Plant Secondary Metabolites Research Articles

Identification of Two R2R3-MYB Genes Involved in Flavan-3-Ols Biosynthesis as Modulated by Salicylic Acid Through RNA-Seq in Grape Berries (Vitis spp.)

Flavan-3-ols are plant secondary metabolites that play important roles in stress resistance. Our previous studies revealed that salicylic acid (SA) activates R2R3-MYB transcription factors, promoting flavan-3-ol biosynthesis. This study identified two R2R3-MYB genes that exhibited positive responses to both exogenous SA and were probably involved in flavan-3-ol biosynthesis through RNA-sequencing, functional enrichment analysis, and qRT-PCR. The results indicated that the contents of total flavan-3-ols and their monomers, (+)-catechin and (−)-epicatechin, in grape berries after exogenous SA application were substantially increased compared to those in the control. A total of 683 differentially expressed genes in response to exogenous SA treatment were identified using RNA-seq. KEGG analysis revealed enrichment of the ‘flavonoid biosynthesis’ and ‘plant hormone signal transduction’ pathways. A specific module highly associated with flavan-3-ol biosynthesis was identified by constructing a co-expression network. Two candidate genes (VvMYB108B and VvMYB145) likely participating in flavan-3-ol biosynthesis were selected using qRT-PCR. Therefore, these two potential genes that respond to SA and putatively participate in flavan-3-ol biosynthesis were identified for the first time. These results lay a solid basis for a more profound understanding of the molecular regulation of flavan-3-ol biosynthesis in grapes.

Maleylpyruvic Acid-Inducible Gene Expression System and Its Application for the Development of Gentisic Acid Biosensors.

Gentisic acid is a secondary plant metabolite, known for its health benefits, not only widely used as a supplement but also implicated as a potential biomarker for cancer-associated metabolism alterations. To advance bioproduction and detection of this compound or its derivatives, cell-based approaches have become of interest in recent years. However, the lack of tools for high-throughput gentisic acid monitoring and compound-metabolizing organism screening limits the progress in this area. Here, we analyzed the gene cluster responsible for gentisic acid metabolism in Cupriavidus necator H16. The transcriptional regulator GtdR-based inducible gene expression system CnGtdR/PgtdA was elucidated, showing that it was activated when C. necator cells were subjected to gentisic acid. Subsequently, a 3-maleylpyruvic acid was identified as a primary inducer for this inducible system. Furthermore, genes gtdA and gtdT, encoding for gentisate 1,2-dioxygenase and MFS transporter, were shown to be essential for inducible system activation in the presence of gentisic acid with GtdA enabling conversion of this phenolic acid into the inducer. The CnGtdRAT/PgtdA-based inducible system was employed to develop a whole-cell biosensor for the intracellular and extracellular detection of gentisic acid. The potential of the 3-maleylpyruvic acid-inducible system was demonstrated by its application in metabolic pathway research, detection of highly unstable 3-maleylpyruvic acid, and development of biosensors for the intracellular or extracellular determination of gentisic acid. In addition, the utility of the biosensor was emphasized by its application for detection of gentisic acid as a potential biomarker for cancer in urine samples.

Antagonistic effects of endophytic fungi from Camellia reticulata pedicels on yeasts: implications for antimicrobial mechanism of nectar

Endophytic fungi are extensive in plant tissues and involved in the defense against stress from harmful microbes. The interaction between pedicel endophytic fungi and nectar yeasts is critical for maintaining nectar homeostasis. This study used Camellia reticulata as the research subject. High-throughput sequencing revealed that the community composition of endophytic fungi in the pedicel is dominated by Ascomycota and Basidiomycota. Their abundance varies at different taxonomic levels, showing sample variability. In total, 27 endophytic fungal isolates were isolated and screened from the pedicel under laboratory conditions. They exhibited antagonistic effects against three nectar yeasts (Metschnikowia reukaufii, Cryptococcus laurentii, and Rhodotorula glutinis) and displayed morphological and physiological diversity. The isolates were classified into the phylum Ascomycota and further categorized into the genera Alternaria, Trichoderma, Fusarium, and Dactylaria. The endophytic fungus D23, which effectively antagonizes nectar yeasts, was identified as Alternaria alternata. This fungus produces various secondary metabolites, including antibiotics such as penicillin G, grandiomycin, and cephalosporin C. The metabolic pathways involved include the biosynthesis of plant secondary metabolites, phenylpropanoids, amino acids, nucleotides, and antibiotics. The endophytic fungal community in C. reticulata pedicel is rich and diverse, making it a valuable material for screening antagonistic strains. This study provides a theoretical basis for the antagonistic effects of endophytic fungal metabolites from the pedicel of C. reticulata against nectar yeasts, highlighting their significance in maintaining nectar stability and reproductive fitness in cross-pollinated plants.

Metabolomic Analysis of Specific Metabolites in Codonopsis pilosula Soil Under Different Stubble Conditions

To investigate the soil-specific metabolites of Codonopsis pilosula under different stubble management practices, this study analyzed differentially abundant metabolites in the rhizosphere soils of rotational (DS) and continuous (LS) cropping systems via liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based metabolomic approaches. The results revealed that 66 metabolites, including amino acids and their derivatives, nucleic acids, alcohols, organic acids, amines, fatty acids, purines, and sugars, were significantly different (p < 0.05) between the DS and LS groups. Under continuous cropping, the levels of amines, fatty acids, organic acids, and sugars in the rhizosphere soil were significantly greater (p < 0.05) than those under rotational cropping, whereas the levels of amino acids and their derivatives, nucleic acids, and purines and pyrimidines were significantly lower (p < 0.05). KEGG pathway enrichment analysis revealed that these differentially abundant metabolites were enriched in metabolic pathways such as amino acid metabolism (e.g., alanine, aspartate, and glutamate metabolism), carbon metabolism, the cAMP signaling pathway, ABC transporter proteins, phenylalanine metabolism, and the biosynthesis of plant secondary metabolites. These metabolic pathways were involved in osmoregulation, energy supply, and resilience in plants. In conclusion, inter-root soil metabolites in rotational and continuous cropping of Codonopsis pilosula were able to influence soil physicochemical properties and microbial populations by participating in various biological processes.

Unveiling the Biological Activities, Pharmacological Potential, and Health Benefits of Sorbifolin: A Comprehensive Review.

Bioactive phytochemicals act as important factors with preventive and therapeutic potential in the pathogenesis of several disorders, often related to oxidative stress. Many dietary plant secondary metabolites could lower these conditions. Sorbifolin is one of these metabolites. This work is the first review of sorbifolin, a flavone detected in various plant matrices as a major compound. The present study discussed the natural sources, extraction, purification, quantification, and assessment of the biological activities of sorbifolin. Several databases including Google Scholar, Web of Sciences, and Science-Direct were consulted for relevant English articles related to sorbifolin, the phytochemical profiles of several medicinal plants containing this compound, and its biological activities, such as antioxidant, anticancer, antimicrobial, anti-inflammatory, and antidiabetic. The positive in vitro and in silico outcomes reported in the literature should be followed by additional in vivo and clinical investigations to further research the mechanisms of action, pharmacokinetic/pharmacodynamic activities, toxicological effects, pharmacological properties, and therapeutic potential of sorbifolin.

Synthesis, characterization and photo catalytic activity of silver nano particle derived from Arachis hypogaea L. seed peel extracts

Abstract The study focuses on the green synthesis of silver nanoparticles (AgNPs) using Arachis hypogaea L. seed peel extract (AHSPE) and evaluates their photocatalytic activity against Rhodamine B and Congo Red dyes. The synthesis involved reducing aqueous silver metal ions with AHSPE, characterized by various techniques including UV–visible spectroscopy, X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR), and FT-Raman spectroscopy. UV–visible spectroscopy indicated characteristic absorption peaks at 428 and 439 nm, confirming the formation of AgNPs. XRD analysis revealed an average particle size of 4.77–5.6 nm. FT-IR spectra identified biomolecules such as amines, peptides, amides, lactones, and polyphenols in the extract, acting as reducing and capping agents, thereby stabilizing the AgNPs. SEM analysis showed pristine silver nanoparticles with diameters ranging from 1 to 10 µm. The biosynthesized AgNPs demonstrated strong photocatalytic activity in degrading Rhodamine B and Congo Red dyes. This method did not use any synthetic reagents, making it an environmentally safe and cost-effective alternative for synthesizing silver nanoparticles. The process aligns with green chemistry principles, offering potential applications in photocatalysis and environmental cleanup. The study underscores the importance of biosynthesized nanoparticles due to their unique biological properties and the role of plant secondary metabolites in facilitating green synthesis.

CSIG-32. MODULATION OF HALLMARKS OF GLIOBLASTOMA MULTIFORME BY BIOACTIVE PHYTOCHEMICALS: A PRECLINICAL MECHANISTIC EVALUATION

Abstract Glioblastoma multiforme (GBM) is an aggressive malignancy with extensive neovascularization, an immunosuppressive tumor microenvironment, and high recurrence rates. Bioactive plant secondary metabolites (phytochemicals), many from fruits and vegetables, have been extensively studied for GBM prevention and treatment. The objective of this study is to determine the effects of phytochemicals on GBM hallmarks and associated molecular mechanisms of action through evaluation of in vitro and in vivo studies. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed to collect articles using PubMed, ScienceDirect and Scopus. While 1,344 articles were screened, 115 in vitro and 36 in vivo studies met our inclusion criteria. The Systematic Review Center for Laboratory Animal Experimentation (SYRCLE) review of bias tool assessed in vivo study bias. In vitro, 49 alkaloids, 10 phenolics, 2 sulfur- compounds, and 5 terpenoids modulated GBM hallmarks of cell growth, invasion, cell cycle progression, neovascularization, and apoptosis. The PI3K/Akt/mTOR pathway was suppressed by 13 phytochemicals. Apoptosis was induced by 15 phytochemicals activating caspase-3, whereas 11 compounds inhibited Bcl-2. Cell cycle modulation occurred through decreased cyclin-dependent kinases and increased p53, p21, and p27. In vivo, 17 alkaloids, 5 phenolics, 2 sulfur-compounds, and 2 terpenoids had effects on GBM hallmarks. Invasion and neovascularization were inhibited through MMP, HIF-1α, and VEGF modulation. Six phytochemicals induced apoptosis in vivo through caspases and Bcl-2. Utilizing the SYRCLE review of bias tool, the average score quality of animal studies is 10% with many indicators of bias unclear. Various phytochemicals show promise to prevent and treat GBM due to their effects on numerous GBM hallmarks. Study limitations include the inclusion of literature reporting effects of single agents and lack of clinical studies. Future research is warranted to fill gaps in our analysis before recommending phytochemicals for prevention and treatment of GBM, a perilous malignancy.

The Effects of FR and UVA Irradiation Timing on Multi-Omics of Purple Lettuce in Plant Factories

The synergistic application of far-red (FR) and ultraviolet A (UVA) irradiation presents a promising approach for enhancing growth and the enrichment of secondary metabolites in plants. However, prolonged exposure to these combined light qualities imposes significant stress on plants, hindering their development. Therefore, an initial period of FR irradiation to promote plant growth, followed by a subsequent period of UVA irradiation to enhance the accumulation of plant quality, constitutes a viable strategy. Our study, focusing on purple lettuce, aims to elucidate the response mechanisms of the lettuce leaf under standard white light in commercial production, with the addition of different durations of FR and UVA irradiation, and to explore the complex dynamic changes at the multi-omics level. The results indicate that the duration of FR exposure is crucial in determining biomass-related phenotypes such as fresh weight, while the duration of UVA exposure significantly influences the accumulation of phenotypic markers like anthocyanins. At the transcriptional level, the most extensive transcriptional regulation was observed when FR was applied throughout the entire growth period, and UVA was applied eight days before harvest, significantly impacting pathways such as MAPK signaling cascades, plant hormone signal transduction, photosynthetic processes, and the biosynthesis of secondary metabolites. Metabolomic analysis corroborated the transcriptomic findings, with particular emphasis on antioxidant activity, photoprotection, and defense mechanisms. Our comprehensive analysis suggests that short-term UVA irradiation prior to harvest, based on full growth period FR irradiation, is feasible. The combined application of FR and UVA irradiation fine-tunes plant growth, developmental trajectories, and stress responses by modulating light signals, hormonal signals, and secondary metabolic pathways. These findings not only reveal the adaptive mechanisms of plants to fluctuating light environments but also provide a scientific basis for optimizing light management strategies in controlled plant production systems and precision agriculture.

The Use of Thyme (Thymus vulgaris) Essential Oil for Controlling Cercospora Leaf Spot (Cercospora beticola) on Sugar Beets (Beta vulgaris)

Decreasing efficacy of fungicides and the withdrawal of further hazardous active ingredients in pesticides from use have prompted the search for alternative methods of crop protection. Essential oils (EOs) are secondary metabolites of plants and have been proven to show antibacterial, antifungal, and pest-repellent properties. This study was undertaken to determine the activity of grapefruit, rosemary, pine, sage, and thyme EOs against the fungus Cercospora beticola, which is the most dangerous pathogen of sugar beet and the causal agent of Cercospora leaf spot. According to the determined Minimum Inhibitory Concentration (MIC), thyme EO was found the most effective against C. beticola. For most of the fungal isolates tested, the MIC of this EO was 0.313 mL/L. Thyme EO also inhibited the growth of multi-resistant isolates. Based on the results obtained, thyme EO was subjected to further testing in field conditions, where its efficiency in controlling C. beticola was also proven. The results indicate that the use of thyme EO may be a promising method for the protection of sugar beets, although it requires further optimization in the context of its inclusion in sustainable protection programs assuming a reduced number of synthetic fungicide treatments.

Metabolomic profiling and 16 S rRNA metabarcoding of endophytes of two Aloe species revealed diverse metabolites

Aloe species are often used interchangeably for medicinal and cosmeceutical applications, presenting a challenge to the biological efficacy consistency of some herbal preparations. Sustainable production of high-quality commonly used medicinal plants remains a limitation for commercialisation. Thus, this study investigated the potential for plant substitution by examining bacterial endophytes capable of producing similar host plant secondary metabolites. The metabolite profiles and endophytic bacterial communities of endangered Aloe lettyae were compared with those of Aloe longibracteata using nuclear magnetic resonance spectroscopy and 16 S rRNA gene sequencing. Only 15 metabolites were significantly different between A. lettyae and A. longibracteata based on metabolite concentrations. However, both plants’ functionality and potential application remain comparable. Phytohormones, including indole-3-acetate and 5-hydroxyindole-3-acetate, were more concentrated in A. lettyae than A. longibracteata. Metabolites such as tyrosine, allantoin, and myo-inositol, with human health benefits, were annotated in both species. Aloe lettyae harboured a phylogenetically diverse bacteria community compared to A. longibracteata, with a higher richness of bacterial species, indicating a likelihood of diverse metabolic capabilities among the bacteria. Dominant endophytes, including Bacillus, Comamonas, and Pseudomonas, possess enzymes contributing to various metabolic pathways. The enzymes have the potential to impact the synthesis, or breakdown of plant metabolites, consequently influencing the overall metabolic composition of the host plant. Therefore, this study supports the interchangeability of A. lettyae and A. longibracteata due to their ability to produce similar metabolites, and although the Aloe species exhibit phylogenetically diverse endophytic communities, the feasibility of utilizing their endophytes as producers of secondary metabolites remains viable.Graphical abstract

Cytotoxic Effects of Plant Secondary Metabolites and Naturally Occurring Bioactive Peptides on Breast Cancer Model Systems: Molecular Mechanisms

Breast cancer is the second leading cause of death among women, and the number of mortal cases in diagnosed patients is constantly increasing. The search for new plant compounds with antitumor effects is very important because of the side effects of conventional therapy and the development of drug resistance in cancer cells. The use of plant substances in medicine has been well known for centuries, but the exact mechanism of their action is far from being elucidated. The molecular mechanisms of cytotoxicity exerted by secondary metabolites and bioactive peptides of plant origin on breast cancer cell lines are the subject of this review.

Genome-wide investigation of MYB gene family in Areca catechu and potential roles of AcTDF in transgenic Arabidopsis.

MYB protein, a crucial transcription factor, holds crucial importance in plant growth, development, stress responses, and secondary metabolite regulation. While MYB proteins have been extensively studied, research on MYBs within the palm family, particularly in Areca catechu, remains limited. This study identified 259 MYB genes in Areca catechu, including 105 1R-MYBs, 150 R2R3-MYBs, 3 3R-MYBs, and 1 4R-MYBs. Physicochemical properties, collinearity, and gene structure of these genes were analyzed. The AcMYB is distributed across 16 chromosomes of A.catechu and has 119 and 195 homologs in Arabidopsis and rice, respectively. Cis-acting elements in the promoter region suggest roles in plant hormones, growth, development, and stress. R2R3-MYB genes were divided into eight groups based on tissue expression profiles. The flowering-related gene AcTDF is highly expressed in male flowers. Overexpression of AcTDF in Arabidopsis promotes early flowering, upregulates AtSOC1 and AtFUL, and enhances tolerance to drought and salt stress. These results provide valuable insights for the identification and analysis of the MYB gene family in Areca catechu and offer a basis for the subsequent verification of its related functions and the role and significance of its role in the evolution of palms.

Phthalate exposure and reproductive effects in rodents: a model for approaches on the protective role of natural products.

This review article summarizes the experimental findings in rodents published between 2014 and 2024 concerning phthalates exposure and reproductive outcomes. Rodents were chosen for this review since most studies that have developmental aspects in different phases of exposure and that address more in-depth reproductive mechanisms have been carried out in mice and rats. The evidence of adverse effects of phthalates on fetal development and human and animal reproduction is extensive, with impacts ranging from gene expression to physiological alterations. Despite the large volume of scientific papers pointing out the harmful effects of exposure to phthalates, isolated or in mixtures, at different developmental periods, most of them are associated with the maternal exposure and long-term effects in the offspring. Regular vegetables, fruits, fish, dairy products, and whole grains intake rich in bioactive compounds can mitigate the adverse effects of EDCs in humans and animals at different developmental periods. Various food bioactive compounds (FBCs) such as genistein, resveratrol, lycopene, vitamin E, curcumin, selenium, and plant secondary metabolites (PSMs) present antioxidant, anti-inflammatory, anti-tumor, and other biological properties with the potential to reduce of deleterious effects of phthalates on the reproductive tract. In this review, we aimed to summarize the main studies produced in the last decade about phthalate exposure and reproductive disorders in males and females (at different developmental critical windows). Additionally, we proposed some FBCs and PSMs that could attenuate the main adverse effects caused by phthalate exposure on male reproduction since there is a lack of studies with females.

Trivalent chromium stress trigger accumulation of secondary metabolites in rice plants: Integration of biochemical and transcriptomic analysis

Trivalent chromium [Cr (III)] is a frequently detected environmental contaminant that negatively affects plant metabolism, growth, and yield. Plant secondary metabolites are non-nutrient compounds involved in diverse physiological functions in response to abiotic stresses. This study performed biochemical and transcriptomic analyses to clarify the protective role and mechanism of secondary metabolites in rice seedlings under Cr(III) stress. The content of measured secondary metabolites i.e., total soluble phenolics, flavonoids, lignin, and anthocyanin in rice tissues was significantly enhanced under Cr(III) exposure. Additionally, the activities of several enzymes including chalcone synthase, phenylalanine ammonialyase, peroxidase, and anthocyanidin synthase, were positively activated. Transcriptomic analysis revealed a number of differentially expressed genes (DEGs) in Cr(III)-treated rice seedlings and their expression patterns are tissue-specific. Additionally, the DEGs involved in secondary metabolism pathways were evident after KEGG enrichment analysis. Cr(III) exposure resulted in distinct genetic regulation strategies in rice tissues, with different DEGs activating various sub-pathways in the secondary metabolism pathways to cope with Cr(III) stress. Furthermore, the integrated correlation analysis of the secondary metabolites and transcriptome data identified key genes involved in different steps of the sub-pathways of secondary metabolism pathway in rice plants under Cr(III) stress. This study indicates that the accumulation of secondary metabolites in rice plants is a survival and detoxification strategy to reduce the adverse effects of toxic compounds. Future research should explore how key genes in secondary metabolism pathways aid in Cr(III) detoxification and enhance crop stress tolerance.

Design strategies of polysaccharide, protein and lipid-based nano-delivery systems in improving the bioavailability of polyphenols and regulating gut homeostasis.

Polyphenols are plant secondary metabolites that have attracted much attention due to their anti-inflammatory, antioxidant, and gut homeostasis promoting effects. However, food matrix interaction, poor solubility, and strong digestion and metabolism of polyphenols cause barriers to their absorption in the gastrointestinal tract, which further reduces bioavailability and limits polyphenols' application in the food industry. Nano-delivery systems composed of biocompatible macromolecules (polysaccharides, proteins and lipids) are an effective way to improve the bioavailability of polyphenols. Therefore, this review introduces the construction of biopolymer-based nano-delivery systems and their application in polyphenols, with emphasis on improving the solubility, stability, sustained release and intestinal targeting of polyphenols. In addition, there are possible positive effects of polyphenol-loaded nano-delivery systems on modulating gut microbiota and gut homeostasis, with particular emphasis on modulating intestinal inflammation, metabolic syndrome, and gut-brain axis. It is worth noting that the safety of bio-based nano-delivery systems still need to be further studied. In summary, the application of the bio-based nano-delivery system to deliver polyphenols provides insights for improving the bioavailability of polyphenols and for the treatment of potential diseases in the future.

Harvesting Light: The Interrelation of Spectrum, Plant Density, Secondary Metabolites, and Cannabis sativa L. Yield

The approaching legalisation and associated increasing demand for medicinal and recreational Cannabis sativa L. will lead to a growing relevance for lighting systems designed for Cannabis sativa L. The interplay between plant density, light spectrum, light distribution, yield, and secondary metabolite distribution within the plant has not yet been studied. To fill this knowledge gap, a CBD-dominant Cannabis sativa L. strain was grown in a greenhouse experiment with two plant densities (2.66 and 12 plants −1 m−2) under two different light spectra. The chosen light spectra were two LED fixtures, Solray385 (SOL) and AP67, with an R: FR ratio of 12.9 and 3.7, respectively. The results indicated that light-induced effects on individual plants can be transferred to the plant stock. A low R: FR ratio induced a 16% increase in dry flower yield in the last ten days of flowering, while a change in the light spectrum could increase the potential maximum plant density per square metre. The two spectra did not affect (CBD + CBDA) yield, as a lower flower yield compensated for a higher concentration. CBDA concentration was not significantly affected by plant density. In contrast, the higher density led to an increased total cannabidiol concentration (CBD + CBDA) and altered the distribution of terpenes. Here, the light distribution over the plant stock is particularly decisive, as a more homogenous illumination led to an increased terpene concentration of up to 41%. A Photon Conversion Efficacy (PCE) of 0.05 g mol−1 under SOL and 0.06 g mol−1 under AP67 was achieved. Plants in the centre under the highest light intensity of 1200 PAR showed up to 48% reduced efficacy. These results strongly suggest that light intensity needs to be fine-tuned to the cultivation system to prevent a reduction in efficacy, resulting in yield and quality losses.

Comprehensive analysis of secondary metabolite biosynthetic gene clusters in Helianthus annuus L.: A bioinformatics approach

Plant secondary metabolite gene clusters are regions within the plant genome that encode enzymes and proteins involved in the biosynthesis of secondary metabolites. Helianthus annuus L. is a significant oilseed plant with economic importance. This study aims to comprehensively identify secondary metabolite biosynthetic gene clusters within H. annuus using bioinformatics tools, shedding light on the functions of the enzymes involved. In this study, the plantiSMASH software was utilized to predict secondary metabolite biosynthetic gene clusters in H. annuus. The results from plantiSMASH were analyzed to identify the biosynthetic gene clusters for secondary metabolites on each chromosome. The biological and molecular functions of the enzymes within these clusters were predicted using data from the relevant articles. According to the obtained data, H. annuus lacks a biosynthetic gene cluster on chromosomes four, seven, and fifteen. The identified gene clusters in this plant are polyketide, saccharide, saccharide-terpene, alkaloid, putative, and terpene. The enzyme categories found in secondary metabolite biosynthetic gene clusters include Methyltransferase, Ketosynthase, Glycosyltransferase, BAHD acyltransferase, Dioxygenase, CoA-ligase, Epimerase, PRISE enzymes, Prenyltransferase, Oxidoreductase, Aminotransferase, Copper amine oxidase, Cytochrome P450, Terpene synthase, and Pictet-Spengler enzyme (Bet v1). This comprehensive analysis provides a foundation for further investigations into the biosynthesis of secondary metabolites in H. annuus, offering valuable insights for researchers exploring the medicinal and pharmaceutical potential of this plant species.

Effect of Cashew Nutshell Extract, Saponins and Tannins Addition on Methane Emissions, Nutrient Digestibility and Feeding Behavior of Beef Steers Receiving a Backgrounding Diet.

The beef industry contributes to greenhouse gas emissions through enteric methane emissions, exacerbating climate change. Anacardic acid in cashew nutshell extract (CNSE), saponins and tannins (ST) are plant secondary metabolites that show promise in methane mitigation via antimicrobial effects, potentially exerting changes in ruminal fermentation patterns. This study examined the impact of CNSE, ST, and their combination on methane emissions, digestibility, intake, and performance of sixteen Angus crossbred steers (347 ± 30 kg) receiving a backgrounding diet (70:30 corn silage: cottonseed burrs). The study used a 4 × 4 Latin square design (4 steers, 4 treatments, 4 periods) with a 2 × 2 factorial arrangement, including the main effects of additive (CNSE or ST) fed individually or combined. Thus, steers received the following treatments: (1) no additive, (2) CNSE only, (3) ST only, or (4) both (CNSEST). Non-supplemented steers registered eight more feedbunk visits/d than ST-steers and spent an extra 10 min/d on the feedbunk. The addition of ST tended to increase dry matter, organic matter, and neutral detergent fiber intake. Additives fed individually reduced CP digestibility. Intake of the carrier containing CNSE only was lesser and coincided with a greater methane yield in that treatment. Digestibility and methane mitigation were improved after CNSEST compared with individual inclusion, suggesting synergistic reactions enhanced methane mitigation effects in fibrous diets without affecting the digestibility of nutrients nor animal growth performance.

Assessment of loliolide extracted from Biserula pelecinus, present during in vitro oocyte maturation, on fertilisation and embryo development in sheep

Context As a ‘duty of care’, it is important to test whether new forage plants for ruminants contain secondary compounds (PSCs) that affect reproductive performance. We have previously observed, a posteriori, that the presence of a methanolic extract of Biserrula pelecinus during maturation of sheep oocytes increased fertilisation rate and blastocyst development. This result needed to be verified a priori and, if the outcome was repeated, we needed to identify the plant secondary metabolite responsible. Aims To test whether PSCs from B. pelecinus, when added to the oocyte maturation medium, improve fertilisation rate and blastocyst development; to test whether loliolide is the active molecule produced by B. pelecinus. Methods Methanol–chloroform extracts of B. pelecinus were fractionated using rapid silica filtration and solvents of increasing polarity. Fractions at final concentrations of 0, 100 or 200 μg mL−1 were added to the medium used to mature sheep cumulus–oocyte complexes (COCs) and effects were determined for maturation, subsequent cleavage rate, blastocyst rate, hatching rate, blastocyst efficiency and total blastocyst cell number (TCN). Results Fraction BP-6 at 100 μg mL−1 reduced blastocyst rate (P < 0.05), but had no effect when the dose was doubled to 200 μg mL−1. Further fractionation using semi-preparative high-performance liquid chromatography showed loliolide as the most abundant compound in BP-6. Supplementation of the in vitro maturation medium with loliolide (0, 2.5, 5, 10 and 25 μg mL−1) did not affect any measure of embryo development. All COCs treated with B. pelecinus fractions reached the final stage of embryo development, blastocyst hatching. Total blastocyst cell number was not affected. Conclusion The presence of fractions of B. pelecinus extract during in vitro oocyte maturation can reduce embryo development. Implications In vitro techniques can detect potential effects of forages on reproduction. Some fractions from an extract of B. pelecinus when present during oocyte maturation can reduce embryo development. The abundant PSC, loliolide, was not responsible. There was no indication that a PSC in B. pelecinus improves outcomes.

Silver Nanoparticles (AgNPs) Act as Nanoelicitors in Melissa officinalis to Enhance the Production of Some Important Phenolic Compounds and Essential Oils

ABSTRACTNanoparticles (NPs) are well‐known biostimulants in plant biotechnology, utilised to enhance the physical properties of plants and exhibit positive effects on them. The important key role is the most suitable type, effective dose and size of NP to be used in plant tissue culture systems. In this study, various concentrations of silver nanoparticles (AgNPs; 0, 25, 50, 75 and 100 μg L−1) were tested as elicitors in callus culture with the aim of enhancing secondary metabolite production in lemon balm (Melissa officinalis L.). According to the results obtained, callus formation rates have shown an increase in all applications compared to the control group. The highest callus formation, weight and diameter were observed in 50 μg L−1 application. In this application, the callus structure was compact and its colour was green. However, the aromatic compounds, neral and geranial increased significantly in 25 μg L−1 application. The maximum increase in phenolic compounds such as caffeic acid, chlorogenic acid, proto‐catechic acid, hesperidin and p‐coumaric acid was observed in the 75 μg L−1 AgNP and the highest increase in rosmarinic acid compound was determined in the 50 μg L−1 application. The study found that AgNP applications are an effective method for increasing the production of secondary metabolites in medicinal and aromatic plants, such as lemon balm, in vitro.