Monnieri Plants Research Articles

Indole-3-acetic acid-producing Pantoea strain MTP 17 and Glomus mosseae synergistically improve the growth, yield and bacoside content of Bacopa monnieri

The present study aimed to evaluate the performance of Indole-3-acetic acid-producing bacteria (IPB) and arbuscular mycorrhizal fungi (AMF) in compatible form for early establishment and higher growth, yield, and secondary metabolite content of vegetatively propagated Bacopa monnieri plants. Four species of Glomus, viz., G. mosseae (Gm), G. aggregatum (Ga), G. fasciculatum (Gf), G. intraradices (Gi), and two Indole-3-acetic acid-producing bacteria (IPB), Bacillus mojavensis (Bm), and Pantoea sp. (Pt) were evaluated individually and in combinations in a pot experiment. Various plant growth parameters like height, biomass, NPK uptake, mycorrhizal colonization, bacterial population in the rhizosphere, phosphatase activity, and bacoside-A content were determined and found to vary with different treatments. Among all the treatments, plants inoculated with Gm + Pt performed better than any other treatment or un-inoculated control. The performance of the consortium of Gm + Pt was further validated in a field experiment on the same parameters. A clear synergistic interaction between Gm and Pt was observed as Pt enhanced rooting and subsequently improved the colonization and spore population of Gm. On the other hand, the presence of Gm improved the population of Pt in the rhizosphere. The presence of both microbes improved the bacoside-A yields in B. monnieri. The results of this experiment indicate the compatibility and synergy between G. mosseae and Pantoea sp. and suggest using this microbial consortium for plant propagation and growth enhancement while improving soil biological health.

Retraction Note: Use of the cryptogein gene to stimulate the accumulation of bacopa saponins in transgenic Bacopa monnieri plants.

Retraction Note: Use of the cryptogein gene to stimulate the accumulation of bacopa saponins in transgenic Bacopa monnieri plants.

A metabolic reprogramming in Bacopa monnieri plants induced by methyl-jasmonate and enhanced biosynthesis of triterpene saponins

The bacoside-A is the main bioactive metabolite present in Bacopa monnieri L. The yield of bacoside-A from plant sources is too less to meet industrial demand. This study investigated the effect of methyl jasmonate (MeJA) on metabolic profiles and bacoside-A biosynthesis in greenhouse-grown plants of B. monnieri. The greenhouse-grown plants were kept inside a closed plastic container and then elicited with MeJA vapour. It was found that MeJA treatment (200 µM for 2 h on day zero) followed by harvesting leaves on day three was the optimum treatment, resulting in an increase of 1.8-fold, 2.2-fold, 3.1-fold, and 2.5-fold in bacoside A3, bacopaside II, bacopaside X, and bacopasaponin C in comparison with control plants, respectively. The transcript levels of squalene synthase (SQS), a key gene involved in bacoside-A biosynthesis was transiently induced by MeJA and reached a peak of up to 3.7-fold at 24 h. The relationships between bacosides content and primary metabolites in plants treated with MeJA were analyzed by gas chromatography-mass spectrometry. MeJA enhances glycolysis and TCA cycles, generating acetyl-CoA, which acts as a precursor for bacosides synthesis, as revealed by untargeted metabolomics. Therefore, in planta application of MeJA appears to have a great impact on enhanced bacoside-A accumulation, in the greenhouse-grown plants of B. monnieri. In this study, farmers and the pharmaceutical industry will have access to a simple and time-saving elicitation strategy for improving bacosides production from greenhouse-grown plants without the need for expensive and time-consuming in vitro culture techniques.

Cnidium monnieri (L.) Cusson Flower as a Supplementary Food Promoting the Development and Reproduction of Ladybeetles Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae).

Predaceous ladybeetles are highly polyphagous predators that ingest supplementary food from flowering plants. Flowering plants widely grown in agroecosystems can sustain multiple natural enemies of agricultural pests, and the pollen and nectar resources from flowering plants may have a positive role in natural enemies. Cnidium monnieri (L.) Cusson, an annual herb with many flowers, blooms from May to July. C. monnieri can support several predatory natural enemies, and the addition of C. monnieri strips increases the density of Harmonia axyridis (Pallas) and improves the biological control of apple aphids in an apple orchard. H. axyridis is also the most important natural enemy in wheat aphid biocontrol and is attracted to healthy and aphid-infested C. monnieri plants. In addition, adult Propylaea japonica Thunberg survives significantly longer on C. monnieri flowers than on a water-only diet. In this study, a laboratory experiment was conducted to assess (i) the effect of nutritional supplements derived from C. monnieri flowers on the development and reproduction of H. axyridis under a wheat aphids-only diet; (ii) the effect of C. monnieri flowers on H. axyridis adult reproduction performance. We compared the larval durations, survival, weight, adult longevity, and reproduction of H. axyridis reared on wheat aphids-only and aphids plus C. monnieri flower diets. The results showed that H. axyridis larvae reared on aphids plus flowers had significantly greater weights and survival rates, shorter larval durations, and produced 1.62 times more eggs than those reared on wheat aphids-only diets. H. axyridis adults ingesting a C. monnieri flowers plus an aphid diet increased egg production 1.44 times compared to the aphids-only diet. Our study demonstrates that C. monnieri flowers as a supplementary food positively affect the survival, development, and reproduction performance of H. axyridis.

Copper and iron metal resistant rhizospheric bacteria boost the plant growth and bacoside A content in Bacopa monnieri under stress conditions.

Bacteria that enhance plant growth and development and are found in the vicinity of roots are referred to as plant growth-promoting rhizobacteria. Some beneficial bacteria help plant tolerance to many hazardous chemical elements. In this context, Cupriavidus basilensis, Novosphingobium humi, Bacillus zanthoxyli, Bacillus sp., Paenibacillus alvei, Ancylobacter aquaticus and Ralstonia syzygii metal-tolerant rhizospheric bacteria were isolated from rhizospheric soil associated with Bacopa monnieri. The beneficial effects of rhizospheric bacteria on B. monnieri plant physiology and biochemical responses were investigated under pot conditions at two levels (100μM and 500μM) of CuSO4 or FeCl3. N. humi, A. aquaticus and R. syzygii bacterial strains were associated with significantly increased height and biomass under normal and stress conditions. An assay for indole acetic acid in isolated rhizospheric bacteria found differential secretion except Bacillus zanthoxyli. Bacoside A is a major phytocompound in B. monnieri with medicinal value; maximum induction was observed in the R. syzygii treatment. High concentration of copper and iron salts negatively influenced height, biomass and photosynthetic pigments; however N. humi, A. aquaticus, Bacilllus sp. and R. syzygii beneficial bacterial helped plants under stress conditions. Moreover, a significant enhancement in chlorophyll a and b was noticed in C. basilensis, B. zanthoxyli, Bacilllus sp., P. alvei and R. syzygii treatments, without much influence on carotenoid levels. Therefore, the present study emphasises the importance of isolating plant growth-promoting rhizobacteria for use in bacopa plants exposed to metals such as copper and iron in soil.

Enhancing secondary metabolite production and antioxidants in Bacopa monnieri grown on tannery sludge contaminated soil

The current study focuses on the stimulation of Bacoside A content of the Bacopa plant that played a potential role in the treatment of Alzheimer’s disease and epilepsy. The objective of the study was to explore the potential of the B. monnieri plant for metal tolerance when cultivated on tannery sludge-contaminated soil without compromising its phytochemical quality. Bacoside content of the plant gets 2 times higher than the control without any chemical additives. Bacosides are the alleged bioactive component and attribute to the neuroprotective function. Export-import bank of India acknowledged Bacopa monnieri as the second most crucial medicine as it has metabolites that can cure mental disorders and amnesia. The effect of tannery sludge on enzymatic and non-enzymatic antioxidant potential along with the effect on metabolite i.e. bacoside A content of the plant. The maximum transfer (known as translocation factor) from root to shoot for Cr, Cd, Co, Ni and Pb was 0.33, 0.28, 0.75, 0.69 and 0.43 respectively. This depicts that here in tannery sludge mixed soil, B. monnieri was not behaving like a hyperaccumulator plant. Although it sequestered a large portion of metal in the root region and showed high vegetative growth in a combination of 75:25 tannery sludge and soil. Superoxide dismutase, Catalase, peroxidase, malondialdehyde, proline & ascorbic acid scaled up with increasing tannery sludge in the treatment whereas in the case of chlorophyll a, b, and carotenoid content, a sharp decline trend was observed at 100% tannery sludge treatment. A similar trend was recorded in bacoside A content of the plant, and the highest was 2% that of control in a 75:25 ratio of tannery sludge and soil. Enzymatic antioxidant along with non-enzymatic antioxidants protects the plant from oxidative stress generated by tannery sludge. Principal components confirm our finding that the translocation factor plays a direct role in the activation of the photosynthetic machinery. Principal component-I contributes 59% of the total variance and Principal component-II contributes 33% of the total variance.

Fingerprinting Profile of Saponin Glycosides in Bacopa Monnieri (L.) Wettst

As a medhya rasayana in Ayurveda formulations, B. monnieri has undergone extensive research into its bioactive chemicals in the last few years. To date, no research has been done on the chemical profiles of different extract components, such as the leaf and stem compounds that are crucial for pre-formulation purposes. Routine B. monnieri testing requires the use of welldefined concentrations of bacopaside II and baposide A3. As a result, the study's goal was to identify the active ingredients found in the leaves and stems of the Bacopa monnieri plant. We want to standardize phytoconstituents in B. monnieri leaves and stems using two different markers and the HPLC technique described here. The HPLC fingerprinting analysis of the extract of B. monnieri was carried out. Phosphoric acid: acetonitrile (65: 53 v/v) was used as the mobile phase, and the absorbance at 240 nm was measured after a 50-minute run time. Saponins found in the leaf extract included apigenin, bacopasaponin A, bacoside A3, and bacoside A1, with retention durations of 21.217, 17.843, 15.241, 14.540, 11.274, and 11.263, respectively. Stem extract, on the other hand, revealed the existence of major compounds such as bacopaside II, bacoside A3, bacopaside I, and bacosaponin C, with retention durations of 21.133, 14.186, 14.021, and 13.970, respectively. Phytoconstituents were found in greater abundance in the leaf extract of the plant than in the stem extract. The standard bacopaside II and bacoside A3 were used to match the phytoconstituents found in leaves and stems. HPLC fingerprinting was used to check the quality of plant extracts and their fractions, such as methanol. Keywords: ,,,,

Isolation and characterization of endophytic fungi having plant growth promotion traits that biosynthesizes bacosides and withanolides under in vitro conditions.

Endophytes are regarded with immense potentials in terms of plant growth promoting (PGP) elicitors and mimicking secondary metabolites of medicinal importance. Here in the present study, we explored Bacopa monnieri plants to isolate, identify fungal endophytes with PGP elicitation potentials, and investigate secretion of secondary metabolites such as bacoside and withanolide content under in vitro conditions. Three fungal endophytes isolated (out of 40 saponin producing isolates) from leaves of B. monnieri were examined for in vitro biosynthesis of bacosides. On morphological, biochemical, and molecular identification (ITS gene sequencing), the isolated strains SUBL33, SUBL51, and SUBL206 were identified as Nigrospora oryzae (MH071153), Alternaria alternata (MH071155), and Aspergillus terreus (MH071154) respectively. Among these strains, SUBL33 produced highest quantity of Bacoside A3 (4093 μg mL−1), Jujubogenin isomer of Bacopasaponin C (65,339 μg mL−1), and Bacopasaponin C (1325 μg mL−1) while Bacopaside II (13,030 μg mL−1) was produced by SUBL51 maximally. Moreover, these aforementioned strains also produced detectable concentration of withanolides—Withaferrin A, Withanolide A (480 μg mL−1), and Withanolide B (1024 μg mL−1) respectively. However, Withanolide A was not detected in the secondary metabolites of strain SUBL51. To best of our knowledge, the present study is first reports of Nigrospora oryzae as an endophyte in B. monnieri with potentials of biosynthesis of economically important phytomolecules under in vitro conditions.Supplementary InformationThe online version contains supplementary material available at 10.1007/s42770-021-00586-0.

In Vitro Propagation, Phytochemical and Neuropharmacological Profiles of Bacopa monnieri (L.) Wettst.: A Review.

Bacopa monnieri has been used as a reputed drug in the Indian traditional ayurvedic system for centuries. This medicinal herb with important phytopharmaceuticals has been popularly known as “Brahmi”. In recent years, B. monnieri has been extensively studied for its bioactive constituents, constituents responsible for memory enhancing effect, and also its diverse other useful effects. It possesses many pharmacological activities such as antioxidant, gastrointestinal, endocrine, antimicrobial, anti-inflammatory etc. The plant has been also used for the treatment of neurological and neuropsychiatric diseases. Due to its multipurpose therapeutic potential, micropropagation using axillary meristems and de novo organogenesis has been extensively studied in the species and is being reviewed. High frequency direct shoot organogenesis can be induced in excised leaf and internode explants in the absence of exogenous phytohormones and the rate of induction is enhanced in the presence of exogenous cytokinins, supplements, growth regulators, etc. Using explants from tissue culture raised plants, direct shoot regeneration leading to production of more than 100 rooted plants/explant within 8–12 weeks period with 85%–100% survival in the field after acclimatization can be expected following optimized protocols. Bioreactor based micropropagation was found to increase the multiplication rate of shoot cultures for the commercial propagation of B. monnieri plants. The maximum content of bacosides has been recorded in shoot biomass using an airlift bioreactor system. Further studies for the biosynthesis of bacosides and other secondary metabolites need to be conducted in the species utilizing untransformed shoot cultures in bioreactors.

Bioremediation of arsenic by soil methylating fungi: Role of Humicola sp. strain 2WS1 in amelioration of arsenic phytotoxicity in Bacopa monnieri L.

Fungi mediated arsenic (As) stress modulation has emerged as an important strategy for the mitigation of As mediated stress management in plants for reducing As contamination to the food chain. In the present study, total of 45 fungal strains were isolated from the three As contaminated sites of West Bengal, India. These strains were morphologically different and inhibited variable As tolerance (10 to 5000mgl-1As). Total 21 fungal isolates, tolerant up to 5000mgl-1 AsV, were investigated for As removal (10mgl-1 As) after 21d of cultivation under laboratory conditions. The As bioaccumulation in fungal biomass ranged between 0.146 to 11.36gkg-1 biomass. Range of volatilized As was between 0.05 to 53.39mgkg-1 biomass. Most promising bioaccumulation and biovolatilization potential were observed in strains viz., 2WS1, 3WS1 and 2WS9. Strain 2WS1 showed highest As biovolatilization (53.39mgkg-1 biomass) and was identified as Humicola sp. using ITS/5.8S rDNA gene sequencing. This is the first report of Humicola sp. having As biomethylation property. Best first 8 As biomethylating fungal strains were further tested for their As remediation and PGP potential in Bacopa monnieri plant grown in As contaminated soil (20mgkg-1) in a pot experiment under greenhouse conditions. The highest leaf stem ratio and lowest As content in leaf tissues were observed in 2WS1 inoculated Bacopa monnieri plants. The presence of arsM gene in 2WS1 strain suggests As biovolatilization as possible bioremediation and As stress mitigation strategy of 2WS1. Therefore, application of this strain of Humicola sp. strain 2WS1 in As contaminated soils could be a potential and realistic mitigation strategy for reducing As contamination to cropping system coupled with enhanced productivity.

Enhancement of secondary metabolites in Bacopa monnieri (L.) Pennell plants treated with copper-based nanoparticles in vivo.

The study aims to document the effect of starch-stabilised copper-based nanoparticles (CuNPs) on the biosynthesis of pharmaceutically valuable secondary metabolites, especially saponins, of the reputed nootropic herb Bacopa monnieri (L.) Pennell. CuNPs were synthesised chemically by the reduction of cupric sulphate pentahydrate with ascorbic acid using starch as the capping agent. They were characterised by UV-visible spectrophotometry, Fourier-transform infra-red spectroscopy, X-ray diffraction, high-resolution transmission electron microscopy and zeta potential. The nanoparticles consisted of cuprous oxide and metallic copper, were approximately spherical, polydispersed with diameter <20 nm. Hydroponically grown B. monnieri plants were treated in vivo with the CuNPs between the concentrations of 0-100 mg l-1. Spectrophotometric estimation of the total contents of saponins, alkaloids, phenolics, flavonoids and DPPH radical scavenging capacity from the methanolic extracts of the whole plants showed a hormetic increase in the content of secondary metabolites in a concentration-dependent manner from 5 mg l-1 until it declined at toxic metabolic concentration. This was accompanied by an increase in ROS markers hydrogen peroxide and malondialdehyde as well as a hormetic effect on activities of phenylalanine ammonia lyase and antioxidant enzymes catalase, ascorbate peroxidase and superoxide dismutase. CuNPs at sub-toxic concentrations were found to enhance secondary metabolism and antioxidant capacity in Bacopa monnieri through ROS-mediated defence response.

Antipruritic Effect of Natural Coumarin Osthole through Selective Inhibition of Thermosensitive TRPV3 Channel in the Skin.

Coumarin osthole is a dominant bioactive ingredient of the natural Cnidium monnieri plant commonly used for traditional Chinese herbal medicines for therapies and treatments including antipruritus and antidermatitis. However, the molecular mechanism underlying the action of osthole remains unclear. In this study, we report that osthole exerts an antipruritic effect through selective inhibition of Ca2+-permeable and thermosensitive transient receptor potential vanilloid 3 (TRPV3) cation channels that are primarily expressed in the keratinocytes of the skin. Coumarin osthole was identified as an inhibitor of TRPV3 channels transiently expressed in HEK293 cells in a calcium fluorescent assay. Inhibition of the TRPV3 current by osthole and its selectivity were further confirmed by whole-cell patch clamp recordings of TRPV3-expressing HEK293 cells and mouse primary cultured keratinocytes. Behavioral evaluation demonstrated that inhibition of TRPV3 by osthole or silencing by knockout of the TRPV3 gene significantly reduced the scratching induced by either acetone-ether-water or histamine in localized rostral neck skin in mice. Taken together, our findings provide a molecular basis for use of natural coumarin osthole from the C. monnieri plant in antipruritic or skin care therapy, thus establishing a significant role of the TRPV3 channel in chronic itch signaling or acute histamine-dependent itch sensation.

Triterpenoid Saponins Investigation and Pharmacological (Cytotoxic and Antioxidant) Properties of Bacopa monnieri L. Cultivated in Iraq

Bacopa monnieri L. (Scrophulariaceae), synoname is Herpestis monniera that provides bioactive compounds, especially triterpenoid saponins (Bacosides) which exhibits an important biological activities, like hypothyroidism, anticonvulsant, memory enhancing and antistress. Because there are no researches about B. monnieri L. plant that grow in Iraq, and there active compounds especially triterpenoid saponin (TS), and there effects. This study was detected the presence of (TS) in, and examined the cytotoxic and the antioxidant activity of these compounds in vitro. The study was included the extraction and identification of TS from the whole parts of B. monnieri L. by using three methods, and the best yield was analyzed by High Performance Liquid Chromatographic (HPLC) to identify bacosides compounds. In vitro, TS was examined the cytotoxic activity against two cancer cell lines, human cervical cancer (Hela), rhabdomyo sarcoma (RD), and rat embryogenic fibroblast (REF) as a normal cell, at concentrations of 62.5, 125, 250, 500 and 1000 ?g/ml at 24, 48, and 72-hr. Free radical 1,1Dyphenyl-2-picrylhydrazyl radical (DPPH) was used for testing the ability of the TS as antioxidant at 20, 40, 60, 80, 100 ?g/ml concentrations. The results revealed that B. monnieri plant had many components of bacosides when detected by HPLC. Cytotoxic study suggested that TS inhibited the growth of cancer cells, and this effect is depending on the extraction of TS concentration. The effect of TS on Hela cell line was more than that for RD, while the highest effect of TS on Hela was at the concentration of 250 ?g/ml after 48 hr. The cytotoxic effect has a significant effect at (P?0.05). The results revealed that TS has high antioxidant influence 99.37% at 100 ?g/ml concentrations, followed by 98.20% in 80 ?g/ml concentration.

Thidiazuron induced in vitro morphogenesis for sustainable supply of genetically true quality plantlets of Brahmi

An effective and rapid method for propagating genetically true plantlets of a pharmaceutically important plant Bacopa monnieri L. was developed from thidiazuron (TDZ) pulse treated nodal segment explants. Explants treated for 24 h with different concentration of 0.0, 5.0, 10, 20 or 40 μM thidiazuron (TDZ) prepared in liquid Murashige and Skoog (MS) medium induces multiple shoot on MS basal medium without growth regulators. The highest frequency 95.3 ± 2.3% of regeneration and maximum number 43.7 ± 2.0 of shoots per explants with an average shoot height of 5.4 ± 0.29 cm were achieved after 08 weeks of culture from the explants treated with 20 μM TDZ. Simultaneously, the regenerated microshoots were rooted on the same growth regulator free media. Elongated shoots with roots were successfully acclimated to ex vitro condition with a 97% survival rate. Genomic stability of in vitro developed plants of B. monnieri was first time determined using flow cytometry and single primer amplification reaction (SPAR) methods viz., random amplified polymorphic DNA (RAPD), intersimple sequence repeat polymorphic DNA (ISSR) and directed amplification of minisatellite DNA (DAMD) and compared with donor plant. Nuclear DNA (nDNA) content of in vitro propagated B. monnieri plants has been estimated as 2.08 pg/2C based on the flow cytometric investigation of nuclei isolated from leaf tissues. No significant variations in nDNA content, plant genome size and ploidy level were observed between micropropagated plants and field grown donor plants. Likewise, the SPAR markers also revealed monomorphic banding profiles in micropropagated plantlets of B. monnieri and similar with to that of the donor plants verifying their genomic stability.

Selective Inhibition of Thermosensitive TRPV3 Channel by Natural Coumarin Osthole for Itch Relief

Natural coumarin osthole is a dominant bioactive ingredient of Cnidium monnieri plant that is widely used as traditional Chinese medicine for remedies such as antipruritus and antidermatitis. However, the underlying molecular mechanism of action for osthole remains unknown. In this study we show that osthole exhibits antipruritic effect by selective inhibition of thermosensitive and Ca2+-permeable TRPV3 channels primarily expressed in the keratinocytes of the skin. We utilized the calcium fluorescent assay in FlexStation 3 96-well format and identified coumarin osthole as a selective inhibitor of TRPV3 channels expressed in HEK293 cells or HaCaT keratinocytes. Whole-cell patch clamp recordings further confirmed that osthole selectively inhibited TRPV3 current activated by TRP channel agonist 2-APB (50 µM) in dose-dependent manner with an IC50 value of 34.1 ± 0.7 μM. Behavioral evaluations revealed that inhibition of TRPV3 by osthole or knockout of TRPV3 gene significantly attenuated mouse scratching induced by either acetone-ether-water (AEW) or histamine in localized rostral neck skin. Taken together, our findings identify osthole as a relatively selective blocker of TRPV3, demonstrating a critical role of TRPV3 in chronic itch and also providing a molecular basis for use of coumarin osthole from C. monnieri in anti-pruritic therapy or skin care.

In Vitro Analysis of Bacosine as Novel Therapeutic Agent for Murine Breast Cancer

The anti-metastatic activity of bacosine (Bacopa monnieri plant) was studied using 4T1 murine breast cancer cell line by targeting matrix metalloproteinases (MMPs) which plays important role in invasion and metastasis. The inhibitory concentration (IC50) of bacosine for 4T1 cell line was 21 μM, which indicates that it is toxic to cancer cell line. The effect of different concentrations (10, 21 and 50 μM) of bacosine on metastatic matrix metalloproteinase-9 (MMP-9) gene expression of 4T1 murine breast cancer cells was studied by real time PCR. At high concentrations of bacosine (21 and 50 μM) MMP-9 gene expression was down-regulated after 24 h of treatment in a concentration dependent manner. The effect of bacosine on apoptosis was evaluated by annexin binding assay which showed anti-apoptotic effect in early and late apoptotic stages of cancer cells. These results suggest that bacosine plays a role in regulating cell metastasis by inhibiting MMP-9 and showing anti-apoptotic effect in breast cancer cells.

Triterpenoid Saponins Investigation and Pharmacological (Cytotoxic and Antioxidant) Properties of Bacopa monnieri L. Cultivated in Iraq

Bacopa monnieri L. (Scrophulariaceae), synoname is Herpestis monniera that provides bioactive compounds, especially triterpenoid saponins (Bacosides) which exhibits an important biological activities, like hypothyroidism, anticonvulsant, memory enhancing and antistress. Because there are no researches about B. monnieri L. plant that grow in Iraq, and there active compounds especially triterpenoid saponin (TS), and there effects. This study was detected the presence of (TS) in, and examined the cytotoxic and the antioxidant activity of these compounds in vitro. The study was included the extraction and identification of TS from the whole parts of B. monnieri L. by using three methods, and the best yield was analyzed by High Performance Liquid Chromatographic (HPLC) to identify bacosides compounds. In vitro, TS was examined the cytotoxic activity against two cancer cell lines, human cervical cancer (Hela), rhabdomyo sarcoma (RD), and rat embryogenic fibroblast (REF) as a normal cell, at concentrations of 62.5, 125, 250, 500 and 1000 ?g/ml at 24, 48, and 72-hr. Free radical 1,1Dyphenyl-2-picrylhydrazyl radical (DPPH) was used for testing the ability of the TS as antioxidant at 20, 40, 60, 80, 100 ?g/ml concentrations. The results revealed that B. monnieri plant had many components of bacosides when detected by HPLC. Cytotoxic study suggested that TS inhibited the growth of cancer cells, and this effect is depending on the extraction of TS concentration. The effect of TS on Hela cell line was more than that for RD, while the highest effect of TS on Hela was at the concentration of 250 ?g/ml after 48 hr. The cytotoxic effect has a significant effect at (P?0.05). The results revealed that TS has high antioxidant influence 99.37% at 100 ?g/ml concentrations, followed by 98.20% in 80 ?g/ml concentration.

Morpho-histological characterization and direct shoot organogenesis in two types of explants from Bacopa monnieri on unsupplemented basal medium

In the present study, high frequency regeneration has been obtained via de novo direct shoot organogenesis from leaf and internode explants in Murashige and Skoog (MS) basal medium without any phytohormone supplementation in Bacopa monnieri, an indigenous traditionally used medicinal herb. Leaves and internodes from different positions were excised from 4-weeks-old in vitro propagated B. monnieri plants and cultured on MS basal medium supplemented with 3% (w/v) sucrose and 0.75% (w/v) agar for 4 weeks. The induction of de novo shoot buds was observed at petiolar cut edges of leaf and both proximal and distal cut ends of internode explants within 10–15 days of culture. The first histological changes could be observed after 4–5 days, with meristematic activity of vascular bundles. Proliferation of epidermal cells gave rise to dome-shaped protuberances followed by shoot apical meristems formation and their vascular connections with explant tissues within 2 weeks of culture. However, a basipetal gradient of shoot regeneration from both types of explants collected along the branch axis was noticed after 4 weeks of culture. Leaf and internode explants near the basal region exhibited significantly higher number of shoot buds and micro shoots (8.8/leaf explant and 15/internode explant). Microshoots (7–12 micro shoots/leaf or internode explants) elongated (shoot length 8–9 cm) within 8 weeks on phytohormone free MS medium. Excised micro shoots rooted (100%) in hormone free MS medium within two weeks of culture. Rooted plants were then acclimatized and transferred to field with 95% survival. This protocol may be used for micropropagation, genetic transformation as well as a model system for evaluation of changes associated with acquisition of competence of differentiated cells in phytohormone free medium.

Optimization of a Bacopa monnieri-based genetic transformation model for testing the expression efficiency of pathway gene constructs of medicinal crops

A fast regenerating Agrobacterium tumefaciens-mediated transformation protocol for Bacopa monnieri (L.) Wettst. was developed as a model system for heterologous expression of terpenoid indole alkaloid pathway genes from Catharanthus roseus (L.) G. Don. The direct regeneration of shoots from leaf explants co-cultured with A. tumefaciens resulted in the integration of a tryptophan decarboxylase (tdc) and strictosidine synthase (str) cassette ( ) in the regenerated progeny. The highest transformation efficiency (83.88%) was achieved when leaf explants were infected on the adaxial laminar surface by manual pricking with 48- to 72-h-old suspensions (OD600 = 0.5–0.6) of A. tumefaciens strain LBA1119 (carrying the binary vector pMOG22). The heterologous expression of tryptophan decarboxylase and strictosidine synthase genes that are otherwise not present in B. monnieri plants was confirmed through semi-quantitative PCR and metabolite quantification assays. The entire protocol duration from co-cultivation through regeneration of transgenic plants to their establishment in the glass house took 40–45 d. The developed B. monnieri model can be used to test expression cassettes carrying genes for plant secondary metabolic pathway engineering, especially those genes that are expressed in differentiated cell, tissue, or organs.

Bioactive isochromenone isolated from Aspergillus fumigatus, endophytic fungus from Bacopa monnieri

Fungal endophytes are a significant reservoir of novel bioactive secondary metabolites. Present communication describes isolation and structure determination of isochromenone, from endophytic microorganism &lt;em&gt;Aspergillus&lt;/em&gt; &lt;em&gt;fumigatus&lt;/em&gt; hosted in &lt;em&gt;Bacopa&lt;/em&gt; &lt;em&gt;monnieri&lt;/em&gt; plant. Further, its biological evaluation revealed it as antioxidant and antitubercular. The methanol extract of &lt;em&gt;A&lt;/em&gt;. &lt;em&gt;fumigatus&lt;/em&gt; inhibits the growth of the virulent strain of &lt;em&gt;Mycobacterium&lt;/em&gt; &lt;em&gt;tuberculosis&lt;/em&gt; H37RV with minimum inhibitory concentration 500 μg/mL. This is the first report of isolation of isochromenone from &lt;em&gt;A. fumigatus.&lt;/em&gt;