Multitrophic Interactions Between Arbuscular Mycorrhizal Fungi, Foliar Endophytic Fungi and Aphids.

Almost all living plants can be simultaneously colonised by arbuscular mycorrhizal fungi in the roots and endophytes in the shoots, while also being attacked by insect herbivores. However, to date, no study has ever examined the multitrophic interactions between these two different fungal groups and insects on any species of forb. Here, we examined the effects of two commercial species mixtures of arbuscular mycorrhizal fungi (AMF) and two foliar endophytes (Colletotrichum acutatum and Cladosporium oxysporum) on the growth of an invasive weed, Impatiens glandulifera, and the aphids that attack it. AMF reduced plant biomass, which was most evident when C. oxysporum was inoculated. Mycorrhizal fungi had few effects on aphids, and these depended on the identity of the endophytes present. Meanwhile, endophytes tended to increase aphid numbers, but this depended on the identity of the AMF inoculum. Throughout, there were differences in the responses of the plant to the two mycorrhizal mixtures, demonstrating clear AMF specificity in this plant. These specific effects were also strongly affected by the endophytes, with a greater number of interactions found between the AMF and endophytes than between the endophytes themselves. In particular, AMF reduced infection levels by the endophytes, while some endophyte inoculations reduced mycorrhizal colonisation. We suggest that both AMF and endophytes could play an important part in future biological control programmes of weeds, but further multitrophic experiments are required to unravel the complexity of interactions between spatially separated parts of the plant microbiome.

Evaluating the use of seaweed extracts against root knot nematodes: A meta-analytic approach.

Plant parasitic nematode (PPN) control has historically relied on the use of synthetic chemical nematicides, however many are toxic to both human health and the environment. The withdrawal of the more harmful nematicides coupled with increases in soil temperatures and increased occurrence of pests and diseases associated with climate change, may enable PPN to increase in numbers and spread globally. The need for sustainable and environmentally friendly management options is necessary while facing future food security scares in order to feed the ever-growing population. Seaweed extracts have been used for decades in agriculture and horticulture as soil biostimulants, however there is a growing body of evidence to suggest that they could be used to reduce the occurrence of damaging PPN infections. Using meta-analysis, we investigated whether seaweed extracts applied to soil could reduce root knot nematode (RKN) abundance and whether there could be confounding factors that influence their efficacy. We found that seaweed extracts reduce RKN performance and that various factors affected the efficacy of seaweed, including the seaweed species itself and the crop the seaweed was applied to. Ascophyllum nodosum extracts were found to be the most effective. Particular RKN species were more sensitive than others to seaweed species used and, in some cases, specific seaweed species only affected particular RKN species. Different life cycle stages were also differentially susceptible to seaweed application, where both egg hatching and population abundance could be reduced via seaweed use. This research indicates that seaweed extracts could potentially be used to help reduce RKN attack on plants.

Meta-analysis of the role of entomopathogenic and unspecialized fungal endophytes as plant bodyguards.

Herbaceous plants harbour species-rich communities of asymptomatic endophytic fungi. Although some of these endophytes are entomopathogenic, many are not, and remarkably little is known about how the presence of these fungi in plant tissues affects phytophagous insects. Here we show through a meta-analysis that both entomopathogenic and nonentomopathogenic endophytes have a negative effect on insect herbivores. Growth and performance of polyphagous and sucking insects are reduced by nonentomopathogenic endophytes, but monophages are unaffected, likely because the latter are better adapted to secondary metabolites produced or induced by the fungi. Furthermore, studies using excised leaves report weaker effects than those with intact plants, likely caused by chemical changes being masked by leaf excision. Most surprisingly, endophyte infection of seeds produces the greatest effect on insect herbivores in subsequent mature plants, even though the usual mode of fungal transmission is infection of leaves by airborne spores. We conclude that these ubiquitous hidden fungi may be important bodyguards of plants. However, in order to fully understand their roles in plant protection, we must be aware that minor differences in experimental design can lead to contradictory results.

Explaining European fungal fruiting phenology with climate variability.

Here we assess the impact of geographically dependent (latitude, longitude, and altitude) changes in bioclimatic (temperature, precipitation, and primary productivity) variability on fungal fruiting phenology across Europe. Two main nutritional guilds of fungi, saprotrophic and ectomycorrhizal, were further separated into spring and autumn fruiters. We used a path analysis to investigate how biogeographic patterns in fungal fruiting phenology coincided with seasonal changes in climate and primary production. Across central to northern Europe, mean fruiting varied by approximately 25 d, primarily with latitude. Altitude affected fruiting by up to 30 d, with spring delays and autumnal accelerations. Fruiting was as much explained by the effects of bioclimatic variability as by their large-scale spatial patterns. Temperature drove fruiting of autumnal ectomycorrhizal and saprotrophic groups as well as spring saprotrophic groups, while primary production and precipitation were major drivers for spring-fruiting ectomycorrhizal fungi. Species-specific phenology predictors were not stable, instead deviating from the overall mean. There is significant likelihood that further climatic change, especially in temperature, will impact fungal phenology patterns at large spatial scales. The ecological implications are diverse, potentially affecting food webs (asynchrony), nutrient cycling and the timing of nutrient availability in ecosystems.

Soil Inoculation with Bacillus spp. Modifies Root Endophytic Bacterial Diversity, Evenness, and Community Composition in a Context-Specific Manner.

The use of microbial inoculants containing plant growth-promoting rhizobacteria as a promoter of plant fitness and health is becoming increasingly popular in agriculture. However, whether and how these bacteria affect indigenous bacterial communities in field conditions is sparsely explored. We studied the effects of seed inoculation and field soil application of ubiquitous soil bacteria, B. cereus, B. subtilis, and B. amyloliquefaciens, on the diversity, evenness, and richness of endophytic bacterial communities in sprouting broccoli roots using high-throughput metagenome sequencing. The multiple operational taxonomic units (OTUs) assigned to different bacterial taxa clearly showed changes in ecological measures and relative abundances of certain taxa between control and treatment groups. The Bacillus inocula, themselves, failed to flourish as endophytes; however, the effects they extended on the endophytic bacterial community were both generic as well as species specific. In each case, Pseudomonadales, Rhizobiales, Xanthomonadales, and Burkholderiales were the most abundant orders in the endosphere. B. amyloliquefaciens drastically reduced the most abundant genus, Pseudomonas, while increasing the relative abundance of a range of minor taxa. The Shannon-Weiner diversity and Buzas and Gibson's evenness indices showed that the diversity and evenness were increased in both B. amyloliquefaciens and mixed treated plants. The UniFrac measurement of beta diversity showed that all treatments affected the specific composition of the endophytic bacterial community, with an apparent interspecies competition in the mixed treatment. Taken together, Bacillus species influenced the diversity, evenness, and composition of the endophytic bacterial community. However, these effects varied between different Bacillus spp. in a context-specific manner.

Developing Soil Microbial Inoculants for Pest Management: Can One Have Too Much of a Good Thing?

Soil microbes present a novel and cost-effective method of increasing plant resistance to insect pests and thus create a sustainable opportunity to reduce current pesticide application. However, the use of microbes in integrated pest management programs is still in its infancy. This can be attributed primarily to the variations in microbial inoculum performance under laboratory and field conditions. Soil inoculants containing single, indigenous microbial species have shown promising results in increasing chemical defenses of plants against foliar feeding insects. Conversely, commercial inoculants containing multiple species tend to show no effects on herbivore infestation in the field. We present here a simple model that endeavours to explain how single and multiple species in microbial inoculants differentially govern insect population dynamics via changes in plant chemical profiles. We discuss further how this knowledge can be applied to manipulate soil microbial species and develop 'tailored' microbial inoculants that could be used in plant protection against antagonists.

Infection by a foliar endophyte elicits novel arabidopside-based plant defence reactions in its host, Cirsium arvense.

Endophytic fungi live asymptomatically within plants. They are usually regarded as nonpathogenic or even mutualistic, but whether plants respond antagonistically to their presence remains unclear, particularly in the little-studied associations between endophytes and nongraminoid herbaceous plants. We investigated the effects of the endophyte Chaetomium cochlioides on leaf chemistry in Cirsium arvense. Plants were sprayed with spores; leaf material from both subsequent new growth and the sprayed leaves was analysed 2 wk later. Infection frequency was 91% and 63% for sprayed and new growth, respectively, indicating that C. cochlioides rapidly infects new foliage. Metabolomic analyses revealed marked changes in leaf chemistry with infection, especially in new growth. Changes in several novel oxylipin metabolites were detected, including arabidopsides reported here for the first time in a plant species other than Arabidopsis thaliana, and a jasmonate-containing galactolipid. The production of these metabolites in response to endophyte presence, particularly in newly infected foliage, suggests that endophytes elicit similar chemical responses in plants to those usually produced following wounding, herbivory and pathogen invasion. Whether endophytes benefit their hosts may depend on a complex series of chemically mediated interactions between the plant, the endophyte, other microbial colonists and natural enemies.

Fungi in a changing world: growth rates will be elevated, but spore production may decrease in future climates.

Very little is known about the impact of climate change on fungi and especially on spore production. Fungal spores can be allergenic, thus being important for human health. The aim of this study was to investigate how climate change influences the responsive ability of fungi by simulating differing environmental regimes. Fungal species with high spore allergenic potential and atmospheric abundance were grown and experimentally examined under a variety of temperatures and different nutrient availability. Each represented the average decadal air temperature of the 1980s, 1990s and 2000s in the UK, along with an Intergovernmental Panel on Climate Change (IPCC) climate change scenario for 2100. All tests were run on six fungal species: Alternaria alternata, Aspergillus niger, Botrytis cinerea, Cladosporium cladosporioides, Cladosporium oxysporum and Epicoccum purpurascens. Mycelium growth rate and spore production were examined on each single species and competitive capacity among species combinations in pairs. All fungal species grew faster at higher temperatures, and this was more pronounced for the temperature projection in 2100. Most species grew faster when there was lower nutrient availability. Exceptions were the species with the highest growth rate (E. purpurascens) and with the highest competition capacity (A. alternata). Most species (except for E. purpurascens) produced more spores in the richer nutrient medium but fewer as temperature increased. C. cladosporioides was an exception, exponentially increasing its spore production in the temperature of the 2100 scenario. Regarding competitive capacity, no species displayed any significant alterations within the environmental range checked. It is suggested that in future climates, fungi will display dramatic growth responses, with faster mycelium growth and lower spore production, with questions risen on relevant allergen potential.

Novel pheromone-mediated reproductive behaviour in the stag beetle, Lucanus cervus.

The iconic European stag beetle (Lucanus cervus) (Coleoptera: Lucanidae) is one of the largest terrestrial beetles in Europe. Due to decreasing population numbers, thought to be a consequence of habitat loss, this beetle has become a near-threatened species across much of Europe, and a reliable monitoring system is required to measure its future population trends. As part of a programme aimed at conserving UK populations, we have investigated the chemical ecology of the beetle, with a view to developing an efficient semiochemical-based monitoring system. Such a scheme will be beneficial not only in the UK but across the European range of the species, where the beetle is of conservation concern. Here, we report on a surprising discovery of a male-produced pheromone, which provokes initial sexual receptivity in females, and which has not been previously identified in the animal kingdom. Furthermore, we assign sex pheromone function to a previously described female-specific compound.

The Invasive Plant Impatiens glandulifera Manipulates Microbial Associates of Competing Native Species.

Impatiens glandulifera or Himalayan balsam is one of the most invasive weeds across Europe and can seriously reduce native plant diversity. It often forms continuous monocultures along river banks, but the mechanisms of this arrested succession are largely unknown. Here, we investigated the effect of arbuscular mycorrhizal (AM) fungi on balsam competitive ability with two native plant species, Plantago lanceolata and Holcus lanatus. We also studied how competition with Impatiens affects colonisation by foliar endophytes and mycorrhizas of two other co-occurring native species, Urtica dioica and Cirsium arvense. Mycorrhizal colonisation reduced balsam growth when the plants were grown singly, but appeared to have little effect when balsam experienced intra- or interspecific competition. Competition with balsam together with the addition of mycorrhizas had no effect on P. lanceolata biomass, suggesting that the fungi were beneficial to the latter, enabling it to compete effectively with balsam. However, this was not so with H. lanatus. Meanwhile, competition with Impatiens reduced endophyte numbers and mycorrhizal colonisation in U. dioica and C. arvense, leading to enhanced susceptibility of these plants to insect attack. Himalayan balsam is known to degrade soil fungal populations and can also reduce foliar beneficial fungi in neighbouring plants. This allows the plant to compete effectively with itself and other native species, thereby leading to the continuous monocultures.

Addition of Arbuscular Mycorrhizal Fungi Enhances Terpene Synthase Expression in Salvia rosmarinus Cultivars.

Culinary herbs are commercially cultivated for their wide range of volatile compounds that give characteristic aromas and tastes. Rosemary (Salvia rosmarinus Spenn.) is an excellent model for assessment of methods improvement of volatile production as cultivars offer a wide variety of aromatic profiles due to the large family of terpene synthase genes. Arbuscular mycorrhizal fungi (AMF) associations have been shown to improve essential oil production in aromatic plants and offer one approach to enhance aroma in commercial herb production. Changes in the expression of seven different terpene synthases were compared in six rosemary cultivars in response to addition of AMF to a peat substrate. Addition of AMF profoundly influenced terpene synthase expression in all cultivars and did so without impacting the optimised plant size and uniformity achieved in these conditions. In addition, two methods for AMF application, developed with the horticultural industry in mind, were tested in this study. Uniform incorporation of AMF mixed into the growing substrate prior to planting of a root plug produced the most consistent root colonisation. Overall, our findings demonstrate the potential for the use of AMF in the improvement of aroma in culinary herbs within a commercial setting but show that outcomes are likely to greatly vary depending on variety.

Differential effects of foliar endophytic fungi on insect herbivores attacking a herbaceous plant.

Foliar endophytic fungi appear to be ubiquitous in nature, occurring in a very wide range of herbaceous plants. However, their ecological role within forbs is very poorly known and interactions with foliar-feeding insects virtually unexplored. In this study, leaves of Cirsium arvense were infected with different combinations of endophyte fungi that had been previously isolated from this plant species. Two months later, leaf material was fed to larvae of a generalist insect, Mamestra brassicae, and adults of a specialist feeder, Cassida rubiginosa. Endophytes had different effects on the two insects; one species, Chaetomium cochliodes, reduced growth of M. brassicae but increased feeding by C. rubiginosa. Another species, Cladosporium cladosporioides, increased beetle feeding also, but had no effect on M. brassicae. Interactions were also seen between fungal species and dual infection with C. cladosporioides and Trichoderma viride greatly reduced beetle feeding. It is concluded that endophytes have significant effects on foliar feeding insects that differ with degree of specialism of the herbivore. We suggest that these effects are due to chemical changes in the host, brought about by fungal infection. These fungi have received remarkably little attention in the study of insect-plant interactions and yet could be important determinants of insect growth and even population dynamics.

Underground networking: the potential for improving yield and quality of pot-grown herbs with mycorrhizas.

With constant pressure on herb growers to perform to a continuous high standard, finding new ways to improve herb quality and or quantity are gaining importance, with arbuscular mycorrhizal fungi (AMF) presenting one possible solution. Viviane Schroeder, Alan Gange and Anthony Stead discuss the introduction of AMF to the herb growth cycle and discuss the benefits and costs that their symbiosis with plants bring to modern agriculture.

Aphids and Mycorrhizal Fungi Shape Maternal Effects in Senecio vulgaris.

Plant performance in any one generation is affected not only by the prevailing environmental conditions, but also by the conditions experienced by the parental generation of those plants. These maternal effects have been recorded in a many plant species, but the influence of external biotic (as opposed to abiotic) factors on shaping maternal effects have been rarely examined. Furthermore, almost all previous studies have taken place over one plant generation, rather than across multiple generations. Here, we studied the influence of insect herbivory and arbuscular mycorrhizal (AM) fungal colonisation on the shaping of maternal effects in the annual forb Senecio vulgaris. We grew plants with and without aphids (Myzus persicae) and AM fungi (hereafter termed 'induction events') over four successive generations, wherein seeds from plants in any one treatment were used to grow plants of the same treatment in the next generation, all in identical environmental conditions. We found strong evidence of maternal effects in the second plant generation, i.e., after one induction event. These plants took longer to germinate, flowered in a shorter time, produced lighter seeds and were shorter and of lower biomass than their parents. Aphid attack tended to enhance these effects, whereas AM fungi had little influence. However, thereafter there was a gradual recovery in these parameters, so that plants experiencing three inductions showed similar life history parameters to those in the original generation. We conclude that experiments investigating maternal effects need to be performed over multiple plant generations and that biotic factors such as insects and mycorrhizas must also be taken into account, along with abiotic factors, such as nutrient and water availability.

Effect of weather conditions, substrate pH, biochar amendment and plant species on two plant growth-promoting microbes on vegetated roofs and facades.

Background: Vegetated building envelopes (VBEs), such as vegetated roofs and facades, are becoming more frequent in urban planning nowadays. However, harsh growing conditions restrain the application of VBEs. Plant growth-promoting microbes (PGPMs) might help ease the stresses, but first, it is necessary to investigate how to ensure their survival and growth under VBE conditions. Methods: We conducted three experiments to test the impact of various factors on the microbial populations of inoculated PGPMs in VBEs, a mycorrhizal fungus Rhizophagus irregularis and a bacterium Bacillus amyloliquefaciens. The first experiment was conducted by inoculating the two PGPMs separately in Sedum roof plots, and the microbial populations associated with Poa alpina was monitored for two consecutive years under local weather conditions. The second experiment was conducted in a laboratory testing the effect of substrate pH (substrates collected from balcony gardens) on R. irregularis population associated with Trifolium repens and Viola tricolor. The third experiment was conducted on a meadow roof testing the effect of biochar amendment on R. irregularis population associated with Thymus serpyllum and Fragaria vesca. Results: In the first experiment, Bacillus was found to associate with P. alpina, but Rhizophagus wasn't. Yet, the fungus induced high Bacillus population density in the Rhizophagus treated plots in the first year. In the second experiment, Rhizophagus abundance in T. repens was higher in the neutral substrate (6-6.5), while V. tricolor was more colonized in acidic substrate (5-5.5), suggesting an important interactive effect of substrate pH and plant species on Rhizophagus abundance. The third experiment suggested a negligible impact of biochar amendment on Rhizophagus abundance for both host plants. Conclusion: Three experiments demonstrate that PGPM inoculation on VBEs is feasible, and various factors and interactions affect the PGPM populations. This paper provides reference and inspiration for other VBE research involving substrate microbial manipulation.

Apple endophyte community in relation to location, scion and rootstock genotypes and susceptibility to European canker.

European apple canker, caused by Neonectria ditissima, is a severe disease of apple. Achieving effective control is difficult with the currently available pesticides. Specific apple endophytes associated with cultivars may partially contribute to the cultivar response to the pathogen and thus could be used for disease management. We sought to determine whether the overall endophyte community differed among cultivars differing in their susceptibility to N. ditissima and to identify specific microbial groups associated with the susceptibility. Using Illumina MiSeq meta-barcoding, we profiled apple tree endophytes in 16 scion-rootstock combinations at two locations and quantified the relative contribution of scion, rootstock and location to the observed variability in the endophyte communities. Endophyte diversity was primarily affected by the orchard location (accounting for 29.4% and 85.9% of the total variation in the PC1 for bacteria and fungi, respectively), followed by the scion genotype (24.3% and 19.5% of PC2), whereas rootstock effects were small (<3% of PC1 and PC2). There were significant differences in the endophyte community between canker-resistant and -susceptible cultivars. Several bacterial and fungal endophyte groups had different relative abundance between susceptible and resistant cultivars. These endophyte groups included putative pathogen antagonists as well as plant pathogens. Their possible ecological roles in the N. ditissima pathosystem are discussed.

Climate change and spring-fruiting fungi.

Most macrofungi produce ephemeral fruit bodies during autumn but some have adapted to spring fruiting. In this study, temporal changes in the time of spring fruiting in Norway and the UK during 1960-2007 have been investigated by statistical analyses of about 6000 herbarium and field records, covering 34 species. Nearly 30 per cent of the temporal variation in fruiting could be ascribed to spatial and species-specific effects. Correcting for these effects, linear trends towards progressively earlier fruiting were detected during the entire period in both Norway and the UK, with a change in average fruiting day of 18 days over the study period. Early fruiting was correlated with high winter temperatures in both countries, indicating that the observed phenological changes are likely due to earlier onset of spring. There were also significant correlations between climatic conditions in one year and timing of fruiting the following year, indicating that below-ground mycelia are influenced by climatic conditions over a longer time period before fruiting. Fruiting dates were, however, not strictly related to changes in vernal accumulated thermal time. Our results indicate that global warming has lead to progressively earlier fruiting of spring fungi in northwest Europe during the last half century.

Effects of mycorrhizal fungi on insect herbivores: a meta-analysis.

Mycorrhizal status of the host plant is often ignored in studies on plant-herbivore interactions, but mycorrhizal colonization is known to induce many morphological, physiological, and biochemical changes in host plants, which in turn may alter plant quality as a host for insect herbivores. Both positive and negative effects of mycorrhizal colonization of the host plant on performance and density of insect herbivores have been reported in previous studies. We have conducted a meta-analysis of 34 published and unpublished studies on this topic in order to find out the sources of variation in mycorrhizae effects on insect herbivores. Effects of mycorrhizae on chewing insects depended upon the parameter measured and the degree of herbivore feeding specialization. Density and consumption of chewing insects were higher on mycorrhizal plants, but this did not lead to greater plant damage, presumably because herbivore survival tended to be lower on mycorrhizal plants. Mono- and oligophagous chewers benefited from mycorrhizal colonization of their host plants, whereas performance of polyphagous chewers was reduced on mycorrhizal plants. Among sucking insects, phloem feeders benefited from mycorrhizal infection, but performance of mesophyll feeders was lower on mycorrhizal plants. The type of mycorrhiza was not important for chewing insects, but performance of sucking insects was increased more by arbuscular mycorrhizal fungi (AM) than by ectomycorrhizae (ECM). Among AM inoculation studies, the most commonly used fungal species, Glomus intraradices, tended to have a negative effect on chewer performance, whereas all other fungal species tended to have a positive effect. There was no significant difference in results between studies using inoculation and fungicides, field and laboratory studies, and published and unpublished studies. Mycorrhizal status of the host plant thus influences insect herbivore performance, but the magnitude and direction of the effect depend upon the feeding mode and diet breadth of the insect and the identity of fungi.

Endophytic actinomycetes from Azadirachta indica A. Juss.: isolation, diversity, and anti-microbial activity.

Endophytic actinomycetes from Azadirachta indica A. Juss. were screened and evaluated for their anti-microbial activity against an array of pathogenic fungi and bacteria. A total of 55 separate isolates were obtained from 20 plants, and 60% of these showed inhibitory activity against one or more pathogenic fungi and bacteria. Actinomycetes were most commonly recovered from roots (54.5% of all isolates), followed by stems (23.6%), and leaves (21.8%). The dominant genus was Streptomyces (49.09% of all isolates), while Streptosporangium (14.5%), Microbispora (10.9%), Streptoverticillium (5.5%), Sacchromonospora sp. (5.5%), and Nocardia (3.6%) were also recovered. Streptomyces isolates AzR 006, 011, and 031 (all from roots) had acute activity against Pseudomonas fluorescens, while AzR027, 032, and 051 (also all from roots) showed activity against Escherichia coli. Meanwhile, an isolate of Nocardia sp. from leaves (AzL025) showed antagonism against Bacillus subtilis. Overall, 32 of the 55 were found to have broad spectrum significant antimicrobial activity, while about 4% of them showed strong and acute inhibition to pathogenic fungi and bacteria. Isolates of Streptomyces AzR031, 008, and 047, Nocardia sp. AzL025, and Streptosporangium sp. AzR 021 and 048 are of particular interest because they showed significant antagonistic activity against root pathogens, including Pythium and Phytophthora sp. Thus, many of the isolates recovered from A. indica in this study may be used in developing potential bio-control agents against a range of pathogenic fungi and bacteria and in the production of novel natural antimicrobial compounds. These results not only further our understanding of plant-microbe interactions but also indicate that there is an untapped resource of endophytic microorganisms that could be exploited in the biotechnological, medicinal, and agricultural industries.