Comparative genetic analysis of trichome-less and normal pod genotypes of Mucuna pruriens (Fabaceae).

Velvet bean (Mucuna pruriens) seeds contain the catecholic amino acid L-DoPA (L-3,4-dihydroxyphenylalanine), which is a neurotransmitter precursor and used for the treatment of Parkinson's disease and mental disorders. The great demand for L-DoPA is largely met by the pharmaceutical industry through extraction of the compound from wild populations of this plant; commercial exploitation of this compound is hampered because of its limited availability. The trichomes present on the pods can cause severe itching, blisters and dermatitis, discouraging cultivation. We screened genetic stocks of velvet bean for the trichome-less trait, along with high seed yield and L-DoPA content. The highest yielding trichome-less elite strain was selected and indentified on the basis of a PCR-based DNA fingerprinting method (RAPD), using deca-nucleotide primers. A genetic similarity index matrix was obtained through multivariant analysis using Nei and Li's coefficient. The similarity coefficients were used to generate a tree for cluster analysis using the UPGMA method. Analysis of amplification spectra of 408 bands obtained with 56 primers allowed us to distinguish a trichome-less elite strain of M. pruriens.

AFLP markers for identification of Swertia species (Gentianaceae).

The genus Swertia is well known for its medicinal properties, as described in the Indian pharmacopoeia. Different members of this genus, although somewhat similar in morphology, differ widely in their pharmacological and therapeutic properties. The most important species of this genus, with maximal therapeutic properties, is S. chirayita, which is often adulterated with other less-potent Swertia spp. There is an existing demand in the herbal drug industry for an authentication system for Swertia spp, in order to enable their commercial use as genuine phytoceuticals. To this end, we used amplified fragment length polymorphism (AFLP) to produce DNA fingerprints for six Swertia species. Nineteen accessions (2 of S. chirayita, 3 of S. angustifolia, 2 of S. bimaculata, 5 of S. ciliata, 5 of S. cordata, and 2 of S. alata) were used in the study, which employed 64 AFLP selective primer pairs. Only 46 selective primer pairs were found to be useful for all the accessions. A total of 5312 fragments were produced by these 46 primer pairs. Species-specific markers were identified for all six Swertia species (131 for S. chirayita, 19 for S. angustifolia, 181 for S. bimaculata, 47 for S. ciliata, 94 for S. cordata, and 272 for S. alata). These AFLP fingerprints of the Swertia species could be used to authenticate drugs made with Swertia spp and to resolve adulteration-related problems faced by the commercial users of these herbs.

Endophytic bacteria from Ocimum sanctum and their yield enhancing capabilities.

Endophytes are beneficial microbes that reside intercellularly inside the plants. Interaction of endophytes with the host plants and their function within their host are important to address ecological relevance of endophyte. Four endophytic bacteria OS-9, OS-10, OS-11, and OS-12 were isolated from healthy leaves of Ocimum sanctum. These isolated microbes were screened in dual culture against various phytopathogenic fungi viz. Rhizoctonia solani, Sclerotium rolfsii, Fusarium solani, Alternaria solani, and Colletotrichum lindemuthianum. Of these, strain OS-9 was found to be antagonistic to R. solani, A. solani, F. solani, and C. lindemuthianum while OS-11 was found antagonistic against A. solani only. The growth-promoting benefits of the endophytes were initially evaluated in the glasshouse by inoculated seeds of O. sanctum. Treatment with endophytes OS-10 and OS-11 resulted in significant enhancement of growth as revealed by increase in fresh as well as dry weight. Further, field trials involving two genotypes OS Purple and CIM-Angana were conducted with strains OS-10 and OS-11. The growth-promoting effect was visible on both the genotypes tested as the endophytes significantly enhanced fresh herbage yield (t/ha). Interestingly, these endophytes increased the content of essential oil particularly in cultivar OS Purple and thereby increasing the total oil yields. Molecular characterization of strain OS-11 indicated the strain to be highly related to the type strain of Bacillus subtilis.

In vitro propagation of Rauwolfia serpentina using liquid medium, assessment of genetic fidelity of micropropagated plants, and simultaneous quantitation of reserpine, ajmaline, and ajmalicine.

Rauwolfia serpentina holds an important position in the pharmaceutical world because of its immense anti-hypertensive properties resulting from the presence of reserpine in the oleoresin fraction of the roots. Poor seed viability, low seed germination rate, and enormous genetic variability are the major constraints for the commercial cultivation of R. serpentina through conventional mode. The present optimized protocol offers an impeccable end to end method from the establishment of aseptic cultures to in-vitro plantlet production employing semisolid as well liquid nutrient culture medium and assessment of their genetic fidelity using polymerase chain reaction based rapid amplification of polymorphic DNA analysis. In vitro shoots multiplied on Murashige and Skoog basal liquid nutrients supplemented with benzo[a]pyrene (1.0 mg/L) and NAA (0.1 mg/L) and in-vitro rhizogenesis was observed in modified MS basal nutrient containing NAA (1.0 mg/L) and 2% sucrose. In-vitro raised plants exhibited 90-95% survival under glass house/field condition and 85% similarity in the plants regenerated through this protocol. Field established plants were harvested and extraction of indole alkaloid particularly reserpine, ajmaline and ajmalicine and their simultaneous quantitation was performed using monolithic reverse phase high-performance liquid chromatography (HPLC).

Toxic effects of oral administration of extracts of dried calyx of Hibiscus sabdariffa Linn. (Malvaceae).

The effects of a 90-day oral administration of water and alcohol extracts of dried calyx of Hibiscus sabdariffa were evaluated in albino rats. Haematological, biochemical and histopathological changes were monitored every 30 days.The death of the animals was preceded by a severe loss in weight, accompanied with diarrhoea in animals on the 2000 mg/kg dose. There was an increase in food intake (g) per kg body weight per day in the aqueous (A) and ethanol (E) 300 mg/kg extract groups. Significant reductions in the erythrocyte count with no difference in total leucocyte count were observed. The activity of aspartate aminotransferase (AST) was enhanced by the administration of aqueous and 50% ethanol extract with a significant increase in its level at higher doses (p < 0.05). Alanine aminotransferase (ALT) and creatinine levels were significantly affected by all the extracts at the different dose levels. However, aqueous extracts exhibited a significant increase in creatinine levels (p < 0.05) at higher doses. The cholesterol levels were generally not significantly affected by the extracts. No significant histopathological changes were observed, although there was a significant reduction in the weight of the spleen of the animals administered with ethanol and water extracts when compared with the control (p < 0.01). Other organs were of the same relative weight.

Modulation of inflammatory mediators by coumarinolignoids from Cleome viscosa in female swiss albino mice.

The effect of coumarinolignoid cleomiscosins A, B and C isolated from the plant Cleome viscosa on inflammatory mediators were studied in female swiss albino mice. A mixture of coumarinolignoid A, B, and C at 10, 30 and 100 mg/kg body weight once a day for 14 consecutive days were administered orally to the mice. Pro-inflammatory mediators such as IL-6, TNF-alpha and nitric oxide were estimated from culture supernatant obtained from peritoneal macrophages stimulated by LPS and anti-inflammatory mediator IL-4 was estimated from culture supernatant obtained from spleenocytes stimulated by Con-A. For further confirmation, expressions of inflammatory mediators from serum and mortality rate were studied in LPS-induced toxicity model in mice. The expression of Pro-inflammatory mediators was significantly (P <0.05) decreased in coumarinolignoids treatment group in dose dependent manner, whereas the anti-inflammatory mediator expression was significantly increased in coumarinolignoids at 10 mg/kg treatment. Mortality rate was also significantly reduced in treatment group in LPS-induced toxicity model. The result of this study concluded that the oral administration of coumarinolignoids inhibited the pro-inflammatory mediators and enhances the production of anti-inflammatory mediator in dose dependent manner.

Anxiolytic and sedative effects of extracts of Hibiscus sabdariffa Linn (family Malvaceae).

Aqueous (A), hydroalcoholic (AE) and ethanolic (E) extracts and fractions of dried calyxes of Hibiscus sabdariffa were evaluated for anxiolytic property using elevated-plus maze (EPM), and sedative properties using ketamine-induced sleep in animal models. The extracts exhibited a dose-dependent increase in the time spent in the open arm with ethanol extracts having the best anxiolytic activity. The extracts at A 5 mg/kg, AE 5 mg/kg and A 50 mg/kg did not cause an increase in time spent in the open arms (p < 0.05) but other doses significantly did when compared with the vehicle control. The fractions of the hydroalcoholic extracts showed no significant anxiolytic activity. Neither the extracts nor the fractions significantly reduced or increased latency to sleep after a single dose except AE 300 (p < 0.01). There was significant reduction in onset of sleep, and increase in sleeping time with A and AE extracts with multiple doses at A 300, AE 50 and AE 300 mg/kg dose groups (p < 0.001, p < 0.01 and p < 0.05 respectively). A reduction in sleeping time after several daily doses of ethanol extracts doses was observed. A single dose of one of the fractions (EAC at 50 mg/kg) caused a significant reduction (p < 0.05) in sleep duration. The study showed that extracts of Hibiscus sabdariffa possess anxiolytic and sedative effects which become more pronounced with administration of repeated doses of the extracts.

Gallic acid-based indanone derivatives as anticancer agents.

Gallic acid-based indanone derivatives have been synthesised. Some of the indanones showed very good anticancer activity in MTT assay. Compounds 10, 11, 12 and 14 possessed potent anticancer activity against various human cancer cell lines. The most potent indanone (10, IC(50)=2.2 microM), against MCF-7, that is, hormone-dependent breast cancer cell line, showed no toxicity to human erythrocytes even at higher concentrations (100 microg/ml, 258 microM). While, indanones 11, 12 and 14 showed toxicities to erythrocytes at higher concentrations.

Immunomodulatory effect of extracts of Hibiscus sabdariffa L. (Family Malvaceae) in a mouse model.

The immunomodulatory activity of water and alcohol extracts (including its fractions) of the dried calyx of the plant was evaluated in mice. The ability of the extracts to inhibit or enhance the production of two cytokines, namely tumor necrosis factor-alpha (TNF- alpha) and interleukin-10 (IL-10), respectively, implicated as proinflammatory and antiinflammatory interleukins were also evaluated. The extracts at doses of 50 mg/kg were found to possess higher immunostimulatory activities in comparison with levamisole (positive control), with significant effects when compared with the vehicle-treated group (p < 0.01). Increased activity was observed with increase in doses of the 50% ethanol and absolute ethanol extracts. The insoluble fraction exhibited a significant dose-dependent immunostimulatory activity (p < 0.05), while the residual water-soluble fraction exhibited activity at 100 mg/kg body weight. The production of tumor necrosis factor-alpha (TNF-alpha), was low in all the extract groups tested, while the production of interleukin 10 (IL-10) was high compared with the control. The production of IL-10 was high in 300 mg/kg aqueous extract. The insoluble fraction exhibited a profound dose-dependent immunostimulatory activity higher than the positive control at 100 mg/kg. This study established the immunoenhancing properties of the extracts of this plant confirming that the immunomodulatory activity is cell mediated and humoral. The insoluble fraction could find use as an immunostimulatory agent in humans.

Recent advances in plant hepatoprotectives: a chemical and biological profile of some important leads.

Medicinal plants have been traditionally used for treating liver diseases since centuries. Several leads from plant sources have been found as potential hepatoprotective agents with diverse chemical structures. Although, a big list of hepatoprotective phytomolecules was reported in the scientific literature, only a few were potent against various types of liver damages. Of which, silymarin, andrographolide, neoandrographolide, curcumin, picroside, kutkoside, phyllanthin, hypophyllanthin, and glycyrrhizin have largely attracted the scientific community. This review focuses discussion on the chemistry, biological activity, mode of action, toxicity, and future prospects of these leads.