Efficacy of leaves extract of Calotropis procera Ait. (Asclepiadaceae) in controlling Anopheles arabiensis and Culex quinquefasciatus mosquitoes

The present study aimed to investigate, the larvicidal, adult emergence inhibition and oviposition deterrent activity of aqueous leaves extract of Calotropis procera against Anopheles arabiensis and Culex quinquefasciatus as natural mosquito larvicide. The larvicidal activity was monitored against 2nd, 3rd and 4th instar larvae of each mosquito species 24 h post-treatment. Adult emergence inhibition activity was tested by exposing 3rd instar larvae of each mosquito species to different concentrations of extracts (200, 400, 600, 800 and 1000 ppm for An. arabiensis and 100, 200, 300, 400, 500 and 600 ppm for Cx. quinquefasciatus). Probit analysis was used to analyze data from bioassay experiments. The oviposition deterrent activity was tested by using three different concentrations of extracts (1000, 500 and 200 for An. arabiensis, and 1000, 500 and 100 for Cx. quinquefasciatus) that caused high, moderate and low larval mortality in the larvicidal experiment against 3rd instar larvae. It was found that, LC50–LC90 values calculated were 273.53–783.43, 366.44–1018.59 and 454.99–1224.62 ppm for 2nd, 3rd and 4th larval instars, respectively, of An. arabiensis and 187.93–433.51, 218.27–538.27 and 264.85–769.13 ppm for 2nd, 3rd and 4th larval instars, respectively, of Cx. quinquefasciatus. Fifty percent of adult emergence inhibition (EI50) was shown at 277.90 and 183.65 ppm for An. arabiensis and Cx. quinquefasciatus, respectively. The pupal stage was not affected till a concentration of 5000 ppm. The extract showed oviposition deterrence and effective repellence against both mosquito species at different concentrations, with the observation on that maximal eggs were laid in low concentration of extract. These results suggest that the leaves extract of C. procera possess remarkable larvicidal, adult emergence inhibitor, repellent and oviposition deterrent effect against both An. arabiensis and Cx. quinquefasciatus, and might be used as natural biocides for mosquito control.     

Development of laticifer cells in callus cultures of Calotropis procera (Ait.) R. Br.

Tissue cultures were established from stem explants of Calotropis procera, a hydrocarbon yielding desert shrub on Murashige and Skoog's medium supplemented with 1.5 mg. 1^–01 2,4-D + 0.5 mg.1^–1 kinetin and polyvinylpyrrolidone. Laticifer cells were not present in young callus but were observed after 4 weeks of callus growth when examined histochemically. These young laticifers were detected in the 5th week of culture and were distinguished from surrounding cells by the presence of characteristic cytoplasm and thin walls. A group of cells with extensive branching was developed after 8 weeks of growth of the callus cultures. These cells were thick walled and contained latex particles in coagulated masses. Positive Liebermann-Burchard test proved the presence of terpenoids in these laticifers.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - KIN Kinetin - PVP Polyvinylpyrrolidone - HHS Heidenhain's Haematoxylin and safranin     

A taxonomic revision of Calotropis (Asclepiadaceae)

The genus Calotropis of the tribe Asclepiadeae of Asclepiadaceae subfamily Ascle-piadoideae is revised throughout its Tange in tropical and subtropical Africa and Asia. The three species recognized are keyed out, illustrated and described with appropriate bibliography, synonymy, notes on ecology, distribution and uses. Distribution maps for the species are provided. The taxonomic position of the genus is discussed and a historical review is provided.     

Hepatoprotective and antioxidant activities of flowers of Calotropis procera (Ait) R. Br. in CCl4 induced hepatic damage

Hepatoprotective activity of 70% ethanolic extract of flowers of C. procera was studied against CCl4 induced hepatic injury in albino rats and mice. In addition, antioxidant activity was studied by in vitro models. Pre-treatment with 70% ethanolic extract (CPA) reduced the biochemical markers of hepatic injury like serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, alkaline phosphatase, bilirubin, cholesterol, HDL and tissue glutathione (GSH) levels. Similarly pretreatment with CPA reduced the CCl4 induced elevation in the pentobarbitone sleeping time. Histopathological observations also revealed that pretreatment with CPA protected the animals from CCl4 induced liver damage. CPA demonstrated dose dependant reduction in the in vitro and in vivo lipid peroxidation induced by CCl4. In addition it showed dose dependant free radical scavenging activity. The results indicate that flowers of C. procera possess hepatoprotective property possibly because of its anti-oxidant activity. This property may be attributed to the quercetin related flavonoids present in the flowers of Calotropis procera.     

Density‐dependent reproduction and pollen limitation in an invasive milkweed,Calotropis procera (Ait.) R. Br. (Apocynaceae)

Calotropis procera (Ait.) R.Br. (Apocynaceae), an invasive woody milkweed, has expanded its range in northern Australia affecting rangeland and pastoral productivity. While self‐compatibility should enhance the species range expansion, spread of C. procera is limited by the availability of larger wasp and bee species that are able to vector its solid pollinia. Pollination efficiency is thus likely dependent on both pollinator abundance and plant density. Calotropis procera flowers year round in Australia but fruiting is limited to the warm months of the year when pollinators are most abundant, indicating that seasonal regulation of reproduction may be due to pollinator limitation. We examine the propositions that C. procera reproduction is regulated by the interaction between plant population density and pollinator pressure and that low pollinator pressure causes low per capita plant fecundity. All pollinators belonged to Order Hymenoptera and pollinator composition was similar at six of the seven sites. Fruit production per plant (fecundity) was lower above and below intermediate densities (350–550 plants ha^?1) of flowering plants with evidence of a weak Allee effect at lower plant density. Pollinator visitation rates per plant were low at high and low plant densities, and greatest at intermediate densities, while pollen supplementation experiments showed that C. procera is pollen limited (Pollen Limitation Index_fruit = 0.9) even at intermediate densities. Pollen limitation caused by low pollinator pressure at low plant densities and pollinator satiation at high plant densities may account for these fruit production trends. Management should be conducted in the colder months when pollinator pressure is low and plants are not reproducing. In addition, where stand eradication cannot be achieved in one attempt, management should reduce flowering plants to below intermediate densities where the fecundity per plant is low.     

Callus culture and in vitro biosynthesis of cardiac glycosides from Calotropis gigantea (L.) Ait

Calotropis gigantea (L.) Ait., belonging to the family Asclepiadaceae, is a source of many cardiac glycosides (CGs) and their steroidal moieties (genins). These CGs have been reported to have anti-proliferative activity on tumor cell lines and are potential targets for cancer chemotherapy. However, the abundance of CGs in wild plants is particularly restricted and it is difficult to isolate the desired compound in required quantities. This study is the first attempt to standardize the induction and proliferation of callus from various explants of C. gigantea specifically for the production of CGs. Callus growth was accompanied by CG measurement using high-performance liquid chromatography-tandem mass spectrometry. Murashige and Skoog (MS) and modified Murashige and Skoog (MMS) media were optimized with various combinations and concentrations of auxin and cytokinin for induction and growth of calli from a range of explant sources. While leaves and stem explants resulted in greatest callus induction, MMS medium was found to be optimal. However, no CG was produced from callus grown on this medium. In contrast, the induction and proliferation of callus on MS medium were optimum at primary stages, but growth slowed during the third subculture. Therefore, calli were transferred to MMS medium to promote callus proliferation and production of CGs. As a result, three CGs and two genins were biosynthesized. Furthermore, the callus induction data in MS medium indicated that among different auxins, 2,4-dichlorophenoxyacetic acid was the best for callus induction compared to 1-naphthylacetic acid and indole-3-acetic acid. The data also revealed that the cytokinin/auxin ratio was critical rather than their independent presence for the induction of callus. Thus, the in vitro biosynthesis of targeted CGs may offer an alternative pathway for new source of anti-proliferative agents in required quantities.     

Morpho-ecological aspects on the pollination ofCalotropis procera (Asclepiadaceae) in Israel

Calotropis procera (Asclepiadaceae) grows in Israel under extremely hot climatic conditions. Flower morphology is adapted towards the concealment of nectar, in order to protect it against robbery by ants and from evaporation. Pollinia while being inserted within the stigmatic chamber, are soaked in the liquid nectar which, by its concentration controls pollen germination. Two Carpenter bees (Xylocopa) are the only pollinators and their behavior on flowers is discussed.     

Antimicrobial,immunomodulatory and cytotoxic activities of green synthesized nanoparticles from Acacia honey and Calotropis procera

Calotropis procera and Somra (Acacia) honey are used in traditional medicine. The benefits of mixing 20% Somra honey and C. procera leaf water extract (CPLWExt) were aimed to be studied. Honey/CPLWExt were utilized to produce silver nanoparticles (AgNPs) separately. AgNPs were characterized via UV/Vis and electron microscope scanning. Bio-molecules in CPLWExt/honey were investigated utilizing FT-IR spectroscopy. Biological activities of CPLWExt and honey were tested. The outcomes showed that CPLWExt and honey have numerous functional groups and could produce AgNPs. CPLWExt, CPLWExt + AgNPs, honey and honey + AgNPs hindered the growth of rat splenocytes, while CPLWExt + honey invigorated it. Antimicrobial power was found in CPLWExt and honey, which increased in the presence of AgNPs. Honey/honey + AgNPs suppressed the proliferation of HeLa and HepG2 cells. In conclusion, honey/CPLWExt could produce AgNPs and showed immunomodulatory and antibacterial power. Somra honey/honey + AgNPs have anticancer power. Somra honey + CPLWExt reflected a good immunostimulatory powers that can be nominated as an immunostimulant.     

Seaweed Liquid Extract as an Alternative Biostimulant for the Amelioration of Salt-stress Effects in Calotropis procera (Aiton) W.T

Scientists consider saltwater one of the effective environmental stress that negatively affects the growth and establishment of trees and shrubs worldwide. Utilizing the potential of Bio-stimulant compounds present in the brown seaweed extract is an alternative strategy to improve crop tolerance to salinity. This study focused on the application of seaweed extract as a Bio-stimulant agent to counteract the salt stress on the growth and some physiochemical aspects of milkweed seedlings. In this experiment, the seedlings were treated with seaweed extract (SWE) of Sargassum angustifolium at four concentrations (non-SWE or control, 0.5, 1.0, and 1.5%) and then exposed to salt stress at four levels (0, 7.5, 15, and 30 dS m^?1 of diluted seawater) in a completely randomized design (four replications per treatment) over a time-span of 3 months. The results indicated that SWE-treated seedlings could tolerate salinity up to 15 dS m^?1 and also increase the survival rate by 69%. The growth parameters like height, specific leaf area, root length and volume, root and shoot dry weight were considerably enhanced by SWE (1%) from 7.5 to 30 dS m^?1. Moreover, gas exchanges and chlorophyll pigments were markedly increased using SWE (0.5%) under salt lower 15 dS m^?1 than control. Also, both SWE and salt stress significantly enhanced antioxidant enzymes over control, but SWE more increased the parameters. SWEs agent at different dosages significantly decreased electrolyte leakage at all salinity levels (except in 7.5 and 15 dS m^?1) compared to control. SWEs (1%) resulted in increasing K⁺ uptake but decreasing Na⁺ uptake and markedly enhancing K^+/Na⁺ ratio in stressed-milkweed versus free-salt stress. Totally, this research illustrates the potential of SWEs (at lower dosages) for elevating milkweed tolerance to moderate salinity stress and highlights the possibility of applying it as Bio-stimulant fertilizer.

     

Anastomosing laticifer in the primary and secondary structures of Calotropis procera (Aiton) W.T.Aiton (Apocynaceae) stems

Protoplasma - An in-depth understanding of the development and distribution of laticifer (latex secretory structure) will be important for the production of both rubber and medicines and will...     

Latex constituents from Calotropis procera (R. Br.) display toxicity upon egg hatching and larvae of Aedes aegypti (Linn.)

Calotropis procera R. Br. (Asclepiadaceae) is a well-known medicinal plant with leaves, roots, and bark being exploited by popular medicine to fight many human and animal diseases. This work deals with the fractionation of the crude latex produced by the green parts of the plant and aims to evaluate its toxic effects upon egg hatching and larval development of Aedes aegypti. The whole latex was shown to cause 100% mortality of 3rd instars within 5 min. It was fractionated into water-soluble dialyzable (DF) and non-dialyzable (NDF) rubber-free materials. Both fractions were partially effective to prevent egg hatching and most of individuals growing under experimental conditions died before reaching 2nd instars or stayed in 1st instars. Besides, the fractions were very toxic to 3rd instars causing 100% mortality within 24 h. When both fractions were submitted to heat-treatment the toxic effects were diminished considerably suggesting low thermostability of the toxic compounds. Polyacrylamide gel electrophoresis of both fractions and their newly fractionated peaks obtained through ion exchange chromatography or desalting attested the presence of proteins in both materials. When submitted to protease digestion prior to larvicidal assays NDF lost most of its toxicity but DF was still strongly active. It may be possible that the highly toxic effects of the whole latex from C. procera upon egg hatching and larvae development should be at least in part due to its protein content found in NDE However the toxicity seems also to involve non protein molecules present in DF.     

Prospects for the classical biological control of Calotropis procera (Apocynaceae) using coevolved insects

Calotropis procera (Apocynaceae), a native of tropical Africa, the Middle East and the Indian subcontinent, is a serious environmental and rangeland weed of Australia and Brazil. It is also a weed in Hawaii in the USA, the Caribbean Islands, the Seychelles, Mexico, Thailand, Vietnam and many Pacific Islands. In the native range C. procera has many natural enemies, thus classical biological control could be the most cost-effective option for its long-term management. Based on field surveys in India and a literature search, some 65 species of insects and 5 species of mites have been documented on C. procera and another congeneric-invador C. gigantea in the native range. All the leaf-feeding and stem-boring agents recorded on Calotropis spp. have wide host range. Three pre-dispersal seed predators, the Aak weevil Paramecops farinosus, the Aak fruit fly Dacus persicus in the Indian subcontinent and the Sodom apple fruit fly Dacus longistylus in the Middle East, have been identified as prospective biological control agents based on their field host range. In Australia and Brazil, where C. procera has the potential to spread across vast areas, pre-dispersal seed predators would help to limit the spread of the weed. While the fruits of C. procera vary in size and shape across its range, those from India are similar to the ones in Australia and Brazil. Hence, seed-feeding insects from India are more likely to be suitable due to adaptation to fruit size and morphology. Future survey efforts for potential biological control agents should focus on North Africa.     

Enzymatic activities and protein profile of latex from Calotropis procera.

The laticifer fluid of Calotropis procera is rich in proteins and there is evidence that they are involved in the pharmacological properties of the latex. However, not much is known about how the latex-containing proteins are produced or their functions. In this study, laticifer proteins of C. procera were pooled and examined by 1D and 2D electrophoresis, masses spectrometry (MALDI-TOF) and characterized in respect of proteolytic activity and oxidative enzymes. Soluble laticifer proteins were predominantly composed of basic proteins (PI>6.0) with molecular masses varying between 5 and 95 kDa. Proteins with a molecular mass of approximately 26,000 Da were more evident. Strong anti-oxidative activity of superoxide dismutase (EC 1.15.1.1) (1007.74+/-91.89 Ug(-1)DM) and, to a lesser extent ascorbate peroxidase (EC 1.11.1.1) (0.117(d)+/-0.013 microMol H(2)O(2)g(-1)min(-1)), were detected. However, catalase (EC 1.11.1.6) was absent. The strong proteolytic activities of laticifer proteins from C. procera were shown to be shared by at least four distinct cysteine proteinases (EC 3.4.22.16) that were isolated by gel filtration chromatography. Serine and metaloproteinases were not detected and aspartic proteinase activities were barely visible. Chitinases (EC 3.2.1.14) were also isolated in a chitin column and their activities quantified. The presence of these enzymatic activities in latex from C. procera may confirm their involvement in resistance to phytopathogens and insects, mainly in its leaves where the latex circulates abundantly.     

Biodiversity of endophytic fungi in different leaf ages of Calotropis procera and their antimicrobial activity

Calotropis procera has many important medicinal properties with proven pharmacological potential. Some of these properties may be mediated by its fungal endophytes. This study analyzed, for the first time, the community of endophytic fungi of C. procera outside its region of origin. A total of 156 fungal isolates distributed across 19 taxa were obtained from 468 fragments of C. procera leaves at different stages of maturation. The rate of endophyte colonization increased with the leaf age/development. The dominant species of endophytic fungi of C. procera introduced in Northeast Brazil were different from those found in studies on the same species and other species of the same genus in native regions. The dominant endophyte was Phaeoramularia calotropidis (63.5 %), followed by Guignardia bidwellii (21.1 %). Six isolates of endophytic fungi showed antimicrobial activity against human pathogenic micro-organisms and one plant pathogenic fungus. The antibacterial activity was more intense than the antifungal activity. The endophytic Curvularia pallescens (URM 6048) stood out inhibited Gram-positive bacteria, Staphylococcus aureus, Streptococcus pyogenes, the plant pathogenic fungus Colletotrichum dematium. Ecological and biotechnological aspects of endophytic mycota are discussed.     

Identification and Sequence-Based Validation of the EST-SSR Markers from Calotropis procera

Plant Molecular Biology Reporter - Calotropis procera (Aiton) Dryand is a medicinally and economically important woody shrub used to cure various diseases. In the present study, for the first time,...     

Molecular cloning and anti-invasive activity of cathepsin L propeptide-like protein from Calotropis procera R. Br. against cancer cells

Cathepsin L of cancer cells has been shown to play an important role in degradation of extracellular matrix for metastasis. In order to reduce cell invasion, cathepsin L propeptide-like proteins which are classified as the I29 family in the MEROPS peptidase database were characterized from Calotropis procera R. Br., rich in cysteine protease. Of 19 candidates, the cloned and expressed recombinant SnuCalCp03-propeptide (rSnuCalCp03-propeptide) showed a low nanomolar K_i value of 2.3?±?0.2?nM against cathepsin L. A significant inhibition of tumor cell invasion was observed with H1975, HT29, MDA-BM-231, PANC1, and PC3 with a 76, 67, 67, 63, and 79% reduction, respectively, in invasion observed in the presence of 400?nM of the rSnuCalCp03-propeptide. In addition, thermal and pH study showed rSnuCalCp03-propeptide consisting of secondary structures was stable at a broad range of temperatures (30–70?°C) and pH (2–10, except for 5 which is close to the isoelectric point of 5.2).     

Phenylpropanoid and flavonoid glycosides from the leaves of Clerodendrum infortunatum (Lamiaceae)

Clerodendrum infortunatum L. (syn.: Clerodendrum viscosum Vent.), a member of the Lamiaceae, yielded one undescribed jasmonic acid derivative, ten acteosides, and two flavonoids. The jasmonic acid derivative was identified as 6'-O-caffeoyl-12-glucopyranosyloxyjasmonic acid. The acteosides were identified as isoacteoside, acteoside, 2''-O-acetyl-martyonside, 3''-O-acetyl-martyonside, martynoside, brachynoside, leucosceptoside A, jionoside C, jionoside D, incanoside C. The flavonoids were identified as apigenin 7-O-glucuronide and acacetin 7-O-glucuronide. The structures of the isolated components have been identified by UHPLC-HRMS, 1D and 2D NMR spectroscopic analyses, spectrometric techniques, and in comparison with published NMR data. The absolute sugar configuration was determined by GLC-MS/MS analysis of the octylated derivative of the sugar moiety after hydrolysis. Among the known compounds, ten are reported for the first time from this species, while the acteoside leucosceptoside A and the two flavonoids have been isolated for the first time from the genus Clerodendrum. The chemophenetic significance of the compounds obtained from C. infortunatum is summarized in comparison to those found in other Clerodendrum species.