Alkaloid Yield Research Articles

Assessment of phytochemicals, antioxidant, antibacterial activity, and profiling of functional molecules in a freshwater fern, Salvinia cucullata Roxb.

Salvinia cucullata is a freshwater weed available abundantly in nature and also a troublesome aquatic weed globally. This study was conducted to know the chances of using invasive waste fern S. cucullata as a useful product in different biological applications and to explore the functional molecules and the biological properties of ethanolic extracts of S. cucullata (EESC) . The highest yield of total phenolic, flavonoid, tannins, alkaloids, and saponins content and maximum antioxidant activity (Qualitative assay) are observed in ethanolic extract of S. cucullata than other solvents used in this study. Hence, EESC was chosen for further quantitative antioxidant activity. The ROS scavenging activity of EESC was found to be dose-dependent in assays like 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6- sulfonic acid) (ABTS), hydroxyl, superoxide anion racial scavenging assays, and the ferric reducing power assay. Antibacterial activity of EESC was tested against various gram-negative and gram-positive bacteria, where the zone of inhibition was recorded against Bacillus subtilis as high (16.76 mm) and lowest (10.96 mm) on methicillin-resistant Staphylococcus aureus (MRSA). The presence of many functional groups including alkanes, aldehydes, esters, and amides in EESC is revealed by Fourier infrared spectroscopy (FTIR) analysis. The volatile phytochemical compounds including pentacosadiynoic acid, diazoprogesterone, neophytadiene, heptacosane, and vitamin E were revealed by Gas Chromatography-Mass spectrometry (GC-MS) analysis. These compounds have been identified to hold pharmacological values accountable for the antibacterial and antioxidant activity of EESC. The findings of the present study are a value-added asset in the path to search of unexplored natural resources as biological agents in clinical applications.

A comprehensive comparison on root yield and content of polysaccharides and alkaloids from ten germplasms of Isatis indigotica Fort. in two growing areas

In order to promote the application of high-quality varieties and explore the diversity of cultivated resources of Isatis indigotica Fort., the source of medicinal Banlangen, 10 germplasms were planted in Heilongjiang (HLJ) and Shanxi (SX) provinces to analyze two-year agronomic traits, yields and active ingredient contents in this study. The results showed that germplasm no. 2 had the highest yields in both HLJ (236.54 kg/mu) and SX (368.68 kg/mu), and it had the highest Epigoitrin (RSG) content in HLJ in 2020 (0.8946 %). In addition, the polysaccharide (IRPS) content of germplasm no. 2 was the highest in HLJ (up to 39.100 mg/g), and ranked second in SX (30.767 mg/g). It was the only germplasm with IRPS content that exceeded 30 mg/g in both areas. The yield and RSG content of germplasm no. 6 was slightly lower than that of germplasm no. 2. The germplasms no. 2 and 6 had remarkable advantages in yield and active component content which were worthy of promotion as excellent cultivars in planting industry. Germplasm no. 5 and 10 had advantages in root width and IRPS content, respectively, which could be used as breeding germplasms. Screening germplasm with excellent traits and promoting its cultivation will help to ensure the quality stability of medicinal materials, and to improve economic efficiency of Banlangen production.

PHOSPHATE FERTILIZER AND NANO-MAGNESIUM FERTILIZATION EFFECTS ON GENE EXPRESSION, GROWTH, AND YIELD TRAITS OF DATURA (DATURA STRAMONIUM L.)

A study on the effects of phosphate fertilization and nano-magnesium application on several genes that control alkaloid synthesis, growth, and yield traits in Datura (Datura stramonium L.) underwent a field experiment in 2021 at the College of Agriculture, the University of Kerbala, Iraq. Using a randomized complete block design, the experiment had two factors with three replications. The first factor used phosphate (P) fertilizer, i.e., 0, 25, 50, and 75 kg P ha-1, while the second factor included a nano-magnesium application by spraying with concentrations of 0, 60, 120, and 180 mg Mg L-1. The results showed that adding 50 and 75 kg P ha-1 caused a decline in the concentration of atropine, hyoscyamine, and scopolamine in Datura leaves (22.77, 81.02, and 68.90 mg g-1) and seeds (40.93, 65.69 and 99.79 mg g-1), respectively. Sequentially, 25 and 50 kg P ha-1 generated the most yields of alkaloids in Datura leaves, with an average of 149.10 and 149.12 kg P ha-1. Nano-magnesium application at the concentration of 180 mg Mg L-1 caused a significant decrease in the concentration of atropine, hyoscyamine, and scopolamine in seeds and leaves, i.e., with average values in leaves (29.50, 90.25, and 71.25 mg g-1) and seeds (46.25, 82.49 and 121.320 mg g-1), respectively. However, nano-magnesium concentrations of 0 and 120 mg Mg L-1 gave the highest yield of alkaloids in the leaves, with average values of 152.30 and 152.81 kg ha-1. The nano-magnesium concentration of 120 mg Mg L-1 contributed the largest yield of alkaloids in seeds, with an average of 78.65 kg ha-1. The results also showed phosphorus addition significantly decreased the PMT, TR1, and H6H gene expressions, whereas nano-magnesium application only reduced the H6H gene expression. High quantities of fertilizers phosphorus and nano-magnesium boost Datura's vegetative growth and production but lowered the alkaloid yield, thus recommending a balanced proportion.

Phytochemical Characterization of Vitex negundo Leaves: a Potent Antiandrogenic and Antioxidant Agent

This study was conducted to characterise phytochemicals and to explore the biological activities of Vitex negundo leaves. The washed, course powder of V. negundo leaves were extracted with different solvents of increasing polarity. All the extracts were characterized and biological activities were compared. The results revealed that the ethanolic and cold water extracts showed the presence of all phytochemicals studied except protein compared to other extracts. Further, the quantitative estimation of phytochemicals showed that the ethanolic extract had highest yield and maximum amount of total polyphenols, flavonoids, and alkaloids with the least amount of tannins compared to other extracts studied. Furthermore, the highest total polyphenol content corresponds with the potent biological activities. Indeed, in vitro antioxidant and antisteroidogenic activities were highest in the ethanolic extract than others. To conclude, the present study is the first to report the characterization and antiandrogenic property of V. negundo leaf extracts. The ethanolic extract of V. negundo leaves can be used as an antioxidant and antiandrogenic agent. Hence, it can be considered for the treatment of hyperandrogenic conditions like polycystic ovary syndrome, etc.

Central composite experimental design for ultrasound‐assisted extraction optimization of alkaloid compounds and antioxidant properties from cocoa pod husk ( Theobroma cacao L.)

The objective of this research was to identify ultrasound-assisted extraction (UAE) parameters for maximizing alkaloid yield and antioxidant activity from cocoa pod husk (CPH). The findings indicated that the optimum UAE parameters were extraction temperature 60°C, extraction time 50 min, and solvent to sample ratio 50 ml/g. Under these UAE parameters, total alkaloid content (TAC), alkaloid extraction efficiency (AEE), DPPH radical scavenging capacity (DRSC), and ferric reducing antioxidant power (FRAP) obtained from the CPH were 45.62 mg AE/g dried sample (DS), 63.28%, 16.57, and 2.15 mg TE/g DS, respectively, which were not significantly different (p < .05) from the predicted values (42.57 mg AE/g DS, 59.04%, 15.06 and 0.99 mg TE/g DS, respectively). The residual moisture, water activity, density, water soluble index, and pH of enriched-alkaloids powder from the CPH were 7.66%, 0.39, 0.13 g/ml, 93.77%, and 5.08, respectively. TAC, total phenolic content (TPC), total flavonoid content (TFC), and total saponin content (TSC) of powder were 655.43 mg AE/g DS, 33.60 mg GAE/g DS, 48.51 mg CE/g DS, and 186.48 mg EE/g DS, respectively. ABTS radical scavenging capacity (ARSC), DRSC, cupric reducing antioxidant capacity (CUPRAC) and FRAP of powder were 84.48, 49.54, 155.01, and 6.01 mg TE/g DS, respectively. The results achieved from this study imply that the powder obtained from the CPH is a rich source of alkaloids. Thus, it is a potential candidate for application in the functional food and nutraceutical industries. Practical applications Cocoa pod husk is generated in huge amounts and is a rich source of alkaloids. Recovery of alkaloids from the CPH for potential application in the functional foods and nutraceuticals not only increases value for cocoa tree but also reduces pollution risk to environment.

Gamma ray–induced tissue responses and improved secondary metabolites accumulation in Catharanthus roseus

The present study investigated the impact of gamma ray irradiation on callus biomass growth and the yield of vincristine and vinblastine of in vitro grown tissues of Catharanthus roseus. The biochemical alteration underlying the synthesis of secondary metabolites has also been studied and a comparison of yield was prepared. The embryogenic tissues were exposed to 20, 40, 60, 80, and 100Gy gamma ray doses and the callus biomass fresh weight, the embryogenesis (the embryo numbers, germination, plant regeneration), the alteration of protein, proline, and sugar attributes at different morphogenetic stages were monitored. The callus biomass growth was maximum (1.65g) in 20Gy exposed tissues and was less in 100Gy treatment (0.33g). The gamma-irradiated embryogenic tissues differentiated into embryos but the embryogenesis % and somatic embryo number per culture reduced with increasing doses. It was least in 80Gy where very low numbers of embryos were formed (3.45 and 3.30 mean torpedo and cotyledonary embryo numbers per callus mass, respectively) which later germinated into plantlets. Protein, proline, sugar, and different antioxidant enzymes, i.e., superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) activities, were investigated as the tissues were exposed to gamma ray elicitation/signaling, evoking cellular stress. Increased 80Gy gamma dose inhibited a 42.73% decrease in protein accumulation at initiation stages of embryogenic tissue. Soluble sugar level also declined gradually being least in 80Gy treated tissues (14.51mg gm-1 FW) compared to control (20.2mg gm-1 FW). Proline content, however, increased with increasing gamma doses, maximum at 80Gy (8.28mg gm-1 FW). The SOD, APX, and CAT activity increased linearly with enhanced level of gamma doses and maximum, i.e., 3.91 EU min-1mg-1, 1.71 EU min-1mg-1, and 4.89 EU min-1mg-1, protein activity was noted for SOD, APX, and CAT, respectively, at 80Gy gamma rays treated tissues. The quantification of vinblastine and vincristine in gamma ray elicitated tissues was made by using high-pressure thin layer chromatography (HPTLC). Somatic embryo-regenerated plant's leaves had the maximum yield of vinblastine (15.13 µgm gm-1 DW) at 40Gy irradiation dose compared to control (13.30 µgm gm-1 DW)-the increased yield % is 13.75. The stem is also rich source producing 11.98 µgm gm-1 DW of vinblastine. Among the various developmental stages of embryos, vinblastine content was highest in germinating stage of embryos (10.14 µgm gm-1 DW) compared to other three, i.e., initiation, proliferation, and maturation embryo stages. Similarly, highest accumulation of vincristine (6.32µg gm-1 DW) was noted at low gamma irradiation dose (20Gy) in leaf tissues. The present study indicates that the synthesis of vinblastine and vincristine was growth- and development-specific and the lower 20-40Gy gamma levels were more effective in enriching alkaloids while higher doses declined yield. KEY POINTS: • Vinblastine and vincristine yield was quantified in in vitro grown tissues and leaves of embryo regenerated Catharanthus roseus after gamma ray treatment. • The accumulation of vinblastine and vincristine was maximum in regenerated leaves; low doses were more efficient in improving yield. • Gamma ray irradiation impacted biochemical profiles, caused cellular stress, and perhaps responsible for improved alkaloid yield.

PHYTOCHEMICAL AND PHARMACOLOGICAL ACTIVITIES OF MUKIA MADERASPATANA

Objective: The study aimed to evaluate the bioactive compounds of Mukia maderaspatana and its properties. In addition, the preliminary phytochemical analysis and antioxidant potential of Mukia maderaspatana were performed. Methods: Extraction of leaves of Mukia maderaspatana made using Soxhlet apparatus, followed by evaluation of phytochemicals such as alkaloids, flavonoids, coumarin, glycosides, terpenoids, tannin, steroid, saponin, phenol, anthraquinone by the standard methodology of Sofowara, Trease and Evans, Harborne. Quantification of phytochemicals were performed accordingly to standard methodology. Antioxidant activity were performed accordingly to methodology of Shimada, Prieto, Zhang and Benzie. Methodology of Hema et al. is followed for detection of bioactive compounds by GC-MS analysis. Results: The Qualitative analysis revealed the presence of higher concentration of phytocompounds in Hydroalcoholic extract. The yield of alkaloid (215.8±4.78 mg AE), flavonoid (307.10±8.73 mg QE), phenol (374.50±6.41 mg GAE), tannin (129.98±5.32 mg TAE) and terpenoids (119.23±4.17 mg LE) are higher in hydroalcoholic extract than aqueous and methanolic extract. IC50 value for DPPH radical scavenging activity, Total antioxidant activity, Hydrogen peroxide scavenging activity and Ferric reducing antioxidant power activity are (61.43µg/ml), (59.68µg/ml), (55.69µg/ml), (64.60µg/ml). GC-MS analysis revealed the presence of about thirty compounds in hydroalcoholic extract of Mukia maderaspatana. Conclusion: The present study reveals that Mukia maderaspatana is an important medicinal plant. We can conclude that Leaves of Mukia maderaspatana are good source of antioxidants, which might cure the disease related to oxidative stress.

Optimization of the extract from flower of Citrus aurantium L. var. amara Engl. and its inhibition of lipid accumulation.

Flower of Citrus aurantium L. var. amara Engl. (CAVA) has been confirmed to have promising anti-obesity effects. However, the regulation of alkaloid extracts from flower of CAVA (Al) on lipid metabolism remain unknown. In this study, Al was optimized by ultrasound-assisted extraction using response surface methodology. The optimal conditions were ultrasonic time 72 min, ethanol concentration 78% and liquid/solid ratio 30 ml/g with the maximum alkaloid yield 5.66%. LC-MS assay indicated that the alkaloid compounds were enriched in Al after optimization. Nine alkaloid compounds were identified in Al by LC-MS assay and stachydrine, caffeine and cathine appeared as the major alkaloid compounds. Bioactivity assay showed that Al treatment significantly increased superoxide dismutase (SOD) activity, and reduced malonaldehyde (MDA) and reactive oxygen species (ROS) levels. Al administration also reversed oleic acid-induced hepatic steatosis in Hep G2 cells by inhibiting the expression of lipogenesis-signaling genes including fatty acid synthase (FAS), peroxisome proliferator-activated receptor subtype γ (PPARγ), uncoupling protein 2 (UCP2), and retinol binding protein (RBP4). However, OA-induced reduction of lipolysis-related gene carnitine palmitoyl transferase 1A (CPT1A) in Hep G2 cells was not improved by Al supplementation. Moreover, the increased SOD activity and decreased MDA and ROS contents were also observed in Caenorhabditis elegans by Al addition. Al intervention exhibited the ability to inhibit lipid accumulation in C. elegans by suppressing expression of lipid metabolism-related genes. These results suggested that the alkaloid extracts from the flower of CAVA showed great potential to regulate lipid metabolism. PRACTICAL APPLICATIONS: The extraction of alkaloid extracts from the flower of CAVA was optimized with a maximum yield of 5.66%. The regulatory effects and mechanisms of Al on lipid metabolism of Hep G2 cells and Caenorhabditis elegans were also investigated. More clinical studies are required to evaluate the potential of using alkaloids from the flower of CAVA as therapeutic agents against lipid metabolic disorders.

An innovative extraction strategy for herbal medicine by adopting p-sulphonatocalix[6]/[8]arenes.

Alkaloids exist in various herbal medicine widely and exhibit diverse biological and pharmacological activities. p-Sulphonatocalix[6]arenes (SC6A) and p-sulphonatocalix[8]arenes (SC8A) are water-soluble supramolecular macrocycles and are applied to the extraction of alkaloids from herbal products. In this study, an innovative method of SC6A/SC8A assisted extraction of the alkaloids from herbs was established. SC6A and SC8A were designed to extract 27 alkaloids from seven herbal medicines. Based on the significant solubilisation and extraction effect, Stephaniae Tetrandrae Radix (Fangji, FJ) was selected to obtain the optimal extraction process by adopting single factor test and orthogonal experiment. Then, the alkaloids and SC6A/SC8A were separated by one-step alkalisation and SCnA were reused. The host-guest complexes between alkaloids and SCnA were determined by competitive fluorescence titration, differential scanning calorimetry (DSC), Fourier-transform infrared (FTIR) and proton nuclear magnetic resonance (1 H-NMR) analysis. The optimum condition for SC6A assisted extraction was 5:1:80 (g/g/mL) for herbs/SC6A/solution ratio, 355-250 μm particle size and ultrasonicate 0.5 h, whilst 10:1:40 (g/g/mL) for herbs/SC8A/solution ratio, 355-250 μm particle size and ultrasonicate 0.5 h for SC8A assisted extraction. The total yield of alkaloids (fangchinoline and tetrandrine) from FJ was increased by 4.87 times and 5.97 times with SC6A and SC8A. Moreover, a good reusability of SC6A/SC8A was achieved by alkalisation dissociation. Host-guest complexes were determined by competitive fluorescence titration at a molar ratio of 1:1 between most alkaloids (25/27, except evodiamine and rutaecarpine) and SC6A/SC8A. The complex structure was proved by DSC, FTIR and 1 H-NMR analysis. The study provided an effective eco-friendly and energy-saving extraction method of alkaloids from herbal medicine.

Simultaneous extraction and enrichment of alkaloids from lotus leaf by in-situ cloud point-reinforced ionic liquid assisted mechanochemical extraction technology

A novel, green and efficient in-situ cloud point-reinforced ionic liquid-assisted mechanochemical extraction (IS-CPR-IL-MCE) technique coupled with ultrahigh performance liquid chromatography was established to simultaneously extract, concentrate and separate five alkaloids (including liensinine, isoliensinine, neferine, O-demethyl nuciferine and nuciferine) from lotus leaf. The parameters affecting the extraction efficiency of target analytes, such as the amount of IL, surfactant and salt, grinding time, pH, enrichment temperature and time were systematically studied and optimized by single-factor and multifactor experiments. The highest extraction yield of alkaloids was obtained by IS-CPR-IL-MCE under the following conditions: 0.23 g of 1-dodecyl-3-methylimidazolium bromide, 0.37 g of Triton X-100, 7.73 of pH, 5 min of milling time, 2.5 g of (NH4)2SO4, 60 ℃ and 20 min of enrichment temperature and time. The developed method exhibited a good linearity in the range of 0.5–100.0 μg/mL with the coefficients of correction more than 0.9992, low limit of detection of 8.0–11.2 ng/mL, acceptable reproducibility below 4.45%, and satisfactory recovery in the range of 80.4–93.7% The results demonstrated that the proposed method was successfully applied to the extraction of alkaloids from lotus leaf.

Plant growth and phytoactive alkaloid synthesis in kratom [Mitragyna speciosa (Korth.)] in response to varying radiance.

Leaves harvested from kratom [Mitragyna speciosa (Korth.)] have a history of use as a traditional ethnobotanical medicine to combat fatigue and improve work productivity in Southeast Asia. In recent years, increased interest in the application and use of kratom has emerged globally, including North America, for its potential application as an alternative source of medicine for pain management and opioid withdrawal syndrome mitigation. Although the chemistry and pharmacology of major kratom alkaloids, mitragynine and 7-hydroxymitragynine, are well documented, foundational information on the impact of plant production environment on growth and kratom alkaloids synthesis is unavailable. To directly address this need, kratom plant growth, leaf chlorophyll content, and alkaloid concentration were evaluated under three lighting conditions: field full sun (FLD-Sun), greenhouse unshaded (GH-Unshaded), and greenhouse shaded (GH-Shaded). Nine kratom alkaloids were quantified using an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Greenhouse cultivation generally promoted kratom height and width extension by 93–114% and 53–57%, respectively, compared to FLD-Sun. Similarly, total leaf area and leaf number were increased by 118–160% and 54–80% under such conditions. Average leaf size of plants grown under GH-Shaded was 41 and 69% greater than GH-Unshaded and FLD-Sun, respectively; however, no differences were observed between GH-Unshaded and FLD-Sun treatments. At the termination of the study, total leaf chlorophyll a+b content of FLD-Sun was 17–23% less than those grown in the greenhouse. Total leaf dry mass was maximized when cultivated in the greenhouse and was 89–91% greater than in the field. Leaf content of four alkaloids to include speciociliatine, mitraphylline, corynantheidine, and isocorynantheidine were not significantly impacted by lighting conditions, whereas 7-hydroxymitragynine was below the lower limit of quantification across all treatments. However, mitragynine, paynantheine, and corynoxine concentration per leaf dry mass were increased by 40%, 35%, and 111%, respectively, when cultivated under GH-Shaded compared to FLD-Sun. Additionally, total alkaloid yield per plant was maximized and nearly tripled for several alkaloids when plants were cultivated under such conditions. Furthermore, rapid, non-destructive chlorophyll evaluation correlated well (r2 = 0.68) with extracted chlorophyll concentrations. Given these findings, production efforts where low-light conditions can be implemented are likely to maximize plant biomass and total leaf alkaloid production.

Improvement of Phytopharmaceutical and Alkaloid Production in Periwinkle Plants by Endophyte and Abiotic Elicitors

Periwinkle plant represents a major source of immensely vital terpenoid indole alkaloids and natural antioxidants which are widely used in cancer chemotherapy. A pot experiment was done to evaluate the role of two periwinkle endophytes (Streptomyces sp. and Bacillus sp.) with or without abiotic elicitors (aluminum chloride, tryptophan, and chitosan) on plant biomass, physio-biochemical attributes, phytopharmaceutical constituents, and alkaloid production. Inoculation with endophyte microbes significantly increased plant growth, nitrogen, phosphorus, potassium, carotenoids, ascorbic acid, and alkaloid yield. It also decreased oxidative biomarkers (hydrogen peroxide and malondialdehyde) and had no significant effects on flavonoids and anthocyanin. In this regard, Streptomyces sp. was more effective than Bacillus sp. Foliar spraying with chitosan significantly increased plant growth, chlorophyll, ions, antioxidant capacity, phytopharmaceutical constituents (total soluble phenols, flavonoids, and anthocyanin), and alkaloid yield, associated with a decline in oxidative biomarkers. Conversely, aluminum chloride application generally increased oxidative biomarkers, which was associated with a decreasing effect on plant growth, chlorophyll, and ions. Application of either tryptophan or chitosan with endophyte microbes increased plant growth, chlorophyll, ions, antioxidants, and alkaloid; meanwhile, it decreased oxidative biomarkers. On the contrary, aluminum chloride with endophytes evoked oxidative damage that was associated with a reduction in plant growth, chlorophyll, ions, and phytopharmaceutical constituents. The current study provides a proof-of-concept of the use of the endophyte Streptomyces sp. with chitosan for enhancing periwinkle plant biomass, phytopharmaceuticals accumulation, and alkaloid production.

The Effect of Acids on Alkaloid Yield in Pressurized Water Extraction of Narcissus Pseudonarcissus

Pressurized water (PW) extraction of galanthamine from Narcissus pseudonarcissus bulbs was performed. The obtained yield was compared with the yield from conventional acidified water extraction and methanolic Soxhlet extraction. Both PW and conventional acidified water extraction were followed by a subsequent purification step for the alkaloids. The PW extraction (70 °C, 150 bar, 45 min) yielded as much galanthamine as methanolic-Soxhlet extraction (ca. 3.50 mg/g). Meanwhile, acid-base extraction with 1% of HBr (v/v) at 65 °C for 3 h gave a lower yield (ca. 2.65 mg/g). A higher PW temperature did not significantly increase the galanthamine yield. Pressure increase is not necessary since more water-soluble compounds such as proteins and polysaccharides are co-extracted, resulting in high viscosity of the water extract solution, which hampers the filtration process. Hence, the acidity of the solution is highly important both in the case of PW extraction and acidified water extraction. Besides galanthamine, the total alkaloid profile following Narcissus alkaloids was also obtained. Lycoramine, O-methyloduline, norgalanthamine, epi-norgalanthamine, narwedine, oduline, haemanthamine, O-methyllycorenine, and a haemanthamine derivate were identified. Although a high yield was obtained from PW extraction, the further purification needs to be improved to obtain an economically feasible industrial extraction process.

Influence of plant growth regulators on morphogenic response, biomass and camptothecin production in the callus cultures of Chonemorpha fragrans (moon) Alston

In vitro morphogenic response of mature seed embryo-derived callus cultures of Chonemorpha fragrans was studied using solid and liquid Murashige and Skoog medium amended with cytokinins or their combinations with naphthalene acetic acid at 0.5 mg L-1. The plant growth regulators (PGRs) combination and concentrations tested could not stimulate organogenesis after three subcultivations of the callus cultures on the same PGRs-amended solid medium and when cultivated in the liquid but, formation of morphogenic callus was observed. Evaluation of biomass and camptothecin production showed that the PGRs influenced biomass and CPT yield of the callus cultures. The alkaloid yield of various explants of 3-4 weeks old axenic seedlings was higher in roots (0.019% CPT) followed by mature seed embryos (0.0053%), cotyledons (0.0039%), hypocotyls (0.0024%) and leaves (0.0017%), and no significant difference was observed in yield of CPT from callus induced from the various explants. Camptothecin yield of morphogenic callus cultures cultivated in liquid medium was lower than that of solid due to extracellular leaching effect of the alkaloid. Amount of synthesized CPT in the callus cultures also varied with PGR type and concentration amended in the cultivation medium, and was association with biomass production. Results of the present study suggest that callus cultures offer alternative tissue source for in vitro CPT yield enhancement through biotechnological approaches, with application in the large-scale production of the alkaloid to conserve the ever-decimated natural population of the medicinal woody climber for CPT.

Independent Evolution of a Lysergic Acid Amide in Aspergillus Species.

Ergot alkaloids derived from lysergic acid have impacted humanity as contaminants of crops and as the bases of pharmaceuticals prescribed to treat dementia, migraines, and other disorders. Several plant-associated fungi in the Clavicipitaceae produce lysergic acid derivatives, but many of these fungi are difficult to culture and manipulate. Some Aspergillus species, which may be more ideal experimental and industrial organisms, contain an alternate branch of the ergot alkaloid pathway, but none were known to produce lysergic acid derivatives. We mined the genomes of Aspergillus species for ergot alkaloid synthesis (eas) gene clusters and discovered that three species, A. leporis, A. homomorphus, and A. hancockii, had eas clusters indicative of the capacity to produce a lysergic acid amide. In culture, A. leporis, A. homomorphus, and A. hancockii produced lysergic acid amides, predominantly lysergic acid α-hydroxyethylamide (LAH). Aspergillus leporis and A. homomorphus produced high concentrations of LAH and secreted most of their ergot alkaloid yield into the culture medium. Phylogenetic analyses indicated that genes encoding enzymes leading to the synthesis of lysergic acid were orthologous to those of the lysergic acid amide-producing Clavicipitaceae; however, genes to incorporate lysergic acid into an amide derivative evolved from different ancestral genes in the Aspergillus species. Our data demonstrate that fungi outside the Clavicipitaceae produce lysergic acid amides and indicate that the capacity to produce lysergic acid evolved once, but the ability to insert it into LAH evolved independently in Aspergillus species and the Clavicipitaceae. The LAH-producing Aspergillus species may be useful for the study and production of these pharmaceutically important compounds. IMPORTANCE Lysergic acid derivatives are specialized metabolites with historical, agricultural, and medical significance and were known heretofore only from fungi in one family, the Clavicipitaceae. Our data show that several Aspergillus species, representing a different family of fungi, also produce lysergic acid derivatives and that the ability to put lysergic acid into its amide forms evolved independently in the two lineages of fungi. From microbiological and pharmaceutical perspectives, the Aspergillus species may represent better experimental and industrial organisms than the currently employed lysergic acid producers of the plant-associated Clavicipitaceae. The observation that both lineages independently evolved the derivative lysergic acid α-hydroxyethylamide (LAH), among many possible lysergic acid amides, suggests selection for this metabolite.

Effect of jasmonic acid on alkaloids content and salinity tolerance of Catharanthus roseus based on morpho-physiological evaluation

PurposeCatharanthus roseus is a vital medicinal plant due to its content of alkaloids vincristine and vinblastine which are known to have a special interest in the pharmaceutical industries. Little is known about the effects of jasmonic acid (JA) on C. roseus plants grown under saline conditions. MaterialsPots experiment was conducted to explore the impacts of four treatments of JA (0, 0.50, 1.0, and 1.50 mM) on the growth of C. roseus plants which were irrigated with saline (EC = 5.27 dS m−1) or tap water (EC = 0.53 dS m−1). ResultsThe growth of C. roseus plants was reduced due to the irrigation with saline water; on the other hand, the salinity induced the accumulation of alkaloids. Salt stress induced a 41% increase in the alkaloid yield of C. roseus plants compared to the control plants irrigated with fresh water. Some defense mechanisms including: osmoregulation, ion compartmentalization, and antioxidants accumulation were activated in C. roseus plants upon salt stress. The foliar application of JA to the C. roseus plants increased the growth parameters and mitigated the salt stress; moreover, it enhanced nutrients uptake. Spraying of C. roseus plants with JA significantly enhanced the biosynthesis of compatible solutes and decreased the activity of pyrogallol peroxidase (PPX) and polyphenol oxidase (PPO). Foliar application of JA increased the alkaloids yield of C. roseus plants. ConclusionJA reduced the negative effects of salt stress in C. roseus plants through increasing osmoregulation compounds and regulating the activity of antioxidant enzymes. Spraying of C. roseus with JA could be used as a sustainable strategy for promotion its medicinal property by augmenting secondary metabolites production.

In vitro antioxidant and antibacterial activities of phenolic and alkaloid extracts of Laurus nobilis

Laurus nobilis one of medicinal and aromatic plant endemic to the Mediter-ranean region. It is highly appreciated for the condiment benefits of its leaves. Due to its richness in bioactive compounds, the leaves are also used in traditional therapy to treat several infectious diseases. Phenolic com-pounds and alkaloids are particularly important because they are used in many pharmaceutical, food supplement, and cosmetic fields. The presence of phenolic compounds and alkaloids in L. nobilis leaves attracted our atten-tion due to their various functions such as antioxidant capacity and antibac-terial properties. The main objective of this study was to determine antioxi-dant and antibacterial activities of phenolic and alkaloid extracts of L. nobilis. The quantification of antioxidant substances allowed obtaining phenolic compounds content of 86.46 EAG mg/g and a total alkaloids yield of 700 μg/g. The results of antioxidant activity show that, although the alkaloid concen-trations are low, their antioxidant power is higher than phenolic compounds. The results of antibacterial activity of L. nobilis extracts revealed that the highest inhibition was recorded for alkaloid extract against staphylococcal strains responsible for food poisoning. However, L. nobilis phenolic com-pounds showed better activity against phytopathogenic strains associated particularly with potato rot.

Alkaloid-rich vs. antioxidant-rich yerba mate (Ilex paraguariensis) extracts: Exploiting the selectivity of supercritical CO2 using hydrated ethanol as co-solvent

Yerba mate (Ilex paraguariensis A.St-Hil.) is an autochthonous plant species from South America whose leaves exhibit beneficial health properties due to the presence of alkaloids and antioxidants. In the present study, the content of alkaloids and antioxidants in yerba mate extracts obtained by supercritical CO2 extraction was analyzed. Extractions were carried out using ethanol and hydrated ethanol as co-solvents at up to 333 K and 26.0 MPa. Bioactive extracts with a high concentration and yield of alkaloids (11.9 wt% and 5.0 g kg−1 of yerba mate, respectively) were obtained when pure ethanol was used as co-solvent. The alkaloid selectivity decreased and the antioxidants extraction capacity increased to a maximum of 8.4 g kg−1 of yerba mate when hydrated ethanol was loaded as co-solvent. Thus, by controlling the solvent composition and the extraction conditions, extracts rich in alkaloids or antioxidants can be obtained for the development of nutraceutical and functional food products.

Optimization of Extraction Process for Total Alkaloids from Maca with Different Color Types

This study was carried out to optimize the acidic extraction of total alkaloids from Maca with different color type. The effects of three extraction parameters including solid-to-liquid ratio, temperature and duration on extraction efficiency were investigated by orthogonal array design. Results were as follows:The order of effect of different factors on extraction efficiency for yellow Maca and black Maca were solid-to-liquid ratio > duration > temperature. The order of effect of different factors on extraction efficiency for purple Maca were duration > solid-to-liquid ratio > temperature. The content of total alkaliods in yellow Maca was 9.89%±0.38% under the optimum extracting conditions:Ratio of solid to liquid 1:30, temperature 75℃, time 5.5 h.The optimal extraction condition of purple Maca were 90℃ temperature for 5.5 h with solid-to-liquid ratio 1:25, and the yield of total alkaloids was 10.36%±0.58%. The optimal extraction parameters for black Maca was as follows:Temperature 85℃, time 6 h, solid-liquid ratio 1:20, under these conditions, the extraction yield of total alkaloids was 10.01%±0.78%. In conclusion, the optimal extraction conditions for total alkaloids in different color types Maca had some differences.

Genetic Variability, Correlation and Path Analysis among Quantitative and Qualitative Traits of Opium Poppy (Papaver somniferum L.)

Background: The poppy seeds commonly known as ‘khus-khus’ or ‘postadana’, obtained from opium poppy plant (Papaver somniferum L.) is known one of the world’s oldest medicinal plant. Its alkaloids, seeds and oil used since time immemorial for therapeutic, food stuffs and economical purpose. An effective breeding program plays an important role for selection, genetic improvement and development of high yielding cultivars/variety in opium poppy. Therefore, a study was conceded out to evaluate the genetic variability, path analysis for different agronomic traits along with their major chemical constituents of opium alkaloids in 30 accessions/landraces of opium poppy (Papaver somniferum L.) collected from different geographical places of India. Methods: The 30 genotypes of Opium poppy including different breeding lines and landraces collected from different places were grown in year (2017-18) in randomized complete-block designs (RCBD) with three replications at the research farm of CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow. Morpho-metric data were recorded on five competitive randomly selected plants in each line for following thirteen traits. Dried husk/straw capsules were chemically analyzed through HPLC. The pooled mean data were statistically analysed by using Windostat statistical software 4.0 versions available at Department of Genetics and Plant Breeding, CSIR-CIMAP Lucknow. Result: Variation among the mean of thirty diverse genetic stocks was highly significant (p less than 0.01) for all thirteen traits. High heritability coupled with high Genetic advance as percent mean (GAM) and Genotypic coefficient of variation (GCV) for the traits Thebaine and Nosacapine. The results of associations studies revealed that the trait capsule/plant was positively and significantly correlated with seed (g)yield/plant (gm) and nosacapine at both genotypic and phenotypic level. According to results, a significant genetic variability was also present among 30 accessions, simple selection can be employed to improve seed yield along with alkaloids yield.