Phytochemical and pharmacological investigation of the non-volatile compounds of Lepidium cartilagineum (J. C. Mayer) Thell. and determination of the essential oil composition of its flowers and fruits.

Eighteen compounds, among them phenylpropanoids (1-2), neolignans (3-9), a megastigmane (10), a phenyl glucoside (11), flavonoids (12-14), and N-containing compounds (15-18) were isolated from the methanolic extract of the whole plant of L. cartilagineum. The structures of the compounds were determined by NMR and MS measurements. The composition of the essential oils prepared from the flowers and fruits of L. cartilagineum was investigated using GC and GC-MS measurements. The essential oils were rich in aliphatic aldehydes and hydrocarbons, but low in sulfur-containing compounds, e.g., isothiocyanates. The extracts prepared from the aerial parts and roots of the plant, the essential oil, and the isolated compounds (1-9) were tested for antiproliferative activity against COLO 205 and COLO 320 cell lines and antibacterial activity on Lactobacillus rhamnosus. Dehydrodiconiferyl alcohol γ'-methyl ether (5) possessed marked antiproliferative activity against both human tumor cell lines. Neither the extracts nor the compounds affected the growth of the bacteria and did not influence the biofilm formation of L. rhamnosus. Based on the results, it can be concluded that L. cartilagineum is non-toxic to the human gut microbiome forming L. rhamnosus.

Investigation of the Allelopathic Effect of Two Invasive Plant Species in Rhizotron System.

A key question in plant invasion biology is why invasive plants are more competitive in their introduced habitat than in their native habitat. Studies show that invasive species exhibit allelopathy, influencing other plants by releasing chemicals. Research on allelopathy uses in vitro tests, investigating effects on seed germination and seedling development. Although soil plays a role in modifying allelopathic effects, observations with soil are rare and almost nothing is known about the root development of test plants developing in soil and the effects of allelopathic compounds on root architecture. Our study evaluates the allelopathic effects of false indigo-bush (Amorpha fruticosa L.) and common milkweed (Asclepias syriaca L.) on oilseed rape growth as a model plant. The rhizotron system was used to study the effect of morphology and root architecture. Leaf-soil mixtures at 0.5%, 1%, and 5% concentrations were used. Shoot and root development was strongly inhibited at 5%. But there was no difference between the allelopathy of the two species, and the application of lower concentrations did not show any effect, demonstrating that soil has a significant modifying effect on their allelopathy. Our results highlight that the development of roots growing in the soil is also worth investigating in connection with allelopathy, which can strengthen the ecological importance of allelochemicals during successful invasions.

A case study on the early stage of Pinus nigra invasion and its impact on species composition and pattern in Pannonic sand grassland.

Alien woody species are successful invaders, frequently used for afforestation in regions like semi-arid lands. Shrubs and trees create important microhabitats in arid areas. Understorey vegetation in these habitats has unique species composition and coexistence. However, the impact of solitary woody species on understorey vegetation is less understood. This study evaluated the effect of native (Juniperus communis) and invasive solitary conifers (Pinus nigra) on surrounding vegetation, where individuals were relatively isolated (referred to as solitary conifers). The field study conducted in Pannonic dry sand grassland in 2018 recorded plant and lichen species presence around six selected solitary conifers. Composition and pattern of understorey vegetation were assessed using 26 m belt transects with 520 units of 5 cm × 5 cm contiguous microquadrats. Compositional diversity (CD) and the number of realized species combinations (NRC) were calculated from the circular transects. Results showed native conifer J. communis created more complex, organized microhabitats compared to alien P. nigra. CD and NRC values were significantly higher under native conifers than invasive ones (p = 0.045 and p = 0.026, respectively). Native species also had more species with a homogeneous pattern than the alien species. Alien conifers negatively affected understorey vegetation composition and pattern: some species exhibited significant gaps and clusters of occurrences along the transects under P. nigra. Based on our study, the removal of invasive woody species is necessary to sustain habitat diversity.

L-Aminoguanidine Induces Imbalance of ROS/RNS Homeostasis and Polyamine Catabolism of Tomato Roots after Short-Term Salt Exposure.

Polyamine (PA) catabolism mediated by amine oxidases is an important process involved in fine-tuning PA homeostasis and related mechanisms during salt stress. The significance of these amine oxidases in short-term responses to salt stress is, however, not well understood. In the present study, the effects of L-aminoguanidine (AG) on tomato roots treated with short-term salt stress induced by NaCl were studied. AG is usually used as a copper amine oxidase (CuAO or DAO) inhibitor. In our study, other alterations of PA catabolism, such as reduced polyamine oxidase (PAO), were also observed in AG-treated plants. Salt stress led to an increase in the reactive oxygen and nitrogen species in tomato root apices, evidenced by in situ fluorescent staining and an increase in free PA levels. Such alterations were alleviated by AG treatment, showing the possible antioxidant effect of AG in tomato roots exposed to salt stress. PA catabolic enzyme activities decreased, while the imbalance of hydrogen peroxide (H2O2), nitric oxide (NO), and hydrogen sulfide (H2S) concentrations displayed a dependence on stress intensity. These changes suggest that AG-mediated inhibition could dramatically rearrange PA catabolism and related reactive species backgrounds, especially the NO-related mechanisms. More studies are, however, needed to decipher the precise mode of action of AG in plants exposed to stress treatments.

Concentration-dependent effects of effusol and juncusol from Juncus compressus on seedling development of Arabidopsis thaliana.

Juncus species are valuable sources of phenanthrene compounds that have been used in traditional Chinese medicine for thousands of years. Effusol and juncusol are the most investigated compounds reported to have antimicrobial and anticancer effects; however, to date, their effects on higher plants have not been investigated. In this study, we examined the effects of effusol and juncusol on the growth and other biochemical parameters of the dicot model plant Arabidopsis thaliana in a concentration-dependent manner with a focus on polyamine metabolism. Phenanthrene induced toxic effects on plant growth and development, while effusol and juncusol induced higher biomass and maintained antioxidant defence mechanisms associated with reduced polyamine degradation. Taken together, our results suggest that these compounds could be good candidates for new biopesticide or biostimulant plant growth regulators in the future.

Short-Term Salicylic Acid Treatment Affects Polyamine Metabolism Causing ROS-NO Imbalance in Tomato Roots.

The phytohormone salicylic acid (SA) can influence the polyamine metabolism in plants. Additionally, polyamines (PAs) can regulate the synthesis of SA, providing an exciting interplay between them not only in plant growth and development but also in biotic or abiotic stress conditions. The effect of SA on polyamine metabolism of leaves is well-studied but the root responses are rarely investigated. In this study, tomato roots were used to investigate the effect of short-term exposition of SA in two different concentrations, a sublethal 0.1 mM and a lethal 1 mM. To explore the involvement of SA in regulating PAs in roots, the degradation of PAs was also determined. As both SA and PAs can induce reactive oxygen species (ROS) and nitric oxide (NO) production, the balance of ROS and NO was analyzed in root tips. The results showed that 0.1 mM SA induced the production of higher PAs, spermidine (Spd), and spermine (Spm), while 1 mM SA decreased the PA contents by activating degrading enzymes. Studying the ROS and NO levels in root tips, the ROS production was induced earlier than NO, consistent with all the investigated zones of roots. This study provides evidence for concentration-dependent rapid effects of SA treatments on polyamine metabolism causing an imbalance of ROS-NO in root tips.

Hypusination, a Metabolic Posttranslational Modification of eIF5A in Plants during Development and Environmental Stress Responses.

Hypusination is a unique posttranslational modification of eIF5A, a eukaryotic translation factor. Hypusine is a rare amino acid synthesized in this process and is mediated by two enzymes, deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). Despite the essential participation of this conserved eIF5A protein in plant development and stress responses, our knowledge of its proper function is limited. In this review, we demonstrate the main findings regarding how eIF5A and hypusination could contribute to plant-specific responses in growth and stress-related processes. Our aim is to briefly discuss the plant-specific details of hypusination and decipher those signal pathways which can be effectively modified by this process. The diverse functions of eIF5A isoforms are also discussed in this review.

Antiproliferative Phenanthrenes from Juncus tenuis: Isolation and Diversity-Oriented Semisynthetic Modification.

The occurrence of phenanthrenes is limited in nature, with such compounds identified only in some plant families. Phenanthrenes were described to have a wide range of pharmacological activities, and numerous research programs have targeted semisynthetic derivatives of the phenanthrene skeleton. The aims of this study were the phytochemical investigation of Juncus tenuis, focusing on the isolation of phenanthrenes, and the preparation of semisynthetic derivatives of the isolated compounds. From the methanolic extract of J. tenuis, three phenanthrenes (juncusol, effusol, and 2,7-dihydroxy-1,8-dimethyl-5-vinyl-9,10-dihydrophenanthrene) were isolated. Juncusol and effusol were transformed by hypervalent iodine(III) reagent, using a diversity-oriented approach. Four racemic semisynthetic compounds possessing an alkyl-substituted p-quinol ring (1-4) were produced. Isolation and purification of the compounds were carried out by different chromatographic techniques, and their structures were elucidated by means of 1D and 2D NMR, and HRMS spectroscopic methods. The isolated secondary metabolites and their semisynthetic analogues were tested on seven human tumor cell lines (A2780, A2780cis, KCR, MCF-7, HeLa, HTB-26, and T47D) and on one normal cell line (MRC-5), using the MTT assay. The effusol derivative 3, substituted with two methoxy groups, showed promising antiproliferative activity on MCF-7, T47D, and A2780 cell lines with IC50 values of 5.8, 7.0, and 8.6 µM, respectively.

Corrigendum: Structural Diversity and Bioactivities of Peptaibol Compounds From the Longibrachiatum Clade of the Filamentous Fungal Genus Trichoderma.

[This corrects the article DOI: 10.3389/fmicb.2019.01434.].

Gerardiins A-L and Structurally Related Phenanthrenes from the Halophyte Plant Juncus gerardii and Their Cytotoxicity against Triple-Negative Breast Cancer Cells.

Species in the Juncaceae accumulate different types of secondary metabolites, among them phenanthrenes and 9,10-dihydrophenanthrenes in substantial amounts. These compounds have chemotaxonomic significance and also possess interesting pharmacological activities. The present study has focused on the isolation, structure determination, and pharmacological investigation of phenanthrenes from Juncus gerardii. Twenty-six compounds, including 23 phenanthrenes, have been isolated from a methanol extract of this plant. Twelve compounds, the phenanthrenes gerardiins A-L (1-12), were obtained as new natural products. Eleven phenanthrenes [effusol (13), dehydroeffusol (14), effususin A (15), compressin A, 7-hydroxy-2-methoxy-1-methyl-5-vinyl-9,10-dihydrophenanthrene, juncusol, 2-hydroxy-7-hydroxymethyl-1-methyl-5-vinyl-9,10-dihydrophenanthrene, 2,7-dihydroxy-5-formyl-1-methyl-9,10-dihydrophenanthrene, effususol A, 2,7-dihydroxy-5-hydroxymethyl-1-methyl-9,10-dihydrophenanthrene, and jinflexin C], 1-O-p-coumaroyl-3-O-feruloyl-glycerol, and the flavones apigenin and luteolin were isolated for the first time from this plant. The cytotoxicity of the 23 isolated phenanthrenes in both mouse (4T1) and human (MDA-MB-231) triple-negative breast cancer cells and in a nontumor (D3, human cerebral microvascular endothelial) cell line was tested using an MTT viability assay. The results obtained showed that the dimeric compounds gerardiins I (9), J (10), K (11), and L (12), derived biogenetically from effusol and dehydroeffusol, were cytotoxic to both tumor and nontumor cell lines, while the monomeric compounds exerted no or very low cytotoxicity. Impedance measurements were consistent with the results of the MTT assays performed.

Survival and regeneration ability of clonal common milkweed (Asclepias syriaca L.) after a single herbicide treatment in natural open sand grasslands.

Invasive species are a major threat to biodiversity, human health, and economies worldwide. Clonal growth is a common ability of most invasive plants. The clonal common milkweed Asclepias syriaca L. is the most widespread invasive species in Pannonic sand grasslands. Despite of being an invader in disturbed semi-natural vegetation, this plant prefers agricultural fields or plantations. Herbicide treatment could be one of the most cost-effective and efficient methods for controlling the extended stands of milkweed in both agricultural and protected areas. The invasion of milkweed stand was monitored from 2011 to 2017 in a strictly protected UNESCO biosphere reserve in Hungary, and a single herbicide treatment was applied in May 2014. This single treatment was successful only in a short-term but not in a long-term period, as the number of milkweed shoots decreased following herbicide treatment. The herbicide translocation by rhizomatic roots induced the damage of dormant bud banks. The surviving buds developing shoots, growth of the milkweed stand showed a slow regeneration for a longer-term period. We concluded that the successful control of milkweed after herbicide treatment depends on repeated management of treated areas to suppress further spreading during subsequent seasons.

Characterization of the Plant Growth-Promoting Activities of Endophytic Fungi Isolated from Sophora flavescens.

Endophytic fungi in symbiotic association with their host plant are well known to improve plant growth and reduce the adverse effects of both biotic and abiotic stresses. Therefore, fungal endophytes are beginning to receive increased attention in an effort to find growth-promoting strains that could be applied to enhance crop yield and quality. In our study, the plant growth-promoting activities of endophytic fungi isolated from various parts of Sophora flavescens (a medicinally important plant in Mongolia and China) have been revealed and investigated. Fungal isolates were identified using molecular taxonomical methods, while their plant growth-promoting abilities were evaluated in plate assays. Altogether, 15 strains were isolated, representing the genera Alternaria, Didymella, Fusarium and Xylogone. Five of the isolates possessed phosphate solubilization activities and twelve secreted siderophores, while all of them were able to produce indoleacetic acid (IAA) in the presence or absence of tryptophan. The endogenous and exogenous accumulation of IAA were also monitored in liquid cultures using the HPLC-MS/MS technique to refine the plate assay results. Furthermore, for the highest IAA producer fungi, the effects of their extracts were also examined in plant bioassays. In these tests, the primary root lengths of the model Arabidopsis thaliana were increased in several cases, while the biomasses were significantly lower than the control IAA treatment. Significant alterations have also been detected in the photosynthetic pigment (chlorophyll-a, -b and carotenoids) content due to the fungal extract treatments, but these changes did not show any specific trends.

Structural Diversity and Bioactivities of Peptaibol Compounds From the Longibrachiatum Clade of the Filamentous Fungal Genus Trichoderma.

This study examined the structural diversity and bioactivity of peptaibol compounds produced by species from the phylogenetically separated Longibrachiatum Clade of the filamentous fungal genus Trichoderma, which contains several biotechnologically, agriculturally and clinically important species. HPLC-ESI-MS investigations of crude extracts from 17 species of the Longibrachiatum Clade (T. aethiopicum, T. andinense, T. capillare, T. citrinoviride, T. effusum, T. flagellatum, T. ghanense, T. konilangbra, T. longibrachiatum, T. novae-zelandiae, T. pinnatum, T. parareesei, T. pseudokoningii, T. reesei, T. saturnisporum, T. sinensis, and T. orientale) revealed several new and recurrent 20-residue peptaibols related to trichobrachins, paracelsins, suzukacillins, saturnisporins, trichoaureocins, trichocellins, longibrachins, hyporientalins, trichokonins, trilongins, metanicins, trichosporins, gliodeliquescins, alamethicins and hypophellins, as well as eight 19-residue sequences from a new subfamily of peptaibols named brevicelsins. Non-ribosomal peptide synthetase genes were mined from the available genome sequences of the Longibrachiatum Clade. Their annotation and product prediction were performed in silico and revealed full agreement in 11 out of 20 positions regarding the amino acids predicted based on the signature sequences and the detected amino acids incorporated. Molecular dynamics simulations were performed for structural characterization of four selected peptaibol sequences: paracelsins B, H and their 19-residue counterparts brevicelsins I and IV. Loss of position R6 in brevicelsins resulted in smaller helical structures with higher atomic fluctuation for every residue than the structures formed by paracelsins. We observed the formation of highly bent, almost hairpin-like, helical structures throughout the trajectory, along with linear conformation. Bioactivity tests were performed on the purified peptaibol extract of T. reesei on clinically and phytopathologically important filamentous fungi, mammalian cells, and Arabidopsis thaliana seedlings. Porcine kidney cells and boar spermatozoa proved to be sensitive to the purified peptaibol extract. Peptaibol concentrations ≥0.3 mg ml-1 deterred the growth of A. thaliana. However, negative effects to plants were not detected at concentrations below 0.1 mg ml-1, which could still inhibit plant pathogenic filamentous fungi, suggesting that those peptaibols reported here may have applications for plant protection.