Sapindus Emarginatus Research Articles

On-water Catalyst-free Synthesis of 3-Alkenyl Oxindoles

3-Alkenyl oxindoles are key structural motifs in organic chemistry with a wide range of applications mainly in the pharmaceutical industry, due to excellent biological activities such as anti-inflammatory, antiviral, antibacterial, and anticancer properties. With the discovery of the importance of oxindole derivatives in the pharmaceutical industry, demand for sustainable protocols for the synthesis increased as most classical methods have their limitations and flaws. In this study on-water, catalyst-free, atom-economical green approach was developed for the synthesis of 3- alkenyl oxindole derivatives using oxindole and aldehyde as starting materials. The reaction between oxindole and benzaldehyde was conducted on water under catalyst-free conditions. Interestingly, 100% conversion was observed when the reaction was carried out for 24 hours at 100° C. As the next step, the applicability of the developed approach was proven for different aldehydes. Furaldehyde, cinnamaldehyde, and vanillin were used as the aldehydes to synthesize 3- alkenyl oxindole derivatives, and the isolated yields were obtained as 77%, 54%, and 51% respectively. The desired product formation was confirmed by comparing the Rf value of the product with that of the previously synthesized well-characterized compounds; 3- benzylideneindolin-2-one, 3-(furan-3-methylene)indolin-2-one, 3-(3-phenylallylidene)indolin-2- one, and 3-(4-hydroxy-3-methoxybenzylidene)indolin-2-one using Thin Layer Chromatography. Further characterization was done by Fourier-transform infrared (FT-IR), Ultraviolet-visible (UVVis), and Nuclear Magnetic Resonance (NMR) spectroscopic techniques. In further optimization steps, the reaction was performed using surfactants as the catalyst. By adhering to the green chemistry principles different sources of biosurfactants were used including Acacia concinna pods, Sapindus emarginatus fruit pericarp, Dillenia retusa fruit pericarp, and Trigonella foenumgraecum seeds. In order to compare the effect of biosurfactants, a synthetic surfactant, sodium dodecyl sulfate (SDS) was used as the catalyst. When the reaction was carried out for 21 hours at 100 °C using fruit pericarp of Sapindus emarginatus as the source of biosurfactant, 100% conversion was observed. Using water as the solvent, readily available starting materials, higher yield, and eco-friendliness and catalyst-free conditions suggest the possible use of the developed method for large-scale preparations of 3-alkenyl oxindole derivatives.
 Keywords: Green protocol, Oxindole derivatives, Surfactant, On water synthesis, Catalyst-free

Role of Sapindus emarginatus Pericarp Extract in Gut Serotonin Level and its Emetic Activity in Animal Models

Background: Emesis is a complex and distressing protective mechanism that helps to remove toxic substances from the stomach and prevent further ingestion. The emetics and cathartics are predominantly used for accidental and intentional ingestion of poisons or toxins. The availability and usage of emetics in humans are limited because of their side effects. Therefore, to treat poisoned people, we need effective medications. Sapindus emarginatus Vahl., often called soapnut, is a member of the Sapindaceae family. They have historically been used as emetic, antipruritic, laxative, antifertility, and anti-inflammatory medicines. Objective: This study aims to assess the gut serotonin level and emetic effect of Sapindus emarginatus hydroethanolic pericarp extract (HESE) by using animal models. Methods: Gravimetric analysis was used to determine the HESE's saponin content. The emetic effect of the HESE at a dose of 250, 500, 1000, and 2000 mg/kg was evaluated by copper sulfateinduced emesis in the chick model and cisplatin-induced emesis in the rat-pica model. The serotonin level in rat intestinal mucosa was measured by spectrofluorimetry. Results: HESE was estimated to contain 11.92% saponin. The extract at high doses of 1000 and 2000 mg/kg showed emetic activity evidenced by increased frequency of retching in chick, increased kaolin intake, and anorexia in the rat-pica model. The extract showed a significant increase in serotonin levels in the proximal part of the small intestine in comparison with normal animals. Conclusion: According to the results of the current investigation, which employed various animal models, the HESE demonstrated appreciable emetic activity. The extract at a high dose showed a significant emetic effect due to increased serotonin levels in the gut. The HESE was discovered to be a strong contender for the treatment of poisoned patients. More research are required to validate their adverse effects of frequent usage.

Characterization and screening of candidate plus trees of Sapindus emarginatus Vahl. for saponin content

Soapnut (Sapindus emarginatus Vahl.) is one of the most traded non timber forest product bioresource in Tamil Nadu. The dried fruit contains higher percentage of saponin, a substance rich in detergent properties. As a measure of sustainable consumption, soapnut harvesting in the wild should be minimized and compensated by agroforestry and industrial plants grown outside of the forests. To identify the superior source in terms of higher fruit yield and saponin percentage the germplasm was assembled and studies were undertaken at Forest College and Research Institute, Mettupalayam. A reconnaissance survey was carried out in Tamil Nadu to select Sapindus emarginatus Candidate Plus Trees (CPTs) based on the morphometric attributes of tree and fruiting output during peak fruiting season. Saponin content of selected Candidate Plus Trees were analysed and was found to range between 10.8-18.2%. Among the sources collected fruits collected from Kunjapanai-1 (TN SE 03) source recorded the highest saponin content (18.2%).

Sapindus emarginatus extract embedded with gold nanoparticles: An antiproliferative agent against MCF7 breast cancer cell line

There are numerous studies reported on the usage of the sapindus emarginatus (SE) fruit in cancer and other treatments in the past few years. In this study, crude SE fruit extract was prepared and it was further used to synthesis gold nanoparticles (Au Nps). The synthesized Au Nps were left embedded in the SE fruit extract. The Au Nps embedded in the SE fruit extract (SE-Au Nps) were characterized using UV-Visiable Spectroscopy, Centrifugal Particle Size analyzer (CPS), Scanning Electron Microscope (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). MTT assay was carried out for both SE fruit extract and SE-Au Nps on MCF7 breast cancer cell line and thus compared. The UV-Visible Absorbance for the SE-Au Nps was obtained at 543 nm. The centrifugal particle size analysis of the Au Nps embedded in SE fruit extract showed the size of the nanoparticles to be widely varying with higher fraction of particles between the size ranges of 15 to 20 nm. The morphology of the Au Nps embedded in SE fruit extract was observed using SEM. The presence of Au Nps in SE fruit extract was confirmed using FTIR. The results of the MTT assay on MCF7 breast cancer cell line proved that the % cell viability was less for SE-Au Nps than that of the SE fruit extract alone. Thus, the antiproliferative activity of the SE fruit extract was significantly enhanced by embedding it with Au Nps and it can be effectively used in therapeutic applications after further studies.

Purification of Bacteriocin from Lactobacillus plantarum KY449289 and its Role in the Preservation of Mixed Fruit Juice

Background:: The aim of this study is to isolate, extract and purify the antibacterial protein from Lactobacillus plantarum KY449289 and to assess its bio preservative potential. Objective:: The present study was conducted to evaluate the laxative activity of Hydroethanolic Pericarp Extract of sapindus emarginatus (HESE) in animal models. Methods:: Bacteriocins are antimicrobial peptides synthesized ribosomally which can be used as bio-preservative reducing the risk of chemical preservative’s effect and also replacing the thermal treatments. This study aims to isolate, purify the antibacterial protein from Lactobacillus plantarum KY449289 and to assess its bio preservative potential. In this study, twenty strains were isolated from a yogurt sample and preliminary characterization was carried out, such as morphological, biochemical, and molecular levels as a preliminary assay. Results:: The characterized strains were assessed for the antibacterial activity by well diffusion assay, whereas all twenty strains were shown to have a antibacterial activity against foodborne pathogens, among which two strains VITMM04 and VITMM05 shows the highest zone of inhibition of 15 mm and 10 mm respectively against Listeria monocytogenes. The potent isolate VITMM05 solely showed a broad spectrum antibacterial activity against the pathogens. These strains were further subjected to optimization, purification, and application studies. A high increase in the inhibition zone was recorded with an optimum temperature of 35°C and its activity was found to be stable up to 85°C, optimum pH 6, and 5 mM of EDTA as an enhancer surfactant. On subjected to RP-HPLC, the purified sample showed a distinct peak at 2.192 min corresponding to the peak at 2.192 min for standard bacteriocin. Conclusion:: The six-fold purified bacteriocin was effective in controlling 87% of the microbial population with 20mm zone of inhibiton against the Listeria monocytogenes and this shows the effective role of purified bacteriocin in inhibiting bacterial growth in a greater extent.

Energy optimization from a binary mixture of non-edible oilseeds pyrolysis: Kinetic triplets analysis using Thermogravimetric Analyser and prediction modeling by Artificial Neural Network

Pyrolysis kinetics and thermodynamic parameters of two non-edible seeds, Pongamia pinnata (PP) and Sapindus emarginatus (SE), and their blend in the ratio of 1:1 (PS) were studied using the thermogravimetric analyzer. Kinetic triplets were determined using both model-free [Starink (STR), Friedman (FRM), Iterative Kissinger-Akahira-Sunose (IT-KAS), Iterative Ozawa-Flynn-Wall (IT-OFW), Vyazovkin (VYZ), and Master plot (MP)] and model fitting Coats-Redfern (CR) methods at three different heating rates 10, 30 and 50 °C/min. Activation energies were 192.66, 179.44, and 163.25 kJ/mol for PP, SE, and PS, respectively. It was found that the blend of the two-biomass (PS) showed promising results with lower activation energy compared to the individual biomass. Thermodynamic parameters (ΔG, ΔS, and ΔH) were obtained using the model-free isoconversional method. The three hidden layers of complex neuron topology are well fitted to the experimental DTG curves by artificial neural network (ANN). The study confirmed that the heating rate had a significant impact on the kinetics and thermodynamic parameters. The reaction mechanism was also in consonance with the experimental data. The study suggests that the PP and SE seeds can be an appropriate feed for pyrolysis, and their blend (PS) can be a viable alternative in optimizing the entire process.

Evaluation of the Laxative Activity of Saponin Enriched Hydroethanolic Pericarp Extract of Sapindus emarginatus in Animal Models

Background: Constipation is a common, predominant, chronic gastrointestinal functional disorder. The drugs available to treat constipation are limited because of their side effects for long term use. Therefore, we need effective drug to treat constipation. Sapindus emarginatus Vahl belongs to the family Sapindaceae, commonly known as soapnut. It is traditionally used as an antipruritic, antifertility, laxative, and anti-inflammatory agent. Objective: The present study was conducted to evaluate the laxative activity of hydroethanolic pericarp extract of Sapindus emarginatus (HESE) in animal models. Methods: The saponin content in the extract was measured by the gravimetric analysis. The laxative activity of hydroethanolic pericarp extract of Sapindus emarginatus was evaluated by the weight of feces matter, charcoal meal hyperperistalsis test, and loperamide induced constipation model. Results: The saponin content of the soapnut pericarp was 13.48% and the extract was found to be 11.92%. The results obtained from these models showed a significant dose-dependent increase in fecal weight, peristalsis index, and moisture content compared to control animals. Conclusion: The present study concluded that the oral administration of HESE showed a significant laxative activity in different animal models. The presence of triterpene saponins is supposed to be responsible for this activity. Further studies are needed to confirm their mechanism behind the laxative effect. The administration of the extract was found to be effective in constipation therapy.

Sapindus emarginatus extract embedded with gold nanoparticles: An antiproliferative agent against MCF7 breast cancer cell line

There are numerous studies reported on the usage of the sapindus emarginatus (SE) fruit in cancer and other treatments in the past few years. In this study, crude SE fruit extract was prepared and it was further used to synthesis gold nanoparticles (Au Nps). The synthesized Au Nps were left embedded in the SE fruit extract. The Au Nps embedded in the SE fruit extract (SE-Au Nps) were characterized using UV-Visiable Spectroscopy, Centrifugal Particle Size analyzer (CPS), Scanning Electron Microscope (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). MTT assay was carried out for both SE fruit extract and SE-Au Nps on MCF7 breast cancer cell line and thus compared. The UV-Visible Absorbance for the SE-Au Nps was obtained at 543 nm. The centrifugal particle size analysis of the Au Nps embedded in SE fruit extract showed the size of the nanoparticles to be widely varying with higher fraction of particles between the size ranges of 15 to 20 nm. The morphology of the Au Nps embedded in SE fruit extract was observed using SEM. The presence of Au Nps in SE fruit extract was confirmed using FTIR. The results of the MTT assay on MCF7 breast cancer cell line proved that the % cell viability was less for SE-Au Nps than that of the SE fruit extract alone. Thus, the antiproliferative activity of the SE fruit extract was significantly enhanced by embedding it with Au Nps and it can be effectively used in therapeutic applications after further studies.

Facile Green Synthesis of Ag@g-C3N4 for Enhanced Photocatalytic and Catalytic Degradation of Organic Pollutant

A simple and easy green synthesis of silver nanoparticles (Ag NPs) was prepared using Sapindus emarginatus stem bark extract and anchored over graphitic carbon nitride (g-C3N4) surface. The synthesized materials characterized by UV–Visible, XRD, FESEM, FETEM, FT-IR and Raman spectroscopy ensured the development of Ag NPs anchored over the g-C3N4 surface. The prepared catalyst was utilized for Methylene Blue (MB) dye degradation through visible light photocatalysis. The optimization of operational parameters, like the effect of catalyst dosage, H2O2 concentration and slurry pH, was performed. The results exhibited that compared to pristine g-C3N4, photocatalytic efficacy was enhanced two-fold in Ag@g-C3N4. Also, the prepared Ag@g-C3N4 showed a faster catalytic MB dye reduction (25 s) in the presence of NaBH4. The catalyst was recovered from the reaction solution by centrifugation and reused for five consecutive cycles. The results showed that there was no significant loss in the efficiency of the catalyst. These results confirmed that the green synthesized Ag@g-C3N4 is likely advantageous for organic pollutants degradation.

Adaptive fitness of Sapindus emarginatus Vahl populations towards future climatic regimes and the limiting factors of its distribution

Sapindus emarginatus Vahl (Sapindaceae) also known as ‘Indian Soap nut’ is significantly important for saponin content in its fruits. However, its current population in India is heavily fragmented due to a lack of sustainable harvesting practices. Moreover, changing climatic regimes may further limit its distribution and possibly compromise the survival of the species in nature. The aim of the present study was to: predict the future distribution range of S. emarginatus; identify the bioclimatic variables limiting this distribution and to evaluate its adaptive fitness and genomic resilience towards these variables. To determine future species distribution range and identify limiting bioclimatic variables, we applied two different ecological niche models (ENMs; BioClim and MaxEnt) on real occurrence data (n = 88 locations). The adaptive fitness of the species was evaluated by quantifying the genetic variability with AFLP markers and marker-environmental associations, using AFLP-associated Bayesian statistics. We found 77% overlap between the baseline (2030) and predicted (2100) species distribution ranges, which were primarily determined by maximum temperature (TMAX) and mean annual precipitation (MAP). The TMAX and MAP contributed 43.1% and 27.1%, respectively to ENM model prediction. Furthermore, AFLP loci significantly associated with bioclimatic variables, and TMAX and MAP represent the lowest proportion (6.15%), confirming to the severe response of the species genome towards these variables. Nevertheless, the very low Linkage disequilibrium (LD) in these loci (4.54%) suggests that the current sensitivity to TMAX and MAP is subject to change during recombination. Moreover, a combination of high heterozygosity (0.40–0.43) and high within-population variability (91.63 ± 0.31%) confirmed high adaptive fitness to maintain reproductive success. Therefore, the current populations of S. emarginatus have substantial genomic resilience towards future climate change, albeit significant conservation efforts (including mass multiplication) are warranted to avoid future deleterious impacts of inbreeding depression on the fragmented populations.

Sapindus emarginatus extract embedded with gold nanoparticles: An antiproliferative agent against MCF7 breast cancer cell line

<p>There are numerous studies reported on the usage of the Sapindus emarginatus (SE) fruit in cancer and other treatments in the past few years. In this study, crude SE fruit extract was prepared and it was further used to synthesis gold nanoparticles (Au Nps). The synthesized Au Nps were left embedded in the SE fruit extract. The Au Nps embedded in the SE fruit extract (SE-Au Nps) were characterized using UV Vis spectroscopy, Centrifugal particle size analyzer (CPS), Scanning Electron Microscope (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). MTT assay was carried out for both SE fruit extract and SE-Au Nps on MCF7 breast cancer cell line and compared. The UV- Vis Absorbance for the SE-Au Nps was obtained at 543nm. The centrifugal particle size analysis of the Au Nps embedded in SE fruit extract showed the size of the nanoparticles to be widely varying with higher fraction of particles between the size ranges of 15 to 20nm. The morphology of the Au Nps embedded in SE fruit extract was observed using SEM. The presence of Au Nps in SE fruit extract was confirmed using FTIR. The results of the MTT assay on MCF7 breast cancer cell line proved that the %cell viability was less for SE-Au Nps than that of the SE fruit extract alone. Thus the antiproliferative activity of the SE fruit extract was significantly enhanced by embedding it with Au Nps and it can be effectively used in therapeutic applications after further studies. </p>

FORMULATION AND IN VITRO EVALUATION OF GEL BASED POLYHERBAL VAGINAL WASH

Vaginal Infection is quite common in India, a multi-city study estimated that more than 90% of women between the ages 25 and 35 suffer with vaginal infection and 93% do not follow the best personal hygiene practices. The vaginal yeast infection is caused by Candida albicans. Which is characterized by Itching, irritation in tissues at the vaginal opening (vulva), burning sensation, redness /swelling of the vulva, vaginal pain ,soreness, vaginal rash, watery vaginal discharge . The objective of the present study was to develop poly-herbal vaginal wash for prevention of Candida albicans infection. Aqueous extracts of Azadirachtaindica leaf, Ocimum Sanctum leaf and Sapindus emarginatus were formulated in an aqueous based carbopol-940 (1%w/w) gel system. Prepared poly herbal vaginal wash formulation was evaluated for their physicochemical properties like texture evaluation, pH determination, viscosity and in vitro Antifungal activity were determined along with short term stability studies. The formulated gel based poly-herbal vaginal wash had acceptable physical parameters that showed that they were compatible and in addition to this, these formulations passed the short-term stability studies. The in vitro antifungal activity studies showed that the formulated gel based vaginal wash showed significantly strong activity against C. albicans. Thus, the present study concludes that the formulated herbal vaginal wash is efficient antifungal formulations for the C. albicans vaginal infection.

Bio inspired synthesis of monodispersed silver nano particles using Sapindus emarginatus pericarp extract – Study of antibacterial efficacy

The synthesis of silver nanoparticles employing aqueous extract obtained from the dried pericarp of “Sapindus emarginatus” is reported. Transmission electron microscopy divulges that the silver nanoparticles are not agglomerated and are moderately mono dispersed. Size of the particle ranges from 5 to 20nm with an average particle size of 10nm. Ultraviolet–visible spectra recorded show typical surface plasmon resonance (SPR) at 400nm. X-ray diffraction analysis reveals the crystalline nature of the synthesized silver nanoparticles with face-centred cubic (FCC) geometry. Silver nanoparticles thus obtained demonstrated remarkable antibacterial activity against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Proteus mirabilis, Proteus vulgaris, Klebsiella pneumonia, Pseudomonas aeruginosa and Vibrio cholerae. Freshly prepared samples and sample containing 6nm silver nanoparticles in particular exhibited enhanced activity against gram positive bacteria. The minimal inhibitory concentration was found to be in the range of 150–250μg/mL.

In vitro Screening of Some Indian Medicinal Plants for Their Activity against Extended Spectrum Beta Lactamases

Context: Extended Spectrum Beta Lactamases (ESBLs) play an important role in pathogenesis of various infections by enabling the bacterial species to be resistant to β-lactam antibiotics including extended-spectrum cephalosporins. Plants were selected on the basis of their traditional applications. Objective: Our investigation screens and evaluates15 Indian medicinal plants for antimicrobial efficacy and synergistic potential against ESBLs producing bacteria. Materials and Methods: 6 bacterial strains were screened for their ability to release ESBLs. Plant extracts in methanol and aqueous solvents were screened for their effect on ESBLs producing bacteria. Antimicrobial-linked ESBLs inhibition activity and minimum inhibitory concentration (MIC) of extracts were evaluated by agar well diffusion and microdilution method. Synergistic interactions between plant extracts exhibiting good antimicrobial activity and extended spectrum cephalosporins were explored by Checkerboard method. Results: Three strains were detected as ESBL positive. The results of susceptibility assay clearly showed strong ESBLs inhibitory effect of Crinum latifolium, Oroxylem indicum, Punica granatum, Original Research Article Malik et al.; BBJ, 8(3): 1-10, 2015; Article no.BBJ.18406 2 Sapindus emarginatus and Terminalia chebula and their MIC values ranged from 1.87-30 mg/ml. In vitro interactions between plant extracts and antibiotics cefotaxime and ceftaizidime evaluated in terms of fractional inhibitory concentration (FIC) indices indicated synergism. Discussion and Conclusion: Higher resistance of ESBLs positive strains to β-lactam antibiotics encourages us to search the novel ESBLs inhibitors. Maximum 10-fold decline in the MIC of antibiotics cefotaxime and ceftaizidime was achieved in combination with plant extracts. Owing to enormous clinical significance of ESBL-producing organisms coupled with limited therapeutic options, the results revealed by present study are of paramount importance.

Scouring Cotton Fabric by Water-Extracted Substance from Soap Nut Fruits and Licorice

This study used water to extract substances from hulls of soap nut fruits (Sapindus Emarginatus) and licorice root (Glycyrrhizaglabra L.) found in Thailand to remove waxes in cotton scouring process. CMC value from soap nut fruits were found to be 6 mg/ml with surface tension of 54.67 mN/m and can be used as wetting agent when the temperature is lower than 70 °C. CMC value from licorice root was 10 mg/ml with surface tension of 51.50 mN/m and can be used as wetting agent at temperature higher than 95°C. The results exhibited that both substances can be used to remove waxes in cotton scouring process. At the 40% of substance powder by fabric weight, the cotton fabric absorbed water in 5 seconds.

Immuno-suppressive effects of aqueous extract of soapnut Sapindus emarginatus on the larvae and pupae of vector mosquito, Aedes aegypti

We recently reported the presence of potent anti-mosquito activity in aqueous kernel extract of the soapnut, Sapindus emarginatus, and demonstrated its impact on marker enzymes in larvae and pupae of the vector mosquito, Aedes aegypti. As a sequel to these findings, the present study elucidates immunotoxicity of this extract with respect to hemocyte-mediated cellular immune responses in fourth instar larvae and pupae as well as cuticular melanization reaction in the larvae of A. aegypti. The exposure of these two developmental stages of the mosquito to the soapnut extract at a lethal threshold concentration neither affected hemocyte viability tested up to 3h in vitro nor did it influence the hemocyte count. By contrast, exposure of the mosquito larvae and pupae to this extract significantly reduced the ability of their hemocytes to bind yeast cells, an important early event in the process of non-self recognition by immune cells. Consequently, the phagocytic activity of these hemocytes against yeast cells was also found to be adversely affected upon exposure of larvae and pupae to the extract. Besides, a perceptible initial delay in melanization reaction at the injured site of the cuticle in the extract-exposed larvae was observed. All these findings demonstrate, for the first time, the immuno-suppressive potential of a botanical biocide in the vector mosquito.

Sapindus emarginatus: Phytochemistry & Various Biological Activities

Sapindus emarginatus is important indigenous plant with lots of traditional importance belongs to family sapindaceae. It is commonly called as Soap nut tree which is found in most of the hilly regions of India. Each and every part of the plant is used traditionally in various ailments. Trees of genus Sapindus are cultivated in many parts of India. The secondary metabolites present in Sapindus emarginatus were found to be alkaloids, carbohydrates, phenols, flavonoids, saponins fixed oils & fats. It possesses various activities such as surfactant, mild detergent, anti-inflammatory, antipruritic, antihyperlipidemic, antimicrobial, CNS, emetic, hair tonic, nasal insufflations. Therefore the aim of the present review is to summarize the taxonomy, chemical constituents and various studies on the biological activities of the plant Sapindus emarginatus. © 2011 IGJPS. All rights reserved.

ISSR analysis of soap nut (Sapindus mukorossi Gaertn.) genotypes in Western Himalaya (India)

Members of the genus Sapindus L. (Sapindaceae), commonly known as soap nut, are commercially important plants, that are found both wild and introduced in tropical and sub-tropical regions of the world, predominantly in Indo-Malayasia. Three species, Sapindus emarginatus Vahl, S. mukorossi Gaertn. and S. trifoliatus L., occur in India. In this study, ISSR markers were used to unravel the genetic diversity in soap nut (S. mukorossi) genotypes of Western Himalaya. A total of 54 S. mukorossi genotypes and one accession of Lepisanthes tetraphylla Vahl. as the out-group were considered. ISSR analysis resulted in 258 bands obtained with 20 primers. Out of these, 152 bands were polymorphic revealing 58.91% polymorphism across all genotypes of S. mukorossi. The genetic distance value varied from 0.02 to 0.37 across different genotypes. The results show that ISSR profiles are sufficiently informative and useful to unravel the genetic variations among soap nut plants in the Western Himalayas. The information on genetic diversity of soap nut could further be utilized in the breeding of genetically divergent and geographically isolated populations to obtain better germplasm in terms of yield and quality.

Genetic variability and population structure in Sapindus emarginatus Vahl from India

Sapindus emarginatus is an economically important tropical tree species sparsely distributed in different geographical provinces like Gangetic Plains, Western Ghats, and Deccan Plateau in India. In the present paper estimation of genetic variability within and among 41 accessions representing five populations was carried out using 3 single primer amplification reaction (SPAR) methods viz. RAPD, DAMD and ISSR. The cumulative data analysis was carried out for all three SPAR methods, and showed 82.32% polymorphism across all the accessions of S. emarginatus. Jaccard's similarity values among 41 accessions ranged from 0.15 to 0.49 with an average value of 0.37. The intra-population genetic diversity revealed highest values of Nei's genetic diversity (0.19,) Shannon information index (0.29) and polymorphic loci (55.18%), among the accessions of Gujarat (GJ) population, while the corresponding lowest values were (0.10), (0.15) and (26.40%) respectively among the accessions of Rajasthan (RJ) population. The maximum inter-population average genetic distance (0.20) was between Karnataka (KA) and RJ, while the corresponding least genetic distance (0.06) was between Allahabad (AL) and Varanasi (VS) populations. The analysis of molecular variance (AMOVA) revealed maximum percentage of variation among individuals of populations (72%) followed by 16% among regions and 12% among populations. Principal coordinate analysis (PCA) of cumulative data also supported the clustering pattern in the UPGMA dendrogram. These results suggest that genetic diversity is corroborating with the geographical diversity. Mantel's test was performed which revealed a highly significant correlation between cumulative vs RAPD, and showed the maximum (0.93) correlation coefficient, followed by cumulative vs ISSR (0.78) and cumulative vs DAMD (0.91) respectively, and this clearly indicates that the SPAR methods (RAPD, DAMD and ISSR) are sufficiently informative and are suitable to analyze the genetic variability within and among the populations of S. emarginatus.

Effects of extract of soapnut Sapindus emarginatus on esterases and phosphatases of the vector mosquito, Aedes aegypti (Diptera: Culicidae)

Our earlier investigations with kernels from the soapnut Sapindus emarginatus revealed it as a new source of botanical biocide with potent antimosquito activity, as evident from the proven unique ability of the aqueous kernel extract to kill all the developmental stages of three important vector mosquito species, Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. This extract was also found to be safe for two non-target aquatic insects. As a sequel to these findings, we have further examined quantitative and qualitative changes in total proteins, esterases, and phosphatases in whole body homogenates of fourth instar larvae and pupae of A. aegypti exposed to this extract at an appropriate threshold time for its lethal effect to gain insights into the impact of the botanical biocide on biochemical characteristics of the target vector mosquito at two distinct developmental stages. The profiles of proteins, esterases (acetylcholinesterse, α- and β-carboxylesterases), and phosphatases (acid and alkaline) exhibited distinct patterns of variation during normal development of fourth instar larvae and pupae, indicating intrinsic difference in biochemical features between these two developmental stages of A. aegypti. Upon exposure of the larvae to the extract, significant reduction in the activities of acetylcholinesterse, β-carboxylesterase, and acid phosphatases were recorded, whereas the total proteins, α-carboxylesterase and alkaline phosphatase activities were unaffected. By contrast, only alkaline phosphatase activity was significantly affected in pupae exposed to the extract. Analysis of these enzymes in native PAGE revealed that they exist in isoforms in both the larvae and pupae. The alterations in the levels of enzymatic activities observed from the quantitative assays of various enzymes were reflected by the respective zymograms with perceptible differences in the intensity and the number of bands detected especially with β-carboxylesterase, acid and alkaline phosphatase activity between the control and exposed test organisms. Despite the fact that the soapnut kernel extract causes mortality of both the larvae and pupae of A. aegypti, the findings of this study demonstrate that the impact of this extract is most pronounced in various enzyme profiles of the larvae rather than the pupae. Such discrepancy implicates the presence of unique biochemical mechanisms in the pupae of mosquito for detoxification of botanical biocides.