Himalayan Plants Research Articles

Exploring the physicochemical profile and the binding patterns of selected novel anticancer Himalayan plant derived active compounds with macromolecular targets

Abstract Plants are vital source of compounds offering plethora of therapeutic effects against various ailments without much side effects. Due to wide spread prevalence and drug resistance in cancer; there is an urgent need for discovery of new anti-cancer drugs. In the present study, selected novel anti-cancer plants derived compounds (cmpd1 to cmpd15) from Himalayan region were docked with defined molecular targets that regulate cell proliferation and apoptosis. The binding energies of best docked compounds ranged between −8.0 kcal/mol and −11.71 kcal/mol. Further analysis revealed critical hydrogen bonds and hydrophobic interactions between compounds and targets. The best docked compounds viz., cmpd15 against cyclin-dependent kinase-2 (CDK-2), cmpd8 against CDK-6 and cmpd9 against Topoisomerase I and II showed higher binding affinities than the native co-crystal ligands. The root mean square deviation (RMSD) and potential energy plot clearly indicates the stability of the complexes during 20 ns molecular dynamics (MD) simulation. The Molecular Mechanics/Poisson Boltzmann Surface Area (MM/PBSA) binding energy analysis revealed Van der Waals energy component which is the principal stabilizing energy for their interactions except CDK-2/cmpd15 complex. The polar solvation energy did not have favorable contribution to their stabilization. The binding energy decomposition analysis revealed per residue contribution for each docked complexes. Physicochemical profile studies showed that majority of the compounds conform to Lipinski's rule of five (ROF) having low to high blood brain barrier (BBB) penetration, human intestinal absorption, plasma binding protein inhibition and P glycoprotein inhibition.

Photoprotective, antioxidant screening and new ester from dry root extracts of Potentilla atrosanguinea (Himalayan cinquefoil)

In vitro photoprotective and antioxidant activities of dried aqueous-methanolic (H2O/MeOH) crude extract and ethyl acetate (EtOAc), n-butanol (n-BuOH) as well as aqueous (H2O) fractions of roots of western Himalayan plant Potentilla atrosanguinea (Himalayan cinquefoil) were evaluated. The ability of protection against UVB region followed the trend EtOAc>n-BuOH>H2O/MeOH>H2O. Total phenol content of crude H2O/MeOH extract was found to be 429.808mg GAE/g. The H2O/MeOH crude extract showed highest antioxidant of DPPH radical scavenging, superoxide anion radical scavenging and cupric ion reducing of 90.04, 78.86 and 88.64% respectively at 200μg/mL. One new compound methyl pentatetraconta-30,32,34,36,38,40,42-heptaenoate (PA-1) along with a known pentadecylbutyrate (PA-2) was isolated by column chromatography. Results indicated the importance of root extracts as photoprotective agents in sunscreen preparation in the pharmaceutical industry and considered as a natural source of antioxidants.

Chemical composition and insecticidal activities of essential oils against diamondback moth, Plutella xylostella (L.) (Lepidoptera: Yponomeutidae)

Five Himalayan plants namely, Acorus calamus, Cedrus deodara, Aegle marmelos, Tagetes minuta and Murraya koenigii were used for the extraction of essential oils through hydrodistillation and the major volatile constituents as identified by GC and GC–MS techniques were β-asarone (91.1%), β-himachalene (45.8%), limonene (59.5%), Z-ocimene (37.9%) and α-pinene (54.2%), respectively. Essential oils were tested for their insecticidal properties against larvae of diamondback moth, Plutella xylostella (L.) (Lepidoptera: Yponomeutidae). Results showed that A. calamus was most toxic (LC50 = 0.29 mg mL−1) to P. xylostella followed by C. deodara (LC50 = 1.08 mg mL−1) and M. koenigii (LC50 = 1.93 mg mL−1) via residual toxicity bioassay. Per cent feeding deterrence index and growth inhibition was significantly higher in A. calamus (42.20 and 68.55, respectively) followed by C. deodara (35.41 and 52.47). In repellent activity studies, C. deodara showed high repellence (64.76%) followed by A. calamus (55.05%).

Callus induction and shoot regeneration from rhizome explants of Rheum webbianum Royle- a threatened medicinal plant growing in Kashmir Himalaya

Rheum webbianum Royle is an important medicinal plant of family Polygonaceae growing in Kashmir Himalaya. It possesses hepatoprotective, spasmolytic, anticholesterolaemic, antitumor, antiseptic, antifungal, anti-microbial, anti-Parkinson’s, anti-proliferative, immuno-enhancing, antiviral and antioxidant properties. During the present study an attempt has been made to standardize a protocol for large scale propagation of this threatened Himalayan plant species using rhizome cuttings as explants. Callus was obtained when the rhizome explants were inoculated on MS medium containing different concentrations of auxins either alone or in combination, 2, 4-D at a concentration of 0.5 mg/l was found to be most effective in callus production in a time period of 30 days. Callus derived from rhizome explants differentiated into shoots on MS medium supplemented with different concentrations of auxins (IAA, IBA) and (BAP). Maximum number of shoots (2.8±0.2) was regenerated on MS medium supplemented with BAP (5.0 mg/l) and IAA (2.0 mg/l) after 16 days.

Cloning and functional characterization of β-1, 3-glucanase gene from Podophyllum hexandrum — A high altitude Himalayan plant

Podophyllum hexandrum is a high-altitude medicinal plant exploited for its etoposides which are potential anticancer compounds. ß-1, 3-glucanase cDNA was cloned from the germinating seeds of Podophyllum (Ph-glucanase). Glucanases belong to pathogenesis related glycohydralase family of proteins, which also play an important role in endosperm weakening and testa rupture during seed germination. Analysis of cloned nucleotide sequence revealed Ph-glucanase with an open reading frame of 852bp encoding a protein of 283 amino acids with a molecular mass of 31kDa and pI of 4.39. In-silico structure prediction of Ph-glucanase showed homology with that of Hevea brasiliensis (3em5B). Structural stability and enhanced catalytic efficiency in harsh climatic conditions possibly due to the presence of glycosyl hydrolase motif (LGIVISESGWPSAG) and a connecting loop towards inner side and well exposed carbohydrate metabolism domain-COG5309, can readily hydrolyse cell wall sugar moieties. Seeds from the transgenic Arabidopsis plants over-expressing Ph-glucanase showed better germination performance against a wide range of temperatures and abscisic acid (ABA) stress. This can be attributed to the accumulation of Ph-glucanase at both transcript and protein levels during the seed germination in transgenic Arabidopsis. Results confirm that the cloned novel seed specific glucanase from a cold desert plant Podophyllum could be used for the manipulation of different plant species seeds against various harsh conditions.

Himalayan Alpine Vegetation, Climate Change and Mitigation

The Himalaya are experiencing the most drastic global climate change outside of the poles, with predicted temperature increases of 5–6°C, rainfall increases of 20–30%, and rapid melting of permanent snows and glaciers. We have established a 1500 km trans-Himalayan transect across Nepal, Bhutan, and the Tibetan Autonomous Prefecture (TAP), China to document the effects of climate change on alpine plants and peoples. Data show that Himalayan alpine plants respond to environmental and climate change variables including elevation, precipitation, and biogeography. People use alpine plants mostly for medicines and grazing. Climate change threatens rare, endemic, and useful Himalayan plant species and is being monitored into the future. Mitigation of climate change in the Himalaya takes place, without conscious reference to climate change, through carbon negative livelihoods informed by traditional ecological knowledge (TEK) including conservation of sacred sites, afforestation, tree crops, and soil carbon sequestration through incorporation of mulch and manure.

Invitro anti-mutagenic potential of potentilla fulgens: A western himalayan plant

Invitro anti-mutagenic potential of different extracts/fractions of Potentilla fulgens has been made by an Ames histidine reversion assay using TA98 and TA100 tester strains of Salmonella typhimurium against the direct-acting mutagens, 4-Nitro-O-phenylenediamine (NPD) and Sodium azide. P. fulgens has been used in traditional system of medicine as anti-hyperglycemic, hypoglycemic, anti-hyperlipidemic, anti-oxidant, anti-inflammatory and anti-ulcerogenic agent. Results indicated that n-butanol and water fractions showed strong inhibition activity against the mutagens (NPD, Sodium azide) in both co-incubation as well as pre-incubation treatments in TA98 and TA100 tester strains.

Himalayan Plants as a Source of Anti-Cancer Agents: A Review

Plants are an important source of synthetic and herbal agents used in several pharmaceutical industries. Some of the prominent plant-derived compounds have played a major role in the development of several clinically useful anticancer agents such as vinblastine, vincristine, topotecan and paclitaxel (taxol). Himalayan plants, grown in high altitude are the rich source of various secondary metabolites such as anthraquinones, flavonoids, tannins, alkaloids, etc. This review article covers the active constituents isolated from several anticancerous plants of Himalaya and their effect against several anticancer cell lines.

In vitroregeneration ofCinnamomum tamalaNees from cotyledonary segments

Cinnamomum tamala Nees., a valuable spice-yielding plant of Himalaya, is vulnerable in its natural habitat due to its over-collection for spice and medicinal purpose. In the present study, a successful attempt was made for in vitro shoot bud induction from cotyledons and plant regeneration. The cultured cotyledons of 10–11 wk after flowering induced meristematic response within 4–5 wk of culture where ∼43% explants responded positively on MS medium for tified with 3% sucrose, polyvinyl pyrollidone (100mgL−1) and 6 μM benzyl adenine wherein ∼5–6 shoot buds developed cotyledon−1. Themicro-shoots/shoot buds, developed on initiationmedium, were maintained on MS medium fortified with 3% sucrose and 3 μM kinetin where shoot buds converted into plantlets and supported culture proliferation. Shoots (4 cm long) with fully open leaves were rooted by maintaining on MS medium enriched with 3 μM α-naphthalene acetic acid where 5–6 roots developed per micro-shoot. The rooted plants were transferred to potting mix and maintained in agro-shade ca75% shading where ∼65% transplants survived after two months of transfer.

Steroid; β- Sitosterol from Corydalis govaniana (Wall) growing Himalayan region of Nepal

The Himalayan plant Corydalis govaniana (Wall) commonly known as Bhutkesi in Hindi on detail chemical investigation led to the isolation of β-sitosterol and its structure was determined by detail spectroscopic analysis. DOI: http://dx.doi.org/10.3126/jncs.v29i0.9253Journal of Nepal Chemical Society Vol. 29, 2012 Page: 62-66 Uploaded date: 12/5/2013

Cryotolerance strategies of Pseudomonads isolated from the rhizosphere of Himalayan plants

The cold stress biology of psychrotrophic Pseudomonas strains isolated from the rhizosphere of Himalayan plants have been explored to evaluate their cryotolerance characteristcs. Pseudomonas strains were examined for stress metabolites, viz., exopolysaccharide (EPS) production, intracellular sugar, polyols and amino acid content, ice nucleation activity, and their freezing survival at -10 and -40°C, respectively. High freezing survival was observed for the Pseudomonas strains that were grown at 4°C prior to their freezing at -10 or -40°C. Increased EPS production was noticed when Pseudomonas strains were grown at lower temperatures, i.e., 4 and 15°C, in comparison with their optimal growth temperature of 28°C. All Pseudomonas strains showed low level of type-III class ice nucleation activity at -10°C after 96 h. Considerable differences were noticed in accumulated contents of various intracellular sugars, polyols, amino acids for all Pseudomonas strains when they grown at two different temperatures, i.e., 4 and 28°C, respectively. The unusual complement of stress protectants especially, raffinose, cysteine and aspartic acid that accumulated in the bacterial cells at low temperature was novel and intriguing finding of this study. The finding that raffinose is a key metabolite accumulated at low temperature is an exciting discovery, and to the best of our information this is first report ever signifying its role in bacterial cold tolerance.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-2-667) contains supplementary material, which is available to authorized users.

Developing Spectral Library of Major Plant Species of Western Himalayas Using Ground Observations

A spectral library of commonly occurring Himalayan plant species has been developed. The 512-channel ASD handheld Fieldspec®Pro, 2000 Spectroradiometer with 3 nm spectral resolution and spectral range of 325 to 1,075 nm has been used for recording the leaf, branch or canopy spectra of plant species. Simultaneous measurements of crop biochemical parameters were estimated following standard methods, and vegetation indices were calculated for all the observations. The photographs of the plant species as well as their general descriptions were also detailed in the library. The spectral library has been developed in .Net programming environment. The GUI of library aids in viewing of all the information related to plant species such as spectral details, spectral graphs, general information of species, observation details, plant photographs, species spectral narrowband indices, species biochemical parameters, export options and help through menus and sub menus. The spectra and other information can be exported for further use by the user. The spectral reflectance can be used during classification of Hyperspectral images. The information provided in the library may also be used to explore the application potential of Hyperspectral images in studying chemical constituents, growth behavior, and ambient ecology of plants on a regional scale in Himalayan region. The basic objective of the work was to standardize the technique for vegetation spectral library development and make the data available for comparison by the user.

Genetic and functional diversity among root-associated psychrotrophic Pseudomonad’s isolated from the Himalayan plants

Out of 534 psychrotrophic bacteria, 12 bacteria were selected on the basis of plant growth promoting activities at 4°C and identified as Pseudomonas genus. These strains showed high level of genetic polymorphisms based on RAPD and rep-PCR fingerprinting. This genetic variability revealed that isolates belonging to same species were as high as the variability among different species. Further inoculation of these Pseudomonas strains significantly improves root/shoot biomass and nutrients uptake of lentil plant as compared to non-bacterized control after 40 days of seed showing. Agglomerative hierarchical clustering analysis of pot assay results revealed that genetically diverse strains showing the same prototype in functional parameter and representing diverse blueprint of plant growth promoting attributes. Results of present findings explain the huge beneficial microbial resources from root zone of hilly crops of Himalayan region that could be effectively exploited as bio-inoculums for cold climatic condition.

Observations on the floristic list of the Mersin collected from the highplateaus of Mersin

Mersin University Research Herbarium (MERA) plants which are collected from Middle Taurus mountains are part of the Alpin-Himalayan system. In this research, the importance of these plants in terms of endemism and rare specimens is emphasized and they are compared with some of the Alpin and Himalayan plants. MERA is presented for the first time. The 5000 sheets of specimens were collected between 1996-2009. Our results indicate that Asteraceae, Lamiaceae, Fabaceae, Poaceae are common families collected from the highplateaus in Gulnar, Bozyazi, Mut, Erdemli, Camliyayla and Tarsus province of Mersin.

Patterns of morphological and genetic diversity of Valeriana jatamansi Jones in different habitats and altitudinal range of West Himalaya, India

Importance to know and understand diversity of Himalayan plants is increasingly recognized considering the fact that various natural and anthropogenic pressures might bring about serious influences to morphological and genetic diversity of the vegetation in the region. In this context, Valeriana jatamansi was investigated in detail, taking into account its importance in various Ayurvedic and modern medicines. Randomly selected mature plants from twenty five different populations (located between 1215m to 2775m asl) of V. jatamansi were analysed for their morphological attributes. Further, ISSR markers were used to detect genetic variation among 151 plants of selected 25 populations. Use of 20 primers yielded 125 reproducible polymorphic loci which were used to estimate different parameters of genetic diversity. These parameters were in turn applied to develop relationships with habitat types and altitude range. Significant variation (p<0.05) in above ground dry weight (AGDW) and below ground dry weight (BGDW) across the populations was observed. Nei's genetic diversity index (He) ranged from 0.25 to 0.37 across the populations, with a mean of 0.31. Genetic diversity exhibited a decreasing trend with increasing altitude, and maximum diversity (He=0.325) was observed in the range of 1201–1500m asl. Among the different habitat conditions, highest genetic diversity (He=0.334; Pp=84.38) was observed in grassland habitats while minimum in mixed forest habitats (He=0.285; Pp=72.433). The genetic diversity (He) had significant negative relationships with AGDW, BGDW and rhizome diameter (Pearson r=−0.359, −0.424 and −0.317, respectively; p<0.05). The genetic characterization of V. jatamansi from the western Himalaya by this study suggests influences of habitat types and the altitudinal range upon genetic diversity, and based on these proposals for conservation strategies in favour of the species are made.

Himalayan Plant Species as Pesticidal Agents

The control of diseases and insect, pests for securing world is important issue for us. A significant way to find a better treatment for crop is the search for insecticidal, pesticidal and antimicrobial constituents. From many years research in biology and biochemistry has made it possible a complete new approach for the protection of plants. As part of our Botanical pesticide research project, numerous compounds from Himalayan plant species of Uttarakhand, India with potential antifeedant activity have been identified. The plants selected for complete investigations were Boenninghausenia albiflora, Skimmia anquetillia, Glycosmis arborea, Vitex negundo, Premna barbata, Callicarpa arborea and Clerodendron indicum and the pests selected were the Plecoptera reflexa, the popular defoliator Clostera cupreta, bamboo leaf roller Crypsiptya coclesalis and polyphagous pest Spodoptera litura. The extracts and pure compounds were also tested against plant pathogens, Agrobacterium tumifaciens, Pseudomonas syringae and Pactobacterium carotovorum for antibacterial activity. In all tested plants B. albiflora and S. anquetillia showed potent insecticidal and antibacterial properties. In addition all investigated plants of Rutaceae and Verbenaceae family were found to be active against S. litura pest. Keywords: Botanical pesticides, Boenninghausenia albiflora, Callicarpa arborea, Clerodendron indicum, Glycosmis arborea, Premna barbata, Skimmia anquetillia, Vitex negundo, Mentha Piperita, Cydia Pomonella, Canavanine, Tridecanone

Highly efficient pollination by bumblebees ensures seed production inPedicularis lachnoglossa(Orobanchaceae), an early‐flowering Himalayan plant

Abstract Pedicularis(Orobranchaceae) is a common high altitude genus of the Himalayas that may be affected by pollination limitation. UsingPedicularis lachnoglossafrom Yulong (Jade Dragon) Snow Mountain in Lijiang (Yunnan Province, southwest China), we investigated the effects of high altitude habitats on the process of pollination and seed production. Floral biology, pollinator foraging behavior, breeding system, and pollination efficiency were examined using observation and exclusionary techniques.Pedicularis lachnoglossawas found to be entomophilous and exclusively pollinated byBombus friseanusandB. yunnanicola. Our results indicated that pollination limitation inP. lachnoglossawas not significant. Under open pollination, approximately 80% of flowers were successfully pollinated and developed to fruits, and about 38% of ovules developed to mature seeds. Bumblebee pollination is highly precise and efficient inP. lachnoglossa, because its flowering phenology and floral characters enhance the foraging of bumblebees on flowers. This study supports that animal pollination plays a crucial role in the outbreeding of the early floweringPedicularis. The evolution of floral specification inPedicularishas the advantages of adaptation to bumblebee pollination in adverse high altitude habitats.

Hypolipidemic effect of methanol fraction of Aconitum heterophyllum wall ex Royle and the mechanism of action in diet-induced obese rats

Aconitum heterophyllum is an endangered Himalayan plant included in “lekhaneyagana,” a pharmacological classification mentioned by Charaka in “Charakasamhita” which means reduce excess fat. The subterranean part of the plant is used for the treatment of diseases like nervous system disorders, fever, diarrhea, obesity, etc. In the present study, we are reporting the hypolipidemic effect of methanol fraction of A. heterophyllum. The methanol extract of A. heterophyllum was orally administered in diet-induced obese rats. After four weeks treatment, blood samples were collected for the estimation of serum lipids and lecithin-cholesterol acyltransferase (LCAT). Liver was collected for the assay of HMG-CoA reductase (HMGR). The fecal samples were also collected to estimate the fecal fat content. The A. heterophyllum treatment markedly lowered total cholesterol, triglycerides and apolipoprotein B concentrations in blood serum. It also showed positive effects (increase) on serum high-density lipoprotein cholesterol (HDL-c) and apolipoprotein A1 concentrations. On the other hand, A. heterophyllum treatment lowered HMGR activity, which helps to reduce endogenous cholesterol synthesis and also activated LCAT, helping increase in HDL-c. An increase in fecal fat content is also an indication of the hypolipidemic effect of A. heterophyllum. The significant hypolipidemic effect of A. heterophyllum may be linked to its ability to inhibit HMGR activity and block intestinal fat absorption. The increase in HDL-c may be linked to its ability to activate LCAT enzyme.

Dark purple nectar as a foraging signal in a bird‐pollinated Himalayan plant

• Some plants secrete coloured nectar to attract pollinators, but little is known about the chemical origins of nectar colouration and its ecological function. Leucosceptrum canum stands out as the only plant with coloured nectar recorded in the Himalayas. Here, we focused on the compound associated with the dark colour of the nectar, as well as its secretion dynamics during the flowering season and its relationship to pollinators. • Fresh nectar was analysed by semi-preparative reversed-phase high-performance liquid chromatography (HPLC), LC-MS and HRESIMS (high resolution electronspray ionization mass spectroscopy) to determine which compound causes the nectar colouration. Behavioural experiments were conducted with birds and honeybees to elucidate the effect of the nectar colour and volume on pollinators. • We identified a purple anthocyanidin, 5-hydroxyflavylium, as a natural nectar product for the first time. Two short-billed birds were found to pollinate this plant, which employs two nectar-based mechanisms to direct bird pollinators to reproductively active flowers, controlling nectar palatability and presenting a foraging signal for birds by altering nectar volume and colour in a developmental stage-specific manner. • 5-Hydroxyflavylium was found to be the cause of the nectar colouration, the function of which is to act as a foraging signal to increase pollination efficiency through nectar visibility and palatability.

EVALUATION OF ANTIOXIDANT ACTIVITY OF THE POLYOXYGENATED XANTHONES FROM SWERTIA CHIRATA BUCH., HAM

Swertia chireta is an indigenous medicinal plant which are used for variety of ailments.Moreover, several phytochemicals are present in this plants.The major bioactive compound within the plant is xanthones.This plants are the mojor source of secondary metabolites, like flavonoides and triterpenoid etc . Naturally, the plant Swertia chireta posses a multiple source of different biological componentwhich are solely responsible for different physiological activities. Naturally, bioassay of this plants were investigated with the hexane.Hexan extract of aerial parts of the Himalayan plant, Swertia chirata Buch.Ham. ex Wall.(Gentianaceae) resulted in the isolation of seven polyoxygenated xanthones, fraction Sc-1 – Sc-7. Out of seven compounds extracted from this plant , five components ( Sc - 3 - Sc-7) exhibited antioxidant activity at different magnitude ofpotency.The paper deals with these facts