Search

Alzheimer's disease is linked with diabetes and cancer, emphasising the need for effective treatments. Plantago lanceolata , recognised as safe by various pharmacopeias, was investigated in this study for therapeutic potential. We examined the effects of its leaf extracts and sub-extracts (methanol, hexane, dichloromethane, ethyl acetate, butanol, aqueous) on AChE, BChE, α-amylase, α-glucosidase enzymes, as well as their impact on HDF-a and U87-MG cancer cells. The phytochemical characterisation was performed using ICP-MS and LC-MS/MS. Cytotoxic effects were evaluated on HDF-a and U87-MG cell lines, along with assessments for nuclear abnormalities. Na and K were detected in extracts, with isoleucine and cyanidin-3-O-glucoside being the most concentrated compounds. Extracts at concentrations exceeding 25 µg/mL significantly increased cytotoxicity in HDF-a cell lines compared to the control group, without inducing nuclear abnormalities. Methanol extract demonstrated moderate inhibition against AChE and BChE at concentrations of 100 µg/mL and 500 µg/mL, respectively. These findings suggest that extracts exhibit potential therapeutic effects.

Rubisco, the most prevalent protein on Earth, catalysers both a reaction that initiates C 3 carbon fixation, and a reaction that initiates photorespiration, which stimulates protein synthesis. Regulation of the balance between these reactions under atmospheric CO 2 fluctuations remains poorly understood. We have hypothesised that vascular plants maintain organic carbon-to-nitrogen homoeostasis by adjusting the relative activities of magnesium and manganese in chloroplasts to balance carbon fixation and nitrate assimilation rates. The following examined the influence of magnesium and manganese on carboxylation and oxygenation for rubisco purified from two ecotypes of Plantago lanceolata L.: one adapted to the elevated CO 2 atmospheres that occur near a natural CO 2 spring and the other adapted to more typical CO 2 atmospheres that occur nearby. The plastid DNA coding for the large unit of rubisco was similar in both ecotypes. The kinetics of rubiscos from the two ecotypes differed more when associated with manganese than magnesium. Specificity for CO 2 over O 2 (S c/o ) for rubisco from both ecotypes was higher when the enzymes were bound to magnesium than manganese. Differences in the responses of rubisco from P. lanceolata to the metals may account for the adaptation of this species to different CO 2 environments., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Enzymes capable of processing a variety of compounds enable plants to adapt to diverse environmental conditions. PRISEs (progesterone-5β-reductase/iridoid synthase-like enzymes), examples of such substrate-promiscuous enzymes, are involved in iridoid and cardenolide pathways and demonstrate notable substrate promiscuity by reducing the activated C=C double bonds of plant-borne and exogenous 1,4-enones. In this study, we identified PRISE genes in Plantago media ( PmdP5βR1 ) and Plantago lanceolata ( PlP5βR1 ), and the corresponding enzymes were determined to share a sequence identity of 95%. Despite the high sequence identity, recombinant expressed Pmd P5βR1 was 70 times more efficient than Pl P5βR1 for converting progesterone. In order to investigate the underlying reasons for this significant discrepancy, we focused on specific residues located near the substrate-binding pocket and adjacent to the conserved phenylalanine "clamp". This clamp describes two phenylalanines influencing substrate preferences by facilitating the binding of smaller substrates, such as 2-cyclohexen-1-one, while hindering larger ones, such as progesterone. Using structural analysis based on templates PDB ID: 5MLH and 6GSD from PRISE of Plantago major , along with in silico docking, we identified positions 156 and 346 as hot spots. In Pl P5βR1 amino acid residues, A156 and F346 seem to be responsible for the diminished ability to reduce progesterone. Moreover, the double mutant Pl P5βR_F156L_A346L, which contains the corresponding amino acids from Pmd P5βR1, showed a 15-fold increase in progesterone 5β-reduction. Notably, this modification did not significantly alter the enzyme's ability to convert other substrates, such as 8-oxogeranial, 2-cyclohexen-1-one, and methyl vinyl ketone. Hence, a rational enzyme design by reducing the number of hotspots selectively, specifically improved the substrate preference of Pl P5βR1 for progesterone.

The aim of this trial was to assess the effectiveness of Sediflù, a medical device containing active herbal ingredients, on nocturnal and diurnal persistent coughs in children, with a duration of 3 to 7 days. Children with a dry and/or productive cough were enrolled in this prospective, interventional, multicenter, placebo-controlled, double-blind, randomized clinical study. Clinical efficacy was assessed through the evaluation of the soothing action of Sediflù against dry and/or productive coughing, both at night and during the day, and other effects of coughing associated with quality of sleep: frequency, child's quality of sleep, parental quality of sleep and severity. Treatment with Sediflù improved both night-time and day-time cough scores from day 2. The diurnal score also improved significantly in the Sediflù group at days 3 and 7. Sediflù syrup can be considered a valid treatment for cough management in younger children with upper respiratory tract infections, shortening the cough duration., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: CC-R and ER have been scientific advisors for Cinfa and speakers outside the objective of this work. CC-R has worked as principal investigator in clinical studies promotes by Cinfa. CN and MCC are principal investigator in this trial. The other author declares that they have no competing interest.

The aim of current experiment was to determine the effect of replacement of alfalfa hay with ribwort plantain (Plantago lanceolata) hay in ruminant diets on the fermentation parameters such as gas production, methane (CH 4 ) production, true digestible dry matter (TDDM), true digestibility (TD), partitioning factor, microbial protein, and efficiency of microbial protein using in vitro gas production technique. The alfalfa hay was replaced with P. lanceolata hay in a diets isocaloric (2650 kcal/kg DM) and nitrogenic (17% CP kg DM) at the ratio of 0, 5, 10 and 15%. Partial substitution of alfalfa hay with P. lanceolata hay had no significant effect on gas and methane (ml/incubated substrate or %) production whereas the partial substitution had a significant effect on TDDM, TD, gas (ml/digested DM), CH 4 (ml ml/digested DM) and microbial MP of diets. The replacement of alfalfa hay with ribwort plantain hay shifted the fermentation pattern from gas and methane production to microbial protein production. Therefore alfalfa hay can be replaced with ribwort plantain hay with high digestibility and anti-methanogenic potential in ruminant diets up to 15% to decrease methane production and improve microbial protein production. However further in vivo experiments are required to determine the effect of replacement on feed intake and animal production., (© 2024. The Author(s).)

Pathogens play a key role in insect population dynamics, contributing to short-term fluctuations in abundance as well as long-term demographic trends. Two key factors that influence the effects of entomopathogens on herbivorous insect populations are modes of pathogen transmission and larval host plants. In this study, we examined tritrophic interactions between a sequestering specialist lepidopteran, Euphydryas phaeton, and a viral pathogen, Junonia coenia densovirus, on its native host plant, Chelone glabra, and a novel host plant, Plantago lanceolata, to explore whether host plant mediates viral transmission, survival, and viral loads. A two-factor factorial experiment was conducted in the laboratory with natal larval clusters randomly assigned to either the native or novel host plant and crossed with either uninoculated controls or viral inoculation (20% of individuals in the cluster inoculated). Diapausing clusters were overwintered in the laboratory and checked weekly for mortality. At the end of diapause, all surviving individuals were reared to adulthood to estimate survivorship. All individuals were screened to quantify viral loads, and estimate horizontal transmission postmortem. To test for vertical transmission, adults were mated, and the progeny were screened for viral presence. Within virus-treated groups, we found evidence for both horizontal and vertical transmission. Larval clusters reared on the native host plant had slightly higher horizontal transmission. Survival probability was lower in clusters feeding on the native host plant, with inoculated groups reared on the native host plant experiencing complete mortality. Viral loads did not differ by the host plant, although viral loads decreased with increased sequestration of secondary compounds on both host plants. Our results indicate that the use of a novel host plant may confer fitness benefits in terms of survival and reduced viral transmission when larvae feeding on it are infected with this pathogen, supporting hypotheses of potential evolutionary advantages of a host range expansion in the context of tritrophic interactions., (© 2024 The Ecological Society of America.)

Host plant consumption and pathogen infection commonly influence insect traits related to development and immunity, which are ultimately reflected in the behavior and physiology of the insect. Herein, we explored changes in the metabolome of a generalist insect herbivore, Vanessa cardui (Lepidoptera: Nymphalidae), in response to both dietary variation and pathogen infection in order to gain insight into tritrophic interactions for insect metabolism and immunity. Caterpillars were reared on two different host plants, Plantago lanceolata (Plantaginaceae) and Taraxacum officinale (Asteraceae) and subjected to a viral infection by Junonia coenia densovirus (JcDV), along with assays to determine the insect immune response and development. Richness and diversity of plant and caterpillar metabolites were evaluated using a liquid chromatography-mass spectrometry approach and showed that viral infection induced changes to the chemical content of V. cardui hemolymph and frass dependent upon host plant consumption. Overall, the immune response as measured by phenoloxidase (PO) enzymatic activity was higher in individuals feeding on P. lanceolata compared with those feeding on T. officinale. Additionally, infection with JcDV caused suppression of PO activity, which was not host plant dependent. We conclude that viral infection combined with host plant consumption creates a unique chemical environment, particularly within the insect hemolymph. Whether and how these metabolites contribute to defense against viral infection is an open question in chemical ecology., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Seed endophytic bacteria are beneficial to plants. They improve seedling growth by enhancing plant nutrient uptake, modulating stress-related phytohormone production, and targeting pests and pathogens with antibiotics. Seed endophyte composition can be influenced by pollination, plant cultivar, and soil physicochemical conditions. However, the effects of plant community richness on seed endophytes are unknown. To investigate the effects of increasing plant species richness on the diversity and composition of the seed microbiome, we made use of a well-established long-term biodiversity experiment in Germany (The Jena Experiment). We sampled seeds from different Plantago lanceolata blossoms in a plant diversity gradient ranging from monoculture to 16 species mixtures. The seeds were surface sterilized to remove seed surface-associated bacteria and subjected to a metabarcoding approach to assess bacterial community structure., Results: Our data indicate a very stable core microbiome, which accounted for more than 90% of the reads and was present in all seeds independent of the plant richness level the seeds originated from. It consisted mainly of reads linked to Pseudomonas rhizosphaerae, Sphingomonas faeni and Pirellulla spp. 9% of the obtained reads were not part of the core microbiome and were only present in plots of specific diversity levels. The number of unique ASVs was positively correlated with plant richness. Interestingly, most reads described as non-core members belonged to the same genera described as the core microbiome, indicating the presence of different strains or species with possibly different functional properties important for seed performance., Conclusion: Our data indicate that Plantago lanceolata maintains a large seeds core microbiome across the plant richness gradient. However, the number of unique ASVs increases alongside the plant community richness, indicating that ecosystem biodiversity also mitigates diversity loss in seed endophytes., (© 2024. The Author(s).)

Pollen production is one plant characteristic that is considered to be altered by changes in environmental conditions. In this study, we investigated pollen production of the three anemophilous species Betula pendula, Plantago lanceolata, and Dactylis glomerata along an urbanization gradient in Ingolstadt, Germany. We compared pollen production with the potential influencing factors urbanization, air temperature, and the air pollutants nitrogen dioxide (NO 2 ) and ozone (O 3 ). While we measured air temperature in the field, we computed concentration levels of NO 2 and O 3 from a land use regression model. The results showed that average pollen production (in million pollen grains) was 1.2 ± 1.0 per catkin of Betula pendula, 5.0 ± 2.4 per inflorescence of Plantago lanceolata, and 0.7 ± 0.5 per spikelet of Dactylis glomerata. Pollen production was higher in rural compared to urban locations on average for B. pendula (+ 73%) and P. lanceolata (+ 31%), while the opposite was the case for D. glomerata (- 14%). We found that there was substantial heterogeneity across the three species with respect to the association of pollen production and environmental influences. Pollen production decreased for all species with increasing temperature and urbanization, while for increasing pollutant concentrations, decreases were observed for B. pendula, P. lanceolata, and increases for D. glomerata. Additionally, pollen production was found to be highly variable across species and within species-even at small spatial distances. Experiments should be conducted to further explore plant responses to altering environmental conditions., (© 2023. The Author(s).)

Six different furanocoumarins were isolated from the aerial parts of Ducrosia anethifolia and tested in vitro for plant cell elongation in etiolated wheat coleoptile. They were also tested for their ability to control three different weeds: ribwort plantain, annual ryegrass, and common purslane. These compounds exhibited strong inhibition of plant cell elongation. In the case of (+)-heraclenin, the IC 50 was lower than 20 μM, indicating a better inhibition than the positive control Logran®. Computational experiments for docking and molecular dynamics revealed for the investigated furanocoumarins bearing an epoxide moiety an improved fitting and stronger interaction with the auxin-like TIR1 ubiquitin ligase. Furthermore, the formed inhibition complex remained also stable during dynamic evaluation. Bidental interaction at the active site, along with an extended hydrogen-bond lifetime, explained the enhanced activity of the epoxides. The in vitro weed bioassay results showed that Plantago lanceolata was the most affected weed for germination, root, and shoot development. In addition, (+)-heraclenin displayed better inhibition values than positive control even at 300 μM concentration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Multiparasite communities inhabiting individual hosts are common and often consist of parasites from multiple taxa. The effects of parasite community composition and complexity on host fitness are critical for understanding how host-parasite coevolution is affected by parasite diversity. To test how naturally occurring parasites affect host fitness of multiple host genotypes, we performed a common-garden experiment where we inoculated four genotypes of host plant Plantago lanceolata with six microbial parasite treatments: three single-parasite treatments, a fungal mixture, a viral mixture, and a cross-kingdom treatment. Seed production was affected by both host genotype and parasite treatment, and their interaction jointly determined the growth of the hosts. Fungal parasites had more consistent negative effects than viruses in both single- and mixed-parasite treatments. These results demonstrate that parasite communities have the potential to affect the evolution and ecology of host populations through their effects on host growth and reproduction. Moreover, the results highlight the importance of accounting for the diversity of parasites as well as host genotypes when aiming to predict the consequences of parasites for epidemics as the effects of multiparasitism are not necessarily additive of single-parasite effects, nor uniform across all host genotypes., (© The Author(s) 2023. Published by Oxford University Press on behalf of The Society for the Study of Evolution (SSE).)

Viruses are a vastly underestimated component of biodiversity that occur as diverse communities across hierarchical scales from the landscape level to individual hosts. The integration of community ecology with disease biology is a powerful, novel approach that can yield unprecedented insights into the abiotic and biotic drivers of pathogen community assembly. Here, we sampled wild plant populations to characterize and analyze the diversity and co-occurrence structure of within-host virus communities and their predictors. Our results show that these virus communities are characterized by diverse, non-random coinfections. Using a novel graphical network modeling framework, we demonstrate how environmental heterogeneity influences the network of virus taxa and how the virus co-occurrence patterns can be attributed to non-random, direct statistical virus-virus associations. Moreover, we show that environmental heterogeneity changed virus association networks, especially through their indirect effects. Our results highlight a previously underestimated mechanism of how environmental variability can influence disease risks by changing associations between viruses that are conditional on their environment., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Secondary plant chemistry mediates a variety of communication signals among species, playing a fundamental role in the evolutionary diversification of communities and ecosystems. Herein, we explored diet-mediated host plant effects on development and immune response of a generalist insect herbivore. Vanessa cardui (Nymphalidae) caterpillars were reared on leaves of three host plants that vary in secondary metabolites, Plantago lanceolata (Plantaginaceae), Taraxacum officinale (Asteraceae) and Tithonia diversifolia (Asteraceae). Insect development was evaluated by larval and pupal viabilities, survivorship, and development rate. Immune response was measured as phenoloxidase (PO) activity. Additionally, chemical profiles of the host plants were obtained by liquid chromatograph-mass spectrometry (LC-MS) and the discriminant metabolites were determined using a metabolomic approach. Caterpillars reared on P. lanceolata exhibited the highest larval and pupal viabilities, as well as PO activity, and P. lanceolata leaves were chemically characterized by the presence of iridoid glycosides, phenylpropanoids and flavonoids. Taraxacum officinale leaves were characterized mainly by the presence of phenylpropanoids, flavones O-glycoside and germacranolide-type sesquiterpene lactones; caterpillars reared on this host plant fully developed to the adult stage, however they exhibited lower larval and pupal viabilities compared to individuals reared on P. lanceolata. Conversely, caterpillars reared on T. diversifolia leaves, which contain phenylpropanoids, flavones and diverse furanoheliangolide-type sesquiterpene lactones, were not able to complete larval development and exhibited the lowest PO activity. These findings suggested that V. cardui have adapted to tolerate potentially toxic metabolites occurring in P. lanceolata (iridoid glycosides), however caterpillars were not able to cope with potentially detrimental metabolites occurring in T. diversifolia (furanoheliangolides). Therefore, we suggest that furanoheliangolide-type sesquiterpene lactones were responsible for the poor development and immune response observed for caterpillars reared on T. diversifolia., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The objective of this study was to determine the effect of grazing plantain-based pastures on urine volume, urination frequency, and urinary nitrogen (UN) concentration of dairy cows under a typical pastoral dairy practice offering approximately 25% supplemented feeds. The experiment was a completely randomised design with three pasture treatments (perennial ryegrass-white clover (RGWC); RGWC + low plantain rate (PLL); and RGWC + high plantain rate (PLH)), five replicate plots, and repeated in two sequential grazing periods. Forty-five lactating Friesian × Jersey cows were randomly assigned into three groups of 15 animals each to graze over six days in adaptation paddocks and three days in experimental plots. Urine flow sensors were used to measure urine volume and urinary frequency, while spot urine sampling was conducted to determine nitrogen (N) concentration in cow urine. The results showed that including 25% plantain in the diet (PLH) increased daily urine volume by 44% and the daily number of urinations by 28%, compared to grazing the RGWC pasture. In addition, N concentration in cow urine was decreased by 18 and 29% when the diet contained 18% and 25% plantain, respectively. In conclusion, under a typical dairy farm practice, incorporating plantain into the RGWC pasture with the proportion of 25% plantain in the diet can increase the number of urine patches and reduce the concentration of N in the urine, thereby providing the opportunity to decrease N leaching from pastoral systems.

Interactions among parasite species coinfecting the same host individual can have far reaching consequences for parasite ecology and evolution. How these within-host interactions affect epidemics may depend on two non-exclusive mechanisms: parasite growth and reproduction within hosts, and parasite transmission between hosts. Yet, how these two mechanisms operate under coinfection, and how sensitive they are to the composition of the coinfecting parasite community, remains poorly understood. Here, we test the hypothesis that the relationship between within- and between-host transmission of the fungal pathogen, Phomopsis subordinaria, is affected by co-occurring parasites infecting the host plant, Plantago lanceolata . We conducted a field experiment manipulating the parasite community of transmission source plants, then tracked P. subordinaria within-host transmission, as well as between-host transmission to naïve recipient plants. We find that coinfection with the powdery mildew pathogen, Podosphaera plantaginis , causes increased between-host transmission of P. subordinaria by affecting the number of infected flower stalks in the source plants, resulting from altered auto-infection. In contrast, coinfection with viruses did not have an effect on either within- or between-host transmission. We then analyzed data on the occurrence of P. subordinaria in 2018 and the powdery mildew in a multi-year survey data set from natural host populations to test whether the positive association predicted by our experimental results is evident in field epidemiological data. Consistent with our experimental findings, we observed a positive association in the occurrence of P. subordinaria and historical powdery mildew persistence. Jointly, our experimental and epidemiological results suggest that within- and between-host transmission of P. subordinaria depends on the identity of coinfecting parasites, with potentially far-reaching effects on disease dynamics and parasite co-occurrence patterns in wild populations., Supplementary Information: The online version contains supplementary material available at 10.1007/s10682-022-10182-9., Competing Interests: Conflict of interestThe authors declare no conflicts of interest., (© The Author(s) 2022.)

Ribwort plantain ( Plantago lanceolata L.), which belongs to the Plantaginaceae family, has been widely used as a herbal plant in traditional medicine across the globe. The present study aimed to investigate the biologically active substances of P. lanceolata root fractions, as well as the cytotoxic and antibacterial activities of extracts. The cytotoxic activity of ethyl acetate, dichloromethane, and n-butanol extracts of P. lanceolata root was evaluated by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. The P.lanceolata root extracts were also evaluated on gram-positive and negative bacteria by disc diffusion and microtiter broth dilution methods. The phytochemical content was also examined by gas chromatography-mass spectrometry. The P.lanceolata root extracts were cytotoxic; IC 50 values against HCT-116 at 72 h were 168.553 μg/mL, 167.458 μg/mL, and 205.004 μg/mL for ethyl acetate, dichloromethane, and n-butanol root extracts, respectively. The dichloromethane extract of P. lanceolata root had the highest inhibitory effect against S. paratyphi (14.00±1.0 mm) at the concentration of 100 mg/mL. The minimum MIC and MBC (5 and 15 mg/mL) were observed for dichloromethane extract of P. lanceolata root against S. paratyphi . The main composition of ethyl acetate extract was 1,2-Benzenedicarboxylic acid and mono(2-ethylhexyl) ester (60.93%). The major compositions in dichloromethane and n-butanol extracts were 1,2-Benzenedicarboxylic acid, mono(2-ethylhexyl) ester (60.64%) and 2-Methyl-1-butanol (.+/-.)- (17.85%). As evidenced by the results of the present research, P. lanceolata extracts are a significant source of bioactive metabolites. Therefore, they can play a prominent role in the production of pharmaceutical materials., Competing Interests: The authors declare that they have no conflict of interest.

While pathogenic and mutualistic microbes are ubiquitous across ecosystems and often co-occur within hosts, how they interact to determine patterns of disease in genetically diverse wild populations is unknown. To test whether microbial mutualists provide protection against pathogens, and whether this varies among host genotypes, we conducted a field experiment in three naturally occurring epidemics of a fungal pathogen, Podosphaera plantaginis, infecting a host plant, Plantago lanceolata, in the Åland Islands, Finland. In each population, we collected epidemiological data on experimental plants from six allopatric populations that had been inoculated with a mixture of mutualistic arbuscular mycorrhizal fungi or a nonmycorrhizal control. Inoculation with arbuscular mycorrhizal fungi increased growth in plants from every population, but also increased host infection rate. Mycorrhizal effects on disease severity varied among host genotypes and strengthened over time during the epidemic. Host genotypes that were more susceptible to the pathogen received stronger protective effects from inoculation. Our results show that arbuscular mycorrhizal fungi introduce both benefits and risks to host plants, and shift patterns of infection in host populations under pathogen attack. Understanding how mutualists alter host susceptibility to disease will be important for predicting infection outcomes in ecological communities and in agriculture., (© 2022 The Authors. New Phytologist © 2022 New Phytologist Foundation.)

This study aimed to elucidate the role of bacteria colonising mycorrhizal hyphae in organically bound sulfur mobilisation, the dominant soil sulfur source that is not directly plant available. The effect of an intact mycorrhizal symbiosis with access to stable isotope organo- 34 S enriched soils encased in 35 µm mesh cores was tested in microcosms with Agrostis stolonifera and Plantago lanceolata . Hyphae and associated soil were sampled from static mesh cores with mycorrhizal ingrowth and rotating mesh cores that exclude mycorrhizal ingrowth as well as corresponding rhizosphere soil, while plant shoots were analysed for 34 S uptake. Static cores increased uptake of 34 S at early stages of plant growth when sulfur demand appeared to be high and harboured significantly larger populations of sulfonate mobilising bacteria. Bacterial and fungal communities were significantly different in the hyphospheres of static cores when compared to rotating cores, not associated with plant hosts. Shifts in bacterial and fungal communities occurred not only in rotated cores but also in the rhizosphere. Arylsulfatase activity was significantly higher in the rhizosphere when cores stayed static, while atsA and asfA gene diversity was distinct in the microcosms with static and rotating cores. This study demonstrated that AM symbioses can promote organo-S mobilization and plant uptake through interactions with hyphospheric bacteria, enabling AM fungal ingrowth into static cores creating a positive feedback-loop, detectable in the microbial rhizosphere communities.

The main objective of this study was to investigate whether the most common wild plant species of urban grassland can be used for phytoremediation of soils polluted with heavy metals. The study was conducted in the city of Varaždin, in northern Croatia. The content of heavy metals (Cd, Cu, Fe, Mn, Ni, Pb, Zn) was determined in soil samples as well as in unwashed and washed plant samples (Taraxacum officinale, Plantago lanceolata, Trifolium repens). The results show that the most polluted site is the railway station, while most sites are polluted by road traffic. The soils are most enriched with Pb, Cu, Zn and Cd. The bioconcentration factors for all three plant species are <1, indicating the relatively low capacity of phytoextraction. A considerable amount of heavy metals is found in the dust deposited on the plant surface, which is confirmed by a statistically significant difference between washed and unwashed plant samples. In addition, the biomass of each plant species that can be removed (in t/ha year), the mass of specific heavy metal that can be removed (in kg/ha), and the years required for phytoremediation are reported. In conclusion, phytoremediation with only common plant species of urban grassland is not possible within a reasonable period of time.

This article assesses the use of under-vine living mulches in Mediterranean vineyards characterized by limited water resources, one of the reasons why this agronomic practice is currently unusual in these environments. The aim of the study was to test whether the use of this alternative method in Mediterranean vineyards could suppress noxious weeds without hindering optimal vineyard development. For this purpose, four native species were selected as living mulches: Festuca ovina , Pilosella officinarum , Plantago coronopus , and Plantago lanceolata . The variables measured during three years in two different experimental farms were: (a) living mulch cover, as a possible predictor of weed suppression success; (b) weed density and weed biomass, with special attention to noxious weed species; and (c) pruning weights, measured in the last year to analyze the cumulative effect of the treatments on the grapevine vegetative growth. Our results revealed that living mulches with high cover rates (average over 70%) also showed weed suppression of up to 95%, significantly controlling the occurrence of noxious weeds such as Erigeron canadensis . No significant effect of the different treatments on vine vegetative growth was found, although further studies would be necessary. Based on these findings, it can be concluded that under-vine living mulches could be an efficient and environmentally friendly method for weed control in Mediterranean vineyards where irrigation is available.

In this study, three types of extracts isolated from leaves of Plantain ( Plantago lanceolata ) were tested for their chemical content and biological activities. The three bioproducts are combinations of polysaccharides and polyphenols (flavonoids and iridoidic compounds), and they were tested for antioxidant, antifungal, antitumor, and prebiotic activity (particularly for polysaccharides fraction). Briefly, the iridoid-enriched fraction has revealed a pro-oxidant activity, while the flavonoid-enriched fraction had a high antioxidant potency; the polysaccharide fraction also indicated a pro-oxidant activity, explained by the co-presence of iridoid glycosides. All three bioproducts demonstrated moderate antifungal effects against Aspergillus sp., Penicillium sp., and dermatophytes, too. Studies in vitro proved inhibitory activity of the three fractions on the leukemic tumor cell line THP-1, the main mechanism being apoptosis stimulation, while the polysaccharide fraction indicated a clear prebiotic activity, in the concentration range between 1 and 1000 µg/mL, evaluated as higher than that of the reference products used, inulin and dextrose, respectively.

We know little about the underlying genetic control of phenotypic patterns of seed traits across large-scale geographic and environmental gradients. Such knowledge is important for understanding the evolution of populations within species and for improving species conservation. Therefore, to test for genetic variation in Plantago lanceolata , we made reciprocal crosses between northern and southern genotypes that span the species' range in Europe. The results provide evidence of transgenerational genetic effects on seed mass and germination timing. Northern mothers produced larger seeds with delayed germination, in contrast to southern mothers, which produced smaller seeds with accelerated germination. A maternal latitude affected both the seed coat, solely maternal tissue, and embryo/endosperm tissues. Thus, latitudinal variation in seed size and germination timing can be explained, in part, by the direct influence of maternal genotype, independent of zygotic genes that parents pass directly to the embryo and endosperm. Data suggest that researchers exploring the existence and evolution of large-scale geographic variation within species test for transgenerational genetic effects. In addition, data suggest that transgenerational control of seed traits should be considered when developing procedures designed to facilitate species conservation and restoration.

Biomonitors are considered a cheap alternative of active air samplers, especially where spatial pattern of air quality is to be monitored, requiring numerous parallel measurements. Of higher plants, Plantago lanceolata L. has been proven a good monitor species with proper accumulation capacity. While biomonitoring studies are difficult to compare due to inherent errors such as the diverse plant material used in different studies, the No. 227 OECD GUIDELINE FOR THE TESTING OF CHEMICALS: Terrestrial Plant Test: Vegetative Vigour Test provides a tool to test extract of aerosol samples under controlled laboratory conditions. In our study, this guideline was followed to experimentally treat Plantago with the aqueous extract of a diesel exhaust sample. Accumulation pattern of polyaromatic hydrocarbons (PAHs) was assessed and compared to samples collected in the field. Unlike most studies reported in the literature, both in the experimentally treated and field Plantago samples, high ratio of high molecular weight PAHs was experienced. Distribution pattern of accumulated PAHs showed strong correlation between the experimentally treated sample and most of the field plantain samples, underlying the usefulness of laboratory treatments for bioaccumulation studies., (© 2021. The Author(s).)

AbstractThe inherently variable nature of epidemics renders predictions of when and where infection is expected to occur challenging. Differences in pathogen strain composition, diversity, fitness, and spatial distribution are generally ignored in epidemiological modeling and are rarely studied in natural populations, yet they may be important drivers of epidemic trajectories. To examine how these factors are linked to epidemics in natural host populations, we collected epidemiological and genetic data from 15 populations of the powdery mildew fungus, Podosphaera plantaginis , on Plantago lanceolata in the Åland Islands, Finland. In each population, we tracked spatiotemporal disease progression throughout one epidemic season and coupled our survey of infection with intensive field sampling of the pathogen. We found that strain composition varied greatly among populations in the landscape. Within populations, strain composition was driven by the sequence of strain activity: early-active strains reached higher abundances, leading to consistent strain compositions over time. Co-occurring strains also varied in their contribution to the growth of the local epidemic, and these fitness inequalities were linked to epidemic dynamics: a higher proportion of hosts became infected in populations containing strains that were more similar in fitness. Epidemic trajectories in the populations were also linked to strain diversity and spatial dynamics: higher infection rates occurred in populations containing higher strain diversity, while spatially clustered epidemics experienced lower infection rates. Together, our results suggest that spatial and/or temporal variation in the strain composition, diversity, and fitness of pathogen populations are important factors generating variation in epidemiological trajectories among infected host populations.

AbstractA central challenge in ecology and evolutionary biology is to understand how variation in abiotic and biotic factors combine to shape the distribution, abundance, and diversity of focal species. Environmental gradients, whether natural (e.g., latitude, elevation, ocean proximity) or anthropogenic (e.g., land-use intensity, urbanization), provide compelling settings for addressing this challenge. However, not all organisms are amenable to the observational and experimental approaches required for untangling the factors that structure species along gradients. Here we highlight herbaceous plants in the genus Plantago as models for studying the ecology and evolution of species interactions along abiotic gradients. Plantago lanceolata and P. major are native to Europe and Asia but distributed globally, and they are established models for studying population ecology and interactions with herbivores, pathogens, and soil microbes. Studying restricted range congeners in comparison with those cosmopolitan species can provide insight into abiotic and biotic determinants of range size and population structure. We highlight one such species, P. rugelii , which is endemic to eastern North America. We give an overview of the literature on these focal Plantago species and explain why they are logical candidates for studies of species interactions across environmental gradients. Finally, we emphasize collaborative and community science approaches that can facilitate such research and note the amenability of Plantago for authentic research projects in science education.

Roots are at the core of plant water dynamics. Nonetheless, root morphology and functioning are not easily assessable without destructive approaches. Nuclear Magnetic Resonance (NMR), and particularly low-field NMR (LF-NMR), is an interesting noninvasive method to study water in plants, as measurements can be performed outdoors and independent of sample size. However, as far as we know, there are no reported studies dealing with the water dynamics in plant roots using LF-NMR. Thus, the aim of this study is to assess the feasibility of using LF-NMR to characterize root water status and water dynamics non-invasively. To achieve this goal, a proof-of-concept study was designed using well-controlled environmental conditions. NMR and ecophysiological measurements were performed continuously over one week on three herbaceous species grown in rhizotrons. The NMR parameters measured were either the total signal or the transverse relaxation time T 2 . We observed circadian variations of the total NMR signal in roots and in soil and of the root slow relaxing T 2 value. These results were consistent with ecophysiological measurements, especially with the variation of fluxes between daytime and nighttime. This study assessed the feasibility of using LF-NMR to evaluate root water status in herbaceous species.

When plants compete what influences that interaction? To answer this we measured belowground competition directly, as the simultaneous capture of soil ammonium and nitrate by co-existing herbaceous perennials, Dactylis glomerata and Plantago lanceolata, under the influence of: species identity; N uptake and biomass of focal and neighbour plants; location (benign lowland versus harsher upland site); N availability (low or high N fertilizer); N ion, ammonium or nitrate production (mineralisation) rate, and competition type (intra- or interspecific), as direct effects or pairwise interactions in linear models. We also measured biomass as an indirect proxy for competition. Only three factors influenced both competitive N uptake and biomass production: focal species identity, N ion and the interaction between N ion and neighbour N uptake. Location had little effect on N uptake but a strong influence on biomass production. N uptake increased linearly with biomass only in isolated plants. Our results support the view that measuring resource capture or biomass production tells you different things about how competitors interact with one another and their environment, and that biomass is a longer-term integrative proxy for the outcomes of multiple separate interactions-such as competition for N-occurring between plants., (© 2020 John Wiley & Sons Ltd.)

New plant pathogen invasions typified by cryptic disease symptoms or those appearing sporadically in time and patchily in space, might go largely unnoticed and not taken seriously by ecologists. We present evidence that the recent invasion of Pyrenopeziza plantaginis (Dermateaceae) into the Pacific Northwest USA, which causes foliar necrosis in the fall and winter on Plantago lanceolata (plantain), the primary (non-native) foodplant for six of the eight extant Taylor's checkerspot butterfly populations ( Euphydryas editha taylori , endangered species), has altered eco-evolutionary foodplant interactions to a degree that threatens butterfly populations with extinction. Patterns of butterfly, larval food plant, and P. plantaginis disease development suggested the ancestral relationship was a two-foodplant system, with perennial Castilleja spp. supporting oviposition and pre-diapause larvae, and the annual Collinsia parviflora supporting post-diapause larvae. Plantain, in the absence of P. plantaginis disease, provided larval food resources throughout all butterfly life stages and may explain plantain's initial adoption by Taylor's checkerspot. However, in the presence of severe P. plantaginis disease, plantain-dependent butterfly populations experience a six-week period in the winter where post-diapause larvae lack essential plantain resources. Only C. parviflora , which is rare and competitively inferior under present habitat conditions, can fulfill the post-diapause larval feeding requirements in the presence of severe P. plantaginis disease. However, a germination timing experiment suggested C. parviflora to be suitably timed for only Washington Taylor's checkerspot populations. The recent invasion by P. plantaginis appears to have rendered the ancestrally adaptive acquisition of plantain by Taylor's checkerspot an unreliable, maladaptive foodplant interaction.

In exploring the roles of phenotypic plasticity in the establishment and early evolution of invading species, little empirical attention has been given to the importance of correlational selection acting upon suites of functionally related plastic traits in nature. We illustrate how this lack of attention has limited our ability to evaluate plasticity's role during invasion and also, the costs and benefits of plasticity. We addressed these issues by transplanting clones of European-derived Plantago lanceolata L. genotypes into two temporally variable habitats in the species' introduced range in North America. Phenotypic selection analyses were performed for each habitat to estimate linear, quadratic, and correlational selection on phenotypic trait values and plasticities in the reproductive traits: flowering onset and spike and scape lengths. Also, we measured pairwise genetic correlations for our "colonists." Results showed that (a) correlational selection acted on trait plasticity after transplantation, (b) selection favored certain combinations of genetically correlated and uncorrelated trait values and plasticities, and (c) using signed, instead of absolute, values of plasticity in analyses facilitated the detection of correlational selection on trait value-plasticity combinations and their adaptive value. Based on our results, we urge future studies on species invasions to (a) measure correlational selection and (b) retain signed values of plasticity in order to better discriminate between adaptive and maladaptive plasticity., Competing Interests: The authors disclose no conflicts., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Maternal effects of pathogen infection on progeny development and disease resistance may be adaptive and have important consequences for population dynamics. However, these effects are often context-dependent and examples of adaptive transgenerational responses from perennials are scarce, although they may be a particularly important mechanism generating variation in the offspring of long-lived species.Here, we studied the effect of maternal infection of Plantago lanceolata by Podosphaera plantaginis , a fungal parasite, on the growth, flower production and resistance of the progeny of six maternal genotypes in nutrient-rich and nutrient-poor environments. For this purpose, we combined a common garden study with automated phenotyping measurements of early life stages, and an inoculation experiment.Our results show that the effects of infection on the mother plants transcend to impact their progeny. Although maternal infection decreased total leaf and flower production of the progeny by the end of the growing season, it accelerated early growth and enhanced resistance to the pathogen P. plantaginis .We also discovered that the effects of maternal infection affected progeny development and resistance through a three way-interaction between maternal genotype, maternal infection status and nutrient availability. Synthesis . Our results emphasize the importance of maternal effects mediated through genotypic and environmental factors in long-living perennials and suggest that maternal infection can create a layer of phenotypic diversity in resistance. These results may have important implications for both epidemiological and evolutionary dynamics of host-parasite interactions in the wild., (© 2020 The Authors. Journal of Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.)

Exposure to reactive oxygen species can easily result in serious diseases, such as hyperproliferative skin disorders or skin cancer. Herbal extracts are widely used as antioxidant sources in different compositions. The importance of antioxidant therapy in inflammatory conditions has increased. Innovative formulations can be used to improve the effects of these phytopharmacons. The bioactive compounds of Plantago lanceolata (PL) possess different effects, such as anti-inflammatory, antioxidant, and bactericidal pharmacological effects. The objective of this study was to formulate novel liquid crystal (LC) compositions to protect Plantago lanceolata extract from hydrolysis and to improve its effect. Since safety is an important aspect of pharmaceutical formulations, the biological properties of applied excipients and blends were evaluated using assorted in vitro methods on HaCaT cells. According to the antecedent toxicity screening evaluation, three surfactants were selected (Gelucire 44/14, Labrasol, and Lauroglycol 90) for the formulation. The dissolution rate of PL from the PL-LC systems was evaluated using a Franz diffusion chamber apparatus. The antioxidant properties of the PL-LC systems were evaluated with 2,2-diphenyl-1-picrylhydrazyl (DPPH) and malondialdehyde (MDA) assessments. Our results suggest that these compositions use a nontraditional, rapid-permeation pathway for the delivery of drugs, as the applied penetration enhancers reversibly alter the barrier properties of the outer stratum corneum. These excipients can be safe and highly tolerable thus, they could improve the patient's experience and promote adherence.

Hairy root induction in Plantago lanceolata was optimized to take advantage of transformed root cultures. The highest frequency of transformation was achieved using leaf explant, A4 strain, pre-cultivation of explant, 150 µM Acetosyringone, 5 min inoculation, half-strength Murashige and Skoog basal medium as co-cultivation, and half-strength Gamborg's basal medium as a selective medium with 3% sucrose. Among the studied compound encompassing gallic acid, catalpol and apigenin, only the production of gallic acid in hairy roots was affected by 20 mg L -1 AgNO 3 and 100 mg L -1 chitosan at 24 hr which yielded 7.63, 4.76-fold increase in its content, respectively. The methanolic extracts of hairy roots elicited by 20 mg L -1 AgNO 3 exhibited anti-bacterial activity (MIC and MBC = 25 mg mL -1 ) against Klebsiella pneumoniae , Proteus vulgaris and Salmonella typhi and anti-bacterial potential of non-elicited hairy roots of P. lanceolata (MIC = 25 mg mL -1 and MBC = 35 mg mL -1 ) were more active against Klebsiella pneumoniae and P. vulgaris than other bacteria. The methanolic extracts of the P. lanceolata hairy roots demonstrated significant cytotoxic activity on colorectal carcinoma cell line (SW-480) with IC 50 = 250.65 ± 6.8 µg mL -1 in comparison to human embryonic kidney (HEK-293) with IC 50 = 5263.65 ± 4.6 µg mL -1 . Plantago lanceolata hairy roots showed important biological activity explaining its role in traditional medicine.

Under the predicted conditions of climate change, the productivity of temperate grasslands may be affected by drought stress, especially in spring and summer. In this scenario, water-deficit-tolerant species such as plantain and chicory are interesting alternatives for use in sheep production systems. In this study, we compared a mixture of plantain and chicory herbage (PCH) with a grass-based permanent sward (GBS) on the weight gain and meat quality of lambs finished on these grasslands. Fifteen weaned lambs (31.3 kg and 4 months of age) were assigned to each treatment for seven weeks in late spring and live weight gain (LWG), carcass and meat quality were evaluated. There was a tendency ( p = 0.09) in final weight (40.3 ± 0.8 kg) and live weight gain (173 ± 10 g/d) to be higher in PCH compared to GBS. Carcass weight, dressing percentage and meat quality in terms of pH, color and tenderness did not differ ( p > 0.05) and were considered to be of good quality. We concluded that both swards result in comparable lamb performance and good meat quality.

Plantain and chicory are interesting forage species since they present good nutritional quality and are more resistant to drought than many temperate grasses. The fatty acid (FA) profile in milk and meat is related to a growing concern for the consumption of healthy foods, that is, with a lower content of saturated FA, higher polyunsaturated FA (PUFA) and a favourable n-6 : n-3 FAs ratio. Our objective was to evaluate the FA content in ewe's milk and lamb's meat fed a plantain-chicory mixture (PCH) or a grass-based permanent sward (GBS) dominated by perennial ryegrass. Eighteen Austral ewes in mid-lactation were allocated to PCH and GBS treatments. Milk samples were obtained during September (spring). Thirty weaned lambs were finished on both treatments from November to December (7 weeks), slaughtered and their meat sampled. Fat from milk and meat samples was extracted and stored until analysed by gas chromatography. Milk fat from GBS was higher than from PCH (P < 0.05) in C18:0 (11 385 v. 5874 mg/100 g FA), 9c-18:1 (15 750 v. 8565 mg/100 g FA), 11 t-18:1 (4576 v. 2703 mg/100 g FA) and 9c,11 t-18:2 (1405 v. 921 mg/100 g FA) and lower in 18:2n-6 (827 v. 1529 mg/100 g FA) and 18:3n-3 (943 v. 1318 mg/100 g FA) FA. Total mono-unsaturated FA was higher in GBS than PCH (P < 0.05). Meat fat from PCH swards presented a higher (P < 0.05) content than GBS for 18:2n-6 (46.8 v. 28.2 mg/100 g FA), linolenic (24.6 v. 14.2 mg/100 g FA), polyunsaturated FA (119.7 v. 73.4 mg/100 g FA), n-6 (65.9 v. 40.8 mg/100 g FA) and n-3 (53.8 v. 32.5 mg/100 g FA), respectively. No effect of treatment (P > 0.05) was detected for 9c-18:1 (283.9 v. 205.8 mg/100 g FA), 11 t-18:1 (26.2 v. 19.3 mg/100 g FA) and 9c,11 t-18:2 (10.1 v. 7.6 mg/100 g FA), for PCH and GBS. These results suggest that grazing a PCH mixture results in a higher concentration of PUFA in ewes' milk and in lambs' fat, as compared to a GBS sward.

Plant species vary in their growth response to arbuscular mycorrhizal (AM) fungi, with responses ranging from negative to positive. Differences in response to AM fungi may affect competition between plant species, influencing their ability to coexist. We hypothesized that positively responding species, whose growth is stimulated by AM fungi, will experience stronger intraspecific competition and weaker interspecific competition in soil containing AM fungi, while neutrally or negatively responding species should experience weaker intraspecific and stronger interspecific competition. We grew Plantago lanceolata, which responds positively to AM fungi, and Bromus inermis, which responds negatively to AM fungi, in an additive response surface competition experiment that varied the total density and relative frequency of each species. Plants were grown in sterilized background soil that had been inoculated with whole soil biota, which includes AM fungi, or a microbial wash, that contained other soil microbes but no AM fungi, or in sterilized soil that contained no biota. The positively responding P. lanceolata was more strongly limited by intraspecific than interspecific competition when AM fungi were present. By contrast, the presence of AM fungi decreased the strength of intraspecific competition experienced by the negatively responding B. inermis. Because AM fungi are almost always present in soil, strong intraspecific competition in positively responding species would prevent them from outcompeting species that respond neutrally or negatively to AM fungi. The potential for increased intraspecific competition to offset growth benefits of AM fungi could, therefore, be a stabilizing mechanism that promotes coexistence among plant species.

Plantago lanceolata L. (buckhorn plantain) is an encroaching winter weed described as one of the most successful noncultivated colonizing species around the world. Control of P. lanceolata in southeastern USA Carya illinoinensis (Wangenh.) K. Koch production has not been studied, nor has the role of temperature on germination using a thermal gradient table. Seed of P. lanceolata collected from a Georgia C. illinoinensis grove were tested for the effects of temperature over time to establish differences in effects on germination using a thermal gradient table. Temperatures ranged from 13.5 to 30.5 °C for 288 h. Cumulative P. lanceolata seed germination was 66% occurring at 17.8 °C at 242 h. Over the 288 h experiment, maximum P. lanceolata germination was 27% occurring at 17.0 °C, 187 h after initiation. Control of P. lanceolata with residual herbicides, or in combination with 2,4-dichlorophenoxyacetic acid (2,4-D) was evaluated in the interrow of C. illinoinensis groves containing Trifolium repens L., and in greenhouse experiments. Pre- and post-emergent herbicides included indaziflam, halosulfuron-methyl, and simazine applied alone, or in combination with 2,4-D in late autumn after P. lanceolata emergence in a C. illinoinensis grove. Indaziflam in combination with 2,4-D controlled P. lanceolata greater than 90% when applied in C. illinoinensis groves and greenhouse experiments. Halosulfuron-methyl and simazine applied alone, or in combination with 2,4-D, provided 67% or less P. lanceolata control in the grove experiments, and 83% or less in greenhouse experiments. Results suggested that herbicide applications should be made during the time when diurnal temperatures are between 15 and 30 °C, while abiding pre-harvest interval restrictions. Post- and pre-emergent herbicides may aid in controlling emerged weeds and reducing further weed emergence during the autumn of that year.

The fungal parasite Podosphaera plantaginis employs both sexual and asexual reproduction to increase its chances of infecting the plant Plantago lanceolata ., Competing Interests: ED, LB No competing interests declared, (© 2019, Decaestecker and Bulteel.)

Background: Understanding the mechanisms by which diversity is maintained in pathogen populations is critical for epidemiological predictions. Life-history trade-offs have been proposed as a hypothesis for explaining long-term maintenance of variation in pathogen populations, yet the empirical evidence supporting trade-offs has remained mixed. This is in part due to the challenges of documenting successive pathogen life-history stages in many pathosystems. Moreover, little is understood of the role of natural enemies of pathogens on their life-history evolution., Results: We characterize life-history-trait variation and possible trade-offs in fungal pathogen Podosphaera plantaginis infecting the host plant Plantago lanceolata. We measured the timing of both asexual and sexual stages, as well as resistance to a hyperparasite of seven pathogen strains that vary in their prevalence in nature. We find significant variation among the strains in their life-history traits that constitute the infection cycle, but no evidence for trade-offs among pathogen development stages, apart from fast pathogen growth coninciding with fast hyperparasite growth. Also, the seemingly least fit pathogen strain was the most prevalent in the nature., Conclusions: We conclude that in the nature environmental variation, and interactions with the antagonists of pathogens themselves may maintain variation in pathogen populations.

Many pathogens possess the capacity for sex through outcrossing, despite being able to reproduce also asexually and/or via selfing. Given that sex is assumed to come at a cost, these mixed reproductive strategies typical of pathogens have remained puzzling. While the ecological and evolutionary benefits of outcrossing are theoretically well-supported, support for such benefits in pathogen populations are still scarce. Here, we analyze the epidemiology and genetic structure of natural populations of an obligate fungal pathogen, Podosphaera plantaginis . We find that the opportunities for outcrossing vary spatially. Populations supporting high levels of coinfection -a prerequisite of sex - result in hotspots of novel genetic diversity. Pathogen populations supporting coinfection also have a higher probability of surviving winter. Jointly our results show that outcrossing has direct epidemiological consequences as well as a major impact on pathogen population genetic diversity, thereby providing evidence of ecological and evolutionary benefits of outcrossing in pathogens., Competing Interests: AL, BB, EN, JS No competing interests declared, (© 2019, Laine et al.)

Within-species diversity is an important driver of ecological and evolutionary processes. Recent research has found that plants can harbour significant epigenetic diversity, but its extent, stability and ecological significance in natural populations is largely unexplored. We analysed genetic, epigenetic and phenotypic variation in a large number of natural grassland populations of Plantago lanceolata, covering a broad geographical and environmental range. Within-population diversity and among-population differentiation were calculated from genetic and epigenetic marker data and from measurements of phenotypic traits, both for plants in the field and for the F 1 generation grown in a common environment. We found weak but significant epigenetic population structure. A large part of the epigenetic population differences observed in the field was maintained in a common environment. Epigenetic differences were consistently related to genetic and environmental variation, and to a lesser degree to phenotypic variation and land use, with more grazed populations harbouring greater epigenetic diversity. Our study demonstrates that epigenetic diversity exists in natural populations of a common grassland species, and that at least part of this epigenetic diversity is stable, nonrandom and related to environmental variation. Experimental and more detailed molecular studies are needed to elucidate the mechanistic basis of these observed patterns., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)

Understanding processes maintaining variation in pathogen life-history traits is a key challenge in disease biology, and of importance for predicting when and where risks of disease emergence are highest. Pathogens are expected to encounter tremendous levels of variation in their environment - both abiotic and biotic - and this variation may promote maintenance of variation in pathogen populations through space and time. Here, we measure life-history traits of an obligate fungal pathogen at both asexual and sexual stages under both single infection and coinfection along a temperature gradient. We find that temperature had a significant effect on all measured life-history traits while coinfection only had a significant effect on the number of sexual resting structures produced. The effect of temperature on life-history traits was both direct as well as mediated through a genotype-by-temperature interaction. We conclude that pathogen life-history traits vary in their sensitivity to abiotic and biotic variation in the environment.

The relative importance of bottom-up versus top-down control of population dynamics has been the focus of much debate. In infectious disease biology, research is typically focused on the bottom-up process of host resistance, wherein the direction of control flows from the lower to the higher trophic level to impact on pathogen population size and epidemiology. However, the importance of top-down control by a pathogen's natural enemies has been mostly overlooked.Here, we explore the effects of, and interaction between, host genotype (i.e., genetic susceptibility to pathogen infection) and infection by a hyperparasitic fungus, Ampelomyces spp., on the establishment and early epidemic growth and transmission of a powdery mildew plant pathogen ( Podosphaera plantaginis ). We used a semi-natural field experiment to contrast the impacts of hyperparasite infection, host-plant resistance and spatial structure to reveal the key factors that determine pathogen spread. We then used a laboratory-based inoculation approach to test whether the field experiment results hold across multiple pathogen-host genetic combinations and to explore hyperparasite effects on the pathogen's later life-history stages.We found that hyperparasite infection had a negligible effect on within-host infection development and between-host spread of the pathogen during the onset of epidemics. In contrast, host-plant resistance was the major determinant of whether plants became infected, and host genotype and proximity to an infection source determined infection severity.Our laboratory study showed that, while the interaction between host and pathogen genotypes was the key determinant of infection outcome, hyperparasitism did, on average, reduce the severity of infection. Moreover, hyperparasite infection negatively influenced the production of the pathogen's overwintering structures. Synthesis and applications . Our results suggest that bottom-up host resistance affects pathogen spread, but top-down control of powdery mildew pathogens is likely more effective against later life-history stages. Further, while hyperparasitism in this system can reduce early pathogen growth under stable laboratory conditions, this effect is not detectable in a semi-natural environment. Considering the effects of hyperparasites at multiple points in pathogen's life history will be important when considering hyperparasite-derived biocontrol measures in other natural and agricultural systems.

The amount of damage that herbivorous insects impose on plants varies as a function of plant ontogenetic trajectories in tissue quality and defenses, and the herbivores' own developmental trajectories in body size, mandible shape and detoxification enzymes, among others. However, little is known about how host plant and herbivore ontogeny interact. Using four ontogenetic stages of Plantago lanceolata (Plantaginaceae) and three to five larval stages of the specialist caterpillar Junonia coenia (Nymphalidae), we evaluated how ontogenies in both of these trophic levels shape: (i) caterpillar feeding choice, (ii) performance, and (iii) sequestration of plant allelochemicals. Plant physical (leaf toughness) and chemical (iridoid glycosides) defenses increased, while nutritional quality (water and nitrogen content) decreased, as plants aged. These plant ontogenetic trajectories strongly altered the behavior and physiology of this specialist herbivore, but the magnitude of the response varied with larval stage. In feeding experiments, while first instar larvae showed little preference among plant stages, older larvae significantly preferred juvenile over reproductive stages. In turn, larval consumption increased and digestive efficiency decreased, potentially explaining their decrease in relative growth rate, as larvae and host plant aged, but differences were greater for younger than older caterpillars. Finally, sequestration of plant allelochemicals increased through plant and larval development; however, the major differences due to diet occurred earlier during larval development. Our results highlight that changes in plant ontogeny most strongly influence early herbivore instars, emphasizing the need to consider the developmental stage of both trophic levels to better understand temporal variation in herbivore damage.

Turkey has highly rich floras of medicinal and aromatic plants because of having various climate conditions in different regions. One of these regions is Middle Black Sea Region, especially Ordu Province. Extracts of 10 edible plants ( Arum maculatum L., Hypericum orientale L., Ornithogalum sigmoideum Freyn et Sint., Silene vulgaris Garcke var. macrocarpa , Plantago lanceolata L., Achillea millefolium L. subsp. pannonica , Rumex crispus L., Rumex acetosella L., Capsella bursa-pastoris L., Coronopus squamatus Asch.), grown in Ordu, Turkey, were prepared with different solvents (hexane, ethanol and water, separately) and their anticholinestrase and antiaflatoxigenic activities were evaluated. Additionally, the cupric reducing antioxidant capacities (CUPRAC) and ABTS cation radical scavenging abilities of the extracts were assayed. The ethanol extract of R. acetosella exhibited the highest antioxidant activity (A 0.5 value of 25.31 µg/mL, for CUPRAC activity; IC 50 value of 23.73 µg/mL, for ABTS activity). The hexane extract of C. bursa-pastoris showed the strongest inhibition against AChE enzyme with IC 50 value of 7.24 µg/mL, and the hexane extract of A. millefolium subsp. pannonica had the highest BChE activity with IC 50 value of 6.40 µg/mL. The ethanol extract of P. lanceolata exhibited the strongest inhibition against aflatoxin with 88% inhibition.

Wound healing requires cells that increase both collagen production as a result of inflammatory events and regeneration of epithelial tissue. The Plantago species of herbs have been used in traditional treatment of skin disorders and infectious diseases, and digestive, respiratory, reproductive and circulatory conditions. We investigated the efficacy of different concentrations of Plantago lanceolata L. extract (PLE) for wound healing owing to its anti-inflammatory, anti-bacterial, anti-fungal, anti-oxidant, anti-ulcerative, analgesic and immunomodulatory properties. We used 72 mice in four groups of 18. An excisional 1 cm wound was created in the skin on the back of the mice in all groups. An ointment containing 10% PLE was applied to the wound in group 1, an ointment containing 20% PLE was applied in group 2 and vaseline was applied in group 3. In group 4, no treatment was applied to the wound. On days 7, 14, and 21 of the experiment, six animals in each group were sacrificed after the wounds were photographed and specimens from the wound sites were examined. On day 14, epithelialization was more prominent in group 2, while vascularization and collagen deposition was more advanced in groups 1 and 2 compared to the other groups. Immunohistochemical examination revealed that TGF-β1 expression was elevated on day 14 in all groups; however, this elevation was more limited in groups 1 and 2 than in groups 3 and 4. Although ANGPT-2 expression increased in groups 1 and 4 on day 14, it decreased significantly in groups 2 and 3. We found that different concentrations of PLE exhibited positive effects on wound healing. Application of 10% PLE ointment may be a useful strategy for wound healing.

The most important components of Plantago lanceolata L. leaves are catalpol, aucubin, and acteoside (=verbascoside). These bioactive compounds possess different pharmacological effects: anti-inflammatory, antioxidant, antineoplastic, and hepatoprotective. The aim of this study was to protect Plantago lanceolata extract from hydrolysis and to improve its antioxidant effect using self-nano-emulsifying drug delivery systems (SNEDDS). Eight SNEDDS compositions were prepared, and their physical properties, in vitro cytotoxicity, and in vivo AST/ALT values were investigated. MTT cell viability assay was performed on Caco-2 cells. The well-diluted samples (200 to 1000-fold dilutions) proved to be non-cytotoxic. The acute administration of PL-SNEDDS compositions resulted in minor changes in hepatic markers (AST, ALT), except for compositions 4 and 8 due to their high Transcutol contents (80%). The non-toxic compositions showed a significant increase in free radical scavenger activity measured by the DPPH test compared to the blank SNEDDS. An indirect dissolution test was performed, based on the result of the DPPH antioxidant assay; the dissolution profiles of Plantago lancolata extract were statistically different from each SNEDDS. The anti-inflammatory effect of PL-SNEDDS compositions was confirmed by the ear inflammation test. For the complete examination period, all compositions decreased ear edema as compared to the positive (untreated) control. It can be concluded that PL-SNEDDS compositions could be used to deliver active natural compounds in a stable, efficient, and safe manner., Competing Interests: The authors declare no conflict of interest.

Medicinal plants accumulate heavy metals from contaminated soil, and their consumption can cause poisoning. Our objective was to determine the levels of Pb, Cd, Zn, Cu, Fe, and Mn in four medicinal plant species (Achillea millefolium, Hypericum perforatum, Plantago lanceolata, and Urtica dioica) and their native soil, all sampled at a former smelter. The highest soil Cd, Pb, and Zn levels surpassed the maximum allowed limit 75-fold, 48-fold, and 14-fold, respectively. Their soil levels correlated with those in the plants, but this was not the case with Cu, Fe, and Mn. Heavy metal accumulation seems to depend on the plant species, yet even so, medicinal herbs should be cultivated and gathered only from controlled (uncontaminated) areas. Polluted areas should be monitored on a regular basis, while further research should investigate the connection between the heavy metal levels in the soil, their levels available for plants, and the levels extractable from plants.

Host plant quality for herbivores is modulated by different factors including symbiosis with soil organisms, such as arbuscular mycorrhiza (AM), as well as plant age. However, the role of the developmental stage of the AM in such plant-microbe-herbivore interactions has been neglected. To investigate the effects of AM stage and plant age on aphid performance, individuals of the generalist Myzus persicae were reared on leaves of non-mycorrhized (NM) or mycorrhized (AM) Plantago lanceolata plants at two time points, on young plants (with a minor established AM) and on older plants (with a well-established AM), respectively. Various performance traits were measured in the first aphid generation. Additionally, the body mass of the offspring was recorded to determine effects of previous infestation on the next generation. At the end of the herbivore experiments, plant carbon (C), nitrogen (N) and phosphorus (P) as well as leaf mass per area (LMA) were analyzed as measures of plant quality. Developmental performance traits of the aphids were either affected by AM and time point (nymph body mass at day 6) or by the interaction of both (relative growth rate). However, body mass at day 10 and reproductive performance traits were lower on older plants, independent of AM treatment. In line with these results, host plant quality changed little due to AM but strongly with age, with decreases in leaf N and P, but increases in C and LMA. Furthermore, nymphs gained a higher body mass when feeding on previously infested plants compared to their parents that started to feed on non-infested hosts, likely due to an aphid-induced modification in host traits. In summary, our results indicate that effects of both the developmental stage of AM and the plant age on aphid performance change throughout aphid development and between generations, attributable to shifts in plant quality., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Profilins are small actin-binding proteins found in eukaryotes and involved in cell development, cytokinesis, membrane trafficking, and cell motility. From an allergenic point of view, profilins are panallergens usually involved in allergic polysensitization, although they are generally recognized as minor allergens. The objectives of this study were to identify and characterize the profilin from Plantago lanceolata pollen and to investigate the cross-reactivity between profilins from different pollen allergenic sources. Profilins from P. lancelolata (Pla l 2) and palm tree pollen (Pho d 2) were purified by affinity chromatography, deeply characterized and identified by mass spectrometry. Pla l 2 allergenicity was confirmed by immunoblot with serum samples from a patient population sensitized to profilin. Immunoblot inhibition was performed to study IgG reactivity between different pollen profilins. IgE cross-reactivity was demonstrated by ImmunoCAP inhibition. Pla l 2 is the second P. lanceolata allergen included in the IUIS Allergen Nomenclature database. Four peptides from purified Pla l 2 were identified with percentages of homology with other pollen profilins between 73 and 86%. Eighty-six percent (21/24) of the patient population recognized Pla l 2. The allergenic relatedness between Pla l 2, Pho d 2 and six pollen profilins was confirmed, and IgE cross-reactivity of Pla l 2 with rBet v 2 and rPhl p 12 was demonstrated. Pla l 2 is the profilin from P. lanceolata. The demonstrated allergenicity of this protein and its cross-reactivity with other pollen profilins support its use in profilin diagnostic assays., (Copyright © 2017 Elsevier Ltd. All rights reserved.)