Seed-borne fungal endophytes constrain reproductive success of host plants under ozone pollution.

Tropospheric ozone is among the global change factors that pose a threat to plants and microorganisms. Symbiotic microorganisms can assist plants to cope with stress, but their role in the tolerance of plants to ozone is poorly understood. Here, we subjected endophyte-symbiotic and non-symbiotic plants of Lolium multiflorum, an annual species widely distributed in temperate grasslands, to high and low (i.e., charcoal-filtered air) ozone levels at vegetative and reproductive phases. Exposure to high ozone reduced leaf photochemical efficiency and greenness in both symbiotic and non-symbiotic plants. However, ozone-induced oxidative damage at biochemical level (i.e., lipid peroxidation) was mostly detected in symbiotic plants. Ozone exposure at the vegetative phase did not affect the reproductive investment in seeds, indicating full recovery from stress. Ozone exposure at the reproductive phase reduced biomass and seed production only in symbiotic plants indicating a symbiont-associated cost. At low ozone, endophyte-symbiotic plants showed a steeper slope in the relationship between seed number and seed weight (i.e., a number-weight trade-off) compared to non-symbiotic plants. However, when plants were treated at the reproductive phase, ozone increased the imbalance between seed number and seed weight in both endophyte-symbiotic and non-symbiotic plants. Plants with endophytes at the reproductive stage produced fewer seeds, which were not compensated by increased seed weight. Thus, fungal mycelium growing within ovaries or ozone-induced antioxidant systems may result in costs that finally depress the fitness of plants. Despite ozone pollution could destabilize plant-endophyte mutualisms and render them dysfunctional, other endophyte-mediated benefits (e.g., resistance to herbivory, tolerance to drought) could over-compensate these losses and explain the high incidence of the symbiosis in nature.

Getting ready for the ozone battle: Vertically transmitted fungal endophytes have transgenerational positive effects in plants.

Ground-level ozone is a global air pollutant with high toxicity and represents a threat to plants and microorganisms. Although beneficial microorganisms can improve host performance, their role in connecting environmentally induced maternal plant phenotypes to progeny (transgenerational effects [TGE]) is unknown. We evaluated fungal endophyte-mediated consequences of maternal plant exposure to ozone on performance of the progeny under contrasting scenarios of the same factor (high and low) at two stages: seedling and young plant. With no variation in biomass, maternal ozone-induced oxidative damage in the progeny that was lower in endophyte-symbiotic plants. This correlated with an endophyte-mediated higher concentration of proline, a defence compound associated with stress control. Interestingly, ozone-induced TGE was not associated with reductions in plant survival. On the contrary, there was an overall positive effect on seedling survival in the presence of endophytes. The positive effect of maternal ozone increasing young plant survival was irrespective of symbiosis and only expressed under high ozone condition. Our study shows that hereditary microorganisms can modulate the capacity of plants to transgenerationally adjust progeny phenotype to atmospheric change.

Ontogenetic and trans-generational dynamics of a vertically transmitted fungal symbiont in an annual host plant in ozone-polluted settings.

Tropospheric ozone is an abiotic stress of increasing importance in the context of global climate change. This greenhouse gas is a potent phytotoxic molecule with demonstrated negative effects on crop yield and natural ecosystems. Recently, oxidative stress has been proposed as a mechanism that could regulate the interaction between cool-season grasses and Epichloë endophytes. We hypothesized that exposure of Lolium multiflorum plants, hosting endophytes to an ozone-polluted environment at different ontogenetic phases, would impact the trans-generational dynamics of the vertically transmitted fungal symbiont. Here, we found that the ozone-induced stress on the mother plants did not affect the endophyte vertical transmission but it impaired the persistence of the fungus in the seed exposed to artificial ageing. Endophyte longevity in seed was reduced by exposure of the mother plant to ozone. Although ozone exposure did not influence either the endophyte mycelial concentration or their compound defences (loline alkaloids), a positive correlation was observed between host fitness and the concentration of endophyte-derived defence compounds. This suggests that fungal defences in grass seeds were not all produced in situ but remobilized from the vegetative tissues. Our study reveals ozone trans-generational effects on the persistence of a beneficial symbiont in a host grass.

Dormancy, a critical trait for weed success in crop production systems.

Agricultural practices exert selective forces on weed populations. As these practices change over time, weed adaptive traits also evolve, allowing weeds to persist in the new environment. However, only weeds having individuals showing the trait with adaptive significance will be able to cope with these changes, thus allowing a sub-population to be selected for persistence. In addition, changes in agricultural practices can select new weed species showing functional traits with characteristics adaptive to the modified system. Seed dormancy has long been recognized as a trait with enormous adaptive value to adjust weed biology to cropping systems. In this paper, we illustrate with examples of success and failure, the value of seed dormancy as a functional trait to cope with long-term changes in crop production systems. We show that successful outcomes are mostly related to the existence of sufficient variability for the functioning of physiological mechanisms that control dormancy characteristics as influenced by the agricultural environment. Presented examples illustrate how knowledge about the relationship that exists between agricultural practices and their selective pressure on seed dormancy can be instrumental in predicting changes in weed biotype dormancy characteristics or foreseeing the appearance of new weed species in future agricultural scenarios. © 2019 Society of Chemical Industry.

Legacy of historic ozone exposure on plant community and food web structure.

Information on whole community responses is needed to predict direction and magnitude of changes in plant and animal abundance under global changes. This study quantifies the effect of past ozone exposure on a weed community structure and arthropod colonization. We used the soil seed bank resulting from a long-term ozone exposure to reestablish the plant community under a new low-pollution environment. Two separate experiments using the same original soil seed bank were conducted. Plant and arthropod richness and species abundance was assessed during two years. We predicted that exposure to episodic high concentrations of ozone during a series of growing cycles would result in plant assemblies with lower diversity (lower species richness and higher dominance), due to an increase in dominance of the stress tolerant species and the elimination of the ozone-sensitive species. As a consequence, arthropod-plant interactions would also be changed. Species richness of the recruited plant communities from different exposure histories was similar (≈ 15). However, the relative abundance of the dominant species varied according to history of exposure, with two annual species dominating ozone enriched plots (90 ppb: Spergula arvensis, and 120 ppb: Calandrinia ciliata). Being consistent both years, the proportion of carnivore species was significantly higher in plots with history of higher ozone concentration (≈3.4 and ≈7.7 fold higher in 90 ppb and 120 ppb plots, respectively). Our study provides evidence that, past history of pollution might be as relevant as management practices in structuring agroecosystems, since we show that an increase in tropospheric ozone may influence biotic communities even years after the exposure.

Arthropod assemblage related to volatile cues in flowering wheat: interaction between aphid herbivory and soil conditions as induction factors.

Volatile cues released by plants play an important role in plant-insect interactions and are influenced by pests or soil conditions affecting plant metabolism. Field microcosm experiments were used to characterize arthropod spontaneous assemblies in homogenous unstressed wheat patches exposed to volatile cues coming from wheat plants with different levels of stress. The design was a factorial completely randomized block design with three replications. Source wheat pots combined two stress factors: 1) soil degradation level: high and low, and 2) aphid herbivory: with (A) and without (B). Eighteen experimental units consisted of source stressed wheat pots, connected by tubes conducting the volatile cues to sink wheat patches. These patches were located at the end of the tubes placed in a flowering wheat field. Arthropod assemblies on wheat sinks were different between years and they were associated to the source cues. Soil condition was the main discriminating factor among arthropods when a clear contrast between high and low soil degradation was observed, whereas aphid herbivory was the main discriminating factor when soil condition effects were absent. Main soil properties related with arthropods assembly were Mg and K in the first year and cation exchange capacity, total nitrogen, and pH in the second year of experiment. According to this study, spontaneous arthropod distributions in the homogeneous, unstressed wheat patch responded to the volatile cues coming from wheat sources growing in particular soil conditions. It is possible to suggest that soil-plant-herbivore interactions change wheat cues and this phenomenon produces significant differences in neighboring arthropod community structure.

Ozone exposure of a weed community produces adaptive changes in seed populations of Spergula arvensis.

Tropospheric ozone is one of the major drivers of global change. This stress factor alters plant growth and development. Ozone could act as a selection pressure on species communities composition, but also on population genetic background, thus affecting life history traits. Our objective was to evaluate the consequences of prolonged ozone exposure of a weed community on phenotypic traits of Spergulaarvensis linked to persistence. Specifically, we predicted that the selection pressure exerted by high ozone concentrations as well as the concomitant changes in the weed community would drive population adaptive changes which will be reflected on seed germination, dormancy and longevity. In order to test seed viability and dormancy level, we conducted germination experiments for which we used seeds produced by S. arvensis plants grown within a weed community exposed to three ozone treatments during four years (0, 90 and 120 ppb). We also performed a soil seed bank experiment to test seed longevity with seeds coming from both the four-year ozone exposure experiment and from a short-term treatment conducted at ambient and added ozone concentrations. We found that prolonged ozone exposure produced changes in seed germination, dormancy and longevity, resulting in three S. arvensis populations. Seeds from the 90 ppb ozone selection treatment had the highest level of germination when stored at 75% RH and 25 °C and then scarified. These seeds showed the lowest dormancy level when being subjected to 5 ºC/5% RH and 25 ºC/75% followed by 5% RH storage conditions. Furthermore, ozone exposure increased seed persistence in the soil through a maternal effect. Given that tropospheric ozone is an important pollutant in rural areas, changes in seed traits due to ozone exposure could increase weed persistence in fields, thus affecting weed-crop interactions, which could ultimately reduce crop production.

Molecular identification of the secondary endosymbiont Hamiltonella defensa in the rose-grain aphid Metopolophium dirhodum.

This is the first report of the association between the rose-grain aphid Metopolophium dirhodum, a potentially important cereal pest and the facultative symbiont Hamiltonella defensa. The infection with this gamma-proteobacterium was determined by PCR in laboratory-reared and field-collected specimens of an Argentinian population of the aphid. Partial bacterial 16S, IGS and 23S rRNA genes were sequenced and compared to other available Hamiltonella sequences by phylogenetic analysis. the present study provides new information on previously unknown M. dirhodum microbiota.

Mutualism effectiveness and vertical transmission of symbiotic fungal endophytes in response to host genetic background.

Certain species of the Pooideae subfamily develop stress tolerance and herbivory resistance through symbiosis with vertically transmitted, asexual fungi. This symbiosis is specific, and genetic factors modulate the compatibility between partners. Although gene flow is clearly a fitness trait in allogamous grasses, because it injects hybrid vigor and raw material for evolution, it could reduce compatibility and thus mutualism effectiveness. To explore the importance of host genetic background in modulating the performance of symbiosis, Lolium multiflorum plants, infected and noninfected with Neotyphodium occultans, were crossed with genetically distant plants of isolines (susceptible and resistant to diclofop-methyl herbicide) bred from two cultivars and exposed to stress. The endophyte improved seedling survival in genotypes susceptible to herbicide, while it had a negative effect on one of the genetically resistant crosses. Mutualism provided resistance to herbivory independently of the host genotype, but this effect vanished under stress. While no endophyte effect was observed on host reproductive success, it was increased by interpopulation plant crosses. Neither gene flow nor herbicide had an important impact on endophyte transmission. Host fitness improvements attributable to gene flow do not appear to result in direct conflict with mutualism while this seems to be an important mechanism for the ecological and contemporary evolution of the symbiotum.

The interplay between the effectiveness of the grass-endophyte mutualism and the genetic variability of the host plant.

Neotyphodium endophytic fungi, the asexual state of Epichloë species, protect cool-season grasses against stresses. The outcomes of Neotyphodium-grass symbioses are agronomically relevant as they may affect the productivity of pastures. It has been suggested that the mutualism is characteristic of agronomic grasses and that differential rates of gene flow between both partners' populations are expected to disrupt the specificity of the association and, thus, the mutualism in wild grasses. We propose that compatibility is necessary but not sufficient to explain the outcomes of Neotyphodium-grass symbiosis, and advance a model that links genetic compatibility, mutualism effectiveness, and endophyte transmission efficiency. For endophytes that reproduce clonally and depend on allogamous hosts for reproduction and dissemination, we propose that this symbiosis works as an integrated entity where gene flow promotes its fitness and evolution. Compatibility between the host plant and the fungal endophyte would be high in genetically close parents; however, mutualism effectiveness and transmission efficiency would be low in fitness depressed host plants. Increasing the genetic distance of mating parents would increase mutualism effectiveness and transmission efficiency. This tendency would be broken when the genetic distance between parents is high (out-breeding depression). Our model allows for testable hypotheses that would contribute to understand the coevolutionary origin and future of the endophyte-grass mutualism.

Imperfect vertical transmission of the endophyte Neotyphodium in exotic grasses in grasslands of the flooding pampa.

Cool-season grasses establish symbioses with vertically transmitted Neotyphodium endophytes widespread in nature. The frequency of endophyte-infected plants in closed populations (i.e., without migrations) depends on both the differential fitness between infected and non-infected plants, and the endophyte-transmission efficiency. Most studies have been focused on the first mechanism ignoring the second. Infection frequency and endophyte transmission from vegetative tissues to seeds were surveyed in two grasses growing in vegetation units that differ in flood and grazing regimes, and soil salinity. Transmission efficiency and infection frequency for tall fescue did not vary significantly and were 0.98 and 1.00, respectively. For Italian ryegrass, transmission efficiency and infection frequency were 0.88 and 0.57 in humid prairies, and 0.96 and 0.96 in the other vegetation units. Only in humid mesophytic meadows, the observed pattern was irrespective of the presence or absence of grazers. Our results suggest that selection forces for endophyte infection are different for both species. Imperfect transmission was only compensated in tall fescue through an increased fitness of infected plants. Interpreting variations of infection frequency only in terms of differential fitness can be misleading, considering that endophyte transmission can be imperfect and variable in nature. Therefore, this study highlights the importance of measuring transmission efficiency.

Neotyphodium endophyte infection frequency in annual grass populations: relative importance of mutualism and transmission efficiency.

Persistence and ubiquity of vertically transmitted Neotyphodium endophytes in grass populations is puzzling because infected plants do not consistently exhibit increased fitness. Using an annual grass population model, we show that the problems for matching endophyte infection and mutualism are likely to arise from difficulties in detecting small mutualistic effects, variability in endophyte transmission efficiency and an apparent prevalence of non-equilibrium in the dynamics of infection. Although endophytes would ultimately persist only if the infection confers some fitness increase to the host plants, such an increase can be very small, as long as the transmission efficiency is sufficiently high. In addition, imperfect transmission limits effectively the equilibrium infection level if the infected plants exhibit small or large reproductive advantage. Under frequent natural conditions, the equilibrium infection level is very sensitive to small changes in transmission efficiency and host reproductive advantage, while convergence to such an equilibrium is slow. As a consequence, seed immigration and environmental fluctuation are likely to keep local infection levels away from equilibrium. Transient dynamics analysis suggests that, when driven by environmental fluctuation, infection frequency increases would often be larger than decreases. By contrast, when due to immigration, overrepresentation of infected individuals tends to vanish faster than equivalent overrepresentation of non-infected individuals.

Glyphosate-resistant weeds of South American cropping systems: an overview.

Herbicide resistance is an evolutionary event resulting from intense herbicide selection over genetically diverse weed populations. In South America, orchard, cereal and legume cropping systems show a strong dependence on glyphosate to control weeds. The goal of this report is to review the current knowledge on cases of evolved glyphosate-resistant weeds in South American agriculture. The first reports of glyphosate resistance include populations of highly diverse taxa (Lolium multiflorum Lam., Conyza bonariensis L., C. canadensis L.). In all instances, resistance evolution followed intense glyphosate use in fruit fields of Chile and Brazil. In fruit orchards from Colombia, Parthenium hysterophorus L. has shown the ability to withstand high glyphosate rates. The recent appearance of glyphosate-resistant Sorghum halepense L. and Euphorbia heterophylla L. in glyphosate-resistant soybean fields of Argentina and Brazil, respectively, is of major concern. The evolution of glyphosate resistance has clearly taken place in those agroecosystems where glyphosate exerts a strong and continuous selection pressure on weeds. The massive adoption of no-till practices together with the utilization of glyphosate-resistant soybean crops are factors encouraging increase in glyphosate use. This phenomenon has been more evident in Argentina and Brazil. The exclusive reliance on glyphosate as the main tool for weed management results in agroecosystems biologically more prone to glyphosate resistance evolution.

Herbicide selection of Italian ryegrass under different levels of UVB radiation.

Ultraviolet-B radiation is an environmental stress for plants and this situation could become aggravated in the next decades. In this study we used Italian ryegrass (Lolium multiflorum Lam.) as a model system to test whether an environmental stress derived from global change, such as UVB, can influence the efficacy of control procedures and evolution toward herbicide resistance. We grew three generations of Italian ryegrass plants with and without UVB light and subjected them to a series of diclofop-methyl [(+/-)-2-[4-(2,4-dichlorophenoxy) phenoxy] propanoic acid, methyl ester] doses. The effect of selection history was tested with herbicide dose response. The effect of herbicide application on plant survival and biomass varied significantly among herbicide doses and with absence or presence of UVB light. In the absence of herbicide, the decrease in individual fecundity with increasing plant density was similar under both no-UVB and UVB light treatments. Only plants growing without UVB light increased production of reproductive structures in response to the decrease in density caused by herbicide application. Our study shows that UVB light was a weak stress factor for the ryegrass plants. However, when herbicide selection pressure was high, UVB light reduced the evolution toward herbicide tolerance. When selection pressure on the parental plants was lower, the two stress factors had a synergistic effect, causing changes in herbicide efficacy that in turn had demographic and evolutionary consequences. In the field, these interactions between stress factors might be of significance for annual weeds in which seed output is a major determinant in fitness.

Coriander essential oil composition from two genotypes grown in different environmental conditions.

The objective was to study the essential oil composition of coriander fruits in plants growing in environments differing in soil conditions and weediness level. Factorial field experiments were conducted in two locations from the Rolling Pampas, Argentina, and two coriander landraces (European and Argentinean) were tested under two levels of nitrogen fertilization and weediness. Data were evaluated with uni- and multivariate techniques. The variation in the oil composition was related to the relative proportion of the constituents and not to the presence/absence of a particular component. Weather conditions in 1997 favored linalool and camphor in both landraces. Location, fertilization, and weediness also affected the chemical profile. The European landrace showed a more stable concentration of the major components than the Argentinean landrace. These results, which show the relationships between some environmental conditions and the essential oil composition, are useful in the development of innovative strategies aimed to improve oil composition and to manage crop pests.

Limits to tree species invasion in pampean grassland and forest plant communities.

Factors limiting tree invasion in the Inland Pampas of Argentina were studied by monitoring the establishment of four alien tree species in remnant grassland and cultivated forest stands. We tested whether disturbances facilitated tree seedling recruitment and survival once seeds of invaders were made available by hand sowing. Seed addition to grassland failed to produce seedlings of two study species, Ligustrum lucidum and Ulmus pumila, but did result in abundant recruitment of Gleditsia triacanthos and Prosopis caldenia. While emergence was sparse in intact grassland, seedling densities were significantly increased by canopy and soil disturbances. Longer-term surveys showed that only Gleditsia became successfully established in disturbed grassland. These results support the hypothesis that interference from herbaceous vegetation may play a significant role in slowing down tree invasion, whereas disturbances create microsites that can be exploited by invasive woody plants. Seed sowing in a Ligustrum forest promoted the emergence of all four study species in understorey and treefall gap conditions. Litter removal had species-specific effects on emergence and early seedling growth, but had little impact on survivorship. Seedlings emerging under the closed forest canopy died within a few months. In the treefall gap, recruits of Gleditsia and Prosopis survived the first year, but did not survive in the longer term after natural gap closure. The forest community thus appeared less susceptible to colonization by alien trees than the grassland. We conclude that tree invasion in this system is strongly limited by the availability of recruitment microsites and biotic interactions, as well as by dispersal from existing propagule sources.