GCDs simultaneously.

Changes in soil fungal community composition depend on functional group and forest disturbance type.

Disturbances have altered community dynamics in boreal forests with unknown consequences for belowground ecological processes. Soil fungi are particularly sensitive to such disturbances; however, the individual response of fungal guilds to different disturbance types is poorly understood. Here, we profiled soil fungal communities in lodgepole pine forests following a bark beetle outbreak, wildfire, clear-cut logging, and salvage-logging. Using Illumina MiSeq to sequence ITS1 and SSU rDNA, we characterized communities of ectomycorrhizal, arbuscular mycorrhizal, saprotrophic, and pathogenic fungi in sites representing each disturbance type paired with intact forests. We also quantified soil fungal biomass by measuring ergosterol. Abiotic disturbances changed the community composition of ectomycorrhizal fungi and shifted the dominance from ectomycorrhizal to saprotrophic fungi compared to intact forests. The disruption of the soil organic layer with disturbances correlated with the decline of ectomycorrhizal and the increase of arbuscular mycorrhizal fungi. Wildfire changed the community composition of pathogenic fungi but did not affect their proportion and diversity. Fungal biomass declined with disturbances that disrupted the forest floor. Our results suggest that the disruption of the forest floor with disturbances, and the changes in C and nutrient dynamics it may promote, structure the fungal community with implications for fungal biomass-C.

Limited evidence of vertical fine-root segregation in a subtropical forest.

Vertical root segregation and the resulting niche partitioning can be a key underpinning of species coexistence. This could result from substantial interspecific variations in root profiles and rooting plasticity in response to soil heterogeneity and neighbours, but they remain largely untested in forest communities. In a diverse forest in subtropical China, we randomly sampled > 4000 root samples from 625 0-30 cm soil profiles. Using morphological and DNA-based methods, we identified 109 woody plant species, determined the degree of vertical fine-root segregation, and examined rooting plasticity in response to soil heterogeneity and neighbour structure. We found no evidence of vertical fine-root segregation among cooccurring species. By contrast, root abundance of different species tended to be positively correlated within soil zones. Underlying these findings was a lack of interspecific variation in fine-root profiles with over 90% of species concentrated in the 0-10 cm soil zone with only one species dominating in the 10-20 cm soil zone. Root profiles exhibited low responsiveness to root neighbours but tended to be shallow in soils with low phosphorus and copper content. These findings suggest that if there is niche differentiation leading to coexistence in this diverse forest, it would be occurring by mechanisms other than vertical fine-root segregation.

Long-Term Warming and Nitrogen Addition Have Contrasting Effects on Ecosystem Carbon Exchange in a Desert Steppe.

Desert steppe, a unique ecotone between steppe and desert in Eurasia, is considered highly vulnerable to global change. However, the long-term impact of warming and nitrogen deposition on plant biomass production and ecosystem carbon exchange in a desert steppe remains unknown. A 12-year field experiment was conducted in a Stipa breviflora desert steppe in northern China. A split-design was used, with warming simulated by infrared radiators as the primary factor and N addition as the secondary factor. Our long-term experiment shows that warming did not change net ecosystem exchange (NEE) or total aboveground biomass (TAB) due to contrasting effects on C4 (23.4% increase) and C3 (11.4% decrease) plant biomass. However, nitrogen addition increased TAB by 9.3% and NEE by 26.0% by increasing soil available N content. Thus, the studied desert steppe did not switch from a carbon sink to a carbon source in response to global change and positively responded to nitrogen deposition. Our study indicates that the desert steppe may be resilient to long-term warming by regulating plant species with contrasting photosynthetic types and that nitrogen deposition could increase plant growth and carbon sequestration, providing negative feedback on climate change.

The effects of livestock grazing on biodiversity are multi-trophic: a meta-analysis.

Anthropogenic disturbance has generated a significant loss of biodiversity worldwide and grazing by domestic herbivores is a contributing disturbance. Although the effects of grazing on plants are commonly explored, here we address the potential multi-trophic effects on animal biodiversity (e.g. herbivores, pollinators and predators). We conducted a meta-analysis on 109 independent studies that tested the response of animals or plants to livestock grazing relative to livestock excluded. Across all animals, livestock exclusion increased abundance and diversity, but these effects were greatest for trophic levels directly dependent on plants, such as herbivores and pollinators. Detritivores were the only trophic level whose abundance decreased with livestock exclusion. We also found that the number of years since livestock was excluded influenced the community and that the effects of grazer exclusion on animal diversity were strongest in temperate climates. These findings synthesise the effects of livestock grazing beyond plants and demonstrate the indirect impacts of livestock grazing on multiple trophic levels in the animal community. We identified the potentially long-term impacts that livestock grazing can have on lower trophic levels and consequences for biological conservation. We also highlight the potentially inevitable cost to global biodiversity from livestock grazing that must be balanced against socio-economic benefits.

Neighbouring plants modify maize root foraging for phosphorus: coupling nutrients and neighbours for improved nutrient-use efficiency.

Nutrient distribution and neighbours can impact plant growth, but how neighbours shape root-foraging strategy for nutrients is unclear. Here, we explore new patterns of plant foraging for nutrients as affected by neighbours to improve nutrient acquisition. Maize (Zea mays) was grown alone (maize), or with maize (maize/maize) or faba bean (Vicia faba) (maize/faba bean) as a neighbour on one side and with or without a phosphorus (P)-rich zone on the other in a rhizo-box experiment. Maize demonstrated root avoidance in maize/maize, with reduced root growth in 'shared' soil, and increased growth away from its neighbours. Conversely, maize proliferated roots in the proximity of neighbouring faba bean roots that had greater P availability in the rhizosphere (as a result of citrate and acid phosphatase exudation) compared with maize roots. Maize proliferated more roots, but spent less time to reach, and grow out of, the P patches away from neighbours in the maize/maize than in the maize/faba bean experiment. Maize shoot biomass and P uptake were greater in the heterogeneous P treatment with maize/faba bean than with maize/maize system. The foraging strategy of maize roots is an integrated function of heterogeneous distribution of nutrients and neighbouring plants, thus improving nutrient acquisition and maize growth. Understanding the foraging patterns is critical for optimizing nutrient management in crops.

The effects of ectomycorrhizal fungal networks on seedling establishment are contingent on species and severity of overstorey mortality.

For tree seedlings in boreal forests, ectomycorrhizal (EM) fungal networks may promote, while root competition may impede establishment. Thus, disruption to EM fungal networks may decrease seedling establishment owing to the loss of positive interactions among neighbors. Widespread tree mortality can disrupt EM networks, but it is not clear whether seedling establishment will be limited by the loss of positive interactions or increased by the loss of negative interactions with surrounding roots. Depending upon the relative influence of these mechanisms, widespread tree mortality may have complicated consequences on seedling establishment, and in turn, the composition of future forests. To discern between these possible outcomes and the drivers of seedling establishment, we determined the relative importance of EM fungal networks, root presence, and the bulk soil on the establishment of lodgepole pine and white spruce seedlings along a gradient of beetle-induced tree mortality. We manipulated seedling contact with EM fungal networks and roots through the use of mesh-fabric cylinders installed in soils of lodgepole pine forests experiencing a range of overstorey tree mortality caused by mountain pine beetle. Lodgepole pine seedling survival was higher with access to EM fungal networks in undisturbed pine forests in comparison with that in beetle-killed stands. That is, overstorey tree mortality shifted fungal networks from being a benefit to a cost on seedling survival. In contrast, overstorey tree mortality did not change the relative strength of EM fungal networks, root presence and the bulk soil on survival and biomass of white spruce seedlings. Furthermore, the relative influence of EM fungal networks, root presence, and bulk soils on foliar N and P concentrations was highly contingent on seedling species and overstorey tree mortality. Our results highlight that following large-scale insect outbreak, soil-mediated processes can enable differential population growth of two common conifer species, which may result in species replacement in the future.

Effects of neighbour location and nutrient distributions on root foraging behaviour of the common sunflower.

Plants regularly encounter patchily distributed soil nutrients. A common foraging response is to proliferate roots within high-quality patches. The influence of the social environment on this behaviour has been given limited attention, despite important fitness consequences of competition for soil resources among plants. Using the common sunflower (Helianthus annuus L.), we compared localized root proliferation in a high-quality patch by plants grown alone to that of plants in two different social environments: with a neighbouring plant sharing equal access to the high-quality patch, and with a neighbouring plant present but farther from the high-quality patch such that the focal individual was in closer proximity to the high-quality patch. Sunflowers grown alone proliferated more roots within high-nutrient patches than lower-nutrient soil. Plants decreased root proliferation within a high-nutrient patch when it was equidistant to a neighbour. Conversely, plants increased root proliferation when they were in closer proximity to the patch relative to a nearby neighbour. Such contingent responses may allow sunflowers to avoid competition in highly contested patches, but to also pre-empt soil resources from neighbours when they have better access to a high-quality patch. We also compared patch occupancy by sunflowers grown alone with two equidistant high-quality patches to occupancy by sunflowers grown with two high-quality patches and a neighbour. Plants grown with a neighbour decreased root length within shared patches but did not increase root length within high-quality patches they were in closer proximity to, perhaps because resource pre-emption may be less important for individuals when resources are more abundant. These results show that nutrient foraging responses in plants can be socially contingent, and that plants may account for the possibility of pre-empting limited resources in their foraging decisions.

Damage to leaf veins suppresses root foraging precision.

PREMISE: Plants generally increase root growth in areas with high nutrients in heterogeneous soils, a phenomenon called foraging precision. The physiology of this process is not well understood, but it may involve shoot-root signaling via leaf veins. If this is true, then damage to leaf veins, but not to nearby mesophyll, would reduce plant foraging precision. METHODS: To test this hypothesis, we imposed two leaf damage treatments on Plantago asiatica and Prunus jamasakura, removing either the tip of each main vein or mesophyll tissue between the veins with a 3-mm-diameter hole punch. After 30 days or 20 weeks of plant growth, we measured root biomass in the soil in response to soil nutrient concentration. RESULTS: When leaf mesophyll was damaged, root biomass of both species was greater in nutrient-rich patches than in nutrient-poor patches. However, when leaf veins were damaged, root biomass was similar between patches. CONCLUSIONS: These results suggest the importance of shoot-root signaling in plants, emphasizing that physiological processes are not necessarily restricted to single organs. The idea that herbivores that damage leaf veins may affect a plant's ability to selectively forage in high-nutrient patches is novel, with implications for natural and managed systems.

A new method for the rapid characterization of root growth and distribution using digital image correlation.

Rapidly determining root growth patterns is biologically important and technically challenging. Current methods focus on direct observation of roots and require destructive excavations or time-consuming root tracing. We developed a novel methodology based on analyzing soil particle displacement, rather than direct observation of roots. This inferred root growth method uses digital image correlation (DIC) analysis, an established and high-throughput method used in many engineering and science disciplines. By applying DIC analyses to repeated images of plants grown in clear window boxes, we produced visually intuitive and quantifiable strain maps, indicating the magnitude and direction of soil movement. From this, we could infer root growth and rapidly quantify root system metrics. Strain measures were closely associated with the spatial distribution of roots and correlated with root length measured using conventional approaches. The method also allowed for the detection of root proliferation in nutrient-enriched soil patches, indicating its suitability for quantifying biological patterns. This novel application of DIC in root biology is effective, scalable, low cost, flexible and complementary to existing technologies. This method offers a new tool for answering questions in plant biology and will be particularly useful in studies involving temporal dynamics of root processes.

Flowering and floral visitation predict changes in community structure provided that mycorrhizas remain intact.

Pollination is critical for plant fitness and population dynamics, yet little attention is paid to the role of flowering and plant-pollinator interactions in structuring plant communities, including community responses to environmental change. Changes in arbuscular mycorrhizal fungi (AMF), nutrient abundances, and plant litter all affect plant access to different resources, and are known regulators of community structure. Each factor can also affect flowering and plant-pollinator interactions, potentially contributing to changes in community structure. To test whether AMF, nutrients, and litter influenced the relationship between pollination and community structure, we conducted a 5-yr field experiment applying fungicide, adding fertilizer, and removing plant litter in native grassland. We measured the distribution of flowers and floral visits among species in year three and linked these measures to changes in plant composition and species richness between years three and five. We hypothesized that an uneven distribution of flowers and visits among species would lead to greater community change, but that the treatments would disrupt this relationship by altering sexual allocation and recruitment. Consistent with our hypothesis, communities with uneven flower distributions exhibited greater changes in community composition and richness under ambient conditions. However, AMF suppression neutralized this relationship and regulated the other treatment effects, highlighting the potential importance of AMF for stabilizing recruitment dynamics. Combined, AMF suppression and nutrient addition caused species losses when few species flowered, likely by compounding stresses for those species. The treatment effects on the relationship between flowering and community composition were more nuanced, but were likely driven by increased competition and altered flowering among species. By contrast, community composition was more stable when visitation rates were uneven among species, irrespective of any treatments. This suggests that some species require high visitation rates to maintain their populations due to greater dependence on sexual reproduction. Combined, these results highlight the importance of flowering and floral visitation to the dynamics of grassland communities. They also suggest that altered recruitment dynamics is a major, yet understudied, mechanism by which environmental change affects communities. Consequently, understanding the effects of environmental change on plant communities will require study of both plant growth and sexual reproduction.

Photosynthetic opportunity cost and energetic cost of a rapid leaf closure behavior in Mimosa pudica.

PREMISE OF THE STUDY: The rapid leaf movement of Mimosa pudica is expected to be costly because of energetic trade-offs with other processes such as growth and reproduction. Here, we assess the photosynthetic opportunity cost and energetic cost of the unique leaf closing behavior of M. pudica. METHODS: In the greenhouse, we employed novel touch-stimulation machines to expose plants to one of three treatments: (1) untouched control plants; (2) plants touch-stimulated to close their leaves during the day to incur energetic costs associated with leaf movement and reduced photosynthesis; (3) plants touched at night to assess the effects of touch alone. M. pudica is nyctinastic and closes its leaves at night; thus, touching at night does not impart additional costs. We directly assessed costs by comparing physical traits. Leaf re-opening response was measured to assess the potential for plant behavioral plasticity to impact photosynthetic opportunity costs. KEY RESULTS: The cost of rapid leaf closure behavior was expressed as a 47% reduction in reproductive biomass accounting for the effect of touch. Touch itself changed physical traits such as biomass, with touched plants being generally bigger. Plants touched at night re-opened their leaflets 26% quicker than plants touched during the day. CONCLUSIONS: We reason that the reproductive allocation costs incurred by M. pudica can be attributed to a combination of photosynthetic opportunity cost and the energetic cost associated with increased stimulation of leaf movement and that behavioral plasticity has the potential to alter photosynthetic opportunity costs.

Maternal experience and soil origin influence interactions between resident species and a dominant invasive species.

Invasive species dominance in invaded communities may not be long-lasting due to regulatory processes, such as plant-soil feedbacks and neighboring species adaptation. Further, the change in species competitive ability may be contingent upon neighbor identity (i.e., specialized response) or consistent across neighbors (i.e., generalized response). Specialized responses can facilitate overall coexistence, while generalized responses may result in competitive exclusion. We set up a greenhouse experiment to test, in three species, the effect of soil conditions (non-invaded vs. invaded soil) and maternal experience (offspring of maternal plants from invaded vs. non-invaded areas) on species competitive ability against the invader Bromus inermis and conspecifics. If changes in species competitive ability against B. inermis were also evident when interacting with conspecifics, it would suggest a generalized increased/decreased competitive ability. Maternal experience resulted in reduced suppression of B. inermis in the three species and no change in tolerance. On the other hand, tolerance to B. inermis was enhanced when plants grew in soil from invaded areas, compared to non-brome soil. Importantly, both the decreased suppression due to maternal experience with B. inermis and the increased tolerance in invaded soil appear to be invader specific, as no such effects were observed when interacting with conspecifics. Specialized responses should facilitate coexistence, as no individual/species is a weaker or stronger competitor against all other neighbors or under all local soil conditions. Further, the negative plant-soil feedback for B. inermis should facilitate native species recovery in invaded areas and result in lower B. inermis performance and dominance over time.

Asynchrony among local communities stabilises ecosystem function of metacommunities.

Temporal stability of ecosystem functioning increases the predictability and reliability of ecosystem services, and understanding the drivers of stability across spatial scales is important for land management and policy decisions. We used species-level abundance data from 62 plant communities across five continents to assess mechanisms of temporal stability across spatial scales. We assessed how asynchrony (i.e. different units responding dissimilarly through time) of species and local communities stabilised metacommunity ecosystem function. Asynchrony of species increased stability of local communities, and asynchrony among local communities enhanced metacommunity stability by a wide range of magnitudes (1-315%); this range was positively correlated with the size of the metacommunity. Additionally, asynchronous responses among local communities were linked with species' populations fluctuating asynchronously across space, perhaps stemming from physical and/or competitive differences among local communities. Accordingly, we suggest spatial heterogeneity should be a major focus for maintaining the stability of ecosystem services at larger spatial scales.

Change in soil fungal community structure driven by a decline in ectomycorrhizal fungi following a mountain pine beetle (Dendroctonus ponderosae) outbreak.

Western North American landscapes are rapidly being transformed by forest die-off caused by mountain pine beetle (Dendroctonus ponderosae), with implications for plant and soil communities. The mechanisms that drive changes in soil community structure, particularly for the highly prevalent ectomycorrhizal fungi in pine forests, are complex and intertwined. Critical to enhancing understanding will be disentangling the relative importance of host tree mortality from changes in soil chemistry following tree death. Here, we used a recent bark beetle outbreak in lodgepole pine (Pinus contorta) forests of western Canada to test whether the effects of tree mortality altered the richness and composition of belowground fungal communities, including ectomycorrhizal and saprotrophic fungi. We also determined the effects of environmental factors (i.e. soil nutrients, moisture, and phenolics) and geographical distance, both of which can influence the richness and composition of soil fungi. The richness of both groups of soil fungi declined and the overall composition was altered by beetle-induced tree mortality. Soil nutrients, soil phenolics and geographical distance influenced the community structure of soil fungi; however, the relative importance of these factors differed between ectomycorrhizal and saprotrophic fungi. The independent effects of tree mortality, soil phenolics and geographical distance influenced the community composition of ectomycorrhizal fungi, while the community composition of saprotrophic fungi was weakly but significantly correlated with the geographical distance of plots. Taken together, our results indicate that both deterministic and stochastic processes structure soil fungal communities following landscape-scale insect outbreaks and reflect the independent roles tree mortality, soil chemistry and geographical distance play in regulating the community composition of soil fungi.

Antifungal genes expressed in transgenic pea (Pisum sativum L.) do not affect root colonization of arbuscular mycorrhizae fungi.

Genetically modified crops have raised concerns about unintended consequences on non-target organisms including beneficial soil associates. Pea transformed with four antifungal genes 1-3 ß glucanase, endochitinase, polygalacturonase-inhibiting proteins, and stilbene synthase is currently under field-testing for efficacy against fungal diseases in Canada. Transgenes had lower expression in the roots than leaves in greenhouse experiment. To determine the impact of disease-tolerant pea or gene products on colonization by non-target arbuscular mycorrhizae and nodulation by rhizobium, a field trial was established. Transgene insertion, as single gene or stacked genes, did not alter root colonization by arbuscular mycorrhiza fungus (AMF) or root nodulation by rhizobium inoculation in the field. We found no effect of transgenes on the plant growth and performance although, having a dual inoculant with both AMF and rhizobium yielded higher fresh weight shoot-to-root ratio in all the lines tested. This initial risk assessment of transgenic peas expressing antifungal genes showed no deleterious effect on non-target organisms.

Ectomycorrhizal fungi mediate indirect effects of a bark beetle outbreak on secondary chemistry and establishment of pine seedlings.

Dendroctonus ponderosae has killed millions of Pinus contorta in western North America with subsequent effects on stand conditions, including changes in light intensity, needle deposition, and the composition of fungal community mutualists, namely ectomycorrhizal fungi. It is unknown whether these changes in stand conditions will have cascading consequences for the next generation of pine seedlings. To test for transgenerational cascades on pine seedlings, we tested the effects of fungal inoculum origin (beetle-killed or undisturbed stands), light intensity and litter (origin and presence) on seedling secondary chemistry and growth in a glasshouse. We also tracked survival of seedlings over two growing seasons in the same stands from which fungi and litter were collected. Fungal communities differed by inoculum origin. Seedlings grown with fungi collected from beetle-killed stands had lower monoterpene concentrations and fewer monoterpene compounds present compared with seedlings grown with fungi collected from undisturbed stands. Litter affected neither monoterpenes nor seedling growth. Seedling survival in the field was lower in beetle-killed than in undisturbed stands. We demonstrate that stand mortality caused by prior beetle attacks of mature pines have cascading effects on seedling secondary chemistry, growth and survival, probably mediated through effects on below-ground mutualisms.