Chemical Characterization of the Essential Oil Compositions of Mentha spicata and M. longifolia ssp. cyprica from the Mediterranean Basin and Multivariate Statistical Analyses.

This present study aims to characterize the essential oil compositions of the aerial parts of M. spicata L. and endemic M. longifolia ssp. cyprica (Heinr. Braun) Harley by using GC-FID and GC/MS analyses simultaneously. In addition, it aims to perform multivariate statistical analysis by comparing with the existing literature, emphasizing the literature published within the last two decades, conducted on both species growing within the Mediterranean Basin. The major essential oil components of M. spicata were determined as carvone (67.8%) and limonene (10.6%), while the major compounds of M. longifolia ssp. cyprica essential oil were pulegone (64.8%) and 1,8-cineole (10.0%). As a result of statistical analysis, three clades were determined for M. spicata: a carvone-rich chemotype, a carvone/trans-carveol chemotype, and a pulegone/menthone chemotype, with the present study result belonging to the carvone-rich chemotype. Carvone was a primary determinant of chemotype, along with menthone, pulegone, and trans-carveol. In M. longifolia, the primary determinants of chemotype were identified as pulegone and menthone, with three chemotype clades being pulegone-rich, combined menthone/pulegone, and combined menthone/pulegone with caryophyllene enrichment. The primary determinants of chemotype were menthone, pulegone, and caryophyllene. The present study result belongs to pulegone-rich chemotype.

Effects of MhMYB1 and MhMYB2 transcription factors on the monoterpenoid biosynthesis pathway in l-menthol chemotype of Mentha haplocalyx Briq.

MAIN CONCLUSION: Transcription factors MhMYB1 and MhMYB2 correlate with monoterpenoid biosynthesis pathway in l-menthol chemotype of Mentha haplocalyx Briq, which could affect the contents of ( -)-menthol and ( -)-menthone. Mentha haplocalyx Briq., a plant with traditional medicinal and edible uses, is renowned for its rich essential oil content. The distinct functional activities and aromatic flavors of mint essential oils arise from various chemotypes. While the biosynthetic pathways of the main monoterpenes in mint are well understood, the regulatory mechanisms governing different chemotypes remain inadequately explored. In this investigation, we identified and cloned two transcription factor genes from the M. haplocalyx MYB family, namely MhMYB1 (PP236792) and MhMYB2 (PP236793), previously identified by our research group. Bioinformatics analysis revealed that MhMYB1 possesses two conserved MYB domains, while MhMYB2 contains a conserved SANT domain. Yeast one-hybrid (Y1H) analysis results demonstrated that both MhMYB1 and MhMYB2 interacted with the promoter regions of MhMD and MhPR, critical enzymes in the monoterpenoid biosynthesis pathway of M. haplocalyx. Subsequent virus-induced gene silencing (VIGS) of MhMYB1 and MhMYB2 led to a significant reduction (P < 0.01) in the relative expression levels of MhMD and MhPR genes in the VIGS groups of M. haplocalyx. In addition, there was a noteworthy decrease (P < 0.05) in the contents of ( -)-menthol and ( -)-menthone in the essential oil of M. haplocalyx. These findings suggest that MhMYB1 and MhMYB2 transcription factors play a positive regulatory role in ( -)-menthol biosynthesis, consequently influencing the essential oil composition in the l-menthol chemotype of M. haplocalyx. This study serves as a pivotal foundation for unraveling the regulatory mechanisms governing monoterpenoid biosynthesis in different chemotypes of M. haplocalyx.

[Treatment of recurrent incarcerated Bochdalek hernia in an adult]. / Recidív, kizáródott, felnottkori Bochdalek-sérv mutéti megoldása.

Az igen ritka felnottkori nem hiatális, azaz nem paraoesophagealis típusú transdiaphragmaticus sérveket - a veleszületett rekeszizom defektusok mintájára - általánosan Bochdalek, ill. Larey-Morgagni-sérveknek nevezik. Etiológia tekintetében a nem diagnosztizált és kezelt veleszületett eredet, a traumás kontúziós-szakadásos, az iatrogen, ill. a recidív típus említendo meg.Esetismertetésünkben egy felnottkori recidív, kizáródott Bochdalek-sérv sikeres mutéti ellátását ismertetjük. A 23 éves férfi beteg kórelozményében 11 éves korában bal oldali Bochdalek-sérv miatt végzett thoracoscopos rekeszizom sutura szerepel. Epigastrialis fájdalmak, hányinger, hányás, akut hasi megbetegedés klinikai tünetei miatt jelentkezett Intézetünkben. Az elvégzett sürgos mellkasi és hasi CT-vizsgálat a bal mellüregben elhelyezkedo, kizáródott, vékonybélkacsokat tartalmazó Bochdalek-sérvet igazolt. Sürgos laparotomia során az életképesnek bizonyult sérvtartalmat (a vékonybéltraktus 2/3 része, a colon flexura lienalisa és a pancreas farok) a hasüregbe reponáltuk, a sérvkaput direkt suturával zártuk, és szövetszeparáló sebészi hálóval fedtük, valamint a mellüreget draináltuk. A postoperatív szak eseménytelenül zajlott. Kontroll-CT-vizsgálat a reconstruált rekeszizom és pleuro-peritonealis rétegek folytonosságát mutatta. A 10. posztoperatív napon panaszmentesen bocsátottuk otthonába.Megbeszélés: Mint minden kizáródott sérv esetében, a diagnózis mihamarabbi felállítása és az idoben elvégzett mutét kulcsfontosságú. A mellkasi drenázs szükségességét minden esetnél körültekintoen mérlegelni kell. A mutét után a mell- és hasüregben kialakult új anatómiai viszonyok miatt cardialis és respiratoricus szövodmények alakulhatnak ki. Álláspontunk szerint a betegség ritkasága miatt centrumban kezelendo. Ezen ritka állapot sikeres gyógyítása többszakmás együttmuködésen alapul, melynek meghatározó eleme a helyesen megválasztott rekeszi felszínt helyreállító mutéti technika alkalmazása.

Allopolyploid origin and diversification of the Hawaiian endemic mints.

Island systems provide important contexts for studying processes underlying lineage migration, species diversification, and organismal extinction. The Hawaiian endemic mints (Lamiaceae family) are the second largest plant radiation on the isolated Hawaiian Islands. We generated a chromosome-scale reference genome for one Hawaiian species, Stenogyne calaminthoides, and resequenced 45 relatives, representing 34 species, to uncover the continental origins of this group and their subsequent diversification. We further resequenced 109 individuals of two Stenogyne species, and their purported hybrids, found high on the Mauna Kea volcano on the island of Hawai'i. The three distinct Hawaiian genera, Haplostachys, Phyllostegia, and Stenogyne, are nested inside a fourth genus, Stachys. We uncovered four independent polyploidy events within Stachys, including one allopolyploidy event underlying the Hawaiian mints and their direct western North American ancestors. While the Hawaiian taxa may have principally diversified by parapatry and drift in small and fragmented populations, localized admixture may have played an important role early in lineage diversification. Our genomic analyses provide a view into how organisms may have radiated on isolated island chains, settings that provided one of the principal natural laboratories for Darwin's thinking about the evolutionary process.

Metal nanoparticles decorated mint-cellulose acetate composite as an efficient catalyst for the reduction of methyl orange.

Water contamination caused by toxic compounds has emerged as one of the most severe challenges worldwide. Biomass-based nanocomposites offer a sustainable and renewable alternative to conventional materials. In this study, a nanocomposite of mint and cellulose acetate (Mint-CA) was prepared and employed as a supportive material for Cu nanoparticles (CuNPs) and Ag nanoparticles (AgNPs). The selectivity of CuNPs@mint-CA and AgNPs@mint-CA was assessed by comparing their performance in the reduction reaction of various dyes solutions. AgNPs@mint-CA exhibited superior catalytic performance, with a removal of 95.2 % for methyl orange (MO) compared to 68 % with CuNPs@mint-CA. The absorption spectra of MO exhibited a distinct peak at 464 nm. The reduction reaction of MO by AgNPs@mint-CA followed pseudo-first-order-kinetic with a rate constant of k = 0.0063 min-1 (R2 = 0.928). The highest removal of MO was achieved under the following conditions: a catalyst weight of 40 mg, an initial MO concentration of 0.07 mM, the addition of 0.5 mL of 0.1 M NaBH4, and a temperature of 25 °C. Furthermore, the AgNPs@mint-CA catalyst exhibited exceptional reducibility even after five use cycles, highlighting its potential for efficiently removing MO.

[Identification and functional analysis of jasmonic acid signaling repressor McJAZ8 gene in Mentha canadensis].

Mentha canadensis, as a plant with medicinal and culinary uses, holds significant economic value. Jasmonic acid signaling repressor JAZ protein has a crucial role in regulating plant response to adversity stresses. The M. canadensis McJAZ8 gene is cloned and analyzed for protein characterization, protein interactions, and expression patterns, so as to provide genetic resources for molecular breeding of M. canadensis for stress tolerance. This experiment will analyze the protein structural characteristics, subcellular localization, protein interactions, and gene expression of McJAZ8 using bioinformatics, yeast two-hybrid(Y2H), transient expression in tobacco leaves, qRT-PCR, and other technologies. The results show that:(1)The full length of the McJAZ8 gene is 543 bp, encoding 180 amino acids. The McJAZ8 protein contains conserved TIFY and Jas domains and exhibits high homology with Arabidopsis thaliana AtJAZ1 and AtJAZ2.(2)The McJAZ8 protein is localized in the nucleus and cytoplasm.(3)The Y2H results show that McJAZ8 interacts with itself or McJAZ1/3/4/5 proteins to form homologous or heterologous dimers.(4)McJAZ8 is expressed in different tissue, with the highest expression level in young leaves. In terms of leaf sequence, McJAZ8 shows the highest expression level in the fourth leaf and the lowest expression level in the second leaf.(5) In leaves and roots, the expression of McJAZ8 is upregulated to varying degrees under methyl jasmonate(MeJA), drought, and NaCl treatments. The expression of McJAZ8 shows an initial upregulation followed by a downregulation pattern under CdCl_2 treatment. In leaves, the expression of McJAZ8 tends to gradually decrease under CuCl_2 treatment, while in roots, it initially decreases and then increases before decreasing again. In both leaves and roots, the expression of McJAZ8 is downregulated to varying degrees under AlCl_(3 )treatment. This study has enriched the research on jasmonic acid signaling repressor JAZ genes in M. canadensis and provided genetic resources for the molecular breeding of M. canadensis.

[Cloning and expression pattern analysis of abscisic acid receptor gene McPYL4 in Mentha canadensis].

Mentha canadensis is a traditional Chinese herb with great medicinal and economic value. Abscisic acid(ABA) receptor PYLs have important roles in plant growth and development and response to adversity. The M. canadensis McPYL4 gene was cloned, and its protein characteristics, gene expression, and protein interactions were analyzed, so as to provide genetic resources for genetic improvement and molecular design breeding for M. canadensis resistance. Therefore, the protein characteristics, subcellular localization, gene expression pattern, and protein interactions of McPYL4 were analyzed by bioinformatics analysis, transient expression of tobacco leaves, RT-qPCR, and yeast two-hybrid(Y2H) techniques. The results showed that the McPYL4 gene was 621 bp in length, encoding 206 amino acids, and its protein had the conserved structural domain of SRPBCC and was highly homologous with Salvia miltiorrhiza SmPYL4. McPYL4 protein was localized to the cell membrane and nucleus. The McPYL4 gene was expressed in all tissue of M. canadensis, with the highest expression in roots, followed by leaves, and it showed a pattern of up-regulation followed by down-regulation in leaves 1-8. In both leaves and roots, the McPYL4 gene responded to the exogenous hormones ABA, MeJA, and the treatments of drought, AlCl_3, NaCl, CdCl_2, and CuCl_2. Moreover, McPYL4 was up-regulated for expression in both leaves and roots under the MeJA treatment, as well as in leaves treated with AlCl_3 stress for 1 h, whereas McPYL4 showed a tendency to be down-regulated in both leaves and roots under other treatments. Protein interactions showed that McPYL4 interacted with AtABI proteins in an ABA-independent manner. This study demonstrated that McPYL4 responded to ABA, JA, and several abiotic stress treatments, and McPYL4 was involved in ABA signaling in M. canadensis and thus in the regulation of leaf development and various abiotic stresses in M. canadensis.

Mentha piperita silver nanoparticle-loaded hydrocolloid film for enhanced diabetic wound healing in rats.

OBJECTIVE: To investigate the role of Mentha piperita silver nanoparticle-loaded carbopol gel for enhanced wound healing in a diabetic rat model. This research further aims to explore bioactive compounds derived from Mentha piperita obtained from high altitude. METHOD: Methanolic extracts of Mentha piperita (MP), Mentha spicata (MS) and Mentha longifolia (ML) were used to synthesise silver nanoparticles (AgNP). AgNP synthesis was confirmed by ultraviolet-visible (UV-Vis) spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The antioxidant activity was assessed by 2, 2-diphenyl-1-picrylhydrazyl (DDPH) assay. Antiglycation potential was determined by measuring the fluorescent advanced glycation end products. The bioactive compound identified in the Mentha piperita methanolic (MPM) fraction through electrospray ionisation tandem mass spectrometric analysis (ESI-MS) was responsible for the highest antiglycation. The effects of MPM and MPM.AgNP-loaded Carbopol (Sanare Lab, India) on wound healing were compared in male, alloxan-induced, diabetic albino rats (200-250g), divided into control and treated groups. Effects on wound healing were assessed via histopathology. RESULTS: UV-Vis and FTIR confirmed NP synthesis with peaks for flavonoids and polyphenols. SEM and XRD explored the cubical, 30-63nm crystalline NP. The maximum antioxidant and antiglycation potential was observed in order of; MP.AgNP>MS.AgNP>ML.AgNP. The highest antioxidant activity was observed by methanolic and aqueous MP.AgNPs (88.55% and 83.63%, respectively) at 2mg.ml-1, and (75.16% and 69.73%, respectively) at 1mg.ml-1, compared to ascorbic acid (acting as a positive control, 90.01%). MPM.AgNPs demonstrated the best antiglycation potential of 75.2% and 83.3% at 1mg.ml-1 and 2mg.ml-1, respectively, comparable to positive control (rutin: 88.1%) at 14 days post-incubation. A similar trend was observed for antimicrobial activity against Bacillus subtilis, Micrococcus luteus and Escherichia coli with an inhibition zone of 21mm, 21.6mm and 24.6mm. Rosmarinic acid was the active compound present in Mentha piperita, as identified by ESI-MS. MPM.AgNP-loaded Carbopol resulted in 100% wound closure compared with control at 20 days post-wounding. In the treatment group, re-epithelialisation was achieved by day 18, compared with 25 days for the positive control group. CONCLUSION: MPM.AgNP-loaded Carbopol demonstrated safer and more effective biological properties, hence accelerating the diabetic excision wound healing process in alloxan-induced diabetic rats.

Transcriptome analysis of waterlogging-induced adventitious root and control taproot of Mentha arvensis.

KEY MESSAGE: The present study reports differentially expressed transcripts in the waterlogging-induced adventitious root (AR) of Mentha arvensis; the identified transcripts will help to understand AR development and improve waterlogging stress response. Waterlogging notably hampers plant growth in areas facing waterlogged soil conditions. In our previous findings, Mentha arvensis was shown to adapt better in waterlogging conditions by initiating the early onset of adventitious root development. In the present study, we compared the transcriptome analysis of adventitious root induced after the waterlogging treatment with the control taproot. The biochemical parameters of total carbohydrate, total protein content, nitric oxide (NO) scavenging activity and antioxidant enzymes, such as catalase activity (CAT) and superoxide dismutase (SOD) activity, were enhanced in the adventitious root compared with control taproot. Analysis of differentially expressed genes (DEGs) in adventitious root compared with the control taproot were grouped into four functional categories, i.e., carbohydrate metabolism, antioxidant activity, hormonal regulation, and transcription factors that could be majorly involved in the development of adventitious roots. Differential expression of the upregulated and uniquely expressing thirty-five transcripts in adventitious roots was validated using qRT-PCR. This study has generated the resource of differentially and uniquely expressing transcripts in the waterlogging-induced adventitious roots. Further functional characterization of these transcripts will be helpful to understand the development of adventitious roots, leading to the resistance towards waterlogging stress in Mentha arvensis.

Morphological and genetic evidence suggest gene flow among native and naturalized mint species.

PREMISE: Cultivation and naturalization of plants beyond their natural range can bring previously geographically isolated taxa together, increasing the opportunity for hybridization, the outcomes of which are not predictable. Here, we explored the phenotypic and genomic effects of interspecific gene flow following the widespread cultivation of Mentha spicata (spearmint), M. longifolia, and M. suaveolens. METHODS: We morphologically evaluated 155 herbarium specimens of three Mentha species and sequenced the genomes of a subset of 93 specimens. We analyzed the whole genomes in a population and the phylogenetic framework and associated genomic classifications in conjunction with the morphological assessments. RESULTS: The allopolyploid M. spicata, which likely evolved in cultivation, had altered trichome characters, that is possibly a product of human selection for a more palatable plant or a byproduct of selection for essential oils. There were signs of genetic admixture between mints, including allopolyploids, indicating that the reproductive barriers between Mentha species with differences in ploidy are likely incomplete. Still, despite gene flow between species, we found that genetic variants associated with the cultivated trichome morphology continue to segregate. CONCLUSIONS: Although hybridization, allopolyploidization, and human selection during cultivation can increase species richness (e.g., by forming hybrid taxa), we showed that unless reproductive barriers are strong, these processes can also result in mixing of genes between species and the potential loss of natural biodiversity.

Efficient nanostructured materials to reduce nutrient leaching to overcome environmental contaminants.

Nutrient leaching is a major reason for fresh and ground water contamination. Menthol is the major bioactive ingredient of Mentha arvensis L. and one of the most traded products of global essential oil market. The indigenous production of menthol crystals in developing countries of the world can prove to be the backbone for local growers and poor farmers. Therefore, present research was designed to check the effects of nano-structured plant growth regulators (PGRs) (28-homobrassinolide and ethephon) with reduced leaching potentials on the essential oil and menthol (%) of Mentha arvensis L. The prepared nano-formulations were characterized by Fourier transform infrared (FTIR) spectroscopy, Laser induced breakdown spectroscopy (LIBS), Differential scanning colorimetry-thermal gravimetric analysis (DSC-TGA), Scanning electron microscopy (SEM), Atomic absorption spectrometry (AAS) and Zeta potential and Zeta size analysis. The menthol (%) was determined by modified spectrophotometric and gas chromatographic (GC) method. The highest essential oil (%) was obtained by the application of 28-homobrassinolide-Zn-NPs-L-II (0.92 ± 0.09%) and ethephon-Ca-NPs-L-III (0.91 ± 0.05%) as compared to the control (0.65 ± 0.03%) and blank (0.62 ± 0.09%). The highest menthol (%) was obtained by applying 28-homobrassinolide-Ca-NPs-L-I (80.06 ± 0.07%), 28-homobrassinolide-Ca-NPs-L-II (80.48 ± 0.09%) and 28-homobrassinolide-Ca-NPs-L-III (80.84 ± 0.11%) and ethephon-Ca-NPs-L-III (81.53 ± 0.17%) and ethephon-Zn-NPs-L-II (81.93 ± 0.26%) as compared to control (67.19 ± 0.14%) and blank (63.93 ± 0.17%).

Depolymerized carrageenan expresses elicitor-like activity on Mentha arvensis L. under arsenic stress: Insights into arsenic resilience and monoterpene synthesis.

Heavy metals contaminate agricultural land by limiting the productivity of crops and making them or their products unfit for consumption. Arsenic (As) is a potentially hazardous metalloid that severely impacts plants' survival. Menthol mint (Mentha arvensis L.) bears volatile compounds that are harshly exaggerated by diverse environmental factors like drought, salinity, heavy metal, temperature, photoperiod, and luminosity stresses. In this study, the phytotoxicity of As was examined in M. arvensis L. and its alleviation through the supplementation of oligomers of carrageenan. Noticeably, scanty information is available regarding the effect of irradiated carrageenan (ICA) on As-stressed plants. In order to observe the same in the case of M. arvensis L., the effect of ICA on As-treated plants was explored. The ICA concentration (foliar-applied) selected for the study was 80 mg L-1, 100 mg L-1 and 120 mg L-1, and that of As (soil-applied) was 80 mg kg-1 soil. Excess accumulation of As resulted in reduced growth, enzymatic activities, and yield and quality parameters of M. arvensis L. under As toxicity. However, the foliage application of ICA strengthens the antioxidant machinery and other physiological and oxidative stress biomarkers of the plant by facilitating the activity of superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), and proline, and, therefore aids in alleviating the toxicity generated by As. Nevertheless, ICA supplementation proves beneficial in enhancing the monoterpene synthesis (essential oil production and its active constituents) of M. arvensis L. by maintaining a steady-state equilibrium between reactive oxygen species (ROS) production and its scavenging process.

Anti-breast cancer activity of the essential oil from grapefruit mint (Mentha suaveolens × piperita).

Grapefruit mint (Mentha suaveolens × piperita) is a hybrid, perennial, and aromatic plant widely cultivated all over the world and used in the food, cosmetics, and pharmaceutical industries mostly for its valuable essential oil. Herein, we evaluated the anticancer activity of the grapefruit mint essential oil, cultivated in Iran. For the chemical composition analysis of essential oil, GC-MS was used. MTT assay was utilized for assessing the cytotoxic activity of the essential oil. The type of cell death was determined by annexin V/PI staining. Essential oil effect on the expression of maternally expressed gene 3 (MEG3), a regulatory lncRNA involved in cell growth, proliferation, and metastasis, was studied using qRT-PCR. Linalool (43.9%) and linalool acetate (40.1%) were identified as the dominant compounds of essential oil. Compared with MCF-7, the MDA-MB-231 cells were more sensitive to essential oil (IC50 = 7.6 µg/ml in MCF-7 and 5.9 µg/ml in MDA-MB-231 after 48 h). Essential oil induced cell death by apoptosis. Wound healing scratch assay confirmed the anti-invasive effect of essential oil. In addition, essential oil upregulated the tumor suppressor MEG3 in breast cancer cells. These results provide new insights into grapefruit mint essential oil potential application as an anticancer adjuvant in combination treatments for breast cancer patients.

Effects of mint, menthol, and tobacco-flavored e-cigarettes on tobacco withdrawal symptoms in adults who smoke menthol cigarettes: A laboratory pilot study.

BACKGROUND: Menthol cigarette smoking has remained stable or increased in certain groups, despite an overall decline in cigarette smoking rates in the U.S. Understanding whether e-cigarettes alter patterns of menthol cigarette use is critical to informing efforts for reducing the public health burden of menthol cigarette smoking. This 2019-2020 laboratory pilot study evaluated whether self-administration of mint-, menthol-, or tobacco-flavored e-cigarettes would differentially impact tobacco withdrawal symptoms in e-cigarette-naïve adults who smoke menthol cigarettes daily. METHODS: Participants (N=17; 35.3% Female; mean age=51.8) attended three laboratory sessions after 16-hours of tobacco abstinence. Participants self-administered a study-provided JUUL e-cigarette (0.7mL with 5% nicotine by weight) at each session in which flavor was manipulated (mint vs. menthol vs. tobacco; order randomized). Participants completed pre- and post-e-cigarette administration self-report assessments on smoking urges, nicotine withdrawal, and positive and negative affect states. Multilevel linear regression models tested differences between the three flavor conditions for individual study outcomes. RESULTS: Following overnight tobacco abstinence, vaping either a mint or menthol (vs. tobacco) flavored e-cigarette led to significantly greater reductions in smoking urges over time; menthol (vs. tobacco) flavored e-cigarettes also suppressed urges to smoke for pleasure. Notably, no differences in nicotine withdrawal, positive affect, or negative affect were observed. CONCLUSIONS: In this laboratory pilot study, mint and menthol (vs. tobacco) flavored e-cigarettes provided some negative reinforcement effects via acute reductions in smoking urges during tobacco abstinence, yet only menthol flavored e-cigarettes demonstrated suppressive effects on smoking urges for pleasure in adults who smoke menthol cigarettes daily.

Abundant incongruence in a clade endemic to a biodiversity hotspot: Phylogenetics of the scrub mint clade (Lamiaceae).

The Scrub Mint clade(Lamiaceae) provides a unique system for investigating the evolutionary processes driving diversification in the North American Coastal Plain from both a systematic and biogeographic context. The clade comprisesDicerandra, Conradina, Piloblephis, Stachydeoma, and four species of the broadly defined genus Clinopodium(Mentheae; Lamiaceae), almost all of which are endemic to the North American Eastern Coastal Plain. Most species of this clade are threatened or endangered and restricted to sandhill or a mosaic of scrub habitats. We analyzed relationships in this clade to understand the evolution of the group and identify evolutionary mechanisms acting on the clade, with important implications for conservation. We used a target-capture method to sequence and analyze 238 nuclear loci across all species of scrub mints, reconstructed the phylogeny, and calculated gene tree concordance, gene tree estimation error, and reticulation indices for every node in the tree using ML methods. Phylogenetic networks were used to determine reticulation events. Our nuclear phylogenetic estimates were consistent with previous results, while greatly increasing the robustness of taxon sampling. The phylogeny resolved the full relationship between Dicerandra and Conradina and the less-studied members of the clade (Piloblephis, Stachydeoma, Clinopodium spp.). We found hotspots of gene tree discordance and reticulation throughout the tree, especially in perennial Dicerandra. Several instances of reticulation events were uncovered between annual and perennial Dicerandra, and within the Conradina + allies clade. Incomplete lineage sorting also likely contributed to phylogenetic discordance. These results clarify phylogenetic relationships in the clade and provide insight on important evolutionary drivers in the clade, such as hybridization. General relationships in the group were confirmed, while the large amount of gene tree discordance is likely due to reticulation across the phylogeny.

Complete genome sequence of a novel robigovirus infecting Mentha arvensis.

The complete genomic sequence of a novel robigovirus, provisionally named "Mentha arvensis robigovirus 1" (MARV1), was determined by combining next-generation sequencing (NGS), reverse transcription polymerase chain reaction (RT-PCR), and rapid amplification of cDNA ends (RACE) PCR. The complete genomic sequence of this new virus is 7617 nucleotides in length, excluding the 3' poly(A) tail. The MARV1 genome encodes a putative replicase, "triple gene block" proteins, and a coat protein. Phylogenetic analysis demonstrated that MARV1 is a member of the genus Robigovirus, with closest relationships to African oil palm ringspot virus (AOPRV). Furthermore, MARV1-derived small interfering RNAs (siRNAs) showed typical patterns of plant-virus-derived siRNAs produced by the host antiviral RNA interference pathway. This is the first report of a plant virus of the genus Robigovirus in M. arvensis.

Larvicidal potential, antimicrobial properties and molecular docking analysis of Egyptian Mint (Mentha rotundifolia) against Culex pipiens L. (Diptera: Culicidae) and Midgut-borne Staphylococcus aureus.

Mosquitoes prefer stagnant areas near hospitals to live and easily spread pathogenic bacteria. Our current study aims to isolate multidrug-resistant (MDR) Staphylococcus aureus isolates from midguts of Mosquito Culex pipiens and study the potential of mint as a biocontrol strategy against C. pipiens larvae and their midgut-borne S. aureus. Samples of the third and fourth larval instars of C. pipiens were collected from water ponds around three Cairo hospitals. Ciprofloxacin, gentamycin and tetracycline, as well as various concentrations of mint leaf extract (MLE) were tested for antibiotic susceptibility. Sixty-five isolates were obtained and showed antibiotic resistance to tetracycline, gentamycin, ciprofloxacin, and undiluted MLE with resistant percentages (%) of 27.69, 30.76, 17.46, and 23.08%, respectively. Undiluted MLE inhibited 61.53% of the multidrug S. aureus isolates, whereas it couldn't inhibit any of these isolates at dilutions less than 50 µg/mL. The MIC of MLE was ≤ 700 µg/mL, while the MIC of the antibiotics ranged from 0.25 to 5.0 µg/mL for the three antibiotics. The most inhibited S. aureus isolate was identified by 16SrRNA sequencing approach and registered in GenBank as S. aureus MICBURN with gene accession number OQ766965. MLE killed all larval stages after 72 h of exposure, with mortality (%) reaching 93.33 and 100% causing external hair loss, breakage of the outer cuticle epithelial layer of the abdomen, and larvae shrinkage. Histopathology of treated larvae showed destruction of all midgut cells and organelles. Gas chromatography (GC) of MLE revealed that menthol extract (35.92%) was the largest active ingredient, followed by menthone (19.85%), D-Carvone (15.46%), Pulegone (5.0579%). Docking analysis confirmed that alpha guanine and cadinol had the highest binding affinity to both predicted active sites of Culex pipiens acetylcholinesterase. As a result, alpha-guanine and cadinol might have a role as acetylcholinesterase inhibitors.

Variability in Chemical Composition and Biochemical Activities of Mentha x piperita L. Essential Oil, in Response to Mycorrhizal Symbiosis and Heavy Metal Stress.

The present paper highlights the effect of Pb/Cd-stress and/or mycorrhizal colonization by Glomus Intraradices on yield, chemical composition, cytotoxicity and antimicrobial activity of Mentha x piperita L. essential oil. Our findings showed that mycorrhizal colonization could be used to improve the essential oil yield of M. x piperita, either in non-stressed or Pb/Cd-stressed plants. GC-MS analysis revealed three chemotypes: linalool/pulegone (32.6/30.8 %) chemotype in essential oils of non-mycorrhizal Pb-stressed plants, menthone/menthyl acetate (30.3/25.1 %) chemotype in essential oils of non-mycorrhizal Cd-stressed plants and menthol (44.6 %) chemotype in essential oils of non-mycorrhizal non-stressed plants, mycorrhizal non-stressed plants and mycorrhizal Pb/Cd-stressed plants. The cytotoxicity of M. x piperita essential oil, evaluated by brine shrimp lethality bioassay, was increased in presence of Pb/Cd-stress (from 379.58 to 72.84 µm/mL) and decreased in mycorrhizal plants (from 379.58 to 482.32 µm/mL). The antimicrobial activity of M. x piperita essential oil, evaluated by disc diffusion method and determination of Minimum Inhibitory Concentration against ten microorganisms, was enhanced by the mycorrhizal colonization and deceased by the Pb/Cd-stress. In conclusion, the inoculation of medicinal plants with mycorrhizal fungi is a real avenue for alleviating abiotic stress and/or increasing the quantity and quality of secondary metabolites in terms of biological activities.

Interrogation and validation of the interactome of neuronal Munc18-interacting Mint proteins with AlphaFold2.

Munc18-interacting proteins (Mints) are multidomain adaptors that regulate neuronal membrane trafficking, signaling, and neurotransmission. Mint1 and Mint2 are highly expressed in the brain with overlapping roles in the regulation of synaptic vesicle fusion required for neurotransmitter release by interacting with the essential synaptic protein Munc18-1. Here, we have used AlphaFold2 to identify and then validate the mechanisms that underpin both the specific interactions of neuronal Mint proteins with Munc18-1 as well as their wider interactome. We found that a short acidic α-helical motif within Mint1 and Mint2 is necessary and sufficient for specific binding to Munc18-1 and binds a conserved surface on Munc18-1 domain3b. In Munc18-1/2 double knockout neurosecretory cells, mutation of the Mint-binding site reduces the ability of Munc18-1 to rescue exocytosis, and although Munc18-1 can interact with Mint and Sx1a (Syntaxin1a) proteins simultaneously in vitro, we find that they have mutually reduced affinities, suggesting an allosteric coupling between the proteins. Using AlphaFold2 to then examine the entire cellular network of putative Mint interactors provides a structural model for their assembly with a variety of known and novel regulatory and cargo proteins including ADP-ribosylation factor (ARF3/ARF4) small GTPases and the AP3 clathrin adaptor complex. Validation of Mint1 interaction with a new predicted binder TJAP1 (tight junction-associated protein 1) provides experimental support that AlphaFold2 can correctly predict interactions across such large-scale datasets. Overall, our data provide insights into the diversity of interactions mediated by the Mint family and show that Mints may help facilitate a key trigger point in SNARE (soluble N-ethylmaleimide-sensitive factor attachment receptor) complex assembly and vesicle fusion.

Impact of induced mutation-derived genetic variability, genotype and varieties for quantitative and qualitative traits in Mentha species.

PURPOSE: The genus Mentha spp. is an aromatic herb from the family 'Lamiaceae'. It is extensively predominant in temperate and sub-temperate regions of the world. The essential oil of this species is enriched with broad aroma constituents extensively utilized in food, beverages, flavor, cosmetics, perfumery, and pharmaceutical enterprises. With the global menthol market size estimated to be worth USD 765 million in 2022, India (accompanied by China and Brazil) is the world's primary manufacturer, consumer, and exporter of Mentha oil. Despite prominent global demand, the crucial bottleneck in mint cultivation is the need for more superior commercial cultivars. Predominant vegetative propagation mode with difficulties in manual emasculation, differential blooming times, sterile/sub-sterile hybrids, and low seed viability are the primary containment in creating genetic variability by classical breeding approaches. Therefore, genetic complications encountered in conventional breeding have led the breeders to apply mutation breeding as an alternative crop improvement approach in Mentha spp. These attempts at mutation breeding have produced some distinctive mutants as genetic pools for plant breeding programs, and some novel mutant mint cultivars have been made available for commercial cultivation. CONCLUSIONS: The prime strategy in mutation-based breeding has proven an adept means of encouraging the expression of recessive genes and producing new genetic variations. The present review comprises a significant contribution of mutation breeding approaches in the development of mutant mint species and its effects on physiological variation, photosynthetic pigment, essential oil content and composition, phytochemical-mediated defense response, pathogen resistivity, and differential expression of genes related to terpenoid biogenesis. Development and diversification have led to the release of varieties, namely Todd's Mitcham, Murray Mitcham, Pranjal, Tushar, and Kukrail in M. piperita L., Mukta, and Pratik in M. cardiaca Baker, Neera in M. spicata L., Kiran in M. citrata Ehrh., and Rose mint in M. arvensis L. that have revolutionized and uplifted mint cultivation leading to economic gain by the farmers and entrepreneurs.