The local medicinal plant knowledge in Kashmir Western Himalaya: a way to foster ecological transition via community-centred health seeking strategies.

BACKGROUND: The mountainous region of Kashmir is a biodiversity hotspot, with diverse local communities and a rich cultural history linked to nature. Mountain ecosystems are highly vulnerable to climate change. This study emphasises the need to record the indigenous ethnoecological knowledge of wild plants used for the treatment of various ailments at higher elevations in remote areas where globalisation poses a threat to this traditional knowledge. METHODS: The field survey was carried out in 2020-2022, to collect data on wild medicinal plants. Informants were selected randomly to collect indigenous medicinal knowledge using semi-structured interviews and group discussions. Various quantitative indices were employed to evaluate ethnomedicinal data. RESULTS: A total of 110 medicinal plants belonging to 49 families were recorded in the study area. These medicinal plants are extensively used by local communities for the treatment of 20 major disease categories. Asteraceae was the dominant family contributing (9.09%) to medicinal plants, followed by Polygonaceae (8.18%), Apiaceae (7.27%), Lamiaceae (5.45%), and Ranunculaceae (5.45%). We observed 166 remedies were used for the treatment of various diseases in humans, and 9 remedies were used for animals. The most frequently used medicinal remedy was tea or decoction (30.91%). Among the medicinal plants, herbs (85.5%) were most frequently used by the local populations of Kashmir, whereas leaves (10.26%) were used for the treatment of various ailments. Out of 110 species, 31 were endemic, 15 of which are endemic to the Kashmir region and 16 to the Western Himalaya. The highest RFC value was reported for Allium humile (0.77), the highest UV value for Fritillaria cirrhosa (1.33), and the highest ICF value for gastro-intestinal/digestive disorders (0.85). CONCLUSIONS: Local communities still rely on wild medicinal plants for primary healthcare. These communities retained valuable indigenous knowledge, which needs to be preserved for the conservation and sustainable utilisation of natural resources. Further field exploration is required to fully explore indigenous knowledge in the mountainous regions of Kashmir, and this knowledge has the potential to support the ongoing ecological transition.

Antimicrobial efficacy of Mentha piperata-derived biogenic zinc oxide nanoparticles against UTI-resistant pathogens.

Misuse of antibiotics leads to the worldwide spread of antibiotic resistance, which motivates scientists to create new antibiotics. The recurring UTI due to antibiotics-resistant microorganism's challenges scientists globally. The biogenic nanoparticles have the potential to meet the escalating requirements of novel antimicrobial agents. The green synthesis of nanoparticles (NPs) gained more attention due to their reliable applications against resistant microbes. The current study evaluates the biogenic ZnO NPs of Mentha piperata extract against resistant pathogens of urinary tract infections by agar well diffusion assay. The biogenic ZnO NPs revealed comparatively maximum inhibition in comparison to synthetic antibiotics against two bacterial strains (Proteus mirabilis, Pseudomonas aeruginosa) and a fungal strain (Candida albicans).The synthesized biogenic ZnO NPs alone revealed maximum activities than the combination of plant extract (PE) and ZnO NPs, and PE alone. The physiochemical features of ZnO NPs characterized through UV-Vis spectroscopy, FTIR, XRD, SEM, and EDX. The UV-Vis spectroscopy revealed 281.85 nm wavelengths; the XRD pattern revealed the crystalline structure of ZnO NPs. The FTIR analysis revealed the presence of carboxylic and nitro groups, which could be attributed to plant extract. SEM analysis revealed spherical hollow symmetry due to electrostatic forces. The analysis via EDX confirmed the presence of Zn and oxygen in the sample. The physiochemical features of synthesized ZnO NPs provide pivotal information such as quality and effectiveness. The current study revealed excellent dose-dependent antimicrobial activity against the pathogenic isolates from UTI-resistant patients. The higher concentration of ZnONPs interacts with the cell membrane which triggers oxidative burst. They may bind with the enzymes and proteins and brings epigenetic alteration which leads to membrane disruption or cell death.

Evaluation of potential inhibitory effects on acetylcholinesterase, pancreatic lipase, and cancer cell lines using raw leaves extracts of three fabaceae species.

The present study examined the biological potential and phytochemicals of Sophora mollis, Mucuna pruriens, and Indigofera atropurpurea methanolic leaf extracts. In vitro anti-acetylcholinesterase and anti-lipase assays were performed using different concentrations of plant extracts, and the IC50 values were determined. The cytotoxic potential of the selected plant extracts was assessed against HeLa, PC3, and 3T3 cell lines using an MTT assay. S. mollis leaf extract displayed the highest inhibition percentage (114.60% ± 19.95 at 1000 µg/mL) for the anti-acetylcholinesterase activity with a prominent IC50 value of 75.9 µg/mL. The anti-lipase potential was highest with the M. pruriens leaf extract (355.5 µg/mL IC50), followed by the S. mollis extract (862.7 µg/mL IC50). Among the cell lines tested, the cytotoxic potential of the I. atropurpurea extract (91.1 ppm IC50) against the PC3 cell line was promising. High-performance liquid chromatography revealed gallic acid, chlorogenic acid, caffeic acid, vanillic acid, rutin trihydrate, and quercetin dihydrate in varying concentrations in all plant species. The concentration of chlorogenic acid (69.09 ppm) was highest in M. pruriens, and the caffeic acid concentration (45.20 ppm) was higher in S. mollis. This paper reports the presence of bioactive therapeutic compounds in selected species of the Fabaceae family that could be micro-propagated, isolated, and utilized in pharmaceutical industries.

Biofabrication of Fe3O4 Nanoparticles from Spirogyra hyalina and Ajuga bracteosa and Their Antibacterial Applications.

Iron oxide nanoparticles (NPs) have attracted substantial interest due to their superparamagnetic features, biocompatibility, and nontoxicity. The latest progress in the biological production of Fe3O4 NPs by green methods has improved their quality and biological applications significantly. In this study, the fabrication of iron oxide NPs from Spirogyra hyalina and Ajuga bracteosa was conducted via an easy, environmentally friendly, and cost-effective process. The fabricated Fe3O4 NPs were characterized using various analytical methods to study their unique properties. UV-Vis absorption peaks were observed in algal and plant-based Fe3O4 NPs at 289 nm and 306 nm, respectively. Fourier transform infrared (FTIR) spectroscopy analyzed diverse bioactive phytochemicals present in algal and plant extracts that functioned as stabilizing and capping agents in the fabrication of algal and plant-based Fe3O4 NPs. X-ray diffraction of NPs revealed the crystalline nature of both biofabricated Fe3O4 NPs and their small size. Scanning electron microscopy (SEM) revealed that algae and plant-based Fe3O4 NPs are spherical and rod-shaped, averaging 52 nm and 75 nm in size. Energy dispersive X-ray spectroscopy showed that the green-synthesized Fe3O4 NPs require a high mass percentage of iron and oxygen to ensure their synthesis. The fabricated plant-based Fe3O4 NPs exhibited stronger antioxidant properties than algal-based Fe3O4 NPs. The algal-based NPs showed efficient antibacterial potential against E. coli, while the plant-based Fe3O4 NPs displayed a higher zone of inhibition against S. aureus. Moreover, plant-based Fe3O4 NPs exhibited superior scavenging and antibacterial potential compared to the algal-based Fe3O4 NPs. This might be due to the greater number of phytochemicals in plants that surround the NPs during their green fabrication. Hence, the capping of bioactive agents over iron oxide NPs improves antibacterial applications.

Biosorption of Cadmium and Lead by Dry Biomass of Nostoc sp. MK-11: Kinetic and Isotherm Study.

Cadmium (Cd) and lead (Pb) are global environmental pollutants. In this study, Nostoc sp. MK-11 was used as an environmentally safe, economical, and efficient biosorbent for the removal of Cd and Pb ions from synthetic aqueous solutions. Nostoc sp. MK-11 was identified on a morphological and molecular basis using light microscopic, 16S rRNA sequences and phylogenetic analysis. Batch experiments were performed to determine the most significant factors for the removal of Cd and Pb ions from the synthetic aqueous solutions using dry Nostoc sp. MK1 biomass. The results indicated that the maximum biosorption of Pb and Cd ions was found under the conditions of 1 g of dry Nostoc sp. MK-11 biomass, 100 mg/L of initial metal concentrations, and 60 min contact time at pH 4 and 5 for Pb and Cd, respectively. Dry Nostoc sp. MK-11 biomass samples before and after biosorption were characterized using FTIR and SEM. A kinetic study showed that a pseudo second order kinetic model was well fitted rather than the pseudo first order. Three isotherm models Freundlich, Langmuir, and Temkin were used to explain the biosorption isotherms of metal ions by Nostoc sp. MK-11 dry biomass. Langmuir isotherm, which explains the existence of monolayer adsorption, fitted well to the biosorption process. Considering the Langmuir isotherm model, the maximum biosorption capacity (qmax) of Nostoc sp. MK-11 dry biomass was calculated as 75.757 and 83.963 mg g-1 for Cd and Pb, respectively, which showed agreement with the obtained experimental values. Desorption investigations were carried out to evaluate the reusability of the biomass and the recovery of the metal ions. It was found that the desorption of Cd and Pb was above 90%. The dry biomass of Nostoc sp. MK-11 was proven to be efficient and cost-effective for removing Cd and especially Pb metal ions from the aqueous solutions, and the process is eco-friendly, feasible, and reliable.

The Key Roles of ROS and RNS as a Signaling Molecule in Plant-Microbe Interactions.

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) play a pivotal role in the dynamic cell signaling systems in plants, even under biotic and abiotic stress conditions. Over the past two decades, various studies have endorsed the notion that these molecules can act as intracellular and intercellular signaling molecules at a very low concentration to control plant growth and development, symbiotic association, and defense mechanisms in response to biotic and abiotic stress conditions. However, the upsurge of ROS and RNS under stressful conditions can lead to cell damage, retarded growth, and delayed development of plants. As signaling molecules, ROS and RNS have gained great attention from plant scientists and have been studied under different developmental stages of plants. However, the role of RNS and RNS signaling in plant-microbe interactions is still unknown. Different organelles of plant cells contain the enzymes necessary for the formation of ROS and RNS as well as their scavengers, and the spatial and temporal positions of these enzymes determine the signaling pathways. In the present review, we aimed to report the production of ROS and RNS, their role as signaling molecules during plant-microbe interactions, and the antioxidant system as a balancing system in the synthesis and elimination of these species.

Molecular Characterization of Bacterial Isolates from Soil Samples and Evaluation of their Antibacterial Potential against MDRS.

Some soil microbes, with their diverse inhabitance, biologically active metabolites, and endospore formation, gave them characteristic predominance and recognition among other microbial communities. The present study collected ten soil samples from green land, agricultural and marshy soil sites of Khyber Pakhtunkhwa, Pakistan. After culturing on described media, the bacterial isolates were identified through phenotypic, biochemical and phylogenetic analysis. Our phylogenetic analysis revealed three bacterial isolates, A6S7, A1S6, and A1S10, showing 99% nucleotides sequence similarity with Brevibacillus formosus, Bacillus Subtilis and Paenibacillus dendritiformis. The crude extract was prepared from bacterial isolates to assess the anti-bacterial potential against various targeted multidrug-resistant strains (MDRS), including Acinetobacter baumannii (ATCC 19606), Methicillin-resistant Staphylococcus aureus (MRSA) (BAA-1683), Klebsiella pneumoniae (ATCC 13883), Pseudomonas aeruginosa (BAA-2108), Staphylococcus aureus (ATCC 292013), Escherichia coli (ATCC25922) and Salmonella typhi (ATCC 14028). Our analysis revealed that all bacterial extracts possess activity against Gram-negative and Gram-positive bacteria at a concentration of 5 mg/mL, efficiently restricting the growth of E. coli compared with positive control ciprofloxacin. The study concluded that the identified species have the potential to produce antimicrobial compounds which can be used to control different microbial infections, especially MDRS. Moreover, the analysis of the bacterial extracts through GC-MS indicated the presence of different antimicrobial compounds such as propanoic acid, oxalic acid, phenol and hexadecanoic acid.

Mentha: Nutritional and Health Attributes to Treat Various Ailments Including Cardiovascular Diseases.

A poor diet, resulting in malnutrition, is a critical challenge that leads to a variety of metabolic disorders, including obesity, diabetes, and cardiovascular diseases. Mentha species are famous as therapeutic herbs and have long served as herbal medicine. Recently, the demand for its products, such as herbal drugs, medicines, and natural herbal formulations, has increased significantly. However, the available literature lacks a thorough overview of Mentha phytochemicals' effects for reducing malnutritional risks against cardiovascular diseases. In this context, we aimed to review the recent advances of Mentha phytochemicals and future challenges for reducing malnutritional risks in cardiovascular patients. Current studies indicated that Mentha species phytochemicals possess unique antimicrobial, antidiabetic, cytotoxic, and antioxidant potential, which can be used as herbal medicine directly or indirectly (such as food ingredients) and are effective in controlling and curing cardiovascular diseases. The presence of aromatic and flavor compounds of Mentha species greatly enhance the nutritional values of the food. Further interdisciplinary investigations are pivotal to explore main volatile compounds, synergistic actions of phytochemicals, organoleptic effects, and stability of Mentha sp. phytochemicals.

Melatonin Function and Crosstalk with Other Phytohormones under Normal and Stressful Conditions.

Melatonin was discovered in plants in the late nineties, but its role, signaling, and crosstalk with other phytohormones remain unknown. Research on melatonin in plants has risen dramatically in recent years and the role of this putative plant hormone under biotic and abiotic stress conditions has been reported. In the present review, we discuss the main functions of melatonin in the growth and development of plants, its role under abiotic stresses, such as water stress (waterlogging and drought), extreme temperature (low and high), salinity, heavy metal, and light-induced stress. Similarly, we also discuss the role of melatonin under biotic stresses (antiviral, antibacterial, and antifungal effects). Moreover, the present review meticulously discusses the crosstalk of melatonin with other phytohormones such as auxins, gibberellic acids, cytokinins, ethylene, and salicylic acid under normal and stressful conditions and reports melatonin receptors and signaling in plants. All these aspects of melatonin suggest that phytomelatonin is a key player in crop improvement and biotic and abiotic stress regulation.

Catalytic potential of endophytes facilitates synthesis of biometallic zinc oxide nanoparticles for agricultural application.

Recent advances fascinated the use of biological resources in combination with metals to obtain high quality biometals and apply its advantages in different fields. Endophytic mediated Zinc oxide nanoparticles (ZnO-NPs) is an economical and ecofriendly way for farmers to avoid Zn deficiency in rice crop and obtain high yield. Here we synthesized ZnO-NPs utilizing endophytic bacterial strain of Enterobacter hormaechei (E. hormaechei). The physiochemical properties of the prepared NPs were determined through UV-Vis spectroscopy, XRD, FT-IR, SEM and TEM. The prepared NPs revealed surface plasmon resonance (SPR) at 320 nm (nm) and crystalline structure with 21 nm average crystalline size. FT-IR spectra showed the presence of carboxylic, alcohol and amine functional groups, which confirm the biometallic assembling of the ZnO and endophytic bacterial functional groups. SEM showed pyramidal symmetry whereas TEM revealed poly dispersed spherical shape with particle size distribution 18-48 nm. Our results showed that prepared NPs possess significant antifungal, antibacterial and antioxidant potential at 25, 50 and 100 µg/mL concentrations. Moreover, Cytotoxic and hemolytic assay showed significant results (less % viability and hemolysis activity) at 50 and 100 µg/mL (ZnO-NP's) concentrations as compared to control. The prepared ZnO-NPs were used as bio fertilizer in various concentrations as a foliar spray, which showed significant enhancement of the rice plant growth, along with chlorophyll, proteins and carotenoid contents. These results recommend that endophytic mediated ZnO-NPs are biocompatible and possess significant potential for agricultural applications.

Foliar micromorphology of selected medicinal Lamiaceae taxa and their taxonomic implication using scanning electron microscopy.

In this research, 25 medicinally used Lamiaceae species belonging to 20 genera have been studied and identified for the nine disorders. We used scanning electron microscopy (SEM) for qualitative and quantitative morphological character identification. The micromorphological characters observed here were important for distinguishing the studied taxa. The highest medicinal values were reported for Vitex negundo and Scutellaria baicalensis for all considered categories except urinary and otorhinolaryngology disorders. The foliar epidermal anatomical characteristics revealed that the micromorphological features of the Lamiaceae species provide taxonomically significant and accurate identification information to delimitate the family species. Moreover, we focused on both qualitative (epidermal cell shape, stomata type, stomatal pore shape, subsidiary cell shape, glandular trichomes, and non-glandular trichome shape) as well as quantitative features (epidermal cell size, stomata size, stomatal pore size, subsidiary cell size, and trichomes size). The trichomes diversity was different in most species' on adaxial and abaxial surfaces. In most species, anomocytic stomata were observed, but other types such as diacytic, paracytic, and tetracytic type stomata were also examined. The diverse pattern of anatomical characters suggests that the studied taxa provide insight evidence for the taxonomic observation of the Traditional Chinese Medicinal plants from the Lamiaceae. This work sets an avenue for future research and taxonomic exploration of medicinal flora through microscopic investigations. RESEARCH HIGHLIGHTS: This research offers a thorough microscopic identification of the family Lamiaceae. Taxonomic information on the trichome characters and types for the accurate authentication. Qualitative and quantitative characterization of 25 medicinally used Lamiaceae taxa.

Identification, Biochemical Characterization, and Safety Attributes of Locally Isolated Lactobacillus fermentum from Bubalus bubalis (buffalo) Milk as a Probiotic.

The demand of functional foods is on the rise, and researchers are trying to develop nutritious dairy products by using well-characterized strains of bacteria. In this study, we identified locally isolated strains of Lactobacillus fermentum from Bubalus bubalis (Nilli Ravi buffalo) milk and evaluated their potential as probiotics in food products like fermented milk. Fifteen Lactobacillus strains were initially isolated, and only four strains (NMCC-2, NMCC-14, NMCC-17, and NMCC-27) were examined for morphological and biochemical characterizations due to their ability of gas production in Durham tubes. Moreover, these strains were selected for further probiotic characterizations due to their extreme morphological resemblance with lactic acid bacteria for their antimicrobial activity, enzymatic potential, autoaggregation capability, hydrophobicity, and acid and bile tolerance. All selected isolates showed significant probiotic potential. However, NMCC-14 and NMCC-17 strains showed maximum probiotic potential. The isolates (NMCC-2, NMCC-14, NMCC-17, and NMCC-27) were identified as Lactobacillus fermentum utilizing 16S rRNA gene sequencing. The in vivo safety study of NMCC-14 (dose: 1010 CFU/day/mice; 21 days, orally) showed no histological dysfunctions in a mouse model. Pathogenic bacterial enzymes reduced the beneficial bacterial load in the host gastrointestinal tract. These results suggest that the NMCC-14 strain is safe and can be potentially used as a probiotic. Moreover, fermented milk was prepared by using the NMCC-14 strain. The results revealed that NMCC-14 strain-based fermented milk had significantly (p < 0.05) higher protein content (4.4 ± 0.06), water-holding capacity (WHC), and dynamic viscosity as compared to non-fermented milk. The results suggest that L. fermentum NMCC-14 is safe and nontoxic; hence, it can be a beneficial supplement to be used for the development of dairy products to be subjected to further clinical testing.

Review-Insights into Off-Label therapeutic strategies against mild and severe COVID-19 infection.

Currently, prevention and control of the coronavirus disease pneumonia epidemic situation are grim globally. To cope with total sheer carriers and patients of COVID-19 requires intensive medical support and adjunctive therapies to overcome the disease. The epidemic can be controlled with the help of both, disease suppression via community health measures and adjunctive therapies for patients suffering from infection. Till date, we do not have any proper anti-COVID-19 therapy. In order to achieve the overall realization of this pandemic, there is a need to identify treatments depending upon their direct or indirect targets; like inhibition of polyprotein synthesis, transmembrane serine protease, inhibition of viral entry and endocytosis. This could be possible by turning the focus in the direction towards the development of numerous tentative drugs, particularly in the severe to badly ill. Though, majority of these off-label adjunctive medicines are being inspected in a lot of clinical trials at different stages, scientific organizations have endeavored to elucidate the situation where these adjunctive drugs might be practiced as off-label, open- label or compassionate. Our review compiles the adjunctive therapies adopted in COVID-19 infected patients according to clinical severity in conjugation with practicing recommendations from existing guidance rules issued by global professional bodies in healthcare.

Molecular Cloning and Functional Analysis of GmLACS2-3 Reveals Its Involvement in Cutin and Suberin Biosynthesis along with Abiotic Stress Tolerance.

Cutin and wax are the main precursors of the cuticle that covers the aerial parts of plants and provide protection against biotic and abiotic stresses. Long-chain acyl-CoA synthetases (LACSs) play diversified roles in the synthesis of cutin, wax, and triacylglycerol (TAG). Most of the information concerned with LACS functions is obtained from model plants, whereas the roles of LACS genes in Glycine max are less known. Here, we have identified 19 LACS genes in Glycine max, an important crop plant, and further focused our attention on 4 LACS2 genes (named as GmLACS2-1, 2, 3, 4, respectively). These GmLACS2 genes display different expression patterns in various organs and also show different responses to abiotic stresses, implying that these genes might play diversified functions during plant growth and against stresses. To further identify the role of GmLACS2-3, greatly induced by abiotic stresses, we transformed a construct containing its full length of coding sequence into Arabidopsis. The expression of GmLACS2-3 in an Arabidopsis atlacs2 mutant greatly suppressed its phenotype, suggesting it plays conserved roles with that of AtLACS2. The overexpression of GmLACS2-3 in wild-type plants significantly increased the amounts of cutin and suberin but had little effect on wax amounts, indicating the specific role of GmLACS2-3 in the synthesis of cutin and suberin. In addition, these GmLACS2-3 overexpressing plants showed enhanced drought tolerance. Taken together, our study deepens our understanding of the functions of LACS genes in different plants and also provides a clue for cultivating crops with strong drought resistance.

Predicting potential medicinal plants with phylogenetic topology: Inspiration from the research of traditional Chinese medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Plants are a dominant source of pharmacological drugs for the treatment and cure of different disorders and diseases. However, selecting the most biologically active plant species for further screening is still challenging. Phylogeny has strong explanatory powers and provides predictive perspectives that are not available in traditional plant classifications. China, which is endowed with a diverse set of therapeutic cures from Mother Nature, represents an ideal environment for the phylogenetic analysis of potential medicinal plants. MATERIALS AND METHODS: Herein, we prepared a database of 7,451 traditional Chinese medicinal (TCM) plants, including species with therapeutic effects grouped in 14 categories. To limit our exploration of novel therapeutic species, we plotted the medicinal effects on the phylogenetic tree of almost 30,000 species of China to find hot nodes of therapeutic effects. We used the net relatedness index (NRI) and the nearest taxon index (NTI) to identify clustering and overdispersion of the phylogenetic distribution of TCM plants. RESULTS: The NRI and NTI analyses highlighted 3,392 hot node species with single therapeutic effects within 507 genera and 89 families on the phylogenetic tree and about 70% of the 14 medicinal categories clusters identified. The general pattern of the hot nodes on the phylogenetic tree indicates that basal angiosperms and basal eudicots radiated for therapeutic effects. CONCLUSIONS: Our study may provide a more targeted way to discover phylogeny-guided drugs in the early screening stage, which may lead to a higher discovery efficiency of new drugs with meaningful biological activities. Phylogenetic studies of plants that are richer in bioactive compounds can set the ground for the identification and discovery of alternative drugs.

PacBio Single-Molecule Long-Read Sequencing Reveals Genes Tolerating Manganese Stress in Schima superba Saplings.

Schima superba (Theaceae) is a subtropical evergreen tree and is used widely for forest firebreaks and gardening. It is a plant that tolerates salt and typically accumulates elevated amounts of manganese in the leaves. With large ecological amplitude, this tree species grows quickly. Due to its substantial biomass, it has a great potential for soil remediation. To evaluate the thorough framework of the mRNA, we employed PacBio sequencing technology for the first time to generate S. Superba transcriptome. In this analysis, overall, 511,759 full length non-chimeric reads were acquired, and 163,834 high-quality full-length reads were obtained. Overall, 93,362 open reading frames were obtained, of which 78,255 were complete. In gene annotation analyses, the Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Genes (COG), Gene Ontology (GO), and Non-Redundant (Nr) databases were allocated 91,082, 71,839, 38,914, and 38,376 transcripts, respectively. To identify long non-coding RNAs (lncRNAs), we utilized four computational methods associated with protein families (Pfam), Cooperative Data Classification (CPC), Coding Assessing Potential Tool (CPAT), and Coding Non-Coding Index (CNCI) databases and observed 8,551, 9,174, 20,720, and 18,669 lncRNAs, respectively. Moreover, nine genes were randomly selected for the expression analysis, which showed the highest expression of Gene 6 (Na_Ca_ex gene), and CAX (CAX-interacting protein 4) was higher in manganese (Mn)-treated group. This work provided significant number of full-length transcripts and refined the annotation of the reference genome, which will ease advanced genetic analyses of S. superba.

Genome-wide comparative analysis of long-chain acyl-CoA synthetases (LACSs) gene family: A focus on identification, evolution and expression profiling related to lipid synthesis.

In plants, Long-chain acyl-CoA synthetases (LACSs) play key roles in activating fatty acids to fatty acyl-CoA thioesters, which are then further involved in lipid synthesis and fatty acid catabolism. LACSs have been intensively studied in Arabidopsis, but its evolutionary relationship in green plants is unexplored. In this study, we performed a comprehensive genome-wide analysis of the LACS gene family across green plants followed by phylogenetic clustering analysis, gene structure determination, detection of conserved motifs, gene expression in tissues and subcellular localization. Our results identified LACS genes in 122 plant species including algae, low land plants (i.e., mosses and lycophytes), monocots, and eudicots. In total, 697 sequences were identified, and 629 sequences were selected because of alignment and some duplication errors. The retrieved amino acid sequences ranged from 271 to 1056 residues and diversified in intron/exon patterns in different LACSs. Phylogenetic clustering grouped LACS gene family into six major clades with distinct potential functions. This classification is well supported by examining gene structure and conserved motifs. Also, gene expression analysis and subcellular localization substantiate with clade division in the phylogeny, indicating that the evolutionary pattern is visible in their functionality. Additionally, experimental analysis of lacs2 mutant validated that LACS2 plays key roles in suberin synthesis. Thus, our study not only provides an evolutionary mechanism underlying functional diversification but also lays the foundation for further elucidation of the LACS gene family.

Palynological characteristics of selected Lamioideae taxa and its taxonomic significance.

Lamioideae comprised the second-largest subfamily in Lamiaceae. Although considerable progress has recently been made in the taxonomic study of Lamioideae, the subfamily remains one of the most poorly investigated subfamily in Lamiaceae. Therefore, the present study was designed with the aim to document the pollen micromorphology of some selected Lamioideae taxa and its taxonomic significance from Pakistan. Pollen micromorphological features were observed using scanning electron microscopy. The pollen grains are monad, tricolpate, radially/bilateral symmetrical. The pollen grains were small to medium-sized having oblate, oblate/subspheroidal, and subspheroidal shape. Exine sculpturing was observed as reticulate, microreticulate, and bireticulate. The colpus surface ornamentation was found as verrucate, gemmate, scabrate, and psilate. There was a considerable variation between the species in the micromorphology, that is, the coarseness of the reticulum, thickness of the muri comprising the reticulum and the number of secondary lumina per primary lumen. Hence, this study documented the pollen morphology of some selected taxa of the subfamily Lamioideae from Pakistan and strengthens the taxonomic identification of subfamily based on pollen characters, which helps in the correct identification, discrimination of the species of Lamioideae at generic and species level.

Taxonomic implication of leaf epidermal anatomy of selected taxa of Scrophulariaceae from Pakistan.

The family Scrophulariaceae consists of taxonomically complex genera and species. The delimitation of the taxa within this family is always challenging. In this paper, we studied leaf epidermis anatomical characteristics and its taxonomic significance of four species belonging to four genera of the family Scrophulariaceae collected from northern Pakistan. The species were examined under light and scanning electron microscopes (LM and SEM). Qualitative and quantitative foliar epidermal anatomical features were examined for both adaxial and abaxial surfaces. Qualitative characters like epidermal cell shape, epidermal cell cover, anticlinal wall, trichomes type, stomata type and stomata position were examined. Quantitative characters like the length and width of leaf epidermis, stomata, stomatal pore, subsidiary cell and trichomes for both adaxial and abaxial surfaces were studied and measured. Stomatal index within the species and between the species was found to be different on adaxial and abaxial surfaces. Diacytic stomata and glandular trichomes on epidermis were only found in Anticharis glandulosa while rest of the taxa has anomocytic type stomata and dendroid trichomes on both surfaces. Based on the micromorphological characters, we did principal component analysis (PCA), and cluster analysis for the species delimitation and identification. A taxonomic key has been provided to delimit and identify the studied taxa based on foliar epidermal characters. The aim of the present research was to elucidate the micromorphological characters to distinguish the studied taxa for taxonomic purposes.