Enhanced Antioxidant, Antifungal, and Herbicidal Activities through Bioconversion of Diosgenin by Yarrowia lipolytica P01a.

This study explores the bioconversion of diosgenin by Yarrowia lipolytica P01a, focusing on enhancing the antioxidant, antifungal, and herbicidal activities of the resulting extracts. The bioconversion process, involving glycosylation and hydroxylation, produced significant amounts of protodioscin and soyasaponin I. The extracts showed superior antioxidant activity, with up to 97.02% inhibition of ABTS· radicals and 33.30% inhibition of DPPH· radicals at 1000 mg L-1 of diosgenin. Antifungal assays revealed strong inhibitory effects against Botrytis cinerea, Alternaria sp., and Aspergillus niger, with maximum inhibition rates of 67.34%, 35.63%, and 65.53%, respectively. Additionally, the herbicidal activity of the bioconverted extracts was comparable to commercial herbicides, achieving 100% inhibition of seed germination in both monocotyledonous and dicotyledonous plants. These findings suggest that the Y. lipolytica P01a-mediated bioconversion of diosgenin could provide a sustainable and eco-friendly alternative for developing natural biofungicides and bioherbicides.

α-costic acid, the main sesquiterpenoid isolated from Dittrichia viscosa (L) Greuter, induces oxidative stress and autophagy in tomato.

Dittrichia viscosa (L.) Greuter, a perennial plant in the Asteraceae, has strong allelopathic activity due to the high content of various secondary metabolites. The bicyclic sesquiterpenoid α-costic acid is the most abundant secondary metabolite of D. viscosa. Its remarkable insecticidal, antiparasitic, and phytotoxic activities point to its potential use as a natural herbicide, but information on its mode of action is lacking. To shed light on the mechanism of action of α-costic acid in plant cells, we investigated the phytotoxicity of α-costic acid in tomato plants (Solanum lycopersicum L.) through in vivo assays, the underlying cellular effects using biochemical assays, and the effect on subcellular organelles using confocal microscopy on tomato protoplasts incubated with organelle-specific fluorescent probes. In vivo tests showed that α-costic acid inhibited the growth of tomato seedlings and induced chlorosis and spot lesions in leaves. Biochemical assays demonstrated that α-costic acid caused ion leakage, chlorophyll depletion, H2O2 overproduction, callose deposition, and membrane lipid peroxidation. Confocal microscopy demonstrated that α-costic acid determined ROS overproduction and network disruption in mitochondria, singlet oxygen overproduction in chloroplasts, vacuole disintegration, and autophagosome formation. Overall, our data are consistent with a model according to which α-costic acid phytotoxicity is related to oxidative stress in mitochondria and chloroplasts that induces extensive membrane damage, ultimately resulting in cell death associated with autophagy.

Alternaria alternata Pathogen from Cuscuta japonica Could Serve as a Potential Bioherbicide.

Dodder (Cuscuta spp.) is a dangerous parasitic plant that causes serious damage to crop production and is challenging to eliminate. Herbicide application is a common strategy to control dodder in the field, but it is costly, ineffective, and further results in hazardous outcomes. Therefore, our study aims to identify the potential pathogens in naturally occurring dodder infections which may provide efficient biocontrol options. In this regard, the pathogens were isolated from the infected plants, their pathogenicity was validated through inoculation, and the optimal culture conditions for their growth were identified by determining the pathogenicity difference. The pathogenicity range was determined in vitro using the leaves of common horticultural plants and crops. Furthermore, a small range of horticultural plants parasitized by Cuscuta reflexa in the field were inoculated with the pathogen to determine their biosafety and biocontrol potential, and the pathogens were identified by morphological and molecular characterization. We found 7 strains that were isolated after pathogen enrichment culture. Among them, Cbp6 and Cbp7 showed the highest pathogenicity against C. reflexa. After testing the inoculation of more than 50 species of plants, only 9 species showed varying degrees of lesions on leaves, which proved the high biosafety for common plants. Field spraying of these pathogens showed a good control effect on C. reflexa after 21 days; the disease severityreached 66.0%, while its host plant did not display obvious symptoms. In conclusion, the pathogens Cbp6 and Cbp7 were identified as Alternaria alternata, and the results of this study provide a theoretical basis for the biological control of dodder.

Enhancing Eritadenine Production in Submerged Cultures of Shiitake (Lentinula edodes Berk. Pegler) Using Blue LED Light and Activated Charcoal. Revealing Eritadenine's Novel In Vitro Bioherbicidal Activity Against Chrysanthemum morifolium.

Eritadenine from shiitake mushroom is a secondary metabolite with hypocholesterolemic, hypotensive and antiparasitic properties, thus promising for pharmaceutical and agricultural applications. Eritadenine is obtained from submerged mycelial cultures of shiitake, but the actual yields remain unsatisfactory to explore potential applications or industrial-scale production. In this study, green and blue LED lights were tested to increase yields of eritadenine in submerged cultures of shiitake. Notably, blue LEDs increased yields by 13-14 times, reaching 165.7 mg/L, compared to darkness (11.2 mg/L) and green light (12.1 mg/L) (p < 0.05, Tukey test). Nitrogen sources yeast extract (YE) and peptone (at 2 g/L) increased eritadenine production. YE promoted 22.6 mg/L, while peptone 18.3 mg/L. The recovery of eritadenine was evaluated using amberlite and activated charcoal (AC) adsorption isotherms. AC demonstrated the highest adsorption rate, with 75 mg of eritadenine per gram of AC, according to the Freundlich isotherm. The desorption rate reached 93.95% at pH 10. The extract obtained from submerged cultures had eritadenine content of 63.31%, corresponding to 87.86% of recovery, according to HPLC analysis. Furthermore, the novel bioherbicidal potential of eritadenine was tested on in vitro Chrysanthemum morifolium plants. The cultures extract containing eritadenine had a detrimental impact on plant development, generating mortality of 100% at 3%, 0.5%, and 0.25%. Moreover, pure eritadenine exhibited a phytotoxic effect similar than glyphosate on leaves, stems and roots. These findings highlight the significant bioherbicidal properties of eritadenine. Further studies are needed to understand the biosynthetic pathway of eritadenine and its bioherbicidal properties on weeds and illicit crops.

Microbial Bioherbicides Based on Cell-Free Phytotoxic Metabolites: Analysis and Perspectives on Their Application in Weed Control as an Innovative Sustainable Solution.

Weeds cause significant agricultural losses worldwide, and herbicides have traditionally been the main solution to this problem. However, the extensive use of herbicides has led to multiple cases of weed resistance, which could generate an increase in the application concentration and consequently a higher persistence in the environment, hindering natural degradation processes. Consequently, more environmentally friendly alternatives, such as microbial bioherbicides, have been sought. Although these bioherbicides are promising, their efficacy remains a challenge, as evidenced by their limited commercial and industrial production. This article reviews the current status of microbial-based bioherbicides and highlights the potential of cell-free metabolites to improve their efficacy and commercial attractiveness. Stirred tank bioreactors are identified as the most widely used for production-scale submerged fermentation. In addition, the use of alternative carbon and nitrogen sources, such as industrial waste, supports the circular economy. Furthermore, this article discusses the optimization of downstream processes using bioprospecting and in silico technologies to identify target metabolites, which leads to more precise and efficient production strategies. Bacterial bioherbicides, particularly those derived from Pseudomonas and Xanthomonas, and fungal bioherbicides from genera such as Alternaria, Colletotrichum, Trichoderma and Phoma, show significant potential. Nevertheless, limitations such as their restricted range of action, their persistence in the environment, and regulatory issues restrict their commercial availability. The utilization of cell-free microbial metabolites is proposed as a promising solution due to their simpler handling and application. In addition, modern technologies, including encapsulation and integrated management with chemical herbicides, are investigated to enhance the efficacy and sustainability of bioherbicides.

Dereplication of Bioactive Agave Saponin Fractions: The Hidden Saponins.

The good phytotoxicity and selectivity against weeds versus tomato or cress make saponin-rich fractions from Agave macroacantha, A. colorata, A. parryi, and A. parrasana attractive candidates as bioherbicides. The saponin contents have only previously been reported for A. macroacantha, and as a consequence, simultaneous dereplication has been performed on saponin-rich fractions from the other plants by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. This strategy enables the identification of a total of 26 saponins, 14 of which have been described previously and 12 of which are proposed as new saponins. They include isomers and a new sugar chain with a ß-d-apiofuranose unit. The method is corroborated by the isolation of eight dereplicated saponins from A. colorata.

Eryngial: An α,ß-Unsaturated Fatty Aldehyde as the Major Phytotoxin in Spiny Coriander (Eryngium foetidum L.) Essential Oil.

Weed species many times possess allelochemicals as a part of their survival strategy. These metabolites can be potential targets in search of natural phytotoxins. This study aims to evaluate the phytotoxic ability of fatty aldehyde-rich essential oil from spiny coriander (Eryngium foetidum) leaves, also known as fitweed or spiritweed and to further identify the active phytotoxins. This oil dose-dependently inhibited the wheatgrass coleoptile and radicle growth in multiple bioassays with half maximal inhibitory concentration (IC50) 30.6-56.7 µg/mL, while exhibiting a less pronounced effect on the germination (IC50 181.8 µg/mL). The phytotoxicity assessment of two oil constituents identified eryngial (trans-2-dodecenal), exclusively major fatty aldehydic constituent as the potent growth inhibitor with IC50 in the range 20.8-36.2 µg/mL during an early phase of wheatgrass emergence. Eryngial-inspired screening of eleven saturated fatty aldehydes and alcohols did not find a significantly higher phytotoxic potency. In an open vessel, eryngial as the supplementation in agar medium, dose-dependently inhibited the growth of pre-germinated seeds of one monocot (bermudagrass) and one dicot (green amaranth) weed species with IC50 in the range 23.8-65.4 µg/mL. The current study identified eryngial, an α,ß-unsaturated fatty aldehyde of coriander origin to be a promising phytotoxic candidate for weed control.

New Perspective on the Use of α-Bisabolol for Weed Control.

The indiscriminate use of synthetic herbicides reduces its effectiveness. Bioherbicides produced with metabolites emerge as an alternative to managing weeds. We aimed to analyze the phytotoxic potential of the essential oil of Vanillosmopsis arborea (EOVA) and the α-bisabolol molecule, its main component. We evaluated the effects of EOVA and α-bisabolol at different concentrations on the germination, growth, antioxidant metabolism, and photosynthesis of different species. EOVA and α-bisabolol showed promising phytotoxic effects on the germination and initial growth of the weed Senna occidentalis, inhibiting the activity of the antioxidant enzymes and increasing lipid peroxidation. α-Bisabolol reduced the weed seedling growth by inducing oxidative stress, which suggests a greater role in postemergence. Moreover, in the weed postemergence, both EOVA and α-bisabolol caused damage in the shoots, reduced the chlorophyll content, and increased lipid peroxidation besides reducing photosynthesis in S. occidentalis. Overall, we suggest the promising action of α-bisabolol and EOVA as bioherbicides for weed control.

Isolation, Characterization, and Bioherbicidal Potential of the 16-Residue Peptaibols from Emericellopsis sp. XJ1056.

Eco-friendly bioherbicides are urgently needed for managing the problematic weed Amaranthus retroflexus. A mass spectrometry- and bioassay-guided screening approach was employed to identify phytotoxic secondary metabolites from fungi for the development of such bioherbicides. This effort led to the discovery of six phytotoxic 16-residue peptaibols, including five new compounds (2-6) and a known congener (1), from Emericellopsis sp. XJ1056. Their planar structures were elucidated through the analysis of tandem mass and NMR spectroscopic data. The absolute configurations of the chiral amino acids were determined by advanced Marfey's method and chiral-phase liquid chromatography-mass spectrometry (LC-MS) analysis. Bioinformatic analysis and targeted gene disruption identified the biosynthetic gene cluster for these peptaibols. Compounds 1 and 2 significantly inhibited the radicle growth of A. retroflexus seedlings, and 1 demonstrated potent postemergence herbicidal activity against A. retroflexus while exhibiting minimal toxicity to Sorghum bicolor. Structure-activity relationship analysis underscored the importance of trans-4-hydroxy-l-prolines at both the 10th and 13th positions for the herbicidal activities of these peptaibols.

Herbicidal proteins from Bacillus wiedmannii isolate ZT selectively inhibit ryegrass (Lolium temulentum L.).

BACKGROUND: The widespread use of chemical herbicides and the growing issue of weed resistance pose significant challenges in agriculture. To address these problems, there is a pressing need to develop biological herbicides based on bacterial metabolites. RESULTS: In this study, we investigated the impact of the cell-free culture filtrate (CFCF) from the ZT isolate, a bacilliform bacterium obtained from diseased wheat seeds, on the germination and seedling growth of various plant species, including wild oat, ryegrass, redroot, wheat, and chickpea. The results revealed that CFCF had a detrimental effect on the fresh and dry weight of stems and roots in most of the studied plants, except chickpeas. The CFCF was further subjected to separation into aqueous and organic phases using chloroform, followed by the division of the aqueous phase into 13 fractions using an alumina column. Notably, both the aqueous phase (20%) and all 13 fractions (ranging from 50% to 83%) displayed the ability to reduce the root length of ryegrass, a monocotyledonous weed. Liquid chromatography-mass spectrometry (LC-MS) analysis identified that fractions 3 and 7, which were effective against ryegrass but not redroot, contained Cry family proteins, including Cry10 Aa, Cry4 Ba, and Cry4 Aa. Additionally, 16s rRNA gene sequencing revealed that the ZT isolate is closely related (98.27%) to Bacillus wiedmannii. CONCLUSION: Conclusively, metabolites from the ZT bacterium hold promise for monocotyledonous weed-targeted herbicides, providing a constructive strategy to confront agricultural issues tied to chemical herbicides and weed resistance. © 2024 Society of Chemical Industry.

Screening of Secondary Metabolites Produced by Nigrospora sphaerica Associated with the Invasive Weed Cenchrus ciliaris Reveals Two New Structurally Related Compounds.

In the search for new alternative biocontrol strategies, phytopathogenic fungi could represent a new frontier for weed management. In this respect, as part of our ongoing work aiming at using fungal pathogens as an alternative to common herbicides, the foliar pathogen Nigrospora sphaerica has been evaluated to control buffelgrass (Cenchrus ciliaris). In particular, in this work, the isolation and structural elucidation of two new biosynthetically related metabolites, named nigrosphaeritriol (3-(hydroxymethyl)-2-methylpentane-1,4-diol) and nigrosphaerilactol (3-(1-hydroxyethyl)-4-methyltetrahydrofuran-2-ol), from the phytotoxic culture filtrate extract were described, along with the identification of several known metabolites. Moreover, the absolute stereochemistry of (3R,4S,5S)-nigrosphaerilactone, previously reported as (3S,4R,5R)-4-hydroxymethyl-3,5-dimethyldihydro-2-furanone, was determined for the first time by X-ray diffraction analysis. Considering their structural relationship, the determination of the absolute stereochemistry of nigrosphaerilactone allowed us to hypothesize the absolute stereochemistry of nigrosphaeritriol and nigrosphaerilactol.

Soil microbiome analysis supports claims of ineffectiveness of Pseudomonas fluorescens D7 as a biocontrol agent of Bromus tectorum.

IMPORTANCE: Cheatgrass is one of North America's most problematic invasive species. Invasion by this annual grass alters ecosystem structure and function and has proven very challenging to remove with traditional approaches. Commercially available bioherbicides, like P. fluorescens D7, are applied with the goal of providing lasting control from a single application. However, experimental results suggest that this bioherbicide has limited efficacy under field conditions. Potential explanations for variable efficacy include a failure of this bioherbicide to establish in the soil microbiome. However, to our knowledge, no data exist to support or refute this hypothesis. Here, we use a deep-sequencing approach to better understand the effects of this bioherbicide on the soil microbiome and screen for P. fluorescens at 18 months post-application.

Host-virus interaction between tobacco mild green mosaic virus strain U2 and tropical soda apple resulting in systemic hypersensitive necrosis and the host range, survival, spread, and molecular characterization of the virus.

BACKGROUND: Tobacco mild green mosaic virus strain U2 (TMGMV-U2) is a registered active ingredient in a bioherbicide to control tropical soda apple (TSA), Solanum viarum, an invasive weed. As required for registration, we developed empirical data on the host-virus interaction and the virus's host range, survival, spread, and genomic sequence. RESULTS: TMGMV-U2 killed TSA plants by causing systemic hypersensitive necrosis (SHN). It elicited local lesions in inoculated leaves which was followed by the plant's wilting and death. It moved from inoculated terminal leaves through the vasculature to roots and then to newly developed leaves. Phloem death was implicated in wilting and plant death. The SHN response was attenuated in plants grown at constant 32 °C. TMGMV-U2 titer in TSA was low compared to a systemically susceptible tobacco. The virus remained infective for up to 6 months in infected dead TSA tissues and in soil in which infected plants had grown. Susceptible tobacco and pepper plants grown in soil that previously had infected dead TSA or in soil amended with the virus remained asymptomatic and virus-free. A susceptible pepper crop grown in a field block following two consecutive crops of TMGMV-U2-infected susceptible tobacco grew disease-free and virus-free and without yield loss. Purified TMGMV-U2 was infective for 1 year when stored at -20 °C or 5 °C and for 1 month at room temperature. No virus spread was found in the field. Genomic analyses confirmed the registered isolate to be a U2 strain and free of satellite TMV. The TMGMV-U2-susceptible species preponderantly belonged to the Solanaceae. A few hosts that were killed belonged to this family. Several new hosts to TMGMV-U2 were found. These data enabled registration of TMGMV-U2. CONCLUSION: TMGMV-U2 can be used safely as a bioherbicide without risks to nontarget plants and the environment. © 2023 Society of Chemical Industry.

Why are there no widely successful microbial bioherbicides for weed management in crops?

Microbial biopesticides to control plant pathogens and insects in crops have had significant success. However, there have been relatively few successes for microbial bioherbicides in crops, despite considerable numbers of publications and commercial product introductions in this area. Marketed microbial bioherbicide products for use in agriculture have been largely unsuccessful. This article covers the potential advantages of successful microbial bioherbicides, as well as the biological and technical issues that have limited their success. Technologies to overcome the problems that have limited the success of these products are discussed. The many advantages of using killed microbial products (e.g. cell-free filtrates) over living microbial products as bioherbicides are detailed. A commercialized mycoherbicide that has been selected for in the laboratory for control of the parasitic weed Striga hermonthica is being used with some success in Africa, indicating that non-transgenic modification of the genetics of bioherbicide microbes for improved efficacy is acceptable to some regulatory authorities. Genetic modifications to improve efficacy and host range, as well as improved application technology to greatly reduce the amount of product needed are two technologies that are likely to expand the use of microbial bioherbicides in the future. © 2023 Society of Chemical Industry.

Evaluation of Bipolaris yamadae as a bioherbicidal agent against grass weeds in arable crops.

BACKGROUND: Weeds are among the most damaging pests of agriculture, causing ≈10% worldwide reduction in crop productivity each year. Over-reliance on synthetic chemical herbicides has caused weeds around the world to evolve resistance. Bioherbicides may be an alternative. However, among their many constraints including strict environmental requirements, complicated mass-production and high product costs, limited pathogenicity and a narrow spectrum of activity are frequently encountered and are major barriers to commercialization. RESULTS: We isolated a pathogenic fungus, HXDC-1-2, from diseased leaves of a gramineous weed, stiltgrass [Microstegium vimineum (Trin.) A. Camus], from the edge of farmland in Guizhou province, China. HXDC-1-2 was identified as the fungal species Bipolaris yamadae based on the morphological characteristics and ITS-GPDH-EF1α multiple primer analysis. Its potential as a bioherbicide was evaluated by determining its weed control efficacy and crop safety. The ED50 and ED90 values of HXDC-1-2 on Echinochloa crus-galli were 3.22 × 103 and 1.32 × 105 conidia mL-1 , respectively. Host range tests revealed that 20 gramineous weeds including Setaria viridis, Leptochloa chinensis, Eleusine indica, Pseudosorghum zollingeri, Leptochloa panicea, Bromus catharticus, E. crus-galli plants, were extremely susceptible whereas 77 crop species from 27 plant families including rice, wheat, barley, corn, soybean and cotton (excluding cowpea and sorghum) were unaffected. CONCLUSION: Bipolaris yamadae strain HXDC-1-2 has great potential to be developed as a commercial broad-spectrum bioherbicidal agent for controlling grass weeds in arable crops. © 2023 Society of Chemical Industry.

Application technology for bioherbicides: challenges and opportunities with dry inoculum and liquid spray formulations.

Bioherbicides offer many potential benefits as part of an integrated weed management system or a totally biological or organic cropping system. A key factor for success is the selection of appropriate formulation and delivery systems for each target weed and cropping/climatic region. For dry inoculum products, we discuss direct implantation as an example for successful control of woody weeds, with benefits in control extending beyond the treated weeds to surrounding weeds. These applications do not require water and will become less labor-intensive with future robotic application platforms. Indeed, all bioherbicide applications are likely to improve and become more cost-effective with the advance of new application platforms with sensors and targeted control at lower application volume rates. Unmanned aerial vehicles, as new application platforms, are one of several such potential progressive application systems for liquid formulations, and we discuss product design to maintain optimum conditioning of the active ingredient(s) and storage stability. The delivery system must not adversely affect the products and the application volume rate must be appropriate for coverage on the target. Where applied with other products, compatibility must be ensured and appropriate mixing orders observed to assure performance and avoid precipitation or settling. Droplet size is important for allowing the active materials to be included in the spray, which may require droplets with diameter >150 µm for some larger particle biologically active agents. However, droplet size should not be too large to achieve target coverage. In some cases, that may be plant stems rather than leaves, or narrow grass weeds which tend to have highest spray collection efficiency for small droplets. Narrow droplet size spectrum nozzles may help optimize droplet size. We propose spray calculators to help optimize performance for coverage, retention and avoidance of drift losses, bounce, shatter and runoff. These include regulatory-supported, validated models. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Phytotoxicity and Phytotoxic Substances in Calamus tenuis Roxb.

Calamus tenuis is a shrub species distributed across South Asia. It grows well in diversified habitats and tends to dominate plants in the surrounding environment. The phytotoxicity of C. tenuis and the action of its phytochemicals against other plant species could explain its dominant behavior. Compounds with phytotoxic activity are in high demand as prospective sources of ecofriendly bioherbicides. Therefore, we investigated the phytotoxicity of C. tenuis. Aqueous methanol extracts of this plant species significantly limited the growth of four test plant species, two monocots (barnyard grass and timothy), and two dicots (alfalfa and cress), in a dose- and species-dependent manner. Bio-directed chromatographic isolation of the C. tenuis extracts yielded two major active substances: a novel compound, calamulactone {(S)-methyl 8-(5-oxo-2,5-dihydrofuran-2-yl) octanoate}, and 3-oxo-α-ionone. Both of the identified compounds exerted strong growth inhibitory effects on cress and timothy seedlings. The concentrations of 3-oxo-α-ionone and calamulactone required to limit the growth of the cress seedlings by 50% (I50) were 281.6-199.5 and 141.1-105.5 µM, respectively, indicating that the effect of calamulactone was stronger with lower I50 values. Similarly, the seedlings of timothy also showed a considerably higher sensitivity to calamulactone (I50: 40.5-84.4 µM) than to 3-oxo-α-ionone (I50: 107.8-144.7 µM). The findings indicated that the leaves of C. tenuis have marked growth-inhibitory potential, and could affect surrounding plants to exert dominance over the surrounding plant community. Moreover, the two identified phytotoxic substances might play a key role in the phytotoxicity of C. tenuis, and could be a template for bioherbicide development. This paper was the first to report calamulactone and its phytotoxicity.

Unveiling the efficacy of pre-emergent application of young Eucalyptus globulus leaves as a weed control strategy: Bridging macroscopic effects and cellular responses.

Allelopathy, the inhibition of neighbouring plant growth by certain plants, can be particularly useful if applied in a targeted way for weed management. So, this study aimed to assess and characterize the herbicidal activity of fresh and dried leaves from young Eucalyptus globulus Labill. trees applied as a soil amendment. For this, fresh and dried leaves (FL and DL, respectively) were incorporated into the soil at different concentrations (0, 1, 5, and 10% w/w), where Portulaca oleracea L. seeds were sown. After 5 weeks of exposure, results revealed that the soil incorporation of DL at 10% (w/w) presented the strongest herbicidal properties, inhibiting seed germination by 63% and inducing the loss of cell viability. To unravel the possible mode of action and the main targets at both cellular and subcellular levels, an in vitro experiment was performed. Purslane seeds were sown in a nutritive medium containing different dilutions of an aqueous extract prepared with dried eucalyptus leaves. After 5 days of exposure, germinated seedlings were processed for transmission electron microscopy and histological analyses as well as for reactive oxygen species (ROS) in vivo detection by confocal laser scanning microscopy. Results revealed that the allelochemical release from DL induced ROS overproduction, resulting in the loss of cell integrity and organization, which was characterized by damage to several cellular sub-structures, along with enhanced accumulation of lipid droplets. Overall, the incorporation of DL into the soil can represent a sustainable alternative to reduce synthetic herbicide application and subsequent environmental contamination.

Synthesis, Herbicidal Activity, and Structure-Activity Relationships of O-Alkyl Analogues of Khellin and Visnagin.

Khellin and visnagin furanochromones were recently reported as potential new bioherbicides with phytotoxic activities comparable to those of some commercially available herbicides. In this study, we examined the effect of O-alkylation and O-arylalkylation of both khellin and visnagin on its effect on herbicidal and antifungal activity. Synthetic analogues included O-demethyl khellin and visnagin, acetylated O-demethyl khellin and visnagin, O-benzylated demethyl khellin and visnagin, four O-demethyl alkylated khellin analogues, and six O-demethyl alkylated visnagin analogues, many of which are reported here for the first time. Both acetate analogues of khellin and visnagin indicated more activity as herbicides on Lemna pausicostata than visnagin, with IC50 values of 71.7 and 77.6 µM, respectively. Complete loss of activity for all O-alkyl analogues with a carbon chain length of greater than 14 carbons was observed. The O-demethyl butylated visnagin analogue was the most active compound with an IC50 of 47.2 µM against L. pausicostata. O-Demethyl ethylated analogues of both khellin and visnagin were as effective as khellin. In the antifungal bioautography bioassay against Colletotrichum fragariae at 100 µg, the only active O-alkyl and O-arylalkyl analogues were O-ethylated, O-butylated, and O-benzylated visnagin analogues with zones of inhibition of 10, 9, and 9 mm, respectively, an effect comparable to that of visnagin and khellin.

Allelopathic studies with furanocoumarins isolated from Ducrosia anethifolia. In vitro and in silico investigations to protect legumes, rice and grain crops.

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 IC50 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.