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A trade‐off between growth and defence against biotic stresses is common in plants. Fungal endophytes of the genus Epichloë may relieve this trade‐off in their host grasses since they can simultaneously induce plant growth and produce antiherbivore alkaloids that circumvent the need for host defence. The Epichloë ability to decouple the growth‐defence trade‐off was evaluated by subjecting ryegrass with and without Epichloë endophytes to an exogenous treatment with gibberellin (GA) followed by a challenge with Rhopalosiphum padi aphids. In agreement with the endophyte‐mediated trade‐off decoupling hypothesis, the GA‐derived promotion of plant growth increased the susceptibility to aphids in endophyte‐free plants but did not affect the insect resistance in endophyte‐symbiotic plants. In line with the unaltered insect resistance, the GA treatment did not reduce the concentration of Epichloë‐derived alkaloids. The Epichloë mycelial biomass was transiently increased by the GA treatment but at the expense of hyphal integrity. The response of the phyllosphere bacterial microbiota to both GA treatment and Epichloë was also evaluated. Only Epichloë, and not the GA treatment, altered the composition of the phyllosphere microbiota and the abundance of certain bacterial taxa. Our findings clearly demonstrate that Epichloë does indeed relieve the plant growth‐defence trade‐off. Summary statement: A debate has been raised if fungal endophytes can relieve the plant growth‐defence trade‐off. Epichloë endophytes decoupled the trade‐off: the growth‐mediated penalisation of plant defences was observed in E− but not in E+ plants. The Epichloë production of alkaloids was likely the decoupling mechanism. [ABSTRACT FROM AUTHOR]

Context: Data are lacking on the effects of selected endophytes of perennial ryegrass (Lolium perenne L.) on ryegrass persistence. Aim: We aimed to determine the impact of Epichloë endophyte on the persistence of mid- and late-heading perennial ryegrass cultivars. Method: Two mid-heading (Samson, Bronsyn) and two late-heading (One50, Rohan) cultivars, infected with selected endophytes (AR37, nea2/6) or with standard toxic endophyte, were established in a replicated plot study grazed by cattle in a subtropical environment of the upper North Island of New Zealand. Persistence characteristics were quantified at least five times per annum, over 4 years. Key results: Endophyte strain had an effect on persistence; infection with standard endophyte resulted in higher ryegrass ground cover percentage, ryegrass content in pasture dry matter and autumn yield than infection with nea2/6 on many occasions, and with AR37 on some occasions. There were negligible impacts on ryegrass tiller density or nutritive value. Trends were dominated by the main effect of endophyte; interactions with heading date were inconsistent. Conclusions: Cultivars were more persistent when infected with standard endophyte than with selected endophytes, although persistence declined over 4 years for all cultivars and irrespective of heading date. Implications: Reliance on selected endophyte is unlikely to prevent persistence decline of perennial ryegrass in a subtropical environment. Other strategies will be required to maintain the persistence of high-quality pastures based on perennial ryegrass. We determined the impact of endophyte strain (standard toxic endophyte, selected strains nea2/6 and AR37), on the persistence of four perennial ryegrass cultivars. Standard endophyte resulted in greater persistence than the selected endophytes, with higher ryegrass ground cover percentage and content in pasture dry matter by the end of the study, and higher herbage yields in autumn. Persistence declined over 4 years for all cultivars, regardless of endophyte. [ABSTRACT FROM AUTHOR]

Context: Data are lacking on the effects of heading date of perennial ryegrass (Lolium perenne L.) on ryegrass persistence. Aim: We aimed to determine the impact of heading date on perennial ryegrass persistence. Method: Two mid-heading (Samson, Bronsyn) and two late-heading (One50, Rohan) cultivars, infected with Epichloë endophytes (AR37, nea2/6 or standard toxic), were established in a replicated plot study grazed by cattle in a subtropical environment of the upper North Island of New Zealand. Persistence characteristics were quantified at least five times per annum, over 4 years. Key results: Late-heading cultivars had higher yield, nutritive value, perennial ryegrass content in pasture dry matter, ground cover and tiller density than mid-heading cultivars. There were large seasonal impacts on all ryegrass characteristics, with a major decline over summer and recovery during late autumn into early spring, with less recovery in the final year. Overall, there was a strong linear decline in ryegrass content measured during spring (2018–21). Conclusions: The two late-heading cultivars were more persistent than the two mid-heading cultivars, although persistence declined over 4 years for all cultivars. Implications: Results support industry recommendations of choosing late-heading cultivars for improved pasture quality and persistence. We determined the impact of heading date on persistence of perennial ryegrass cultivars infected with a range of endophytes, in a plot study grazed by cattle. The two late-heading cultivars were more persistent than the two mid-heading cultivars, with higher yield, nutritive value, perennial ryegrass content in pasture dry matter, ground cover and tiller density, although persistence declined over 4 years for all cultivars. [ABSTRACT FROM AUTHOR]

Both adults and larvae of Listronotus bonariensis damage ryegrass (Lolium spp.), a dominant component of New Zealand pasture. Damage can be reduced by Epichloë festucae var. lolii, a biotrophic fungal symbiont of ryegrass, through the production of alkaloids with deterrent and/or toxic effects to insect herbivores including L. bonariensis. Here we report results from two pot trials and a field trial comparing endophyte strains with different alkaloid profiles that have been introduced to reduce or eliminate adverse effects on stock. Adult L. bonariensis feeding scars, number of eggs and the proportion of tillers with larval damage and the severity of that damage were recorded. Samples of pseudostem and leaf blades were analysed for alkaloids. AR1 endophyte reduced adult feeding, oviposition, and larval damage whereas AR37 had no effect on adults but was highly effective in reducing larval damage. Reduction in adult feeding and oviposition by different NEA strains was governed by concentrations of peramine and ergovaline. For plants infected with AR37, alkaloid concentrations that determined the extent of larval damage were influenced by host species and ploidy; tetraploid L. perenne was more severely damaged than its diploid equivalent; L. multiflorum cultivars were more susceptible than the L. hybridum and L. perenne cultivars. [ABSTRACT FROM AUTHOR]

Many temperate grasses are both hyper‐accumulators of silicon (Si) and hosts of Epichloë fungal endophytes, functional traits which may alleviate environmental stresses such as herbivore attack. Si accumulation and endophyte infection may operate synergistically, but this has not been tested in a field setting, nor in the context of changing environmental conditions. Predicted increases in atmospheric CO2 concentrations can affect both Si accumulation and endophyte function, but these have not been studied in combination.We investigated how elevated atmospheric CO2 (eCO2), Si supplementation, endophyte‐presence and insect herbivory impacted plant growth, stoichiometry (C, N, P and Si), leaf gas exchange (rates of photosynthesis, stomatal conductance, transpiration rates) and endophyte production of anti‐herbivore defences (alkaloids) of an important pasture grass (tall fescue; Lolium arundinaceum) in the field.eCO2 and Si supplementation increased shoot biomass (+52% and +31%, respectively), whereas herbivory reduced shoot biomass by at least 35% and induced Si accumulation by 24%. Shoot Si concentrations, in contrast, decreased by 17%–21% under eCO2. Si supplementation and herbivory reduced shoot C concentrations. eCO2 reduced shoot N concentrations which led to increased shoot C:N ratios. Overall, shoot P concentrations were 26% lower in endophytic plants compared to non‐endophytic plants, potentially due to decreased mass flow (i.e. observed reductions in stomatal conductance and transpiration). Alkaloid production was not discernibly affected by any experimental treatment. The negative impacts of endophytes on P uptake were particularly strong under eCO2.We show that eCO2 and insect herbivory reduce and promote Si accumulation, respectively, incorporating some field conditions for the first time. This indicates that these drivers operate in a more realistic ecological context than previously demonstrated. Reduced uptake of P in endophytic plants may adversely affect plant productivity in the future, particularly if increased demand for P due to improved plant growth under eCO2 cannot be met. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

Aphids are major pests of cereal and pasture grasses throughout the world, vectoring disease and reducing plant production. There are few control options other than insecticides. Epichloë endophytes that produce loline alkaloids in their hosts provide a possible mechanism of control, with both meadow fescue and tall fescue naturally infected with loline-producing endophytes showing a resistance to Rhopalosiphum padi. We screened Elymus spp. naturally infected with endophytes that produced loline alkaloids at concentrations known to affect aphids on fescue but found no effect on these insects infesting Elymus. A synthetic loline-producing endophyte association with rye also had no effect on the aphids. After hypothesizing that the lolines were being translocated in the xylem in Elymus and rye rather than the phloem, we tested the rye and meadow fescue infected with loline-producing endophytes against a xylem feeding spittlebug. The endophyte in rye inhibited the feeding of the insect and reduced its survival, whereas the endophyte-infected meadow fescue had no effect on the spittlebug but reduced the number of aphids. Lolines applied to the potting medium of endophyte-free and endophyte-infected rye, ryegrass, and tall fescue resulted in a decrease in the aphid populations on the endophyte-free pasture grasses relative to the untreated controls but had no effect on aphid numbers on the rye. We tentatively conclude that lolines, produced in both natural and synthetic association with Elymus and rye, are partitioned in the xylem rather than the phloem, where they are inaccessible to aphids. [ABSTRACT FROM AUTHOR]

Genotyping by sequencing (GBS) was used to reveal the inherent genetic variation within the haploid fungi Sarocladium zeae isolated from diverse Zea germplasm, including modern Zea mays and its wild progenitors—the teosintes. In accordance with broad host relationship parameters, GBS analysis revealed significant host lineages of S. zeae genetic diversity, indicating that S. zeae genetic variation may associate with different evolutionary histories of host species or varieties. Based on a recently identified PKS-NRPS gene responsible for pyrrocidine biosynthesis in S. zeae fungi, a novel PCR assay was developed to discriminate pyrrocidine-producing S. zeae strains. This molecular method for screening bioactive strains of S. zeae is complementary to other approaches, such as chemical analyses. An eGFP-labelled S. zeae strain was also developed to investigate the endophytic transmission of S. zeae in Z. mays seedlings, which has further improved our understanding of the transmission modes of S. zeae endophytes in maize tissues. [ABSTRACT FROM AUTHOR]

In identifying endophytes for use in pastures in New Zealand there have been two strategies used, either exclusion of ergopeptine and lolitrem alkaloids or, retaining some ergopeptine expression to enhance the stability of natural defences against invertebrate pests. Both have their strengths and weaknesses. It is a matter of balancing these to ensure the end-user has access to a product that satisfies their needs – a persistent pasture with low to nil animal toxicity in terms of animal production and welfare. The range and intensity of ergot alkaloids in grazed pasture on both pasture and ruminants is reviewed, with emphasis on New Zealand. Ergot alkaloids associated with pasture are produced by associations between certain Epichloë endophyte strains and temperate grasses. Ergot alkaloids have been shown to improve persistence of pasture through providing resistance/deterrence to insect pests as well as deterring grazing animals. However, ergovaline is toxic to grazing animals. Some commercially available ryegrass-endophyte associations can produce ergovaline concentrations close to those found in associations between ryegrass and the standard endophyte. It is feasible to eliminate ergot alkaloids from pasture grasses in New Zealand as endophyte strains are commercially available that do not express ergovaline and yet still provide excellent pest resistance. [ABSTRACT FROM AUTHOR]

Grasses have developed a wide range of morphological and physiological mechanisms to resist herbivory. For instance, they accumulate silicon (Si) in tissue, as physical defence, and associate symbiotically with foliar Epichloë‐endophytes that provide chemical defence via antiherbivore alkaloids. Recent evidence showed that some Epichloë‐endophytes increase foliar Si in forage grasses; however, whether this impacts insect herbivores is unknown. Furthermore, while Si is primarily a physical defence, it also affects production of plant defensive secondary metabolites; Si supply might therefore affect Epichloë‐alkaloids, although this remains untested.We grew endophyte‐free (Nil) and Epichloë‐infected tall fescue and perennial ryegrass in a factorial combination with or without Si supplementation, in the absence or presence of Helicoverpa armigera. Epichloë‐endophyte strains were AR584 for tall fescue, and AR37, AR1 or Wild‐type (WT) for perennial ryegrass. We assessed how Si supply and Epichloë‐endophytes in interaction with herbivory affected foliar Si and mutualist‐derived alkaloid concentrations. Subsequently, their effects on H. armigera relative growth rates (RGRs) were evaluated.Endophytes generally increased Si concentrations in Si‐supplied plants. In tall fescue AR584 and perennial ryegrass AR37, endophytes increased constitutive (herbivore‐free) and induced (herbivore‐inoculated) Si concentrations by at least 25%; in contrast, in perennial ryegrass, the AR1 endophyte only increased constitutive levels. Si supply did not affect alkaloids produced by AR584 or AR1/WT endophytes; however, in the presence of herbivory, Si supply decreased the induction of alkaloids produced by AR37 endophytes by 33%. For tall fescue, Si supply reduced H. armigera RGR by at least 76%, regardless of endophytic status, whereas, endophyte‐alkaloids played a secondary role only reducing herbivore growth in the absence of Si supply. Conversely, in perennial ryegrass, both Si and endophyte‐alkaloids (regardless of Si supply) reduced herbivore RGR although not synergised.Novel interactions between constitutive and induced Si‐ and alkaloid‐based antiherbivore defences in grasses were observed. Overall, Si had a greater effect on the folivore than endophytes in both grasses. Endophyte defences contributed more to herbivore resistance in perennial ryegrass than tall fescue. We demonstrate that Si and endophytes were not antagonistic and highlight that the protective nature of their interaction varies with the grass‐endophyte species tested. A free Plain Language Summary can be found within the Supporting Information of this article. [ABSTRACT FROM AUTHOR]

The aphid Aploneura lentisci is widespread in Australia and New Zealand, living all year round on roots of its secondary grass hosts. The fungal endophyte (Epichloë festucae var. lolii), strain AR37 in Lolium perenne is known to greatly reduce populations and was a likely reason for the superior growth and persistence of this association previously observed in the field. Aphid populations were quantified in a field trial near Ballarat, comparing yields of perennial ryegrass infected with eight different endophyte strains and an endophyte-free (Nil) control in a common ryegrass background (Grasslands Samson (G. Samson)). AR37 and another endophyte strain, AR5, had significantly fewer aphids than all other endophytes. These differences were significantly related to yield increases taken before and after sampling that persisted until the end of the trial. In a pot trial comparing commercially available ryegrass-endophyte combinations with equivalent Nil controls, aphid numbers were lower on G. Samson AR37 and Banquet II with AR5 (Endo®5) than on all other cultivar-endophyte combinations. Compared with Nil controls, the common toxic strain in G. Samson, and two strains in Trojan also reduced aphid numbers. The AR5 endophyte produces the alkaloid ergovaline but high concentrations of this in roots of potted plants could not account for differences in root aphid numbers. Root concentrations of epoxyjanthitrems, the only known alkaloids produced by AR37, were low and unlikely to be the cause of resistance to A. lentisci. An aphid, Aploneura lentisci , that feeds on grass roots is widespread in pastures in New Zealand and Australia. This paper reports data confirming that perennial ryegrass infected with a strain of Epichloë fungal endophyte, AR37, is highly resistant to this aphid and shows that another endophyte strain, AR5, gives the same level of control. Suppression of high aphid populations by these endophytes in the field at Ballarat resulted in a 37% increase in pasture production over endophyte-free ryegrass over a 5-month period. [ABSTRACT FROM AUTHOR]

In a field study during two growing seasons in northern Sweden, we assessed meadow fescue (Festuca pratensis Huds.) with and without Neotyphodium uncinatum to determine whether the endophyte affects plant performance regarding tiller and biomass production. In addition, we measured loline alkaloids produced in infected grasses. Approximately 1000 plants grew at two different levels of nitrogen fertilization: the normal amount applied in this region and one-half that amount. Increasing the amount of nitrogen increased the total number of tillers on the plants. Endophyte-infected plants produced more tillers per plant than plants without the endophyte; there was, however, no significant difference in proportion of reproductive tillers, winter survival, or total biomass production between endophyte-infected and noninfected plants. Infected plants produced loline alkaloids in different quantities without indication of being influenced by different nitrogen fertilization levels. Key words: endophyte, forage, nitrogen fertilization. Nous avons evalue la fetuque des pres (Festuca pratensis Huds.) dans une etude sur le terrain realisee au cours de deux saisons de croissance dans le nord de la Suede, en presence ou en absence de Neotyphodium uncinatum, afin de determiner si 'endophyte affecte la performance de la plante sur le plan de la production de talles et de biomasse. Nous avons de plus mesure es alcaloides loline produits par l'herbe infectee. Pres d'un millier de plantes ont pousse sous deux conditions de fertilisation a 'azote : la quantite normale appliquee dans cette region et la moitie de cette quantite. L'augmentation de la quantite d'azote ccroissait le nombre total de talles sur les plantes. Les plantes infectes par l'endophyte produisaient davantage de talles par lante que les plantes sans endophyte; cette difference n'etait toutefois pas significative en proportion des talles reproductives, e la survie a l'hiver ou de la biomasse totale entre les plantes infectes et non infectes. Les plantes infectes produisaient des lcaloides loline en quantites differentes, sans indication que cela ait ete influence par les differents niveaux de fertilisation a 'azote. [Traduit par la Redaction] Mots-cles: endophyte, forage, fertilisation a l'azote., Introduction Meadow fescue (Festuca pratensis Huds.) is a perennial pooid grass that grows widely in Northern Europe and parts of Asia. In the Nordic and Baltic countries, it is an [...]

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. Mother plant exposure to ozone did not affect the growth and the transmission of fungal endophytes into host seeds. Seed alkaloid concentration correlated with endophyte mycelial concentration and host fitness. However, ozone‐induced stress on mother plants did impair the longevity of the endophytes in the seeds. [ABSTRACT FROM AUTHOR]

Background. Plants possess a sophisticated immune system to defend from herbivores. These defence responses are regulated by plant hormones including salicylic acid (SA) and jasmonic acid (JA). Sometimes, plant defences can be complemented by the presence of symbiotic microorganisms. A remarkable example of this are grasses establishing symbiotic associations with Epichloë fungal endophytes. We studied the level of resistance provided by the grass' defence hormones, and that provided by Epichloë fungal endophytes, against an introduced herbivore aphid. These fungi protect their hosts against herbivores by producing bioactive alkaloids. We hypothesized that either the presence of fungal endophytes or the induction of the plant salicylic acid (SA) defence pathway would enhance the level of resistance of the grass to the aphid. Methods. Lolium multiflorum plants, with and without the fungal endophyte Epichloë occultans, were subjected to an exogenous application of SA followed by a challenge with the aphid, Sipha maydis. Results. Our results indicate that neither the presence of E. occultans nor the induction of the plant's SA pathway regulate S. maydis populations. However, endophytesymbiotic plants may have been more tolerant to the aphid feeding because these plants produced more aboveground biomass. We suggest that this insect insensitivity could be explained by a combination between the ineffectiveness of the specific alkaloids produced by E. occultans in controlling S. maydis aphids and the capacity of this herbivore to deal with hormone-dependent defences of L. multiflorum. [ABSTRACT FROM AUTHOR]

Aims The leaves of almost all terrestrial plant species are colonized by endophytic fungi. Compared to agronomic grasses, which usually harbor few endophytes, native grasses generally possess greater endophyte species diversity. Existing studies examining endophyte effects on natural grasses under competition normally considered the infection status (infected or uninfected), and rarely considered endophyte species. Methods We examined the effects of endophyte infection and of endophyte species on the interspecific competitive ability of a subdominant species, Achnatherum sibiricum , at two nitrogen levels (high nitrogen and low nitrogen). Achnatherum sibiricum plants infected by two different species of endophyte (Epichloë sibirica and E. gansuensis) and uninfected plants were grown in monoculture and binary mixtures with a dominant species, Stipa grandis (six individuals per species for monocultures and three + three individuals of each species in mixtures). Shoot and root biomass, tiller number and total phenolic concentration were measured after 3 months. Moreover, the aggressivity index was calculated to compare the competitive ability of A. sibiricum relative to S. grandis. Important Findings Both E. gansuensis (Eg)- and E. sibirica (Es)-infected A. sibiricum plants showed a greater competitive ability than the uninfected plants under high nitrogen supply, while the opposite result occurred under low nitrogen supply. At high nitrogen levels, Eg plants had a higher tiller number and a greater shoot biomass inhibitory effect on S. grandis than Es plants had when growing in mixture, while Es plants showed better root growth performance than Eg and uninfected plants under mixture conditions at all nitrogen levels. A higher concentration of phenolic compounds in Eg plants than in Es plants might contribute to the higher inhibitory effect of Eg plants on competing plants. Our study indicates that the interaction between endophyte infection and nitrogen availability can alter the competitive ability of the host plant A. sibiricum but that these two endophyte species work in different ways, which may influence the coexistence of A. sibiricum with the dominant species. [ABSTRACT FROM AUTHOR]

Annual ryegrass (Lolium rigidum Gaud.) is a troublesome invasive plant in southern Australian cropping systems and is typically infected with a fungal endophyte, Epichloë occultans. Potential links between herbicide resistance and endophyte infection were investigated in this study. We surveyed 391 ryegrass populations from geographically distinct, naturalised pasture and cropping areas across southern Australia and compared frequencies of seed-borne endophyte infection in the collected seed samples from mature annual ryegrass. Data on herbicide resistance from similar seedlots were cross-referenced with endophyte infection frequency to examine the potential relationship between herbicide resistance and endophyte infection. Seeds from cropping paddocks exhibited a significantly higher frequency of endophyte infection than pasture paddocks sampled from the same region. Frequencies of endophyte infection in annual ryegrass declined across Australia during 2001--09, whereas the opposite trend was observed during 2009-12. Impacts of the Australian Millennium Drought, the correspondingly altered rainfall patterns and environmental selection pressures have a role in determining observed endophyte infection frequencies. However, there was no significant correlation between endophyte frequency and herbicide resistance in the populations evaluated. Differences in endophyte infection frequencies were associated with farming practices, and require further examination to determine causative selection pressures encountered in Australian field settings. [ABSTRACT FROM AUTHOR]

Abstract: The plant hormone salicylic acid (SA) is recognized as an effective defence against biotrophic pathogens, but its role as regulator of beneficial plant symbionts has received little attention. We studied the relationship between the SA hormone and leaf fungal endophytes on herbivore defences in symbiotic grasses. We hypothesize that the SA exposure suppresses the endophyte reducing the fungal‐produced alkaloids. Because of the role that alkaloids play in anti‐herbivore defences, any reduction in their production should make host plants more susceptible to herbivores. Lolium multiflorum plants symbiotic and nonsymbiotic with the endophyte Epichloë occultans were exposed to SA followed by a challenge with the aphid Rhopalosiphum padi. We measured the level of plant resistance to aphids, and the defences conferred by endophytes and host plants. Symbiotic plants had lower concentrations of SA than did the nonsymbiotic counterparts. Consistent with our prediction, the hormonal treatment reduced the concentration of loline alkaloids (i.e., N‐formyllolines and N‐acetylnorlolines) and consequently decreased the endophyte‐conferred resistance against aphids. Our study highlights the importance of the interaction between the plant immune system and endophytes for the stability of the defensive mutualism. Our results indicate that the SA plays a critical role in regulating the endophyte‐conferred resistance against herbivores. [ABSTRACT FROM AUTHOR]

Earthworms are not a direct pest of turf grass but they are considered a problem on many sports fields, disrupting playability and aesthetics due to the castings they deposit on the playing surface. Also, a number of slug species are well-known foliage destroying pests of a number of agriculturally important crop species. Perennial ryegrass and tall fescue cultivars associated with selected Epichloë endophytes, originally developed for bird management at airports, were assessed to determine their deterrent properties towards worms and slugs. Plots sown with endophyte-free ryegrass had significantly higher numbers of worms and slugs than plots containing the same grass cultivar infected with the endophyte strain AR95. Also, plots sown with endophyte-free tall fescue had significantly higher numbers of slugs (but not worms) than plots containing the same grass cultivar infected with the endophyte strain AR601. Although more research is required on the exact mechanism of action, these results suggest selected novel endophyte-infected grass associations, such as those including the endophyte AR95, may substantially reduce populations of worms and slugs in areas where these grasses are sown. [ABSTRACT FROM AUTHOR]

Context. Small mammalian herbivores, such as European rabbits (Oryctolagus cuniculus), can have significant unwanted impacts on agriculture and horticulture and can attract birds of prey and avian scavengers to airports, increasing the risk of bird strike. Sustainable wildlife management tools that have high efficacy and animal welfare are needed to mitigate these impacts. Aim. We assessed perennial ryegrass (Lolium perenne) and tall fescue (Festuca arundinacea) cultivars associated with selected Epichloë endophytes, originally developed for bird management at airports, to determine their feeding-deterrent properties towards invasive rabbits. Methods. A pellet feed trial using caged domestic rabbits and a pen trial using wild rabbits were conducted to assess rabbit responses to food pellets with novel endophyte-infected or uninfected grass seed and to plots planted with endophyte-infected or uninfected grasses, respectively. Key results. Caged rabbits ate significantly less food pellets containing endophyte-infected grass straw than pellets containing endophyte-free grass straw. Wild rabbits consumed significantly less herbage from plots planted with endophyte-infected grasses than those of the equivalent endophyte-free grass cultivars. Temporal patterns of rabbit feeding behaviour support the hypothesis that deterrence in rabbits resulted from post-ingestion feedback associated with grass-endophyte secondary metabolites. Conclusions. Although more research is required on the exact mechanism of action in rabbits, our results suggest novel endophyte-infected grass associations may substantially reduce rabbit feeding behaviour and possibly rabbit numbers in areas where these grasses are sown. Implications. Novel grass-endophyte associations have great potential for deterring problem wildlife at airports and other amenity areas. [ABSTRACT FROM AUTHOR]

To improve Neotyphodium endophyte-mediated resistance to black cutworm Agrotis ipsilon (Hufnagel) (BCW), a series of experiments was conducted by using several different cultivars of tall fescue, Schedonorus arundinaceus (Schreb.) Dumort. in combination with several different haplotypes of the endophyte Neotyphodium coenophialum (Morgan-Jones & Cams) (plant cultivar X endophyte haplotype = plant line), each producing unique alkaloid profiles. BCW settling response, survival at 5 and 10 d, and larval biomass varied significantly among plant lines. In general, greater variation BCW performance was observed within a single plant cultivar infected with different endophyte haplotypes than among different plant cultivars infected with the same endophyte haplotype, but comparisons among the former were far more numerous. Although five endophytemediated alkaloids representing three alkaloid classes were quantified in the plants, the pyrrolizidine alkaloid N-acetyl norloline was consistently the single best predictor of BCW performance. BCW settling response, 5-d survival, and 10-d survival decreased as levels of the alkaloid N-acetyl norloline increased. The same three response variables also decreased with increasing levels of peramine, but increased with increasing levels of ergovaline. Minor variation in endophyte infection levels occurring among infected plant lines bad no significant influence on BCW performance. Results indicate a potentially important role for N-acetyl norloline and peramine in providing resistance to black cutworm whereas ergovaline appears to be much less important. Therefore, endophyte haplotypes expressing high levels of N-acetyl norloline and peramine may be of particular importance for developing 'friendly' endophyte-enhanced turf and pasture grasses that resist challenging lepidopteran pests, although remaining safe for wildlife and grazing mammals. [ABSTRACT FROM AUTHOR]

Abstract: [1-Phenyl-2-[(E)-3-phenylprop-2-en-1-oyl-κO]ethenyl-κC 1]tetracarbonylmanganese (1a) reacts with PhCCH in CCl4 at room temperature to form [2,4-diphenyl-6-(2-phenylethenyl)pyranyl-η5]tricarbonylmanganese (2a), whose X-ray crystal structure is reported to complement that of its isomer [6-oxo-2,4,7-triphenylcyclohepta-1,4-dienyl-1,2,3,4,5-η]tricarbonylmanganese (3a), previously obtained from the reaction under reflux; but for 1a and PhCCPh the pyranyl complex cannot be isolated before rearrangement to the 3a analogue occurs. More forcing reaction conditions for 1a with Me3SiCCH and for [1-(2-trifluoromethylphenyl)-2-[(E)-3-(2-trifluoromethylphenyl)prop-2-en-1-oyl-κO]ethenyl-κC 1]tetracarbonylmanganese (1b) with Me3SiCCH and PhCCH give new analogues of 3a where previously only 2a analogues had been isolated. The reaction in CCl4 under reflux of PhCCH and the β-deuterio analogue of 1a, [1-phenyl-2-[(E)-3-phenylprop-2-en-1-oyl-3d-κO]ethenyl-κC 1]tetracarbonylmanganese, gave deuteriated 3a with exo-D at the α-carbon, C7. This is inconsistent with the Mn-mediated Ph migration mechanism originally proposed to accommodate the endo position of Ph in 3a, and instead it implicates a cyclopropyl carbonyl-addition intermediate or a cyclopropyl acyl-substitution transition state in the key rearrangement step for 2a → 3a. [Copyright &y& Elsevier]

Reaction of cyclomanganated aryl ketones with CS2 proceeds with insertion into the Mn–C bond to give η2-dithiocarboxylato–Mn(CO)4 compounds. With other cyclomanganated substrates such as that from Ph3P&z.dbnd6;S and also with Mn2(CO)10, CS2 gives (μ3-CS3)2Mn4(CO)16 with bridging trithiocarbonate ligands. [Copyright &y& Elsevier]

The root-feeding scarab insect Costelytra giveni causes severe damage to pasture ecosystems in New Zealand. Loline alkaloids produced by some Epichloë endophytes deter this insect. In two experiments, tall fescue infected with E. coenophiala, strain AR584, and endophyte-free (Nil) controls were subjected to pulse drought stress (DS) or well-watered conditions (WW). The second experiment also included meadow fescue infected with E. uncinata. After 4–6 weeks exposure to the different conditions, roots were excised and fed to C. giveni larvae for 7 days. Relative root consumption (RC), frass production, and relative weight change (RWC) of larvae were measured and the loline content of roots determined. RC and frass output were higher for larvae feeding on Nil DS tall fescue than WW and reduced by AR584. RWC was also greater on DS than on WW Nil plants but reduced by endophyte only in DS plants. RC, frass output, and RWC of larvae were reduced by endophyte in DS and WW meadow fescue, but the effect was greater for WW plants. Loline alkaloid concentration in roots was significantly higher in DS than WW tall fescue in Experiment I but higher in WW than DS meadow fescue in Experiment II. These experiments have demonstrated that moisture status interacts with endophyte to differentially affect root herbivory in tall fescue and meadow fescue. [ABSTRACT FROM AUTHOR]

Epichloë festucae var. lolii and Epichloë sp. Lp TG-3 are filamentous fungal endophytes of perennial ryegrass ( Lolium perenne ) that have a substantial impact on New Zealand's agricultural economy by conferring biotic advantages to the host grass. Overall, Epichloë endophytes contribute NZ$200 million to the economy annually, with strain AR37 estimated to contribute NZ$3.6 billion to the New Zealand economy over a 20-year period. This strain produces secondary metabolites, including epoxyjanthitrems, which are a class of indole diterpenes, associated with the observed effects of AR37 on livestock and insect pests. Until very recently, AR37 was intractable to genetic modification but this has changed with the application of CRISPR-Cas9 based gene editing techniques. In this paper, gene inactivation by CRISPR-Cas9 was used to deconvolute the genetic basis for epoxyjanthitrem biosynthesis, including creating an AR37 strain that has been edited to remove the biosynthesis of all indole diterpenes. We show that gene editing of Epichloë can be achieved without off-target events or introduction of foreign DNA (footprint-less) through an AMA1-based plasmid that simultaneously expresses the CRISPR-Cas9 system and selectable marker. Genetic modification events in these transformants were investigated through genome sequencing and in planta chemistry., Competing Interests: The authors declare that this study received funding from Grasslanz Technology Ltd and PGG Wrightson Seeds Ltd. The funders were not involved in the study design, collection, analysis, interpretation of dat or the writing of this article. They were however involved in the decision to submit it for publication., (Copyright © 2022 Miller, Hudson, Johnson, Singh, Mace, Forester, Maclean, Voisey and Johnson.)

This study investigated effects of seed aging and imbibition on sugar metabolite concentrations in Epichloë endophyte-infected and endophyte-free seed of tall fescue ( Festuca arundinacea Schreb.). Two treatments, namely, accelerated aging and imbibition, were applied to the seeds, with embryo and endosperm tissues analyzed separately. Gas chromatography with flame ionization detection was employed for analysis of sugar metabolites within the seed tissues. Mannitol, ribitol, and trehalose were more abundant in embryo than endosperm tissues and were identified at consistently higher concentrations within endophyte-infected compared to endophyte-free seeds. The ratio of raffinose to sucrose significantly increased with seed aging in both endophyte-free and endophyte-infected embryo tissues, while significantly lower concentrations of trehalose were detected in tissues dissected from aged-seed regardless of endophyte status. This research provides fundamental insight into the metabolic details of endophyte survival in seed and provides a first evaluation of key carbohydrates present in the fungal-plant symbiosis.

Epichloë fungal endophytes of cool season grasses are well-known to produce a range of alkaloids of benefit to the host. Some of these compounds are advantageous to agriculture due to qualities that promote pasture persistence (e.g., the loline class of alkaloids confer insect protection) while others are detrimental to the well-being of grazing livestock. The ergot alkaloids (e.g., ergovaline), produced in ryegrass and tall fescue associations, causes poor animal health in farming regions in many countries around the world and further study is required to improve our knowledge on this class of compounds. Here we present the application of a quantitative LC-MS/MS (liquid chromatography coupled to mass spectrometry) method measuring eight ergot alkaloids (chanoclavine, agroclavine, elymoclavine, lysergol, lysergic acid, ergine, lysergyl-alanine, ergovaline) produced by endophyte infected grasses, to monitor levels in individual tillers from multiple plants of a single cultivar of perennial ryegrass (Lolium perenne cv. "Grasslands Samson") infected with a common toxic endophyte strain (Epichloë festucae var. lolii). Monitoring the expression in individual tillers allows an estimation of the variability within a plant (between tillers) as well as between plants. The study showed that there is significant variation in the concentration of the ergot alkaloids between tillers of a single plant, at or exceeding the level of variation observed between individual plants of a population. This result emphasizes the fundamental importance of robust experimental design and sampling procedures when alkaloid expression assessment is required and these need to be rigorously tailored to the hypothesis being tested.