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Plumbago zeylanica L. 1753 is a medicinally-important herb in family Plumbaginaceae. In this study, we assembled and reported the complete chloroplast genome of P. zeylanica. The plastome of P. zeylanica was 169,178 bp, including a large single-copy region of 92,135 bp, a small single-copy region (SSC) of 13,455 bp and a pair of inverted repeat regions (IRs) of 31,794 bp. It contained 124 genes, including 79 protein-coding genes, 37 tRNA genes and eight rRNA genes. Phylogenetic analysis showed that P. zeylanica formed a close relationship with P. auriculata in Plumbago. The first complete chloroplast genome report of P. zeylanica providing an opportunity to explore the genetic diversity, and would be also helpful in the species identification and conservation. [ABSTRACT FROM AUTHOR]

Plumbago europaea L. belongs to the family Plumbaginaceae, which has many medicinal and nutritional applications. It is among the significant medicinal herbs used widely in folk medicine in Syria to treat skin diseases such as eczema and psoriasis. There are no previous studies of the aerial parts' essential oils of this plant. So, herein we report on determining the chemical composition of essential oils extracted from the aerial parts of P. europaea L., which were collected from a mountainous area in Latakia Province, Syria, by utilising the GC-MS technique. The major compounds identified in the flower's essential oils were: plumbagin (32.4%), hexadecanoic acid (13.79%), heneicosane (9.03%) and 2,6-di-tert-butyl-4-ethylphenol (8.40%). While in the leaves' essential oils were phytol (17.97%), dibutyl phthalate (15.45%), heneicosane (6.91%) and hexahydrofarnesyl acetone (6.72%). In addition, according to the published literature, most of the identified compounds have been reported to exhibit significant biological activities. [ABSTRACT FROM AUTHOR]

Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease that can reduce quality of life. Currently, the goal of therapy is to achieve remission and prevent joint damage and disability. Acalypha indica L. and Plumbago zeylanica L. are known to be involved in rheumatoid pathogenesis. Objective: This study aimed to determine the compounds in Acalypha indica L. and Plumbago zeylanica L. that correlate with target proteins and anti-rheumatoid arthritis mechanisms. Methods: Plant compound data were collected from the KNApSAcK and IMPPAT databases, target protein data were collected using the KEGG pathway, validated using UniProt, and protein-protein interactions were analyzed using STRING. Target protein prediction using SwissTarget Prediction and SEA. Visualization of network pharmacology profiles using Cytoscape software based on the correlation between plant compounds and target proteins. Results: Acalypha indica L., which correlates with target proteins, contained quinine, gallotannin, 1,4 benzoquinone, chrysin, and kaempferol. For Plumbago zeylanica L., the compounds were vanillic acid, cinnamic acid, plumbagin, isoaffinetin, isoorientin, isovitexin, methylnaphthazarin, l-tryptophan, beta-sitosterol, stigmasterol, ficusin, suberosin, and quercetin 3-ol-rhamnoside. Conclusion: Network pharmacology visualization results showed that both Acalypha indica L. and Plumbago zeylanica L. correlated with disease target proteins in their respective rheumatoid arthritis signaling pathways. [ABSTRACT FROM AUTHOR]

Plumbago indica L. contains a valuable bioactive compound called plumbagin. Elicited regenerated shoots grown in vitro could be another source of high-yielding plumbagin. The purpose of this investigation was to examine the effects of elicitor type and concentration, as well as elicitation period, on plumbagin content in in vitro-regenerated shoots of P. indica. Nodal explants were cultured on Murashige and Skoog (MS) medium containing 1 mg/L benzyladenine (BA) in combination with 0–150 mg/L yeast extract or 50–150 µM salicylic acid for four weeks. Plumbagin levels of 3.88 ± 0.38% and 3.81 ± 0.37% w/w g dry extract were achieved from the 50 and 100 mg/L yeast extract-elicited shoots, which were higher than the value obtained for the control. However, the addition of salicylic acid did not increase the plumbagin content. In the elicitation period experiment, nodal explants were cultured on MS medium supplemented with 1 mg/L BA and 50 mg/L yeast extract for durations of three, four and five weeks. The 4-week yeast extract-elicited shoot had a maximum plumbagin content of 3.22 ± 0.12% w/w g dry extract, greater than that of the control. In summary, the plumbagin content of the in vitro P. indica shoots was enhanced by 4-week elicitation using 50 mg/L yeast extract. [ABSTRACT FROM AUTHOR]

Plumbago indica L. is a remedial herb species in the genus of Plumbago described by Linnaeus of the species of Plantarum, extensively in the hot and humid areas to use in the conventional structure of herbs. The distinct division of that plant is used to heal rheumatism, menstrual irregularities, tumors, leprotic infections, acquired immune disease, numbness, immobility, swollen glands, eczema, conjunctivitis, flatulence, extravagated, piles, anorexia, etc. The plant contains a variety of Phytoconstituents including sugars, alkaloids, flavonoids, steroids, phenols, and gums. Numerous categories of research were supported out in distinct kingdoms on the pharmacology of that remedial herb. Methodologically and biologically demonstrate data were concluded on that paper to clear the way for well-founded convenience detailed sources and invigorate further research. [ABSTRACT FROM AUTHOR]

Ethylene is a signalling factor that plays a key role in the response of plants to abiotic stresses, such as cold stress. Recent studies have shown that the exogenous application of 1-aminocyclopropane-1-carboxylate (ACC), an ethylene promoter, affects plant cold tolerance. The cold-responsive specific gene DREB plays a crucial role in enhancing cold tolerance in plants by activating several cold-responsive (COR) genes. However, how the ethylene biosynthesis pathway regulates this gene in the cold response of thermophilic plants has yet to be fully elucidated. In this study, the thermophilic plant Plumbago indica L. was used as an example. Physiological experiments and transcriptomic analyses revealed that cold stress treatment induced the synthesis of endogenous ACC and regulated the ethylene signalling activator PiERF1, and cold signalling also activated PiDREB1A. Spray experiments also revealed that ACC-induced upregulation of the PiERF1 gene reduced the cold tolerance of P. indica and decreased the expression level of the PiDREB1A gene. These results indicate that ethylene signalling directly regulates the downstream gene PiERF1 and initiates the DREB‒COR cold-responsive signalling pathway to regulate cold tolerance, resulting in the negative regulation of cold tolerance in thermophilic plants. [ABSTRACT FROM AUTHOR]

The genus Plumbago (family Plumbaginaceae), commonly known as leadwort, is a sub-tropical shrub that produces secondary metabolite plumbagin, which is employed by pharmaceutical companies and in clinical research. Plumbagin is a potent pharmaceutical because of its anti-microbial, anti-malarial, antifungal, anti-inflammatory, anti-carcinogenic, anti-fertility, anti-plasmodium, antioxidant, anti-diabetic, and other effects. This review documents the biotechnological innovations used to produce plumbagin. The use of modern biotechnological techniques can lead to a variety of benefits, including better yield, increased extraction efficiency, mass production of plantlets, genetic stability, increased biomass, and more. Large-scale in vitro propagation is necessary to minimize over-exploitation of the natural population and allow the use of various biotechnological techniques to improve the plant species and secondary metabolite production. During in vitro culture, optimum conditions are requisites for explant inoculation and plant regeneration. In this review, we provide information on various aspects of plumbagin, depicting its structure, biosynthesis, and biotechnological aspects (both conventional and advanced) along with the future prospects. Key Points: • Critical assessment on in vitro biotechnology in Plumbago species • In vitro propagation of Plumbago and elicitation of plumbagin • Biosynthesis and sustainable production of plumbagin [ABSTRACT FROM AUTHOR]

Background: Plumbago indica L. (PI) is a Thai traditional medicine that has been reported to improve health benefit including anticancer property. Nevertheless, the effects of PI and this mechanism are still less data in lung cancer. Aim: To investigate and compare the effects of PI root extract with high and low active compound, plumbagin, on the growth inhibition and apoptotic induction in A549 lung cancer cells with underlying mechanism. Materials and Methods: Antiproliferative effects of PI were examined by Sulforhodamine B (SRB) and colony formation method. Migratory suppression was studied by Wound healing and Matrigel migration assay. Apoptotic induction, ROS formation, and mitochondrial function were explored by flow cytometric method. Western blotting was used to measure the protein expression. Results: PI extract with high plumbagin level reduced cell viability by dose- and time-dependent than low plumbagin extract and the data was correlated with colony formation assay. Further, the results of migration assay indicated that 50 µg/mL of high plumbagin also caused induction of anti-migratory effects than low plumbagin. For apoptosis, both of PI extract activated the late apoptosis, especially at high dose of PI extract (50 µg/mL) along with stimulation of ROS formation and mitochondrial dysfunction. The mechanism of apoptosis was observed through the induction of expression of active-caspase 3 level correlated with reduction of pro-caspase 3 expression. Conclusion: Therefore, these anti-cancer activities of the PI extract with high plumbagin could be served as the potential beneficial effects on lung cancer than low plumbagin. Additionally, the study of PI effects in vivo study is now required to support this in vitro study information. [ABSTRACT FROM AUTHOR]

Metabolite stuffing may depend upon the session of the collection, soil condition, and geographical regions of the collection site, and this impacts the quantitative production and biological potential of the plant species. A total of 14 accessions of Plumbago zeylanica were collected from different locations in Central India to cover the entire range of topography from the foothills up to the highest peak. The instrumental thin-layer chromatography‒ultraviolet detection (instrumental TLC‒UV) method was used to quantify plumbagin and unweighted pair group method with arithmetic mean (UPGMA) cluster analysis to identify elite sources. The present investigation aims to identify the best quality planting material of P. zeylanica, in terms of elite chemotypes collected from different locations of Central India. UPGMA cluster analysis revealed the variation of the plumbagin content (0.0038‒0.6091% dry weight basis) among the collected accessions of P. zeylanica. The cluster analysis of plumbagin content depicted the distinctness in plant metabolite contents between the individuals as they were clustered by difference between accessions. This study will also promote the use of P. zeylanica in herbal drug development and will aid in the site-specific exploration of elite chemotype(s) with validated pharmacological action to meet the medicinal and commercial demands. [ABSTRACT FROM AUTHOR]

Simple Summary: Diabetic retinopathy is a severe complication of diabetes that can lead to vision loss due to the abnormal growth of blood vessels in the retina. This study investigates how two traditional medicinal plants, Solanum xanthocarpum and Plumbago zeylanica, might be used to treat this condition. We focused on identifying the active compounds in these plants and understanding how they interact with proteins related to diabetic retinopathy. Using advanced computer simulations, we found that these plant compounds effectively bind to and influence key proteins involved in the progression of the disease. Specifically, our research highlighted that compounds from Solanum xanthocarpum and Plumbago zeylanica could target several crucial pathways and proteins associated with diabetic retinopathy. These findings suggest that these plant-derived compounds have significant potential as therapeutic agents for diabetic retinopathy. By offering a new approach to treatment, this research could help improve the vision and overall quality of life of individuals suffering from this challenging condition. (1) Background: Diabetic retinopathy (DR) is a major complication of diabetes, marked by abnormal angiogenesis, microaneurysms, and retinal hemorrhages. Traditional Ayurvedic medicine advocates multi-target strategies for DR management. However, the mechanisms by which Solanum xanthocarpum (SX) and Plumbago zeylanica (PZ) exert therapeutic effects are not well understood; (2) Methods: To investigate these mechanisms, we employed network pharmacology and molecular docking techniques. Phytochemicals from SX and PZ were identified using the IMPPAT database and Swiss Target Prediction tool. DR-related protein targets were sourced from the GeneCards database, and common targets were identified through Venn diagram analysis. STRING and Cytoscape were used to construct and analyze protein–protein interaction networks. Pathway enrichment was performed with Gene Ontology and KEGG databases; (3) Results: We identified 28 active phytoconstituents, targeting proteins such as EGFR, SRC, STAT3, AKT1, and HSP90AA1. Molecular docking and dynamics simulations confirmed the strong binding affinities of these compounds to their targets; (4) Conclusions: The study highlights the multi-target activity of SX and PZ, particularly in pathways related to EGFR tyrosine kinase inhibitor resistance and PI3K–AKT signaling. These findings provide valuable insights into their therapeutic potential for DR, suggesting the effective modulation of key molecular pathways involved in the disease. [ABSTRACT FROM AUTHOR]

Naturally synthesized secondarymetabolites in plants are considered an important source of drugs, food additives, etc. Among them, research on natural plant medicinal components and their synthesis mechanisms has always been of high concern. We identified a novel medicinal floral crop, Plumbago auriculata L., that can be treated with methyl jasmonate (MeJA) for the rapid or sustainable production of natural bioactives from hairy roots. In the study, we globally analyzed the changes in the accumulation of plumbagin and others in the hairy roots of Plumbago auriculata L. hairy roots (PAHR) 15834 in P. auriculata L. based on 100 mmol/L of MeJA treatment by RNA-seq profiling, and we found that there was a significant increase in the accumulation of plumbagin and saponin before 24 h. To explain the principle of co-accumulation, it showed thatMeJA induced JA signaling and the shikimic acid pathway, and the methylvaleric acid (MVA) pathway was activated downstream subsequently by the Mfuzz and weighted gene coexpression analysis. Under the shared metabolic pathway, the high expression of PAL3 and HMGR promoted the activity of the "gateway enzymes" phenylalanine ammonia lyase (PAL) and 3-hydroxy-3-methylglutaryl CoA reductase (HMGR), which respectively induced the high expression of key reaction enzyme genes, including chalcone synthase (CHS), isopentenyl diphosphate (IPP), and farnesyl pyrophosphate synthase (FPS), that led to the synthesis of plumbagin and saponin. We speculated that large amounts of ketones and/or aldehydes were formed under the action of these characteristic enzymes, ultimately achieving their coaccumulation through polyketone and high-level sugar and amino acid metabolism. The study results provided a theoretical basis for carrying out the factory refinement and biosynthesis of plumbagin and saponins and also provided new ideas for fully exploiting multifunctional agricultural crops and plants and developing new agricultural by-products. [ABSTRACT FROM AUTHOR]

The global potential of therapeutic plants in treating microbial illnesses is evident. This study focuses on the Plumbago zeylanica (P.zeylanica)plant and aims to isolate and investigate a bioactive compound with antifungal, antioxidant, and cancer-fighting properties. The compound identified as 2,4-Ditert butyl phenol (2,4-DTBP) was successfully extracted from P. zeylanica using Ethyl acetate and confirmed through GC–MS analysis. The antioxidant effects of 2,4-DTBP are well-known as it effectively suppresses oxidation, preventing material degradation and disintegration. Its ability to neutralize free radicals and reduce reactive oxygen species production makes it valuable in stabilizing various compounds. Notably, it enhances the durability and endurance of plastics, rubber, and polymers. The isolation and purification of 2,4-DTBP were accomplished using HPLC, and its structure was characterized as C14H22O through 1H NMR analysis. The fungicidal activity of 2,4-DTBP against Botrytis cinerea was proven through agar plate assay, and its cytotoxic activity was evaluated using MCF-7 cells (a breast carcinoma cell line) with an identified IC50 value of 5 μg/ml. This study provides insight into the medicinal properties of 2,4-DTPB and its effectiveness against cancer cells. Overall, further investigation and exploration of 2,4-DTBP's potential in cancer research would contribute to its potential application as a novel therapeutic agent in the future. [ABSTRACT FROM AUTHOR]

This study aimed to develop and evaluate a niosomal formulation loaded with Plumbago zeylanica extract (PZE). The solvent evaporation method was used to prepare niosomes and optimized using the Box–Behnken design. The optimized formulation (PZE-Ns-Opt) was characterized for drug release, DPPH assay, α-amylase inhibition assay, α-glucosidase inhibition assay, and confocal laser scanning microscopy (CLSM) study. PZE-Ns-Opt showed a vesicle size of 253.6 nm, PDI of 0.108, entrapment efficiency of 62.4%, and drug release of 84.01%. The CLSM image of the rat's intestine suggested that the rhodamine red B-loaded PZE-Ns-Opts showed superior penetration compared to the control. Further, the antioxidant activity of the prepared formulation was exhibited as 89.46%±0.016 as compared to PZE (78.10%±0.005). In addition to this, α- amylase activity was inhibited by 95.11%±4.62, 85.88%±2.56, and 89.87%±3.65 by acarbose, PZE, and PZE-Ns-Opt, respectively. In comparison, the α-glucosidase activity was inhibited by 88.47%±1.04, 81.07%±0.50 and 85.78%±0.71 by acarbose, PZE, and PZE-Ns-Opt, respectively. To conclude, the development of PZE-Ns-Opt formulation and its characterization showed the establishment's authenticity. The in vitro antidiabetic, antioxidant studies expressed the protective effect from oxidative stress, CLSM of rat's intestine ability to penetrate to great extent and could be a promising candidate for the management of diabetes mellitus. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L. (Plumbaginaceae) commonly known, as chitrak is pharmacologically important plant. Various studies have been undertaken to assess the pharmacological potential of different parts of the plant namely like roots, stem, flower, and leaves as antimicrobial, hepatoprotective, anticancer, antifertility, antiulcer, antifungal and wound healing. The intention of the present review is to deliver a concise account on its ethnobotanical uses, phytochemistry with an in-depth study of its phytoconstituents, facts and prospects of its potential pharmacological activities of this golden plant. An extensive literature survey was undertaken through different online platforms viz. Google Scholar and online databases namely PubMed, Science Direct and Springer. All papers based on traditional medicinal uses and pharmacological properties were included. Sixty three research articles and review articles were found to be apt for inclusion into the review. About 150 articles were retrieved for the purpose. The elaborative results vindicated that Plumbago zeylanica L. holds significant prospects in major health conditions such as diabetes, cardiovascular disorders, ulcer, liver problems, obesity, wound healing, cancer etc. [ABSTRACT FROM AUTHOR]

In 2021, Plumbago indica plants with necrotic spots on their leaves were observed in Beijing, China. Through high-throughput sequencing, we discovered a putative novel member of the genus Cytorhabdovirus, which was provisionally named "plumbago necrotic spot-associated virus" (PNSaV). The full-length negative-sense single-stranded RNA genome of this virus is 13,180 nucleotides in length and contains eight putative open reading frames (ORFs), in the order 3′ leader-N-(P′)-P-P3-M-G-P6-L-5′ trailer. Phylogenetic analysis and pairwise comparisons suggested that PNSaV is most closely related to pastinaca cytorhabdovirus 1, with 59.2% nucleotide sequence identity in the complete genome and 56.4% amino acid sequence identity in the L protein. These findings suggest that PNSaV should be considered a new member of the genus Cytorhabdovirus. [ABSTRACT FROM AUTHOR]

Under salt stress, recretohalophyte Plumbago auriculata tetraploids enhance salt tolerance by increasing selective secretion of Na+ compared with that in diploids, although the mechanism is unclear. Using non-invasive micro-test technology, the effect of salt gland Ca2+ content on Na+ and K+ secretion were investigated in diploid and tetraploid P. auriculata under salt stress. Salt gland Ca2+ content and secretion rates of Na+ and K+ were higher in tetraploids than in diploids under salt stress. Addition of exogenous Ca2+ increased the Ca2+ content of the salt gland in diploids and is accompanied by an increase in the rate of Na+ and K+ secretion. With addition of a Ca2+ channel inhibitor, diploid salt glands retained large amounts of Ca2+, leading to higher Ca2+ content and Na+ secretion rate than those of tetraploids. Inhibiting H2O2 generation and H+-ATPase activity altered Na+ and K+ secretion rates in diploids and tetraploids under salt stress, indicating involvement in regulating Na+ and K+ secretion. Our results indicate that the increased Na+ secretion rate of salt gland in tetraploids under salt stress was associated with elevated Ca2+ content in salt gland. [ABSTRACT FROM AUTHOR]

Plumbago indica L. is a perennial herb with ornamental and anticancer medicinal functions widely distributed in the tropics. It is affected by temperature and cannot bloom normally in colder subtropical regions, which seriously affects its ornamental value. To create low-temperature resistance mutants and enrich new germplasm resources, this study used tissue culture and chemical reagent (0.5 mmol/L NaN3) and low-temperature stress (0°C, full darkness for 48h) induction to target and screen for cold-resistance mutants. The results showed that the ISSR band polymorphism ratio of the 24 suspected mutant materials was 87.5%. The DNA profiles of the 9 mutants initially identified were altered. The content of plumbagin in the stems and leaves of the mutants was examined, and it was found that the accumulation in the leaves of the mutant SA24 could be as high as 3.84 times that of the control, which was 0.5991%. There were significant differences in the anatomical structures of roots, stems and leaves. The mutants mostly exhibited reduced root diameter (only 0.17-0.69 times that of CK), increased stem diameter (up to 2.19 times that of CK), enlarged mesophyll cells, increased thickness (up to 1.83 times that of CK) and high specificity, which are thought to be important for the different cold resistance obtained by the mutants. In the cold resistance experiment, four cold-tolerant mutants were successfully screened according to their morphological characteristics and physiological indexes, and the mutagenesis efficiency could be as high as 2.22% and did not affect the accumulation of plumbagin in their stems and leaves, even higher than CK. The responses of the screened mutants SA15, SA19, SA23 and SA24 to low temperature showed slower leaf wilting, higher light energy conversion efficiency, less accumulation of MDA content, increased enzymatic activities of antioxidant enzymes (SOD, CAT, POD) and more accumulation of soluble sugars and proline content. These characteristics are consistent with the response of cold-resistance plants to low temperatures. The cold- resistance mutants cultivated in soil were observed of agronomic and ornamental traits for one year, mainly manifested as delayed flowering and delayed entry into the senescence stage. This study provides a more rapid and accurate technique for identifying and screening cold-tolerant mutants, and lays the foundation for future experiments on the creation of new coldresistant varieties. [ABSTRACT FROM AUTHOR]

In this study, NMR and GC-MS based comparative metabolomic profiling of the roots of three different species namely, Plumbago indica, P. auriculata and P. zeylanica were investigated followed by multivariate statistical analyses and their antioxidant activity. Also, two alkylated phenols i.e., 2,6-di-tert-butyl phenol and 2,4-di-tert-butyl phenol not reported earlier from this taxon were isolated from P. indica. This metabolic study resulted in the identification of 25 and quantification of 18 metabolites. Principal component analysis showed the clear distinction among the three species. The antioxidant activity in the extracts was tested by free radical scavenging method. The three Plumbago species revealed interesting antioxidant potential, in particular, P. indica, which was rich in naphthoquinones, coumarins, alkylated phenols, sterols, triterpenes, fatty acids showed lowest IC50 value. The results highlighted the role of P. indica in the management of oxidative stress especially when they are utilized in the formation of fermented food products. [ABSTRACT FROM AUTHOR]

In plants, heteromorphic self-incompatibility (HetSI) is a strategy for avoiding self-pollination and promoting outcrossing, and during this process, numerous protein-protein interaction events occur between the pistil and pollen. Previous studies in Primula and Fagopyrum that focused on HetSI systems have provided interesting insights; however, the molecular mechanism underlying HetSI remains largely unknown. In this study, we profiled the proteome of Plumbago auriculata stigmas before and after self-incompatible (SI) and self-compatible (SC) pollination. Comparative analyses were conducted by 4D-DIA (Four-dimensional data independent acquisition), a promising technology that increases the sensitivity and reduces the spectral complexity of proteomic analysis by adding a fourth dimension, ion mobility. The results revealed 33387 peptides and 5311 proteins in all samples. The pathways in which the differentially expressed proteins (DEPs) identified in the P × P (Pin style self-pollinated with pin pollen) vs. PS (Pin style) and T × T (Thrum style self-pollinated with thrum pollen) vs. TS (Thrum style) comparisons were significantly enriched were biosynthesis of secondary metabolites and pentose and glucuronate interconversions. In the P × T (Pin style cross-pollinated with thrum pollen) vs. PS and T × P (Thrum style cross-pollinated with pin pollen) vs. TS comparison, the top three pathways were biosynthesis of secondary metabolites, pentose and glucuronate interconversions, and phenylpropanoid biosynthesis. The phenylpropanoid biosynthesis, cutin, suberine and wax biosynthesis, and flavonoid biosynthesis pathways were enriched in the P × T vs. P × P comparison, and starch and sucrose metabolism, glycerophospholipid metabolism, and alpha-linolenic acid metabolism were abundant in the T × T vs. T × P comparison. The enriched pathways between PS and TS were the biosynthesis of secondary metabolites, phenylpropanoid biosynthesis, and pentose and glucuronate interconversion. Self-incompatibility protein S1 (SI S1), Mitogen-activated protein kinase 3/4 (MPK3/4), Mitogen-activated protein kinase kinase 2/3 (M2K2/3), Exocyst complex component EXO70A1 (E70A1) and Thioredoxin H1/2 (TRXH1/2) were found to be HetSI-related candidates, and O-fucosyltransferase 23 (OFT23), 3- ketoacyl-CoA synthase 6 (KCS6), Receptor-like protein kinase FERONIA (FERON), Fimbrin-5 (FIMB5), Pollen-specific leucine-rich repeat extensin-like protein 4 (PLRX4), Transcription initiation factor IIB-2 (TF2B2) and Pectinesterase 1 (AL11A), etc., were identified as other regulatory transducers. These findings combined with our morphological and reactive oxygen species (ROS) intensity analyses indicate that P. auriculata has typical dry-stigmas and that the HetSI mechanism might differ between the pin and thrum. SI S1 might be the key factor in HetSI, and ROS are overexpressed during SC pollination to rapidly activate the mitogen-activated protein kinase (MAPK)-mediated phosphorylation of E70A1 to maintain stigma receptivity in plants with HetSI. [ABSTRACT FROM AUTHOR]

“Heteromorphic self-incompatibility” (HetSI) in plants is a mechanism of defense to avoid self-pollination and promote outcrossing. However, the molecular mechanism underlying HetSI remains largely unknown. In this study, RNA-seq was conducted to explore the molecular mechanisms underlying selfcompatible (SC, “T×P” and “P×T”) and self-incompatible (SI, “T×T” and “P×P”) pollination in the two types of flowers of Plumbago auriculata Lam. which is a representative HetSI plant. By comparing “T×P” vs. “T×T”, 3773 (1407 upregulated and 2366 downregulated) differentially expressed genes (DEGs) were identified, 1261 DEGs between “P×T” and “P×P” (502 upregulated and 759 downregulated). The processes in which these DEGs were significantly enriched were “MAPK (Mitogen-Activated Protein Kinases-plant) signaling pathway”, “plant-pathogen interaction”,“plant hormone signal transduction”, and “pentose and glucuronate interconversion” pathways. Surprisingly, we discovered that under various pollination conditions, multiple notable genes that may be involved in HetSI exhibited distinct regulation. We can infer that the HetSI strategy might be unique in P. auriculata. It was similar to “sporophytic self-incompatibility” (SSI) but the HetSI mechanisms in pin and thrum flowers are diverse. In this study, new hypotheses and inferences were proposed, which can provide a reference for crop production and breeding. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L. is commonly known as chitrak, consumed since long time owing to its potent medicinal benefits. It is a major source of the yellow crystalline naphthoquinone called plumbagin, which is highly acclaimed for its anticancerous activities on different cancers i.e. prostrate, breast, ovarian, etc. The growing urges for this compound make this plant extremely demanding in the global market; hence, the plant is indiscriminately harvested from its very natural habitat. Therefore, in vitro biomass production of this plant can be a sustainable alternative for plumbagin production. In this present study, it has been found that, compared to other cytokinins, biomass production was enhanced by using aromatic cytokinin meta-topolin (mT). The highest shoot buds produced by mT (1 mg/l) was 13.60 ± 1.14 after 14 d of culture establishment. After 84 d in the same medium, 129.8 ± 2.71 shoots were produced, and the fresh weight of the total biomass was 19.72 ± 0.65 g. The highest number of roots was induced (37.80 ± 0.84) with 1.0 mg/l Indole-3-butyric acid (IBA). The well rooted plantlets were acclimatized in field condition with 87%survival. The regenerated plants' genetic fidelity was accessed through molecular markers i.e. Inter simple sequence repeat (ISSR), Start codon targeted (SCoT) and cytology studies. The monomorphic bands amplified by the primers across in vivo and in vitro plants confer the genetic homogeneity of the regenerants. The plumbagin content from different parts of the in vitro grown plants in vivo mother plant was quantified through High-Performance Liquid Chromatography (HPLC), and found that they do not differ significantly. Even all parts of the in vitro plants produce plumbagin, roots contain the maximum amount (14.67 ± 0.24 mg/g dry weight basis). [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L., a wild shrub, is a vital natural source of plumbagin, a potent 1,4-naphthoquinone renowned for its anti-cancer properties, notably effective against breast, prostate, and ovarian cancers. Traditional plumbagin extraction, involving root uprooting and plant destruction, raises ecological concerns. The primary objective of this study is to enhance plumbagin production by incorporating the elicitation process into in vitro cultivation with regenerated plants that retain all of their intact organs. Seven different elicitors categorized into three distinct groups were employed to stimulate plumbagin content. Among the various elicitors used, this study marks the first application of biogenic silver nanoparticles (AgNPs) from Curcuma amada in stimulating plumbagin production in this plant. The maximum plumbagin content, recorded at 8.98 ± 0.24 mg/g dry weight basis, was found in the roots when elicited with AgNPs at a concentration of 15 mg/l. In addition to that, biotic elicitors (yeast extract, chitosan and casein hydrolysate) and heavy metals (lead, cobalt and nickel) also successfully elicit plumbagin in the root and aerial parts of the plants, quantified through High Performance Liquid Chromatography (HPLC). In our study, we found that certain elicitors induced root browning and tissue necrosis, as confirmed by propidium iodide (PI) staining. The most significant browning effects were observed with chitosan from biotic sources and lead from heavy metals, while no such effects were associated with AgNPs at any concentration. Utilizing intact, entire plants as the subjects for elicitation in our study is a valuable aspect. This approach closely replicates the natural process occurring in intact plants, enhancing the relevance of our findings to practical situations.Key message: Utilizing silver nanoparticles alongside diverse biotic and heavy metal elicitors applied to entire plants in liquid cultures facilitates the sustainable production of plumbagin from various plant organs. [ABSTRACT FROM AUTHOR]

Plumbago auriculata Lam. (Plumbaginaceae) is a medicinal flowering shrub which produces plumbagin that shows diverse pharmacological attributes. P auriculata accumulates a very important napthaquinone, plumbagin which exhibits a wide range of medicinal properties. The present study is aimed at in vitro propagation of P auriculata and a comparative quantification of plumbagin in in vitro and in vivo grown plants. Direct shoot regeneration was induced from the nodal explants of P auriculata. Murashige and Skoog medium supplemented with different combination of 6-Benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA). Best response was obtained with BAP 1.5 mg/L and NAA 2.0 mg/L after 15 days in in vitro culture. Maximum number of roots were induced on ½ strength MS medium supplemented with 2% sucrose (w/v) and 0.7 mg/L Indole-3-acetic acid. Plumbagin was extracted by microwave-assisted extraction method and content was estimated from stem, leaf and root of both propagated plants by HPTLC. It was found that the roots of both in vivo (0.01% w/w) and in vitro (0.066% w/w) grown plants contain the highest amount of plumbagin. In vitro grown roots produced five-fold more plumbagin as compared to in vivo regenerated roots. Maximum increase of plumbagin production was observed in the in vitro grown stem i.e., seven-fold as compared to in vivo induced stem. In vitro propagation of plant provides a rapid and continuous supply of plumbagin while preserving the natural resource. [ABSTRACT FROM AUTHOR]

Chagas disease (CD) caused by the protozoan Trypanosoma cruzi affects more than six million people worldwide. Treatment is restricted to benznidazole (Bz) and nifurtimox (Nf) that display low activity in the later chronic stage besides triggering toxic events that result in treatment abandonment. Therefore, new therapeutic options are necessary. In this scenario, natural products emerge as promising alternatives to treat CD. In the family Plumbaginaceae, Plumbago sp. exhibits a broad spectrum of biological and pharmacological activities. Thus, our main objective was to evaluate, in vitro and in silico, the biological effect of crude extracts of root and of aerial parts of P. auriculata, as well as its naphthoquinone Plumbagin (Pb) against T. cruzi. The phenotypic assays revealed potent activity of the root extract against different forms (trypomastigote and intracellular forms) and strains (Y and Tulahuen), with a compound concentration that reduced 50% of the number of the parasite (EC50) values ranging from 1.9 to 3.9 µg/mL. In silico analysis showed that Pb is predicted to have good oral absorption and permeability in Caco2 cells, besides excellent probability of absorption by human intestinal cells, without toxic or mutagenic potential effects, not being predicted as a substrate or inhibitor of P-glycoprotein. Pb was as potent as Bz against intracellular forms and displayed a superior trypanosomicidal effect (about 10-fold) in bloodstream forms (EC50 = 0.8 µM) as compared to the reference drug (8.5 µM). The cellular targets of Pb on T. cruzi were evaluated using electron microscopy assays and the findings on bloodstream trypomastigotes showed several cellular insults related to the autophagic process. Regarding toxicity in mammalian cells, the root extracts and the naphthoquinone present a moderate toxic profile on fibroblasts and cardiac cell lines. Then, aiming to reduce host toxicity, the root extract and Pb were tested in combination with Bz, and the data showed additive profiles with the sum of the fractional inhibitory concentration indexes (ΣFICIs) being 1.45 and 0.87, respectively. Thus, our work reveals the promising antiparasitic activity of Plumbago auriculata crude extracts and its purified naphthoquinone Plumbagin against different forms and strains of Trypanosoma cruzi in vitro. [ABSTRACT FROM AUTHOR]

Cultivation of hairy roots is an effective method to get high in plumbagin secondary metabolites. With different genotypes, setting up a stable genetic transformation system is peculiar and difficult. The plumbagin is found in the roots of Plumbago auriculata L., a secondary metabolite with significant medicinal value. Still, the natural root grows slowly, and their accumulation period is lengthy (2–6 years). In this paper, we first explored the most effective Agrobacterium rhizogene-mediated (A4, ATCC 15834, and LBA 9402) genetic transformation to induce hairy root of P. auriculata. We compared the difference of the plumbagin concentration in the root between tissue culture and seedling. The results showed that the leaves were soaked with bacterial solution for 25–30 min and then transferred to 1/2 MS + acetosyringone 100 µM agar solidified medium for 2-5d. After co-culture, we transferred the leaves to 1/2 MS + Cefotaxime sodium of different concentrations sterilized for 7d. Under this scheme, three strains can induce hairy roots, with ATCC 15834 having the highest hairy root transformation efficiency (86.78 ± 0.74%) and the earliest root emergence time (8.33 ± 0.58 d). 1-month-grown hairy root showed an increase in plumbagin compared with the untransformed roots of 1-year-old live seedlings, with ATCC 15834 induced roots having the highest content of 38.95 mg·g−1 DW, which was 72.13 times higher than the untransformed roots and 3.95 times higher than that of 1-year-old live seedlings. This is an important experimental basis for biosynthesis mechanism of plumbagin and the feasibility of subsequent commercial production. Key message: A genetic transformation system was established for hairy root of P. auriculata by A. rhizogene. A reliable and rapid method was developed to obtain the plumbagin with high medicinal value. [ABSTRACT FROM AUTHOR]

This study aimed at investigating the chemical composition and the hepatoprotective activities of Plumbago indica L. and P. auriculata Lam. LC–MS/MS analyses for the hydroalcoholic extracts of the aerial parts of the two Plumbago species allowed the tentative identification of thirty and twenty-five compounds from P. indica and P. auriculata, respectively. The biochemical and histopathological alterations associated with thioacetamide (TAA)-induced liver fibrosis in rats were evaluated in vivo where rats received the two extracts at three different dose levels (100, 200 and 400 mg/kg p.o, daily) for 15 consecutive days with induction of hepatotoxicity by TAA (200 mg/kg/day, i.p.) at 14th and 15th days. Results of the present study showed a significant restoration in liver function biomarkers viz. alanine transaminase (ALT), aspartate transaminase (AST), gamma glutamyl transferase and total bilirubin. The liver homogenates exhibited increased levels of antioxidant biomarkers: reduced glutathione (GSH) and catalase (CAT), accompanied with decline in malondialdehyde (MDA). Furthermore, treated groups exhibited a significant suppression in liver inflammatory cytokines: tumor necrosis factor-α (TNF-α) and interlukin-6 (IL-6), and fibrotic biomarker: alpha smooth muscle relaxant. Histopathological examination of the liver showed normality of hepatocytes. Noteworthy, P. indica extract showed better hepatoprotective activity than P. auriculata, particularly at 200 mg/kg. To sum up, all these results indicated the hepatoprotective properties of both extracts, as well as their antifibrotic effect was evidenced by reduction in hepatic collagen deposition. However, additional experiments are required to isolate their individual secondary metabolites, assess the toxicity of the extracts and explore the involved mechanism of action. [ABSTRACT FROM AUTHOR]

Plumbagin is the major secondary metabolite found in Plumbago species, it exhibit a wide variety of pharmaceutical activities. The initial focus of this study was to establish a suitable protocol for direct shoot regeneration from nodal explants on Murashige and Skoog (MS) medium fortified with 6-benzylaminopurine (BAP) and α-naphthalene-acetic acid (NAA). Single shoot regenerated from the nodal explants when cultured on BAP alone, whereas multiple shoots regenerated with BAP and NAA. The maximum number of shoots were obtained on medium fortified with 5.5 µM BAP and 3.0 µM NAA. Furthermore, the shoot cultures were treated with biotic elicitors (lysates of Agrobacterium rhizogenes and Trichoderma viride and yeast extract). The application of elicitors was optimized by the Response Surface Methodology coupled with Box-Behnken design (RSM-BBD). The experimental and predicted values were analyzed and the elicitors were found to be stastically significant. Plumbagin was quantified by liquid chromatography/electrospray ionization tandem. The highest plumbagin yield was found to be 18-fold higher as compared to the control when treated with 1.5% A. rhizogenes lysate, 2.25% T. viride lysate and 2% yeast extract. The best model for optimum production of plumbagin was experimentally verified and the correlation between the predicted (1.751%) and actual values (1.732%) was found to be 98.7% for plumbagin production. The present study suggest that the elicitors used enhance the production of plumbagin considerably. It is the first ever study to report the successful utilization of lysates enhancing plumbagin production in the shoot cultures. Key message: In vitro production of plumbagin from Plumbago zeylenica. Analysis of plumbagin by LC-MS. Application of RSM to optimize the enhanced plumbagin yield by using eliciters. [ABSTRACT FROM AUTHOR]

This study aimed at investigating the chemical composition and the hepatoprotective activities of Plumbago indica L. and P. auriculata Lam. LC–MS/MS analyses for the hydroalcoholic extracts of the aerial parts of the two Plumbago species allowed the tentative identification of thirty and twenty-five compounds from P. indica and P. auriculata, respectively. The biochemical and histopathological alterations associated with thioacetamide (TAA)-induced liver fibrosis in rats were evaluated in vivo where rats received the two extracts at three different dose levels (100, 200 and 400 mg/kg p.o, daily) for 15 consecutive days with induction of hepatotoxicity by TAA (200 mg/kg/day, i.p.) at 14th and 15th days. Results of the present study showed a significant restoration in liver function biomarkers viz. alanine transaminase (ALT), aspartate transaminase (AST), gamma glutamyl transferase and total bilirubin. The liver homogenates exhibited increased levels of antioxidant biomarkers: reduced glutathione (GSH) and catalase (CAT), accompanied with decline in malondialdehyde (MDA). Furthermore, treated groups exhibited a significant suppression in liver inflammatory cytokines: tumor necrosis factor-α (TNF-α) and interlukin-6 (IL-6), and fibrotic biomarker: alpha smooth muscle relaxant. Histopathological examination of the liver showed normality of hepatocytes. Noteworthy, P. indica extract showed better hepatoprotective activity than P. auriculata, particularly at 200 mg/kg. To sum up, all these results indicated the hepatoprotective properties of both extracts, as well as their antifibrotic effect was evidenced by reduction in hepatic collagen deposition. However, additional experiments are required to isolate their individual secondary metabolites, assess the toxicity of the extracts and explore the involved mechanism of action. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L. (Pz) is one of the most important medicinal plant belonging to the family Plumbaginaceae. It is a perennial shrub, growing throughout India and most places of Sri Lanka. It contains various bioactive compounds like alkaloids, flavonoids, naphthoquinones, glycoside, saponins, steroids, tri-terpenoids, coumarins, phenolic compounds etc. Of all the chemical constituents, plumbagin is the principal active compound. Plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone-C11H8O3) is primarily present in roots in higher amounts with only about 1% in the whole plant. The literature reveals that the root and root bark have a wider claim in traditional medicines against various diseases as a memory enhancer, anti-inflammatory, anti-microbial, wound healing, anti-malarial, anti-infertility, anticancer, blood coagulation, and anti-oxidant activities. The present study aims to evaluate the anti-inflammatory and antimicrobial properties of this plant. [ABSTRACT FROM AUTHOR]

The plant Plumbago rosea Linn., belonging to the Plumbaginaceae family, is an important medicinal herb distributed in part of Southeast Asia, and there are many reports of its pharmacological properties. However, the allelopathic activities of P. rosea have not been examined. Thus, the present study was conducted to assess the allelopathic activity of P. rosea and to identify its allelopathic substances. The aqueous methanol stem extract of P. rosea significantly suppressed the seedling growth of barnyard grass (Echinochloa crus-galli L. P. Beauv.), Italian ryegrass (Lolium multiflorum Lam.), timothy (Phleum pretense L.), cress (Lepidium sativum L.), lettuce (Lactuca sativa L.), and alfalfa (Medicago sativa L.). The extract of P. rosea was then purified through chromatographic steps, and two active substances were isolated and determined as 7,4′,5′-tri-O-methyl dihydroquercetin and 7,4′,5′-tri-O-methylampelopsin. The two compounds significantly inhibited the seedling growth of cress, with 7,4′,5′-tri-O-methylampelopsin showing a greater inhibitory effect than 7,4′,5′-tri-O-methyl dihydroquercetin. This result may be due to the 3′-OH group in 7,4′,5′-tri-O-methylampelopsin. The effective concentrations of both compounds required for 50% growth inhibition (EC50 values) of cress seedlings were 0.24 mM and 0.59 mM for root and shoot, and 0.07 mM and 0.21 mM, respectively. These findings suggest that the two compounds may contribute to the allelopathic effect of P. rosea and could be used as a natural source of allelopathic substances. [ABSTRACT FROM AUTHOR]

A high-performance thin-layer chromatography (HPTLC) method has been developed for the quantitative determination and validation of plumbagin in the methanolic fraction of Plumbago zeylanica. For achieving good separation, the mobile phase of toluene‒ethyl acetate (9:2, V/V) for plumbagin was used. Densitometric determination of plumbagin was carried out at 270 nm in reflection/absorption mode; the calibration curves were linear in the range of 100‒600 ng/spot. During analysis, the methanolic fraction of P. zeylanica L. showed the presence of plumbagin. The method is specific, simple, precise, and accurate. The obtained data can be used for the routine analysis of the reported biomarkers in crude drugs and extracts. The quantification and the method validation of these biomarkers have not yet been reported in P. zeylanica which can be utilized for the proper standardization of the plant. [ABSTRACT FROM AUTHOR]

Main Conclusion: Novel cytochrome P450s, CYP81B140 and CYP81B141 from Plumbago zeylanica were functionally characterized to understand their involvement in polyketide plumbagin biosynthesis. Further, we propose 3-methyl-1-8-naphthalenediol and isoshinanolone as intermediates for plumbagin biosynthesis. Plumbago zeylanica L. (P. zeylanica) is a medicinally important plant belonging to the family Plumbaginaceae. It comprises the most abundant naphthoquinone plumbagin having anti-cancer activity. Only the polyketide synthase (PKS) enzyme has been identified from the biosynthetic pathway which catalyzes iterative condensation of acetyl-CoA and malonyl-CoA molecules. The plumbagin biosynthesis involves hydroxylation, oxidation, hydration and dehydration of intermediate compounds which are expected to be catalyzed by cytochrome P450s (CYPs). To identify the CYPs, co-expression analysis was carried out using PKS as a candidate gene. Out of the eight identified CYPs, CYP81B140 and CYP81B141 have similar expression with PKS and belong to the CYP81 family. Phylogenetic analysis suggested that CYP81B140 and CYP81B141 cluster with CYPs from CYP81B, CYP81D, CYP81E and CYP81AA subfamilies which are known to be involved in the hydroxylation and oxidation reactions. Moreover, artificial microRNA-mediated transient individual silencing and co-silencing of CYP81B140 and CYP81B141 significantly reduced plumbagin and increased the 3-methyl-1-8-naphthalenediol and isoshinanolone content. Based on metabolite analysis, we proposed that 3-methyl-1-8-naphthalenediol and isoshinanolone function as intermediates for plumbagin biosynthesis. Transient silencing, over-expression and docking analysis revealed that CYP81B140 is involved in C-1 oxidation, C-4 hydroxylation and [C2–C3] hydration of 3-methyl-1-8-naphthalenediol to form isoshinanolone, whereas CYP81B141 is catalyzing [C2–C3] dehydration and C-4 oxidation of isoshinanolone to form plumbagin. Our results indicated that both CYP81B140 and CYP81B141 are promiscuous and necessary for plumbagin biosynthesis. This is the first report of identification and functional characterization of P. zeylanica-specific CYPs involved in plumbagin biosynthetic pathway and in general hexaketide synthesis in plants. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica shows the presence of many phytochemical constituents of which plumbagin, a type of naphthoquinone, is vital and has major biological effects. Plumbagin is majorly present in the root of the plants, with a minimal concentration in other parts of the plant. However, extracting plumbagin from the roots is destructive harvesting. Thus, micropropagation could be a viable alternative to reduce the collection of plant from wild. Study from micropropagated leaf calli was considered as a steady and sustainable source of phytochemical. P. zeyalnica callus was subcultured in an appropriate medium with plant growth regulators to initiate root cultures. A comparative study was performed to estimate the quantities of plumbagin obtain from the wild plant root part and root created by callus culture differentiation. [ABSTRACT FROM AUTHOR]

The Indian leadwort (Plumbago indica L.) of the family Plumbaginaceae is a plant with high pharmaceutical value, as it contains plumbagin, a naphthoquinone with antibacterial, antifungal and anticancer properties. Among the propagation methods for the Indian leadwort, in vitro propagation is considered an effective method in producing disease-free transplants in a short period of time with high propagation rate. When plants grown in vitro are transferred to ex vitro condition, the environmental factors in the nursery house such as light, temperature, humidity and microorganism in the soil will affect their growth. Characteristics of transplants themselves is also critical for the subsequent growth. It is, thus, essential to establish the standards to evaluate and qualify in vitro plants for transplanting to ex vitro condition. Among these standards, the culture age of in vitro plants affects the maturations of their root, stem and leaves, which can in turn influence the acclimating ability and growth of in vitro plants after transplantation. The purpose of this study is to investigate the effects of the culture age of in vitro Indian leadwort plants on their performance during ex vitro stage. For this purpose, three different culture ages of uniform in vitro plants, 35, 42 and 49 day-old, were studied. After 28 days of cultivation in the nursery house under the light intensity of 70 ± 10 µmol m-2 s-1, temperature of 35 ± 4 °C and relative humidity (RH) of 60 ± 10 %, all three treatments achieved 100% survival rate. Increased fresh and dry weights and percentage of dry matter after cultivation in ex vitro condition were not statistically different between 42 day-old and 49 day-old in vitro plants, but were significantly different between these plants and 35 day-old in vitro plants. The development of shoot and root in ex vitro stage of 42 day-old and 49 day-old in vitro plants was more balanced, as shown by the higher ratio of shoot/root dry weight, than 35 day-old in vitro plants. The results of this study showed that for this Plumbago species, bigger in vitro plants led to better growth during ex vitro stage. These results also indicated that it was possible to transfer in vitro Plumbago plants to ex vitro condition after 5 weeks of in vitro culture stage. [ABSTRACT FROM AUTHOR]

A new spodumene-rich pegmatite was recently discovered on the north side of Plumbago Mountain, Oxford County, Western Maine. The site is about 1.5 km northwest of the famous Dunton gem tourmaline pegmatite. It is an albite-quartz-spodumene pegmatite containing gigantic crystals of spodumene and montebrasite. Some spodumene crystals are more than 11 m in length. The upper portions of the pegmatite contain up to 50 wt% spodumene, which makes it a potentially significant new lithium resource. Preliminary results indicate that Plumbago North Pegmatite contains about 10 Mt of ore with an average Li2O content of 4.68 wt% which is a higher average lithium content than any of the ten top spodumene-producing deposits in the world. [ABSTRACT FROM AUTHOR]

In this study, an efficient method for in vitro regeneration of Plumbago europaea was developed using direct and indirect organogenesis. Accordingly, micropropagation and regeneration were obtained on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators. The effects of explant type and plant growth regulators on shoot organogenesis of P. europaea were evaluated. For the nodal explants, MS medium containing 0.5 mg/l TDZ (11.62 shoots per node) was the best medium for high frequency of micropropagation. In comparison, the highest percentage of direct organogenesis (70%) and number of shoots per explants (14.6) were acquired for the internode explants using 0.5 mg/l TDZ and 0.1 mg/l IAA. The obtained data revealed that TDZ is the most effective cytokinin for the direct shoot organogenesis. The highest indirect organogenesis rate was observed using 2 mg/l BA and 0.1 mg/l NAA for the internode explant. The maximum number of roots was distinguished on ½ MS medium containing 0.5 mg/l IBA (6.42). The rooted plantlets were gradually hardened and acclimatized under ex vitro conditions. As an important outcome, the active compound plumbagin was found mainly in the root tissues of the micro-propagated and regenerated plantlets. Taken all together, this study achieved a successful protocol for in vitro regeneration of P. europrea and could be considered for large-scale multiplication of this important medicinal plant. [ABSTRACT FROM AUTHOR]

Main conclusion: Enhanced secretion of Na+ and Cl− in leaf glands and leaf vacuolar sequestration of Na+ or root retention of Cl−, combined with K+ retention, contribute to the improved salt tolerance of tetraploid recretohalophyte P. auriculata. Salt stress is one of the major abiotic factors threatening plant growth and development, and polyploids generally exhibit higher salt stress resistance than diploids. In recretohalophytes, which secrete ions from the salt gland in leaf epidermal cells, the effects of polyploidization on ion homeostasis and secretion remain unknown. In this study, we compared the morphology, physiology, and ion homeostasis regulation of diploid and autotetraploid accessions of the recretohalophyte Plumbago auriculata Lam. after treatment with 300 mM NaCl for 0, 2, 4, 6, and 8 days. The results showed that salt stress altered the morphology, photosynthetic efficiency, and chloroplast structure of diploid P. auriculata to a greater extent than those of its tetraploid counterpart. Moreover, the contents of organic osmoregulatory substances (proline and soluble sugars) were significantly higher in the tetraploid than in the diploid, while those of H2O2 and malondialdehyde (MDA) were significantly lower. Analysis of ion homeostasis revealed that the tetraploid cytotype accumulated more Na+ in stems and leaves and more Cl− in roots but less K+ loss in roots compared with diploid P. auriculata. Additionally, the rate of Na+ and Cl− secretion from the leaf surface was higher, while that of K+, Mg2+, and Ca2+ secretion was lower in tetraploid plants. X-ray microanalysis of mesophyll cells revealed that Na+ mainly accumulated in different cellular compartments in the tetraploid (vacuole) and diploid (cytoplasm) plants. Our results suggest that polyploid recretohalophytes require the ability to sequester Na+ and Cl−(via accumulation in leaf cell vacuoles or unloading by roots) and selectively secrete these ions (through salt glands) together with the ability to prevent K+ loss (by roots). This mechanism required to maintain K+/Na+ homeostasis in polyploid recretohalophytes under high salinity provides new insights in the improved maintenance of ion homeostasis in polyploids under salt stress. [ABSTRACT FROM AUTHOR]

Plumbago zeylanica L., commonly known as leadwort or Ki Encok plant in Indonesia, has some beneficial effects on human health, such as anticancer, anti-microbial, anti-inflammatory, wound healing, and anti-diabetic. Therefore, this experimental study aims to evaluate the chloroform extracts of P. zeylanica L. leaves for the isolation of the phytoconstituents. The crushed form of the leaves from South Sumatera (6.5 g) was macerated in 65 ml of chloroform for 5 days, while a total of 80 g leaves powder from Central Java was dissolved in 800 ml of chloroform. The extracts were filtered and evaporated to dryness using a rotary evaporator. Subsequently, the identification of secondary metabolites was carried out using Gas Chromatography-Mass Spectrometry (GC-MS). In conclusion, the leaves of P. zeylanica L. contain β -sitosterol, lupeol, plumbagin, phytol, and lauric acid. [ABSTRACT FROM AUTHOR]

Selection of elite accessions is important to get the maximum quantity of bioactive compounds in medicinal plants. In this study, thirteen accessions of Plumbago zeylanica L., a medicinal herb, was studied for phytochemical and genetic diversity analysis. Phytochemical analysis showed that accession number IC-524441 contains the highest amount of flavonoids, phenolic, tannin and plumbagin content, it also possesses maximum antioxidant activity. Genetic diversity analysis showed that 15 SCoT (Start codon targeted) and 20 CBDP (CAAT Box Derived Polymorphism) primers produced 86 and 110 amplicons in total and average were 5.73 and 5.5 amplicons/primer respectively. PIC values ranged from 0.12 to 0.37 and average is 0.25/primer in the case of SCoT marker whereas, in the case of CBDP marker it was 0.15 to 0.59 with average of 0.26/primer. Cluster analysis showed that SCoT marker provides three clusters where cluster 1 and cluster 2 have only 2 accessions while cluster 3 contains nine accessions. Similarly, CBDP marker provides three clusters where cluster 1 contains only 1 accession and cluster 2 and cluster 3 contains six accessions in each. Further AMOVA analysis of SCoT marker possesses maximum variation of 8% among agro-ecological regions whereas in case of CBDP marker its 5%. From the present investigation, it was found that IC-524441 is an elite accession and contains highest amount of bioactive compound. Our results also demonstrate that both SCoT and CBDP markers are informative and can be utilized for evaluation of genetic relationships among Plumbago zeylanica L. accessions. [ABSTRACT FROM AUTHOR]