Diversity of Medicinal Plants Used by the Local Communities of the Coastal Plateau of Safi Province (Morocco).

Traditional herbal medicine is still used for basic healthcare by a significant portion of the population in developing countries. This study aimed to explore the medicinal plant's diversity and to document related traditional knowledge in the Safi region of Morocco. We used semi-structured questionnaires to interview 222 informants living in the study area. To perform data analysis, we used quantitative indices like use value (UV), family use value (FUV), fidelity level (FL), the relative popularity level (RPL), rank of order priority (ROP), and informant consensus factor (ICF). We reported the ethnomedicinal uses of 144 medicinal plants belonging to 64 families. According to the findings, the dominating families were Lamiaceae (17 taxa), Asteraceae (15 taxa), and Apiaceae (12 taxa). The most commonly utilized plant part (48%) was leaves. The decoction was reported as the main preparation method (42%). Highly cited plant species were Marrubium vulgare (UV = 0.56), Salvia rosmarinus Spenn. (UV = 0.47), Thymus serpyllum (UV = 0.32), and Dysphania ambrosioides (UV = 0.29). Papaveraceae (FUV = 0.26), and Urticaceae (FUV= 0.23), Geraniaceae (FUV = 0.17), Oleaceae (FUV = 0.17), Lamiaceae (FUV = 0.17) had the highest family use-values. Gastrointestinal disorders (88%), respiratory diseases (85%), and anemia (66%) have the greatest ICF values. This study reveals the indigenous people's reliance on plant-derived traditional medicine to prevent, alleviate, and treat a broad range of health concerns. Our findings will provide a scientific basis for ethnomedicinal legacy conservation and further scientific investigations aimed at new natural bioactive molecules discovery.

Tinnea gombea (Lamiaceae), a new species from the Sudanian savanna region, Nigeria based on integrative evidence.

Tinnea gombea, endemic to the Sudan savanna grasslands in northern Nigeria, is described and illustrated. We used integrative evidence from morphological characters, ecology and molecular phylogenetic data. The new species is morphologically and ecologically similar to T. barteri and T. aethiopica, but can be readily delimited from these taxa by unique characters including a subshrub growth habit, leaves alternate to subopposite, blades lanceolate, apically acuminate, inflorescences raceme, bearing solitary flowers in upper leaf and bract axils, lilac to purplish dusky flowers and the inflated fruits dehiscent. The distribution and habitat of T. gombea are also distinctive, being restricted to the Sudan savanna, while the two most similar species are widespread in tropical Africa. Additionally, molecular phylogenetic assessments using nrITS and chloroplast trnL-F, matK and rbcL support the placement of T. gombea as a distinct species. Tinnea gombea is here assessed as Critically Endangered due to its small population size and restriction to a small area lacking conservation prioritization.

Anti-Candida and Antibiofilm Activity of Selected Lamiaceae Essential Oils.

BACKGROUND: Candidiasis is a common oral and vaginal infection. Some papers have presented that the essential oils of Lamiaceae plants can have antifungal activity. This study aimed to investigate the activity of 7 essential oils of the Lamiaceae family with known phytochemical compositions against Candida fungi. METHODS: Forty-four strains belonging to six species were tested: C. albicans, C. glabrata, C. guilliermondii, C. krusei, C. parapsilosis, and C. tropicalis. During this investigation, the following methods were used: determination of the minimal inhibitory concentrations (MICs), biofilm inhibition studies, and in silicotoxicity tests. RESULTS: Essential oils of lemon balm (Melissa officinalis) and oregano (Origanum vulgare) showed the best anti-Candida activity, with MIC values below 3.125 mg/mL. Lavender (Lavandula stoechas), mint (Mentha × piperita), rosemary (Rosmarinus officinalis), and thyme (Thymus vulgaris) essential oils were also very active (0.39 to 6.25 or 12.5 mg/mL). Sage (Salvia officinalis) essential oil presented the lowest activity, with MIC values ranging from 3.125 to 100 mg/mL. In an antibiofilm study using MIC values, oregano and thyme essential oils showed the greatest effect, followed by lavender, mint, and rosemary oils. The weakest antibiofilm activity was observed with the lemon balm and sage oils. In silico toxicity research suggests that most of main compounds of Lamiaceae essential oils probably do not exhibit carcinogenicity, mutagenicity, or cytotoxicity. CONCLUSIONS: The obtained results showed that Lamiaceae essential oils have anti-Candida and antibiofilm activity. Further research is required to confirm the safety and efficacy of essential oils in the topical treatment of candidiasis.

Toxicity and Physiological Effects of Nine Lamiaceae Essential Oils and Their Major Compounds on Reticulitermes dabieshanensis.

The volatile metabolites of Salvia sclarea, Rosmarinus officinalis, Thymus serpyllum, Mentha spicata, Melissa officinalis, Origanum majorana, Mentha piperita, Ocimum basilicum and Lavandula angustifolia were determined by gas chromatography-mass spectrometry. The vapor insecticidal properties of the analyzed essential oils and their compounds were screened using Reticulitermes dabieshanensis workers. The most effective oils were S. sclarea (major constituent linalyl acetate, 65.93%), R. officinalis (1,8-cineole, 45.56%), T. serpyllum (thymol, 33.59%), M. spicata (carvone, 58.68%), M. officinalis (citronellal, 36.99%), O. majorana (1,8-cineole, 62.29%), M. piperita (menthol, 46.04%), O. basilicum (eugenol, 71.08%) and L. angustifolia (linalool, 39.58%), which exhibited LC50 values ranging from 0.036 to 1.670 µL/L. The lowest LC50 values were recorded for eugenol (0.060 µL/L), followed by thymol (0.062 µL/L), carvone (0.074 µL/L), menthol (0.242 µL/L), linalool (0.250 µL/L), citronellal (0.330 µL/L), linalyl acetate (0.712 µL/L) and 1,8-cineole (1.478 µL/L). The increased activity of esterases (ESTs) and glutathione S-transferase (GST) were observed but only alongside the decreased activity of acetylcholinesterase (AChE) in eight main components. Our results indicate that S. sclarea, R. officinalis, T. serpyllum, M. spicata, M. officinalis, O. marjorana, M. piperita, O. basilicum and L. angustifolia essential oils (EOs) and their compounds, linalyl acetate, 1,8-cineole, thymol, carvone, citronellal, menthol, eugenol and linalool could be developed as control agents against termites.

Essential Oil Composition and Enantioselective Profile of Agastache urticifolia (Lamiaceae) and Monardella odoratissima (Lamiaceae) from Utah.

Two species within the Lamiaceae (mint) family, Agastache urticifolia and Monardella odoratissima, are aromatic plants that are native to the Intermountain Region (USA). Essential oil produced through steam distillation was examined to establish the essential oil yield and both the achiral and chiral aromatic profiles of both plant species. The resulting essential oils were analyzed by GC/MS, GC/FID, and MRR (molecular rotational resonance). For A. urticifolia and M. odoratissima, achiral essential oil profiles were largely composed of limonene (71.0%, 27.7%), trans-ß-ocimene (3.6%, 6.9%), and pulegone (15.9%, 4.3%), respectively. Between the two species, eight chiral pairs were analyzed and, interestingly, the dominant enantiomer (calculated as ee%) of limonene and pulegone switched between the two species. Where enantiopure standards were not commercially available, MRR was used as a reliable analytical technique for chiral analysis. This study verifies the achiral profile for A. urticifolia and, for the first time to the authors' knowledge, establishes the achiral profile for M. odoratissima and chiral profile for both species. Additionally, this study confirms the utility and practicality of using MRR for determining chiral profiles in essential oils.

Environmentally-Friendly Pesticidal Activities of Callicarpa and Karomia Essential Oils from Vietnam and Their Microemulsions.

There is an ongoing interest to identify alternative pesticidal agents to avoid the chronic problems associated with synthetic pesticides. Essential oils have shown promise as botanical pest control agents. In the present study, the essential oils of four members of the Lamiaceae (Callicarpa candicans, C. erioclona, C. macrophylla, and Karomia fragrans; Vietnamese names: Nàng nàng, Tu châu lông mem, Tu châu lá to and Cà dien, respectively), obtained from wild populations in Vietnam, have been obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. The essential oils were formulated into microemulsions and the essential oils and their microemulsions were screened for mosquito larvicidal activity against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and for molluscicidal activity against Pomacea canaliculata. Atractylone and (E)-caryophyllene dominated the volatiles of C. candicans (CCEO) and C. erioclona (CEEO), while the major component in C. macrophylla (CMEO) and K. fragrans (KFEO) was (E)-caryophyllene. The essential oils and microemulsions of both C. candicans and C. erioclona exhibited excellent larvicidal activity against all three mosquito species (Ae. aegypti, Ae. albopictus, and Cx. quinquefasciatus) with LC50 values <10 µg/mL. Additionally, the larvicidal activity of the microemulsions were significantly improved compared with their free essential oils, especially for C. candicans and C. erioclona. All four essential oils and their microemulsions showed excellent molluscicidal activity with LC50 <10 µg/mL. In most cases, the essential oils and microemulsions showed greater pesticidal activity against target organisms than the non-target freshwater fish, Oreochromis niloticus. The in silico studies on physicochemical and ADMET properties of the major components in the studied essential oils were also investigated and most of the compounds possessed a favorable ADMET profile. Computational modeling studies of the studied compounds demonstrated a favorable binding interaction with the mosquito odorant-binding protein target and support atractylone, ß-selinene, and caryophyllene oxide as potential inhibitors. Based on the observed pesticidal activities of the essential oils and their microemulsions, the Callicarpa species and K. fragrans should be considered for potential cultivation and further exploration as botanical pesticidal agents.

Aryltetralin lignans from Hyptis brachiata inhibiting T lymphocyte proliferation.

Increased activation and proliferation of T lymphocytes plays an essential role in the development of chronic inflammation and autoimmune diseases. Currently used immunosuppressive drugs often do not provide long-lasting relief of symptoms and show a gradual loss of efficacy over time, and are accompanied by various side effects. Therefore, novel immunosuppressive lead substances are needed. For this purpose, an in-house library consisting of 600 extracts of plants from Panama was screened for inhibition of human T lymphocyte proliferation. As one of the hits, an ethyl acetate extract from the aerial parts of Hyptis brachiata (Lamiaceae) exhibited strong inhibitory effects. Subsequent investigation resulted in the isolation of seven aryltetralin lignans, five arylnaphthalene lignans, two flavonoids, three triterpenes, and cinnamyl cinnamate. Aryltetralin lignans inhibited T lymphocyte proliferation in a concentration-dependent manner without induction of apoptosis. No relevant inhibition was observed for the arylnaphthalene lignans, flavonoids, and triterpenes. Additional cell cycle arrest investigations revealed that isolated aryltetralin lignans potently inhibited cell division in G2/M phase similarly to podophyllotoxin. Multifluorescence panel analyses of the extract also showed weak suppressive effects on the production of IL-2 and TNF-α. Therefore, preparations made out of H. brachiata could be further explored as an interesting herbal alternative in the treatment of autoimmune diseases.

The anti-leukemic activity of a luteolin-apigenin enriched fraction from an edible and ethnomedicinal plant, Elsholtzia stachyodes, is exerted through an ER stress/autophagy/cell cycle arrest/ apoptotic cell death signaling axis.

BACKGROUND: Elsholtzia is a genus in the family Lamiaceae, and some species in this genus are commonly used for food and in ethnomedicinal formulations by some ethnic groups of China and Thailand. Despite their apparent utility, few studies have been conducted to evaluate their potential as sources of medicinally active agents. PURPOSE: We aimed to investigate the cytotoxicity of ethanolic extracts from three selected edible plant species of the genus Elsholtzia and the most promising extract was further characterized for the bioactive constituents and signaling mechanisms associated with the anti-leukemic activity. MATERIALS AND METHODS: Ethanolic extracts were screened for cytotoxicity using flow cytometry. HPLC and LC-MS were used to analyze the chemical constituents of the most potent fraction from E. stachyodes. The relevant mechanism of action was assessed by western blot and multispectral imaging flow cytometry (MIFC). RESULTS: The most potent anti-leukemic activity was observed with the ethanolic extract from E. stachyodes. Luteolin and apigenin were characterized as the major constituents in the fraction from E. stachyodes. Mechanistically, the luteolin-apigenin enriched fraction (LAEF) induced the UPR, increased autophagic flux, induced cell cycle arrest and apoptotic cell death. LAEF showed significantly less cytotoxicity towards peripheral blood mononuclear cells (PBMCs) as compared to leukemia cell lines. CONCLUSION: This study is the first to report E. stachyodes as a new source of luteolin and apigenin which are capable of triggering leukemic cell death. This could lead to a novel strategy against leukemia using ethnomedicinal plant extracts as an alternative or supplemental anti-cancer agent.

Conserved chloroplast genome sequences of the genus Clerodendrum Linn. (Lamiaceae) as a super-barcode.

BACKGROUND: The plants of the genus Clerodendrum L. have great potential for development as an ornamental and important herbal resource. There is no significant morphological difference among many species of the genus Clerodendrum, which will lead to confusion among the herbs of this genus and ultimately affect the quality of the herbs. The chloroplast genome will contribute to the development of new markers used for the identification and classification of species. METHODS AND RESULTS: Here, we obtained the complete chloroplast genome sequences of Clerodendrum chinense (Osbeck) Mabberley and Clerodendrum thomsoniae Balf.f. using the next generation DNA sequencing technology. The chloroplast genomes of the two species all encode a total of 112 unique genes, including 80 protein-coding, 28 tRNA, and four rRNA genes. A total of 44-42 simple sequence repeats, 19-16 tandem repeats and 44-44 scattered repetitive sequences were identified. Phylogenetic analyses showed that the nine Clerodendrum species were classified into two clades and together formed a monophyletic group. Selective pressure analyses of 77 protein-coding genes showed that there was no gene under positive selection in the Clerodendrum branch. Analyses of sequence divergence found two intergenic regions: trnH-GUG-psbA, nhdD-psaC, exhibiting a high degree of variations. Meanwhile, there was no hypervariable region identified in protein coding genes. However, the sequence identities of these two intergenic spacers (IGSs) are greater than 99% among some species, which will result in the two IGSs not being used to distinguish Clerodendrum species. Analysis of the structure at the LSC (Large single copy) /IR (Inverted repeat) and SSC (Small single copy)/IR boundary regions showed dynamic changes. The above results showed that the complete chloroplast genomes can be used as a super-barcode to identify these Clerodendrum species. The study lay the foundation for the understanding of the evolutionary process of the genus Clerodendrum.

Activity of essential oils from leaves, flower buds and stems of Tetradenia riparia on Rhipicephalus (Boophilus) microplus larvae.

Around the world, the main problems of livestock are caused by ectoparasites, however, commercial acaracide are toxic to the environment and detrimental to One Health. Therefore, research has increasingly focused on development of natural products as alternatives for tick control. The purpose of this study was to evaluate the larvicidal effect on Rhipicephalus (Boophilus) microplus, through use of essential oils (EOs) extracted from the leaves, flower buds and stems of Tetradenia riparia. The chemical composition of these EOs was determined through gas chromatography coupled to mass spectrometry (GC-MS). They were tested on larvae at concentrations of 100.000 to 40 µg/mL, using the larval packet test and under semi-natural conditions. The main class of compounds in the chemical composition was sesquiterpenes (both oxygenates and hydrocarbons), whereas the predominant compounds in the leaves, flower buds and stems were 14-hydroxy-9-epi-caryophyllene, T-cadinol and 6-7-dehydroroyleanone, respectively. The leaves proved to be the most effective, with highest larvicidal activity (LC99.9 = 83.53 µg/mL). When tested under semi-natural conditions, the oils obtained efficiency above 98% in all compound tests. The results indicated that these EOs were effective against R. (B.) microplus larvae in vitro and ex-situ, proving that this plant has bioactive molecules with significant larvicidal activity.

The total iridoid glycoside extract of Lamiophlomis rotata Kudo induces M2 macrophage polarization to accelerate wound healing by RAS/ p38 MAPK/NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Lamiophlomis rotata (Benth.) Kudo (L. rotata), a Tibetan medicinal plant, is used to treat "yellow-water diseases", such as skin disease, jaundice and rheumatism. Our previous study showed that the iridoid glycoside extract of L. rotata (IGLR) is the major constituent of skin wound healing. However, the role of IGLR in the biological process of trauma repair and the probable mechanism of the action remain largely unknown. AIM OF THE STUDY: To investigate the role of IGLR in the biological process of trauma repair and the probable mechanism of the action. MATERIALS AND METHODS: The role of IGLR in wound healing was investigated by overall skin wound in mice with Hematoxylin and Eosin (H&E) and Masson trichrome staining. The anti-inflammatory, angiogenesis-promoting and fibril formation effects of IGLR were visualized in wound skin tissue by immunofluorescence staining, and the proinflammatory factors and growth factors were assayed by real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Macrophages, dermal fibroblasts, and endothelial cells were cultured to measure the direct/indirect interaction effects of IGLR on the proliferation and migration of cells, and flow cytometry was employed to assess the role of IGLR on macrophage phenotype. Network pharmacology combined with Western blot experiments were conducted to explore possible mechanisms of the actions. RESULTS: IGLR increased the expression of CD206 (M2 markers) through the RAS/p38 MAPK/NF-κB signaling pathway during wound injury in vivo and in vitro. IGLR suppressed the inflammatory cytokines iNOS, IL-1ß and TNF-α in the early stage of wound healing. During the proliferation step of wound repair, IGLR promoted angiogenesis and fibril formation by increasing the expression of VEGF, CD31, TGF-ß and α-SMA in wound tissue, and similar results were verified by RT-PCR and ELISA. In a paracrine mechanism, the extract promoted the proliferation of dermal fibroblasts, and endothelial cells were founded by the conditioned medium (CM). CONCLUSION: IGLR induced M2 macrophage polarization in the early stage of wound healing; in turn, IGLR played a key role in the transition from inflammation to cell proliferation during the biological process of wound healing.

A chromosome-level genome assembly reveals that a bipartite gene cluster formed via an inverted duplication controls monoterpenoid biosynthesis in Schizonepeta tenuifolia.

Biosynthetic gene clusters (BGCs) are regions of a genome where genes involved in a biosynthetic pathway are in proximity. The origin and evolution of plant BGCs as well as their role in specialized metabolism remain largely unclear. In this study, we have assembled a chromosome-scale genome of Japanese catnip (Schizonepeta tenuifolia) and discovered a BGC that contains multiple copies of genes involved in four adjacent steps in the biosynthesis of p-menthane monoterpenoids. This BGC has an unprecedented bipartite structure, with mirrored biosynthetic regions separated by 260 kilobases. This bipartite BGC includes identical copies of a gene encoding an old yellow enzyme, a type of flavin-dependent reductase. In vitro assays and virus-induced gene silencing revealed that this gene encodes the missing isopiperitenone reductase. This enzyme evolved from a completely different enzyme family to isopiperitenone reductase from closely related Mentha spp., indicating convergent evolution of this pathway step. Phylogenomic analysis revealed that this bipartite BGC has emerged uniquely in the S. tenuifolia lineage and through insertion of pathway genes into a region rich in monoterpene synthases. The cluster gained its bipartite structure via an inverted duplication. The discovered bipartite BGC for p-menthane biosynthesis in S. tenuifolia has similarities to the recently described duplicated p-menthane biosynthesis gene pairs in the Mentha longifolia genome, providing an example of the convergent evolution of gene order. This work expands our understanding of plant BGCs with respect to both form and evolution, and highlights the power of BGCs for gene discovery in plant biosynthetic pathways.

Uncovering a miltiradiene biosynthetic gene cluster in the Lamiaceae reveals a dynamic evolutionary trajectory.

The spatial organization of genes within plant genomes can drive evolution of specialized metabolic pathways. Terpenoids are important specialized metabolites in plants with diverse adaptive functions that enable environmental interactions. Here, we report the genome assemblies of Prunella vulgaris, Plectranthus barbatus, and Leonotis leonurus. We investigate the origin and subsequent evolution of a diterpenoid biosynthetic gene cluster (BGC) together with other seven species within the Lamiaceae (mint) family. Based on core genes found in the BGCs of all species examined across the Lamiaceae, we predict a simplified version of this cluster evolved in an early Lamiaceae ancestor. The current composition of the extant BGCs highlights the dynamic nature of its evolution. We elucidate the terpene backbones generated by the Callicarpa americana BGC enzymes, including miltiradiene and the terpene (+)-kaurene, and show oxidization activities of BGC cytochrome P450s. Our work reveals the fluid nature of BGC assembly and the importance of genome structure in contributing to the origin of metabolites.

Antioxidant, antibacterial, and antileishmanial potential of Micromeria nervosa extracts and molecular mechanism of action of the bioactive compound.

AIMS: This study aimed to determine the antibacterial and antileishmanial potential of Micromeria nervosa extracts. The identification of the antileishmanial compound and the study of its molecular mechanism of action have also been undertaken. METHODS AND RESULTS: Ethanol extract showed high polyphenol content and diethyl ether extract exhibited high DPPH scavenging and low beta-carotene bleaching activity (IC50 = 13.04 ± 0.99 and 200.18 ± 3.32 µg mL-1, respectively). However, diethyl ether extract displayed high antibacterial activity against Gram-positive strains including methicillin-resistant Staphylococcus aureus (MIC = 31.25 µg mL-1), Staph. aureus ATCC6538 (MIC = 62.5 µg mL-1), and Listeria monocytogenes ATCC 19115 (MIC = 125 µg mL-1), as well as high antileishmanial activity against the promastigote forms of L. infantum and L. major (IC50 = 11.45 and 14.53 µg mL-1, respectively). The active compound was purified using bioassay-guided fractionation and thin layer chromatography, and identified as ursolic acid using high-performance liquid chromatography coupled with a photodiode array and mass spectrometry. The purified compound was strongly inhibitory against the promastigote and amastigote forms of L. infantum and L. major (IC50 = 5.87 and 6.95 µg mL-1 versus 9.56 and 10. 68 µg mL-1, respectively) without overt cytotoxicity against Raw 264.7 macrophage cells (SI = 13.53 and 11.43, respectively). The commercial compound (ursolic acid) showed similar activity against amastigotes and promastigotes forms of L. infantum and L. major. Moreover, its molecular mode of action against leishmaniasis seems to involve the expression of the ODC and SPS genes involved in thiol pathway. CONCLUSION: Extracts of M. nervosa can be considered as a potential alternative to antimicrobial and antileishmanial drugs.

Investigation of Dracocephalum extract based on bulk and nanometer size as green corrosion inhibitor for mild steel in different corrosive media.

In recent years, green corrosion inhibitors derived from natural plant resources have garnered much interest. In the present work, at first, we investigated the corrosion behavior of mild steel (st-37) in the presence, and absence of Dracocephalum extract based on bulk size as a corrosion inhibitor in two widely used acidic environments (0.5 M H2SO4, and 1.0 M HCl), at room temperature. Then, we used Dracocephalum extract based on nanometer size to reduce the optimal concentration of inhibitor, increase the corrosion resistant, and efficiency. Dracocephalum extract does not contain heavy metals or other toxic compounds, and also good characteristics such as low cost, eco-friendly, and widespread availability, make it suitable nature candidate as an environmentally safe green inhibitor. The anticorrosive behavior was assessed using electrochemical impedance spectroscopy (EIS), and potentiodynamic polarization (PP). In all of the studies, the inhibitory efficiency (IE%) increased as the extract dose was increased. But by using nano extract, in addition to maintaining high efficiency, the amount of inhibitor was reduced significantly. The highest IE% is 94% at the best dose of nano extract (75 ppm), but the highest IE% is 89% at the best dose of the bulk extract (200 ppm) in H2SO4 solution. Also, for the HCl solution, the highest IE% is 88% at the best dose of nano extract (100 ppm), but the highest IE% is 90% at the best dose of the bulk extract (400 ppm), by polarization method. The PP results suggest that this compound has an effect on both anodic, and cathodic processes, and that it adsorbs on mild steel surface according to the Langmuir adsorption isotherm. Optical microscopy, scanning electron microscopy (SEM) analysis, and a solid UV-Visible reflection spectrum were used to investigate the alloys' surface morphology.

A chromosome-level genome assembly reveals that tandem-duplicated CYP706V oxidase genes control oridonin biosynthesis in the shoot apex of Isodon rubescens.

The ent-kaurenoids (e.g., oridonin and enmein) from the Isodon genus (Lamiaceae) are one class of diterpenoids with rich structural diversity and intriguing pharmaceutical activity. In contrast to the well-established gibberellin pathway, oxidative modifications diversifying the ent-kaurene skeleton in Isodon have remained undetermined for half a century. Here we report a chromosome-level genome assembly of I. rubescens, a well-recognized oridonin producer long favored by Asian people as a traditional herb with antitumor effects. The shoot apex was confirmed to be the actual region actively producing ent-kaurene diterpenoids. Through comparative genomics and phylogenetic analyses, we discovered a cluster of tandem-duplicated CYP706V oxygenase-encoding genes located on an ancient genomic block widely distributed in eudicots, whereas almost exclusively emerged in Isodon plants. In the shoot apex, IrCYP706V2 and IrCYP706V7 oxidized the ent-kaurene core in the initial stage of oridonin biosynthesis. Loss of CYP706Vs in other Lamiaceae plants offered an explanation for the specific kaurenoid production in Isodon plants. Moreover, we found that the Isodon genomes encode multiple diterpenoid synthases that are potentially involved in generating diterpenoid diversity. These findings provided new insights into the evolution of the lineage-specific diterpenoid pathway and laid a foundation for improving production of bioactive ent-kaurene-type diterpenoids by molecular breeding and synthetic biology approaches.

Phytochemistry and Polypharmacological Potential of Colebrookea oppositifolia Smith.

BACKGROUND: Colebrookea oppositifolia Smith. is a valuable traditional therapeutic plant belonging to the family Lamiaceae. It is a dense and wool-like shrub that is mostly found in subtropical regions of some countries of Asia, such as China and India. It has been widely used for the mitigation of nervous system disorders like epilepsy. The active constituents of the plant have exhibited antioxidant, anti-microbial, and antifungal properties, which are considered due to the presence of polyphenols and flavonoids as chief chemical constituents. Flavonoids like quercetin, landenein, chrysin, and 5, 6, 7-trimethoxy flavones cause protein denaturation of the microbial cell wall. OBJECTIVES: To comprehend and assemble the fragmented pieces of evidence presented on the traditional uses, botany, phytochemistry, and pharmacology of the plant to reconnoiter its therapeutic perspective and forthcoming research opportunities. METHODS: The available information on Colebrookea oppositifolia has been established by electronically searching peer-reviewed literature from PubMed, Google Scholar, Springer, Scopus, Web of Science, and Science Direct over the earlier few years. RESULTS: The plant has been greatly used for the preparation of many herbal medicines which are used for treating traumatic injuries, fever, rheumatoid arthritis, headache, and gastric problems. From the aerial parts of the plant, a phenylethanoid glycoside named acteoside has been isolated and evaluated for its therapeutic potential viz. immunomodulatory, neuroprotective, hepatoprotective, analgesic, anti-tumour, antispasmodic, antioxidant, antibacterial, free radical scavenger, and improving sexual function. Acteoside showed neuroprotective activities against Aß-peptide, which is neurotoxic and causes apoptosis. The petroleum ether extract of the plant leaves offers many active compounds like sitosterol, n-triacontane, hydroxydotriacontyl ferulate, acetyl alcohol, and 3,7,4,2-tetramethoxyflavones which have shown hepatoprotective potential. CONCLUSION: The plant should be evaluated further for the estimation of some other health benefits. The consequences of restricted pharmacological screening and reported phytomolecules of Colebrookea oppositifolia Smith. advocate that there is still an exigent requisite for in-depth pharmacological studies of the plant.

Diversity of medicinal plants and their therapeutic usages of Kachin people (Jinghpaw) in the central part of Kachin State, Myanmar.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional medical system plays a major role in healthcare in Kachin State, Myanmar, where long-term political instability persists and conventional healthcare facilities are inadequate. A knowledge of the traditional medicinal plants therefore benefits the Kachin people, yet documentation and records of the uses of these plants are rare. In this study, we attempt to answer the questions on what medicinal plants and how they are used by the Kachin people. AIM OF THE STUDY: We aimed to document knowledge of the traditional medicinal plants and to identify those most frequently used by the Kachin people. MATERIALS AND METHODS: Eighty-two informants from eight villages in three townships were interviewed, and their knowledge of medicinal plants was recorded. The reported ailments were classified to the standard categories of the International Classification of Primary Care-2 (ICPC-2) system. Use reports (UR) were employed to evaluate the knowledge consensus of the informants. RESULTS: We recorded a total of 117 species used as medicinal plants, of which 22 are newly recorded medicinal plant species for Myanmar. The plants belonged to 103 genera in 52 families, and were used to treat a total of 72 ailments from 17 ICPC-2 disease categories. Fabaceae and Lamiaceae were the most highly represented families of medicinal plants, with eleven and eight species used, respectively. The most cited species based on URs were Tinospora cordifolia (Willd.) Hook.f. & Thomson (URs = 39), Oroxylum indicum (L.) Kurz (URs = 28), Aquilaria malaccensis Lam. (URs = 26), Chromolaena odorata (L.) R.M.King & H.Rob. (URs = 24), and Chloranthus elatior Link. (URs = 22). Digestive system disorder was the most prevalent disease category, and was treated with 47 different medicinal plants (URs = 142). Leaves were the most commonly used plant part; decoction was the dominant method of preparation; and oral consumption was the most frequent method of administration. CONCLUSION: Our study documented a list of 117 medicinal plants and their uses in traditional medicine based on the local knowledge of the Kachin people. The study also identified the five most frequently cited species and found that the plants investigated are used to treat a total of 72 diseases. The 642 therapeutic reports we collected showcase a rich and diverse living knowledge of medicinal plant use by the Kachin people. Moreover, we present 22 new medicinal records, enriching the list of known medicinal plants in Myanmar. This exploratory study has enabled us to assemble the local knowledge of the Kachin people into solid dataset that will allow further scientific validation and will potentially contribute to better integration of medicinal plants into the healthcare provision for Kachin people in Myanmar.

Eco-friendly synthesis of Ag-NPs using Endostemon viscosus (Lamiaceae): Antibacterial, antioxidant, larvicidal, photocatalytic dye degradation activity and toxicity in zebrafish embryos.

Nanotechnology is a multidisciplinary area of study that has grown significantly in serving many functions and impacting human society. New fields of science have been facilitated by the clean, non-toxic, and biocompatible nature of plant-derived nanoparticles. The present study deals with the first green synthesis of silver nanoparticles (Ag-NPs) using Endostemon viscosus, and their synthesized Ag NPs were characterized by different spectral methods (UV-vis Spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction Spectroscopy (XRD), Transmission Electron Microscopy (TEM) and Energy dispersive X-ray Spectroscopy (EDAX). The change initially observed the production of Ag-NPs in color from green to ash and then confirmed by SPR band at 435 nm in UV-vis spectral analysis. The FTIR findings indicate that many functional groups belong to the pharmaceutically useful phytochemicals, which interact as reducing, capping, and stabilizing agents in synthesizing silver nanoparticles. The predominant peaks in the XRD pattern belong to the planes 210°, 111°, 200°, 241°, and 311° and thus demonstrated the Ag-NPs FCC crystal structure. TEM analysis exhibited spherical-shaped particles with an average size of 13 nm, and the EDAX band showed a distinctive metallic silver peak at 3.0 keV. The antibacterial activity of Ag-NPs tested to show a maximum zone of inhibition of 19 mm for Staphylococcus aureus and 15 mm for Escherichia coli at 100 µg/mL, respectively. Bio-fabricated Ag-NPs were assessed for antioxidant activity (DPPH with % inhibition 57.54% and FRAP with % inhibition 70.89%). The biosynthesized Ag-NPs demonstrated potential larvicidal efficacy against Aedes aegypti with more than 90% at 250 µg/mL. Histological profiles were altered while treating with Ag-NPs at 250 µg/mL. The photocatalytic activity of synthesized E. viscosus Ag-NPs was tested against methylene blue (MB) and crystal violet (CV), and the maximum degradation efficiency was found as 90 and 94%, respectively. Furthermore, the toxicity test on zebrafish embryos demonstrated that aberrations have only been induced at concentrations higher than 500 µg/mL. We conclude that the greenly produced Ag-NPs may find use in biomedical applications based on bacteria and cost-effective industrial wastewater treatment.

Controlled Production of Carnosic Acid and Carnosol in Cell Suspensions of Lepechinia meyenii Treated with Different Elicitors and Biosynthetic Precursors.

Lepechinia meyenii is a medicinal plant specialized in the biosynthesis of different types of antioxidants including the diterpenes carnosic (CA) acid and carnosol (CS). Herein we present the results of plant tissue culture approaches performed in this medicinal plant with particular emphasis on the generation and evaluation of a cell suspension system for CA and CS production. The effect of sucrose concentration, temperature, pH, and UV-light exposure was explored. In addition, diverse concentrations of microbial elicitors (salicylic acid, pyocyanin, Glucanex, and chitin), simulators of abiotic elicitors (polyethylene glycol and NaCl), and biosynthetic precursors (mevalonolactone, geranylgeraniol, and miltiradiene/abietatriene) were evaluated on batch cultures for 20 days. Miltiradiene/abietatriene obtainment was achieved through a metabolic engineering approach using a recombinant strain of Saccharomyces cerevisiae. Our results suggested that the maximum accumulation (Accmax ) of CA and CS was mainly conferred to stimuli associated with oxidative stress such as UV-light exposure (Accmax , 6.2 mg L-1 ) polyethylene glycol (Accmax , 6.5 mg L-1 ) NaCl (Accmax , 5.9 mg L-1 ) which simulated drought and saline stress, respectively. Nevertheless the bacterial elicitor pyocyanin was also effective to increase the production of both diterpenes (Accmax , 6.4 mg L-1 ). Outstandingly, the incorporation of upstream biosynthetic precursors such as geranylgeraniol and miltiradiene/abietatriene, generated the best results with Accmax of 8.6 and 16.7 mg L-1 , respectively. Optimized batch cultures containing 100 mg L-1 geranylgeraniol, 50 mg L-1 miltiradiene/abietatriene (95 : 5 %) and 5 g L-1 polyethylene glycol treated with 6 min UV light pulse during 30 days resulted in Accmax of 26.7 mg L-1 for CA and 17.3 mg L-1 for CS on days 18-24. This strategy allowed to increase seven folds the amounts of CA and CS in comparison with batch cultures without elicitation (Accmax , 4.3 mg L-1 ).