Multipurpose Medicinal Plant Research Articles

Increasing the planting density of Cryptolepis sanguinolenta (Lindl.) Schlt increased root biomass and cryptolepine yield

Cryptolepis sanguinolenta (Lindl.) Schlt. is an important multipurpose medicinal plant used for the treatment of ailments such as malaria. Despite the ongoing efforts in domesticating the herb, the ideal planting density and its benefits are unknown. A study was conducted to determine the influence of six C. sanguinolenta accessions and three planting densities (15, 30 and 45 plants/1.8 m2) on root biomass, cryptolepine concentration and cryptolepine yield. Also, benefit-cost ratios were determined for each plant density across the four cultivation periods (9, 12, 15 and 18 months). The cultivation of C. sanguinolenta at the highest planting density (45 plants/1.8 m2) increased root biomass (value), cryptolepine content (2.08 mg/100 mg dry root) and cryptolepine yield (23.31 mg mg/1.8 m2) compared to those cultivated at lower planting densities (15 and 30 plants/1.8 m2). The duration for growing C. sanguinolenta had a more significant influence on cryptolepine yield but not the cryptolepine content. Plants cultivated for 15 months gave the maximum cryptolepine yield (10.33 g/bed), indicating 15 months as the optimum time to harvest the roots. The benefit-cost analysis revealed that growing the plant at a density of 45 plants/1.8 m2 (25,920 plants/acre) for 18 months was a more profitable venture with a benefit-cost ratio of 3.45. Commercial cultivation of C. sanguinolenta at 45 plants per bed area of 1.8 m2 (25,920 plants/acre) for 15–18 months is recommended as the most profitable and promising cropping practice to ensure the sustainable supply of planting material.

Bioprospecting for Rhizobacteria with the Ability to Enhance Drought Tolerance in Lessertia frutescens.

Lessertia frutescens is a multipurpose medicinal plant indigenous to South Africa that is used for the management of cancer, stomach ulcers, wounds, etc. The use and demand for the raw materials from this plant have been increasing steadily over the years, putting strain on the dwindling wild populations. Although cultivation may provide relief to the strained supply, the persistent drought climate poses a threat to the plant's growth and productivity. This study explored three plant-growth-promoting rhizobacteria isolates, TUTLFNC33, TUTLFNC37 and TUTLFWC74, obtained from the root nodules of Lessertia frutescens as potential bioinoculants that can improve yield, biological activities and the production of secondary metabolites in the host plant. Isolate TUTLFNC37 was identified as the most promising isolate for inoculation of Lessertia frutescens under drought conditions as it induced drought tolerance through enhanced root proliferation, osmolyte proline accumulation and stomatal closure. Superior biomass yield, phenolics, triterpenes and antioxidant activity were evident in the extracts of Lessertia frutescens inoculated with TUTLFNC37 and under different levels of drought. Furthermore, the metabolomics of the plant extracts demonstrated the ability of the isolate to withstand drastic changes in the composition of unique metabolites, sutherlandiosides A-D and sutherlandins A-D. Molecular families which were never reported in the plant (peptides and glycerolipids) were detected and annotated in the molecular networks. Although drought had deleterious effects on Lessertia frutescens, isolate TUTLFNC37 alleviated the impact of the stress. Isolate TUTLFNC37 is therefore the most promising, environmentally friendly alternative to harmful chemicals such as nitrate-based fertilizers. The isolate should be studied to establish its field performance, cross infectivity with other medicinal plants and competition with inherent soil microbes.

Conservation implications of mapping the potential distribution of an Ethiopian endemic versatile medicinal plant, Echinops kebericho Mesfin.

Echinops kebericho is a narrow-range multipurpose medicinal plant confined to Ethiopia. Intense land use change and overharvesting for traditional medicine have resulted in narrow distributions of its populations. It is a threatened species with a decreasing population trend. This study aims to map its potential distribution, which is key to guide conservation efforts and sustainable use. We modeled the potential distribution of E. kebercho using the maximum entropy model (MaxEnt) employing 11 less correlated predictor variables by calibrating the model at two complexity levels and replicating each model 10 times using a cross validation technique. We projected the models into the whole of Ethiopia and produced binary presence-absence maps by classifying the average map from both complexity levels applying three threshold criteria and ensembling the resulting maps into one for the final result. We mapped suitable habitat predicted with high certainty and identified local districts where E. kebericho can be cultivated or introduced to enhance its conservation. We estimated that E.kebercho has about 137,925 km2 of suitable habitat, mainly concentrated in the western highlands of the Ethiopian mountains. Our models at both complexity levels had high average performances, AUC values of 0.925 for the complex model and 0.907 for the simpler model. The variations in performance among the 10 model replicates were not remarkable, an AUC standard deviation of 0.040 for complex and 0.046 for simple model. Although E. kebericho is locally confined, our models predicted that it has a remarkably wider potential distribution area. We recommend introducing E. kebericho to these areas to improve its conservation status and tap its multiple benefits on a sustainable basis. Locally confined threatened plants and animals likely have wider potential distributions than their actual distributions and thus similar methodology can be applied for their conservation.

Cytotoxic effect of various solvent extracts of Acacia nilotica pods on human erythrocyte cells

Acacia nilotica is a multipurpose medicinal plant. Different parts of this plant are used to alleviate or treat various diseases. In the current study, an attempt was made to study the cytotoxicity of Acacia nilotica pod extracts towards normal human erythrocytes. The pods of A. nilotica were ground, then three different solvents were used in different ratios to prepare crude extracts, which were screened for in vitro hemolytic activity. UV-visible spectrophotometric method was used for the quantification of the hemolytic effect. The hemolysis percentages of the different extracts were found to be very minimal (Methanol extract: 4.65±0.24%; Ethanol extract: 3.77±0.48% and Aqueous extract: 2.69±0.51%), while Triton X-100 produced total hemolysis at the lowest concentration. These findings suggested that this plant could be considered as safe to human erythrocytes. However, further studies through in vivo toxicological tests should be considered in the future in order to best justify its safety.

The Potential of Rhizobacteria to Mitigate Abiotic Stress in Lessertia frutescens

Lessertia frutescens is a multipurpose medicinal plant indigenous to South Africa. The curative ability of the medicinal plant is attributed to its rich phytochemical composition, including amino acids, triterpenoids, and flavonoids. A literature review of some of the phytochemical compounds, particularly amino acids, in L. frutescens shows a steady decrease in concentration over the years. The reduction of the phytochemical compounds and diminishing biological activities may be attributed to drought and salt stress, which South Africa has been grappling with over the years. Canavanine, a phytochemical which is associated with the anticancer activity of L. frutescens, reduced slightly when the plant was subjected to salt stress. Like other legumes, L. frutescens forms a symbiotic relationship with plant-growth-promoting rhizobacteria, which facilitate plant growth and development. Studies employing commercial plant-growth-promoting rhizobacteria to enhance growth and biological activities in L. frutescens have been successfully carried out. Furthermore, alleviation of drought and salt stress in medicinal plants through inoculation with plant growth-promoting-rhizobacteria is well documented and effective. Therefore, this review seeks to highlight the potential of plant-growth-promoting rhizobacteria to alleviate the effect of salt and drought in Lessertia frutescens.

Toxicological evaluation of different solvent fractions of Waltheria indica linn. root in male wistar rats

Waltheria indica Linn. root (WILR) is a multipurpose medicinal plant with abundance of phytochemical compounds. The optimal solvents for extraction of a particular bioactive compound depends not only on the yield of the compound but also on the toxic effect of the solvent used. Therefore, this study was designed to evaluate the toxic effects of hexane, dichloromethane and ethyl acetate solvent fractions of WILR in male Wistar rats. Three doses (200, 500, or 1000 μg/kg BW) of hexane, dichlorometh ane and ethyl acetate solvent fractions of WILR were used. Male Wistar rats (n=5) were administered with 200, 500, or 1000 μg/kg of hexane, dichloromethane, and ethyl acetate soluble extracts of WILR, while control received distilled water, daily for 15 days. The rats were thereafter sacrificed, blood samples were collected and serum separated. Haematological serum biochemical parameters were determined according to standard procedure. The result showed non-significant effect on relative organ weight and haematological parameters while the serum ALT was significantly increased by administration of hexane, dichloromethane, and ethyl acetate soluble extracts of WILR. Blood Urea Nitrogen was also increased significantly (p < 0.05) at 200 and 500 μg/kg of hexane and dichloromethane respectively. The ethyl acetate and hexane fraction also caused severe periportal cellular infiltration in the liver while the hexane fraction caused glomerular necrosis. The hexane, dichloromethane and ethyl acetate soluble extracts of (WILR) have adverse effect on the integrity liver and kidney. Hence, these extracts should be used with caution in patient with hepatic and renal impairment.

Toxicological evaluation of different solvent fractions of Waltheria indica linn. root in male wistar rats

Waltheria indica Linn. root (WILR) is a multipurpose medicinal plant with abundance of phytochemical compounds. The optimal solvents for extraction of a particular bioactive compound depends not only on the yield of the compound but also on the toxic effect of the solvent used. Therefore, this study was designed to evaluate the toxic effects of hexane, dichloromethane and ethyl acetate solvent fractions of WILR in male Wistar rats. Three doses (200, 500, or 1000 μg/kg BW) of hexane, dichlorometh ane and ethyl acetate solvent fractions of WILR were used. Male Wistar rats (n=5) were administered with 200, 500, or 1000 μg/kg of hexane, dichloromethane, and ethyl acetate soluble extracts of WILR, while control received distilled water, daily for 15 days. The rats were thereafter sacrificed, blood samples were collected and serum separated. Haematological serum biochemical parameters were determined according to standard procedure. The result showed non-significant effect on relative organ weight and haematological parameters while the serum ALT was significantly increased by administration of hexane, dichloromethane, and ethyl acetate soluble extracts of WILR. Blood Urea Nitrogen was also increased significantly (p < 0.05) at 200 and 500 μg/kg of hexane and dichloromethane respectively. The ethyl acetate and hexane fraction also caused severe periportal cellular infiltration in the liver while the hexane fraction caused glomerular necrosis. The hexane, dichloromethane and ethyl acetate soluble extracts of (WILR) have adverse effect on the integrity liver and kidney. Hence, these extracts should be used with caution in patient with hepatic and renal impairment.

A Review on Traditionally Used African Medicinal Plant Annickia chlorantha, Its Phytochemistry, and Anticancer Potential.

Annickia chlorantha Setten & P.J.Maas belongs to the Annonaceae family and is a multi-purpose medicinal plant, which has been extensively used for the traditional treatment option for malaria in western and central Africa. Its phytochemical composition is dominated particularly by various biologically active protoberberines and acetogenins. This review aims to provide a comprehensive review on the traditional uses, phytochemical profiles, and the toxicology of this plant from a myriad of available publications. Even after its tremendous applications against several different human ailments, this plant has been underestimated for its anticancer potential. Herein, based on the phytochemical composition, we discuss the probable mode of mechanism for its antiproliferative activity, which highlights its importance for cytotoxicity screenings against cancer cells. Additionally, this article discusses several research questions and suggests the future directions of its applications in medicinal plant-based anticancer research.

Ethnobotany of traditional medicinal plants and associated indigenous knowledge in Dawuro Zone of Southwestern Ethiopia

BackgroundThe study aimed at documenting the indigenous and local knowledge and use of traditional medicinal plants for treating human and livestock ailments in Dawuro Zone of Ethiopia.MethodsA survey was conducted among traditional healers and native administrators through discussion, interviews, and field observations. The snowball sampling technique was used to select 384 traditional healers in purposefully selected 50 villages spanning seven districts for face-to-face individual interviews. The chi-square test was applied to establish associations between traditional healers’ demographics, the distance between the village site and the nearest natural forest and a health center, and SPSS V.20 software was used for the analysis.ResultsThe traditional healers of the study area reported the use of 274 traditional medicinal plant species belonging to 217 genera and 82 families. Asteraceae (11.68%), Fabaceae (9.49%), and Lamiaceae (9.12%) were the foremost frequently used families. Herb species (54.8%) and leaves (65%) were predominantly sourced from the wild environment. The quantity of medicinal plants used (x2 = 278.368, df = 20, P = 0.000) and years of (experience in) traditional healing using herbs (x2 = 76.358, df = 10, P = 0.000) varied with distance from the natural forests. The service charge for healing had strong positive association (x2 = 24.349, df = 5, P = 0.000) with healer’s age (x2 = 309.119, df = 184, P = 0.000) and educational level (x2 = 851.230, df = 598, P = 0.000) with distance of traditional healer's residence from the medical institution. The agricultural activities, urbanization, low or no charge for the healing service, the secrecy and oral transfer of the knowledge, and the demand for medicinal and other multiple purposes species were some of the factors threatening the resource and the associated knowledge as well as the service in the study area.ConclusionThere are diversified traditional medicinal plants applied for healthcare of the community and domestic animals of the study area. The source of remedies mostly depends on herbs of natural forests, and the leaf was the most frequently used plant part. Developing conservation intervention and sustainable systems of utilization is needed for multipurpose medicinal plants. Finally, integrating with modern system and formalizing, legalizing, and capacitating the traditional medicine practitioners are needed for access of primary healthcare systems to rural communities.

Fatty acid composition, acute toxicity and anti-inflammatory activity of the n-hexane extract from Ranunculus macrophyllus Desf. roots

Ranunculus macrophyllus Desf. is a multipurpose edible medicinal plant, whose roots are used to treat feminine infertility and to increase weight. The aim of this work was to study for the first time the acute toxicity and anti-inflammatory activity of their n-hexane extract. Gas chromatography was used to study the chemical composition of the extract, while xylene induced ear edema was used to assess its anti-inflammatory activity. In silico methods were used to determine the possible anti-inflammatory mechanism of action and pharmacokinetic behavior of the major compounds. Gas chromatographic analysis led to identification of 19 fatty acids, including linoleic, α-linolenic and palmitic acids as major compounds. The n-hexane extract did not induce any visible signs of toxicity or mortality to mice at 2000 mg/kg body weight, and successfully inhibited the xylene induced ear inflammation (67% at 200 mg/kg). This activity may be attributed to linoleic and α-linolenic acids that were found to be potent inhibitors of cyclooxygenase 2 and phospholipase A2 in the in silico study. The n-hexane extract exhibited strong anti-inflammatory activity with no acute toxicity, which opens the doors for the use of R. macrophyllus roots and their n-hexane extract in food, nutraceutical and pharmaceutical industries.

Pomegranate (Punica granatum L.) and Metabolic Syndrome Risk Factors and Outcomes: A Systematic Review of Clinical Studies.

Pomegranate (Punica granatum L.) can be considered a multipurpose medicinal and dietary plant due to its anti-inflammatory and antioxidant actions. Pomegranate can be used to prevent or treat metabolic syndrome (MetS) risk factors. Although previously published reviews addressed the effects of pomegranate on different diseases, there is no systematic review that exclusively focuses on clinical trials related to all MetS-related risk factors. In view of this limitation, the objective of this up-to-date, comprehensive, and systematic review is to critically evaluate the potential of pomegranate (P. granatum) on various MetS risk factors on the basis of clinical studies. PubMed, EMBASE, MEDLINE, Google Scholar, COCHRANE, and Clinical Trials.gov databases were searched on 15 October 2021. The Preferred Reporting Items for a Systematic Review and Meta-Analysis guidelines were followed, and the bias risk evaluation was performed according to the Cochrane Handbook for Systematic Reviews of Interventions. We identified 5683 studies in the databases. After removing the duplicates, 3418 studies remained. Of these, 147 studies met the eligibility criteria, and finally, only 20 were included in the qualitative analysis. The included studies suggest that pomegranate can be beneficial to reduce body weight, blood pressure, glycemia, triglycerides, total cholesterol, and low-density lipoprotein cholesterol. Moreover, it can augment high-density lipoprotein cholesterol levels and improve insulin resistance. Although relevant effects were observed, additional well-designed clinical trials are needed to determine the correct formulations and doses to be used to prevent or treat MetS components.

Micropropagation, morpho-anatomical characterization, and genetic stability studies in Lippia javanica (Burm.f.) Spreng: a multipurpose medicinal plant

Micropropagation, morpho-anatomical characterization, and genetic stability studies in Lippia javanica (Burm.f.) Spreng: a multipurpose medicinal plant

PERBANDINGAN KADAR BESI (Fe) PADA DAUN KELOR (Moringa oleifera) YANG TUMBUH DI DATARAN TINGGI DAN DATARAN RENDAH SECARA SPEKTROFOTOMETRI SERAPAN ATOM (SSA)

Moringa leaves are a multipurpose medicinal plants commonly consumed as vegetables. Moringa leaves are rich in nutrients, one of which is iron (Fe) as much as 7 mg / 100g. Iron is useful for preventing anemia. Moringa grows in the highlands and lowlands with different soil conditions, so this allows for differences in iron levels in the leaves. This study aims to determine whether there are significant differences between Moringa leaves that grow in the highlands and lowlands. Samples were obtained from one of the residents' yards in kec. Gisting (highland) and kec. Teluk Betung (lowland). The sampling technique was carried out by purposive sampling. The tools used for iron analysis are Atomic Absorption Spectrophotometry at a wavelength of 248.40 nm. Linear regression line equation is obtained, which is y = 0.010483 + 0.09074x with a correlation coefficient (r) that is 0.9985. The average level of iron from Moringa leaves that grew in the highlands and lowlands respectively 6.26 mg / 100g and 6.16 mg / 100g. The results of the calculation of the t test found that tcount = 1.06. This tcount value will be compared with ttable with a 99% confidence level of 4.60. If tcount is smaller than ttable then Ho is accepted and Ha is rejected so that there is no significant difference between iron in moringa leaves that grow in the highlands and lowlands.

Potent Roots on the Move

Calumba <i>(Jateorhiza calumba</i> / <i>J. palmata)</i> and abutua <i>(Cissampelos pareira)</i> are multi-purpose medicinal plants, whose roots have been used in Eastern, Southern, and West-Central Africa for a considerable time. In the early modern era, the Portuguese adapted the roots, which became commercially traded in the Portuguese empire across the Indian and Atlantic Oceans. By the early nineteenth century, the roots were recognized also by the British and North American pharmacopoeias, but there was confusion and secrecy surrounding their origins and quality. Engaging with recent historiography about African and imperial plant medicines, we demonstrate that calumba and abutua had long, dynamic, and mobile continuities on the continent—first in Portuguese imperial settings and then in appropriation by British botanists and colonial officials in East Africa. Exploring African agency in the making of knowledge about calumba and abutua is problematic, however. While in the longue durée, calumba and abutua maintained their value and meanings within Africa, in certain moments, these plants became objects of scientific curiosity and heightened commercial interest in the West.

Cryopreservation of aromatic ginger Kaempferia galanga L. by encapsulation-dehydration

Kaempferia galanga L. is an endangered multi-purpose medicinal plant in Family Zingiberaceae, the rhizomes of which are used for several ayurvedic formulations. Encapsulation-dehydration (ED) method was optimized for cryopreservation of shoot tips of K. galanga. Shoot tips (STs) bearing the apical meristem dissected from the established in vitro shoot cultures were preconditioned in MS+0.4 M sucrose prior to encapsulation in calcium alginate and the beads subsequently transferred to MS liquid+0.3 M sucrose for 3 days afterward dehydration inside the laminar airflow for 4 hours upon rapid freezing in LN and rapid thawing produced maximum 62.2% survival and 46.7% regeneration rates. Shoot regeneration was observed from the apical meristems exclusive of intermediary callus phase. The plantlets regenerated from cryopreserved STs transferred to the field were phenotypically analogous with the mother plant.

Phytochemical Profile, Antioxidant Activity of Leaf of Hoslundia opposita Vahl and Docking of its Phytochemical Components with Uropathogenic Strain F11(ID:3NRP) and NFeoB from Escherichia coli BL21(ID:5FH9)

Hoslundia opposita Vahl is a multi-purpose medicinal plant used traditionally to treat gonorrhea, cystitis, cough, fever, snake bites, convulsion in many parts of Africa. The objectives of this research are to evaluate the phytochemical profile, antioxidant activity of the leaves of Hoslundia opposita Vahl and to dock its phytochemical components with uropathogenic Escherichia coli strain F11(ID:3NRP) and NFeoB from Escherichia coli BL21(ID:5FH9). Harborne’s method was used for the identification of the class of phytochemicals while GC-MS was used to identify the type of phytochemicals. Folin-Ciocalteau method was used to determine the total phenolic content while aluminum colorimetric assay was used to estimate the total flavonoid content. 1, 1-diphenyl-2-picrylhydrazyl (DPPH) was used to evaluate the antioxidant activity. The molecular docking simulation method was used to investigate the interactions of the phytochemicals with the Escherichia coli receptors. In the phytochemical studies, flavonoids, terpenoids, cardiac glycosides and saponins were identified to be present in the methanolic leaf extract while steroids, alkaloids and anthraquinones were absent. The GC-MS analysis of the methanolic extract revealed presence of 17 compounds out of which 14 were identified. The compounds with appreciable quantity in the leaf extract were 1, 2, 3-benzetriol (38.11%), n-hexadacanoic acid (13.52%) and catechol (9.98%). Assessment of the antioxidant activity using DPPH gave a scavenging activity of 79.86% at 500ug/ml compared with ascorbic acid having a scavenging activity of 90.85% at the same concentration. Evaluation of the total phenolic and flavonoid contents at 500 ug/ml gave the values of 262.54 mg gallic acid equivalent/g extract and 6.24 mg quercetin equivalent/g extract respectively compared with gallic acid and quercetin with maximum concentrations of 385.12 and 12.46 respectively. This shows that the extract has significant antioxidant activity and can be explored as a valuable source of natural antioxidants. GC-MS analysis of the methanolic extract of Hoslundia opposita showed the presence of trans-3-(trifluoromethyl)cinnamic acid, octadecyl ester which was active against Escherichia coli. Docking results with 3NRP and 5FH9 showed binding affinities of -6.1, -6.6, 7.2and -7.0,7.4, 8.5 kJ/mol with the commercial drugs : ciprofloxacin, levofloxacin and tetracycline respectively while that of trans 3-(trifluoromethyl)cinnamic acid, octadecyl ester identified in the leaves were 5.6 and 5.9 kJ/mol.

Traditional uses, phytochemistry, pharmacology and other potential applications of Vitellaria paradoxa Gaertn. (Sapotaceae): A review

Vitellaria paradoxa Gaertn. is a multipurpose medicinal plant of the family Sapotaceae, and it has been widely used usually in the clinical traditional medicine as remedy for a wide range of diseases for several decades. In addition, the plant has also found applications in confectionery, cosmetics and soaps, and pharmaceuticals both locally and internationally. V. paradoxa, which has been identified with >150 phytoconstituents, is rich in oleanane-type triterpene acids and glycosides, such as paradoxosides A-E, tieghemelin A, parkiosides A-C, bassic acid, as well as flavonoids such as quercetin and catechin-type compounds. The extracts and the active constituents of V. paradoxa have been investigated for various pharmacological activities, including but not limited to anticancer, melanogenesis-inhibitory, antibacterial, anti-diabetic, antioxidant, anti-inflammatory, anti-diarrhoeal, and antifungal activities. Additionally, V. paradoxa has also been utilized in nanoparticles (NPs) synthesis. These NPs among other things have shown significant antinociceptive and antiedematogenic activities as well as environmental friendly adsorptive properties for the removal of pollutants from pharmaceutical effluents. Overall, this review comprehensively examines the traditional uses, phytochemistry, pharmacology, toxicology, clinical studies, and nanoparticles synthesized from V. paradoxa and their applications.

High-frequency in vitro propagation and assessment of genetic uniformity and micro-morphological characterization of Origanum majorana L. –A highly traded aromatic herb

Origanum majorana (L.) (family Lamiaceae) is a multipurpose medicinal plant with high demand across the pharmaceutical, cosmetic, food, beverage, and perfume industries that has raised concern for its mass propagation through in vitro culture techniques. The present report describes an effective in vitro propagation system for an aromatic medicinal plant Origanum majorana L. The Murashige and Skoog's (MS) medium supplemented with 1.0 mg L-1 6-benzylaminopurine (BAP) initiated a maximum of 4.4 ± 0.22 micro-shoots from the nodal segment explants. The regenerated shoots were further proliferated (98.0 ± 0.22 shoots per explant/culture vessel) on MS medium augmented with 0.5 mg L-1 BAP and 0.25 mg L-1 α-naphthalene acetic acid (NAA). Individual shootlets were rooted on a half-strength MS medium with 1.0 mg L-1 indole-3 butyric acid (IBA) (4.8 ± 0.20 roots/shoots). The rooted plantlets were transplanted into eco-friendly paper cups containing soilrite®: manure; sand (2:1:1) and maintained in the greenhouse for hardening. Inter simple sequence repeats (ISSR) molecular markers were utilized to investigate the somaclonal variations between in vitro developed plants and the mother plant of O. majorana. The band patterns of regenerated plantlets have shown genetic uniformity with the donor plant. The foliar micro-morphological features under in vitro and ex vitro conditions were compared with the mother plant and elucidated morphologically stable plantlets development through gradual acclimatization. This is the foremost report on ISSR molecular marker-based genetic fidelity studies and foliar micro-morphological evaluation of micropropagated plants of O. majorana.

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We identified the phytochemicals in Withania somnifera L, a multipurpose medicinal plant of the Himalayan región using TLC, FTIR and HPLC. Eleven formulations were made by mixing in different ratios of Withania leaf extract, wood ash and distilled wáter. Wood chips and twigs of 3-years old Pinus roxburghii tree were completely burnt till ashes was used as Wood ash. The 11-formulationss were i.e. formulation I (Withania leaf extract 100%)), formulation II (75:25 concentration (Withania leaf extract: Distilled water)), formulation III (50:50 concentration (Withania leaf extract: Distilled water), formulation IV (25:75 concentration (Withania leaf extract: Distilled water)), formulation V (75:25 concentration (Withania leaf extract: Wood ash)), formulation VI (50:50 concentration (Withania leaf extract: Wood ash)), formulation VII (25:75concentration (Withania leaf extract: Wood ash)), formulation VIII (Wood ash 100%)), formulation IX (75:25 concentration (Wood ash: Distilled water)), formulation X (50:50 concentration (Wood ash:Distilled water)), formulation XI (25:75concentration (Wood ash: Distilled water)). The herbicidal activity of formulations was earlier studied against Cannabis sativa L. (banned narcotic plant). Their herbicidal activity was tested on seeds germination and seedlings growth of Cannabis sativa L weed and 2 crops: Hordeum vulgare L and Cicer arietinum L. In Pot culture, the formulation V spray caused maximum reduction in root length of Cannabis (53 %) &gt; Hordeum (23 %) and Cicer (22 %) than control. The formulations were also tested on crops for their herbicidal effects. In Petri plate bioassy, the formulation V (75:25 concentration (Withania leaf extract: Wood ash) showed maximum reduction in seed germination and seedling growth of C. sativa weed, but had little effect on growth of test crops. The reduction in stem length was maximum (55 %) in Cannabis &gt; Hordeum (20 %) &gt; Cicer (19 %) than control. Glyphosate decreased the stem length of Cicer (80 %) &gt; Hordeum (58 %) and Cannabis (16 %) over control. While the reduction in root length was in Cicer (75 %) &gt; Hordeum (73 %) &gt; Cannabis (18 %) than control. The root and stem extract formulations were more phytotoxic to Cannabis sativa. Formulation V (75:25 concentrations (Withania leaf extract: wood ash)) reduced the number of leaves, number of shoot, root branches and stem and root length of Cannabis sativa.

In vivo bioactivity-guided isolation of antifertility fraction of Waltheria indica Linn. root in male Wistar rats

BackgroundWaltheria indica is a multipurpose medicinal plant with abundance of phytochemical compounds. Antifertility effect of Waltheria indica Linn. root and leaves have been reported. However, the fraction responsible for this antifertility effect needs to be isolated for possible male contraceptive purpose. Therefore, this research was designed to isolate the antifertility fraction of Waltheria indica Linn. root (WILR) in an in vivo model using male Wistar rats. Crude ethanol extract of WILR was sequentially dissolved in hexane, dichloromethane, and ethyl acetate. Rats (n = 5) were administered with 200, 500, or 1000 μg/kg of hexane, dichloromethane, and ethyl acetate soluble extracts of WILR, while control received distilled water, daily for 15 days to determine the soluble extract with most antifertility effect. Thereafter, fractions were separated from dichloromethane soluble WILR extract by column and thin-layer chromatography. Rats (7 groups, n = 5) were administered with each of the fractions (DF1 to DF7; at 1000 μg/kg) to determine the fraction with the highest antifertility. Rats were thereafter sacrificed, and sperm parameters, reproductive hormones, testicular cholesterol, and protein were determined according to standard procedure. Histology of the testis was also done. Data were analyzed using ANOVA at p ≤ 0.05.ResultsDichloromethane soluble fraction (500 μg/kg) significantly decreased sperm concentration (137.00 ± 9.85 to 107.00 ± 13.08 × 106 cells/mL), levels of testosterone (2.90 ± 0.65 to 1.50 ± 0.37 ng/mL), and FSH (0.08 ± 0.08 to 0.99 ± 0.08 IU/L). The dichloromethane soluble fraction also caused the loss of testicular interstitium and spermatogenic cells. DF5 significantly reduced sperm motility (92.00 ± 2.74 to 76.00 ± 5.48%) and LH (2.86 ± 0.52 to 1.47 ± 0.18 IU/L). DF5 also significantly increased levels of prolactin (1.22 ± 0.10 to 1.88 ± 0.48 ng/mL), testicular total protein (7.36 ± 0.35 to 8.54 ± 1.06 g/dL), and testicular cholesterol (34.17 ± 3.65 to 55.76 ± 6.08 mg/mL).ConclusionThe results indicate that the DF5 is the bioactive fraction of WILR responsible for its antifertility effect. The possible antifertility mechanisms are through the reduction in sperm parameters, reproductive hormones, and histological changes in the testis.