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Background/Objectives: The increase in incidences of multidrug resistance exacerbates tuberculosis-related global health challenges and underscores a call for more efforts for development of new antitubercular drugs, including the use of medicinal plants, especially those that have been used for generations by traditional healers. Despite reports of antimicrobial activity and chemical profiling of Kirkia wilmsii (K. wilmsii) extracts, chemical structures of the bioactive agents have not been elucidated. Here, we used a combination of bioactivity-guided fractionation, mass spectrometry, and nuclear magnetic resonance to purify and elucidate the chemical structure of antimycobacterial agents contained in leaf and twig extracts for K. wilmsii. Results: After overnight extraction with acetone and 90 g of dry twigs and leaves produced 5.38 g (6%) and 4.56 g (5%) of product, which displayed moderate antimycobacterial activity of 0.5 and 1 mg/mL, respectively. The antimycobacterial activity was increased six- and three-fold, respectively, after the crude extracts were subjected to solvent–solvent partitioning. Due to many bioactive fractions being obtained after silica gel chromatography purification, fraction 5 of twig extract was prioritized for further purification due to its low minimum inhibitory concentration (MIC) (0.25 mg/mL) and cytotoxicity (20%, in THP-1 cells). Sequential purification of the fraction 5 (twig extract) extracts through the C18 cartridge and high-performance liquid chromatography (HPLC) produced four fractions, which were subjected to structural elucidation. The high-resolution mass spectrometric analyses revealed that the first two eluting peaks had the same mass ion of 441.0822 m/z (M − H⁻), which corresponded to catechin monogallate, and so were the last two eluting peaks, which had a mass ion of 539.0932 m/z (M − H⁻), corresponding to catechin digallate. Further analyses by ¹H, ¹³C, and 2D NMR confirmed the chemical structures of compounds eluting in the first two peaks on HPLC as structural isomers of catechin 3′-monogallate and catechin 4′-monogallate (MIC not determined). Similarly, compounds eluting in the last two peaks were identified as structural isomers catechin 3′-digallate and catechin 4′-digallate, with an MIC of 250 µg/mL against Mycobacterium smegmatis and Mycobacterium tuberculosis H37Rv and an MBC of 500 μg/mL against M. smegmatis. Conclusions: To the best of our knowledge, this study is the first to report the structure of catechin 3′- and 4′-digallate, their antimycobacterial activity, and the existence of acyl migration involving galloyl 3′ and 4′-hydroxyl groups of catechin ring B. Full article

Megaplatypus mutatus, a major poplar pest in South America, tunnels into the xylem, weakening trunks and reducing wood quality. Volatile organic compounds (VOCs) are key mediators of plant–insect interactions and may reflect genotype-specific defence strategies. This study analysed VOC profiles of young and adult Populus deltoides cv. Harvard and P. × canadensis cv. Conti 12 under natural M. mutatus infestation. Gas chromatography–mass spectrometry putatively annotated 31 VOCs, including green leaf volatiles (GLVs), pentyl leaf volatiles (PLVs), terpenes, alcohols, aromatics and phenolics, 12 of which, to our knowledge, have not been previously reported in Populus VOC profiles. Harvard trees showed ~14.5-fold higher total VOC abundance than Conti trees. In Conti, constitutive VOC emissions remained stable regardless of infestation status or age. In contrast, under infestation, Harvard trees emitted10-fold higher constitutive VOCs than non-infested Harvard trees and ~52-fold higher than Conti, a pattern consistent with increased defensive activity. GLVs and PLVs relatively dominated both genotypes, although Harvard showed higher emissions. Terpenes were not detected in young Conti trees under our analytical conditions but were abundant and diverse in infested Harvard trees, which may indicate a stronger terpene-associated response in this clone. Several compounds were detected only under specific genotype–condition combinations in our dataset and therefore represent candidate volatiles for future behavioural and functional studies. These results are consistent with differences in VOC emission patterns between genotypes and age classes, improve our understanding of putative chemical cues in the interaction between Populus and M. mutatus, and provide a basis for future work towards sustainable pest management strategies. Full article

Although Simarouba glauca DC. has been recognized for its therapeutic properties, its anticancer effects against oral cancer have not been adequately investigated. The present study aimed to evaluate the activity of S. glauca leaf extracts against oral squamous cell carcinoma (OSCC). S. glauca leaves were extracted using solvents of increasing polarity, and the resulting fractions were evaluated for their phytochemical composition, antioxidant activity, and cytotoxic effects. Among all extracts, the S. glauca hexane extract (SGHE) exhibited the most potent anticancer activity against cell lines representing OSCC (CAL-27), cervical cancer (HeLa), and mouse mammary tumors (4T1). Bioactivity-guided fractionation identified D-erythro-Sphinganine as a major constituent present in hexane extract, possibly contributing to anticancer activity. But since the anticancer activity of crude hexane extract is superior compared to isolated D-erythro-Sphinganine, we predict a synergistic interaction among the multiple bioactive compounds present in the crude hexane extract. Hence, further studies were carried out with crude hexane extract. Mechanistic studies have shown that the anticancer activity of hexane extract is due to its ability to (a) alter cell cycle progression, (b) trigger apoptosis, and (c) inhibit cell migration in CAL-27 cells. Overall, these findings indicate that the hexane extract of S. glauca leaf possesses multi-target anticancer potential and warrants further mechanistic and in vivo investigations. Full article

Sugar beet (root) is primarily used by industry as a raw material for sugar production, and its large-scale cultivation is closely linked to the sugar industry. Currently, sugar beet leaf (SBL) is not processed and is typically left on the field as green fertilizer after mechanical harvesting. This represents an underutilized biomass stream with potential bioactive compounds. The aim of this study was to evaluate the distribution of polyphenol and proteins in the leaf blade and petioles of different sugar beet cultivars harvested at various time points. Total polyphenols were quantified using vitexin as a reference standard, and the phenolic profile of methanolic extracts was characterized using complementary HPLC-DAD and LC-MS methods. The protein content in leaf blades ranged from 19% to 29% (dry weight) and was significantly influenced by cultivar and harvest date. Petioles contained significantly lower protein content, ranging from 4.9% to 9.5% (dry weight). The total polyphenol content (TPC) varied with cultivar and harvest time, ranging from 7.8 to 11.0 mg/g DW in leaf blades and from 0.8 to 2.7 mg/g DW in petioles. Leaf blades also contained substantially higher concentrations of vitexin derivatives (mean 7.4 ± 2.3 mg/g DW) than petioles (1.1 ± 0.6 mg/g DW). The percentage contribution of vitexin derivatives to TPC was high in both tissues (>70%) and decreased with later harvest dates. The results provide a detailed characterization of polyphenolic and protein distribution in blades and petioles of sugar beet leaves and can support further evaluation of their potential use in value-added applications. Full article

Blossom-end rot (BER) in tomatoes is a physiological disorder primarily caused by the disruption of calcium absorption and transport. This study cultivated tomatoes using a trough-based vermiculite system. Two treatments were established: a calcium-deficient nutrient solution and a calcium-deficient nutrient solution supplemented with 0.1 mg/L BR (n = 40 plants per treatment). The activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD), as well as the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), were measured in the leaves. Calcium ion content was also determined in various plant parts. Statistical analysis of differences was performed using Duncan’s multiple range test at a significance level of p < 0.01. Concurrently, transcriptome sequencing of root, stem, and leaf tissues was conducted via high-throughput sequencing technology. The results showed that foliar application of BR under calcium deficiency significantly reduced the incidence of BER (from 26.67% to 6.67%) and effectively increased calcium ion content in leaves, stems, and roots. At the physiological level, BR treatment markedly enhanced the activities of CAT, POD, and SOD in leaves (by 105.70%, 117.12%, and 82.77%, respectively), while reducing H₂O₂ and MDA contents (by 36.90% and 16.38%, respectively). This indicates that BR alleviates membrane lipid peroxidation damage by enhancing the antioxidant defense system. Gene Ontology (GO) enrichment analysis revealed that the differentially expressed genes (DEGs) were primarily involved in biological processes, such as secondary metabolic processes, response to oxygen-containing compounds, and cell wall organization. KEGG pathway analysis further indicated significant enrichment in pathways, including phenylpropanoid biosynthesis, plant hormone signal transduction, and plant–pathogen interaction. Additionally, key genes, such as the cytochrome c oxidase (COX) gene (Solyc03g013460.1), exhibited a gradient up-regulation pattern (root > stem > leaf) in the oxidative phosphorylation pathway. In conclusion, BR likely enhances tomato tolerance to calcium deficiency stress and effectively reduces BER incidence through multiple pathways: regulating calcium absorption and distribution, activating the antioxidant system, modulating hormone signaling pathways, and enhancing energy metabolism. These findings provide a theoretical basis for the application of BR in agricultural production. Full article

The codling moth (Cydia pomonella L.) is a major pest of pome fruits worldwide, guided by semiochemicals to locate hosts and oviposition sites. Quince (Cydonia oblonga Mill.), although less studied, is also affected by this pest. This study aimed to identify behaviorally active compounds for codling moth by characterizing the volatilome of quince cultivars. Volatile profiles were analyzed across four phenological stages (flowering, unripe, growth, and ripe fruit) using solid-phase microextraction and GC–MS. The cultivars evaluated were Champion, INTA 37, INTA 117, and INTA 147. Female oviposition behavior and neonate larval host choice were also assessed. Identified volatiles included esters, sesquiterpenes, monoterpenes, alcohols, aldehydes, and norisoprenoids. Among monoterpenes, limonene, consistently detected across all cultivars and stages, emerged as a key kairomone. Volatile composition varied across phenological stages, with the fruit growth stage exhibiting the highest diversity and abundance of compounds previously reported as behaviorally active. This pattern coincided with peak female oviposition and larval host selection. Females oviposited mainly on leaf surface, whereas during ripening, eggs were deposited on fruit lacking pubescence. Overall, INTA 147 was the most preferred cultivar. These findings highlight quince volatiles, particularly Limonene, as potential candidates for the development of semiochemical-based tools to improve codling moth management. Full article

Canopy ammonia (NH₃) exchange is a major contributor to agricultural NH₃ emissions and is closely linked to nitrogen-use efficiency. Glutamine synthetase (GS) mediates plant NH₃ assimilation, yet the specific roles of different GS isoenzymes in regulating wheat canopy NH₃ exchange remain unclear. This study aimed to clarify the functional differences of wheat TaGS isoenzymes in modulating canopy–atmosphere NH₃ exchange dynamics using two wheat cultivars (Yumai 49-198 and Xinong 509) under two nitrogen application levels (120 and 225 kg N ha⁻¹). Field experiments combined with FTIR-based NH₃ flux measurement, biochemical assays, and molecular analyses were conducted at anthesis and 16, 24, and 30 days after anthesis (DAA). Results showed that the leaf NH₃ compensation point, determined by apoplastic NH₄⁺ concentration, is a key factor influencing canopy NH₃ exchange. Leaf NH₃ sources exhibited distinct temporal specificity: photorespiration and nitrate reduction dominated at anthesis to 16 DAA, whereas nitrogenous compound degradation prevailed at 24–30 DAA. This temporal partitioning was highly coordinated with TaGS isoenzyme expression: TaGS2 was highest in early grain filling, potentially supporting assimilate NH₃ from photorespiration/nitrate reduction, while TaGS1;1 expression increased progressively, aligning with the scavenging of NH₃ from organic nitrogen degradation. These coordinated patterns suggest that the TaGS isoenzymes play differentiated roles in influencing wheat canopy NH₃ exchange. This study thus provides correlative insights that point to potential molecular targets for breeding nitrogen-efficient wheat cultivars and mitigating agricultural NH₃ emissions sustainably. Full article

Heracleum sosnowskyi is considered to be a dangerous invasive plant species that has successfully naturalized within a variety of plant communities across numerous countries. As a result of its superior competitiveness, the alien species is able to displace the indigenous species from their native habitats, thus changing the ecosystems and decreasing biodiversity. The phytochemicals present in the H. sosnowskyi aqueous extracts were revealed using GC/MS and HPLC/DAD/TOF techniques. Isopsoralen, methoxsalen, (iso)pimpinellin and/or bergapten were determined to be major compounds in the leaf, inflorescence and root extracts. Glutaric, quinic, linolenic, (iso)chlorogenic and other polyphenolic acids were identified in the extracts. Furthermore, a number of furanocoumarins, including hermandiol, bakuchicin, candinols (A and C) and candibirin F, and coumarins, umbelliferone and yunngnins (A and B), were identified in the roots. Additionally, the presence of flavonoids, including astragalin, quercetin 7,3,4-trimethyl ether, nicotiflorin and rutin, has been detected in the flower and leaf extracts. Allelopathic effects of H. sosnowskyi aqueous extracts were tested on four model plants, lettuce (Lactuca sativa L.) and three native Lithuanian meadow herbs, perennial ryegrass (Lolium perenne L.), timothy (Phleum pratense L.) and white clover (Trifolium repens L.), using the Petri dish method. H. sosnowskyi flower and leaf extracts demonstrated the strongest inhibitory effects on the germination and growth of the tested plant seeds. At the highest relative concentrations, 0.5 and 1.0, extracts of Sosnowsky’s hogweed inflorescences inhibited timothy seedling growth by 95.47% (from 19.64 ± 2.57 mm to 0.89 ± 0.73 mm) and 100%, respectively. The leaf extracts exhibited the strongest inhibitory effects on white clover seedlings. The highest relative concentrations tested (0.5 and 1.0) suppressed clover seedling growth by 94.66% (from 41.22 ± 2.53 mm to 2.20 ± 0.63 mm) and 100%, respectively. Additionally, the germination rate and vigor index of model plants were assessed. The research is of significance for the regulation and monitoring of the spreading of aggressive H. sosnowskyi plants. Moreover, it is important for the development of natural herbicides based on active phytotoxic compounds from these plants. Full article

Breast cancer remains one of the leading causes of cancer-related mortality among women worldwide. Although cisplatin is widely used in chemotherapy, its clinical efficacy is often limited by adverse effects and resistance. Thus, natural bioactive compounds are gaining attention as complementary therapeutic agents. This study aimed to evaluate the anti-tumor effects of Annona muricata leaf extract on murine breast cancer 4T1 cells, used alone or in combination with cisplatin. Cisplatin induced intrinsic apoptosis through mitochondrial membrane disruption, up-regulation of the Bax gene and inhibition of the PI3K/AKT/mTOR signaling pathway. Cisplatin also promoted hypoxia by HIF1α gene expression, inflammation by TNFα and IL-6 gene expression, and induced cell cycle arrest at the sub-G1 phase by down-regulation of cyclin D1 and cyclin E1 genes. Annona muricata leaf extract triggered autophagy-mediated 4T1 cell death through mainly mTOR down-regulation and increased expression of Beclin1 and LC3 genes. It also induced cell cycle arrest at sub-G1 and S phases in a concentration- and time-dependent manner. When, combined with cisplatin, Annona muricata extract shifts the cell death pathway from intrinsic apoptosis toward autophagy by reduced caspase-3 gene expression and activity and enhanced LC3-I to LC3-II conversion. Moreover, Annona muricata extract attenuated cisplatin-induced inflammation by inhibiting TNFα and IL-6 gene expression and reinforced cell cycle arrest through suppression of the cyclin D1 gene. In conclusion, our results suggest that Annona muricata leaf extract exerts significant anti-tumor activity in breast cancer cells and may enhance cisplatin efficacy by shifting the signaling pathway from intrinsic apoptosis toward autophagy, and attenuating inflammation-related effects, supporting its potential use as a complementary therapeutic strategy. Full article

Apple cultivation faces soil acidification and pollution due to excessive fertilization, compounded by a scarcity of potassium (K) fertilizers. Polyhalite, a natural multi-nutrient mineral, offers a potential sustainable alternative. Therefore, a three-year field experiment was conducted, comprising a no-potassium control (CK), two conventional potassium fertilizers (sulfate of potash-based and muriate of potash-based), and six polyhalite compound fertilizer treatments (with different basal and topdressing strategies), to evaluate their effects on apple growth and soil fertility. Results showed that the single topdressing application of potassium chloride-type polyhalite compound fertilizer (T6) achieved the highest yield in the final year, which was 10.11–28.03% higher than the other potassium-applied treatments. It also achieved the highest fruit vitamin C and soluble solids content (9.53 mg 100 g⁻¹ and 13.27%, respectively). The T6 treatment demonstrated the best performance in terms of agronomic efficiency and partial factor productivity of potassium fertilizer, reducing fertilizer waste and loss. Furthermore, the T6 treatment effectively increased soil pH, available potassium, and exchangeable calcium levels, thereby improving soil fertility. Thus, polyhalite proves effective in replacing conventional K fertilizers, with the single topdressing of MOP-type polyhalite compound fertilizer (T6) offering the most comprehensive agronomic and environmental benefits. Full article

Adventitious shoot regeneration in woody species is regulated by interactions between plant growth regulators, endogenous hormone metabolism, and environmental cues such as light quality. Here, we investigated the effects of thidiazuron (TDZ) and the cytokinin oxidase/dehydrogenase (CKX) inhibitors INCYDE and phenyladenine (PA), in combination with different light spectra, on morphogenesis in Melia volkensii leaf explants. TDZ induced the highest frequencies of callus formation and adventitious shoot regeneration, particularly under white light. INCYDE promoted localized regeneration responses, including activation of dormant meristematic regions in secondary leaf axils, whereas PA showed limited regeneration efficiency. Light quality significantly influenced morphogenesis, with white and blue light favoring organized shoot development, while red and far-red light suppressed shoot regeneration and promoted callus formation. Cytokinin profiling revealed treatment-dependent shifts in endogenous cytokinin composition, most notably in isopentenyladenine (iP)-type cytokinins, which is consistent with altered cytokinin degradation dynamics. Cis-zeatin-type cytokinins were abundant across treatments, likely reflecting regulation associated with in vitro culture conditions. These findings indicate that cytokinin metabolism and light quality jointly influence organogenic competence in Melia volkensii Gürke, providing a physiological basis for optimizing regeneration strategies in woody plants. This study provides the first integrated analysis of cytokinin-modulating compounds and light spectra on adventitious shoot regeneration in Melia volkensii. The findings establish a physiological basis for improving regeneration protocols in recalcitrant woody species and support future biotechnological applications, including genetic improvement and advanced propagation strategies. Full article

During a late-winter field survey in Baihuashan National Nature Reserve, Beijing, several noctuid moths were observed flying during the daytime at low temperatures and being actively preyed upon by Marsh tits, which removed the heads and wings of captured individuals. These observations indicate that adults of this noctuid lineage are active in late winter, providing a critical nutritional resource for insectivorous birds during the ecologically constrained, food-limited winter period. Here, we formally describe this lineage as a new genus, Shoudus gen. nov., based on a new species, S. baihuashanus sp. nov., collected from Baihuashan reserve, including three specimens retrieved during active interception of tit predation, along with detached wings and heads recovered from the snow. The new genus is placed in the tribe Orthosiini Guenée, 1837, primarily based on adult external morphology, including large compound eyes with long interfacetal hairs and bipectinate male antennae, as well as forewing patterning similar to certain orthosiine genera such as Perigrapha and Clavipalpula. Notably, the dark reddish-brown forewings with sharply contrasting pale markings, as seen in the new genus and these related genera, appear well adapted for camouflage against bark, leaf litter, and exposed soil in their habitats—potentially functioning as both background matching and disruptive coloration. To further assess its phylogenetic placement, we conducted a molecular analysis based on mitochondrial COI sequences (13 newly generated and 6 retrieved from BOLD/NCBI). The resulting maximum likelihood and Bayesian trees consistently support the monophyly of the new genus and reveal a close phylogenetic relationship with Orthosia, the type genus of Orthosiini. This integrative evidence strongly supports the recognition of Shoudus as a distinct lineage within Orthosiini. Full article

Leaves of Cucumis callosus, traditionally employed in Ayurvedic medicine for the treatment of urinary disorders, were investigated in depth for their therapeutic potential against bacterially induced urinary tract infection (UTI) for the first time. The present work is the first to explore the antibacterial activity of C. callosus leaf fractions with an integrative in silico, in vitro, and in vivo approach. Through bioassay-guided fractionation, the chloroform fraction (F1) was identified as the most active, exhibiting potent activity against Uropathogenic Escherichia spp. species. Liquid chromatography–mass spectrometry (LC-MS) analysis of F1 revealed the presence of bioactive compounds, including stigmasterol, 1,2,3,4-tetrahydroisoquinoline, lactose, hydroxy(mesityl)acetic acid, and 2,4-di-tert-butylphenol. Molecular docking studies validated the strong binding affinities of these compounds for bacterial resistance enzymes, including AmpC β-lactamase and carbapenemases, thereby providing plausible mechanisms of antimicrobial action. In vivo studies carried out on female rats infected with Escherichia spp. species revealed a dose-dependent reduction in bacterial load, with a significant decrease in urinary tract inflammation upon F1 administration. Histopathological evaluation confirmed the protective effect, with reduced epithelial damage and inflammation in bladder tissues. These findings indicate significant antibacterial and tissue-protective effects of the C. callosus leaf fraction F1, supporting its ethnomedicinal use and establishing it as a promising phytotherapeutic agent for the treatment of urinary tract infections. Full article

Plant-derived phytotoxins are widely investigated as sustainable alternatives to synthetic herbicides; however, a major limitation is the insufficient chemical characterization of active constituents in many promising candidate species, including Afzelia xylocarpa (Kurz) Craib. In this study, the phytotoxicity of A. xylocarpa leaves and their phytotoxic compounds were investigated to evaluate their potential value as a bioherbicide. The results showed the A. xylocarpa leaf extracts suppressed the seedling growth of Lepidium sativum L., Lactuca sativa L., and Lolium multiflorum Lam. Six compounds were obtained from the A. xylocarpa leaf extracts using bio-guided fractionation and were identified as (+)-dehydrovomifoliol (1), (3R,6R,7E)-3-hydroxy-4,7-megastigmadien-9-one (2), (+)-3-hydroxy-β-ionone (3), (S)-N-(1-hydroxy-3-phenylpropan-2-yl) benzamide (4), isololiolide (5), and (+)-lariciresinol (6). Compounds 1 to 6 significantly reduced seed germination, seedling growth, and dry biomass accumulation into different extents (p < 0.05). L. sativum roots were more susceptible to all the obtained compounds than other growth parameters, except for compound 4. Based on the doses of six compounds required for 50% growth inhibition (defined as EC₅₀ value), compound 3 (EC₅₀ values = 227.4 to 582.3 µM) and compound 5 (EC₅₀ values = 53.8 to 200.8 µM) were the most toxic against all the growth parameters of L. sativum and may be the principal active compounds of the A. xylocarpa leaf extracts. Such phytotoxic effects indicate that these six compounds could be candidates for bioherbicides. Full article

The essential oils obtained by steam distillation of leaves of Piper species have found several applications in bioeconomy due to their various biological properties. Nevertheless, the analysis of essential oils does not provide information regarding the ecologically relevant volatile organic compounds (VOCs) emitted by metabolically active leaves under real-world conditions, challenged or not by herbivore damage. In this study, P. arboreum growing in a highly diverse area was observed as the host of two generalist caterpillars—Gonodonta maria (Erebidae) and Dysodia spissicornis (Thyrididae)—and one Piper-specialist from the genus Eois (Geometridae). The effect of the leaf attack caused by the three different caterpillars on VOCs emission indicated significant and herbivore-specific changes in leaf-induced responses. The profiles of undamaged leaves showed that the two generalist herbivores induced a higher number of single VOCs and of total VOCs emissions by P. arboreum when compared to the herbivory of the specialist caterpillar. Many of the VOCs emitted by herbivore-damaged leaves contained terpenoids that have been previously shown to attract parasitoids, such as (E)-β-ocimene, linalool, DMNT and (E)-β-caryophyllene. All three herbivores significantly altered the VOC profile of P. arboreum leaves compared to undamaged controls, but specific composition signatures were observed, highlighting the complexity of chemical communication at multitrophic levels. Full article