Yashbir S. Bedi

A cross-cultural analysis of Jammu, Kashmir and Ladakh (India) medicinal plant use.

ETHNOPHARMACOLOGICAL RELEVANCE Jammu & Kashmir (J&K) is a predominantly Himalayan state in the north-western part of India. It has three geographically distinct divisions viz., Jammu, Kashmir and Ladakh, which are immensely rich in their biological and cultural diversity. Medicinal plants are an important element of indigenous medical system of the region. The main goal of the present article is to examine the use of ethnomedicinal plants in three divisions of J&K and to discuss cross-cultural consensus on the use of medicinal plants in these divisions. The article also discusses the gaps in the current state of knowledge on ethnomedicinal plants of the region and gives recommendations for the future studies. MATERIALS AND METHODS Scientific literature on ethnomedicinal field studies conducted in J&K state of India available in the journals, edited books and other scientific databases viz., CAB international, DOAJ, Google Scholar, PubMed, Science direct, SciFinder, Scopus and Web of Science were searched. Only field based ethnomedicinal surveys from last four decades up to December 2013 reporting first hand information on the medicinal plants used to treat human health related ailments by indigenous communities of J&K were included in this study. Venn diagram was used to analyze the cross-cultural consensus on the use of ethnomedicinal plants in the three divisions of J&K. RESULTS A total of 948 plant taxa (923 angiosperms, 12 gymnosperms and 13 pteridophytes) belonging to 129 families, 509 genera, 937 species and 11 varieties have so far been reported to have a traditional medicinal use by indigenous communities of J&K. Asteraceae (60 genera, 132 spp.) was the most frequently used family followed by Fabaceae (32 genera, 50 spp.) and Lamiaceae (27 genera, 55 spp.). 514, 415 and 397 medicinal plants were used in Jammu, Kashmir and Ladakh divisions, respectively. Sixty eight plant taxa were used in all the three divisions, whereas 95 plants were common between Ladakh and Jammu, 127 plants between Ladakh and Kashmir, and 216 plants between Jammu and Kashmir. Maximum numbers of plant taxa were used for treating dermatological problems (321), followed by cold, cough and throat related ailments (250), fever (224), joint and muscle related ailments (215), gastrointestinal disorders (210), urogenital ailments (199), respiratory ailments (151), body pain (135) and gynecological disorders (127). CONCLUSIONS This is the first study from the J&K state, which has examined the medicinal plant use in three divisions of J&K and discussed the promising medicinal plant species with cross-cultural consensus. The analysis of the data suggested that while large numbers of plants are used medicinally in each division, there is a low interregional consensus and high variation between medicinal plants used in these divisions, which is due to both cultural divergence as well as biological distinctness. The issues related to current status of knowledge on medicinal plants used by indigenous communities of J&K have been discussed and some recommendations have been made for future studies on medicinal plants in J&K region.

Changing trends in biotechnology of secondary metabolism in medicinal and aromatic plants

AbstractMain conclusionMedicinal and aromatic plants are known to produce secondary metabolites that find uses as flavoring agents, fragrances, insecticides, dyes and drugs. Biotechnology offers several choices through which secondary metabolism in medicinal plants can be altered in innovative ways, to overproduce phytochemicals of interest, to reduce the content of toxic compounds or even to produce novel chemicals. Detailed investigation of chromatin organization and microRNAs affecting biosynthesis of secondary metabolites as well as exploring cryptic biosynthetic clusters and synthetic biology options, may provide additional ways to harness this resource. Plant secondary metabolites are a fascinating class of phytochemicals exhibiting immense chemical diversity. Considerable enigma regarding their natural biological functions and the vast array of pharmacological activities, amongst other uses, make secondary metabolites interesting and important candidates for research. Here, we present an update on changing trends in the biotechnological approaches that are used to understand and exploit the secondary metabolism in medicinal and aromatic plants. Bioprocessing in the form of suspension culture, organ culture or transformed hairy roots has been successful in scaling up secondary metabolite production in many cases. Pathway elucidation and metabolic engineering have been useful to get enhanced yield of the metabolite of interest; or, for producing novel metabolites. Heterologous expression of putative plant secondary metabolite biosynthesis genes in a microbe is useful to validate their functions, and in some cases, also, to produce plant metabolites in microbes. Endophytes, the microbes that normally colonize plant tissues, may also produce the phytochemicals produced by the host plant. The review also provides perspectives on future research in the field.

Molecular cloning and functional characterization of an antifungal PR-5 protein from Ocimum basilicum

Pathogenesis-related (PR) proteins are involved in biotic and abiotic stress responses of plants and are grouped into 17 families (PR-1 to PR-17). PR-5 family includes proteins related to thaumatin and osmotin, with several members possessing antimicrobial properties. In this study, a PR-5 gene showing a high degree of homology with osmotin-like protein was isolated from sweet basil (Ocimum basilicum L.). A complete open reading frame consisting of 675 nucleotides, coding for a precursor protein, was obtained by PCR amplification. Based on sequence comparisons with tobacco osmotin and other osmotin-like proteins (OLPs), this protein was named ObOLP. The predicted mature protein is 225 amino acids in length and contains 16 cysteine residues that may potentially form eight disulfide bonds, a signature common to most PR-5 proteins. Among the various abiotic stress treatments tested, including high salt, mechanical wounding and exogenous phytohormone/elicitor treatments; methyl jasmonate (MeJA) and mechanical wounding significantly induced the expression of ObOLP gene. The coding sequence of ObOLP was cloned and expressed in a bacterial host resulting in a 25kDa recombinant-HIS tagged protein, displaying antifungal activity. The ObOLP protein sequence appears to contain an N-terminal signal peptide with signatures of secretory pathway. Further, our experimental data shows that ObOLP expression is regulated transcriptionally and in silico analysis suggests that it may be post-transcriptionally and post-translationally regulated through microRNAs and post-translational protein modifications, respectively. This study appears to be the first report of isolation and characterization of osmotin-like protein gene from O. basilicum.

Analysis of SSR dynamics in chloroplast genomes of Brassicaceae family.

Simple sequence repeats (SSRs) are present abundantly in most eukaryotic genomes. They affect several cellular processes like chromatin organization, regulation of gene activity, DNA repair, DNA recombination, etc. Though considerable data exists on using nuclear SSRs to infer phylogenetic relationships, the potential of chloroplast microsatellites (cpSSR), in this regard, remains largely unexplored. In the present study we probe various nucleotide repeat motifs (NRMs) / types of SSRs present in chloroplast genomes (cpDNA) of 12 species belonging to Brassicaceae family. NRMs show a non-random distribution in coding and non-coding compartments of cpDNA. As expected, trinucleotide repeats are more common in coding regions while other repeat motifs are prominent in non-coding DNA. Total numbers of SSRs in coding region show little variation between species while considerable variation is exhibited by SSRs in non-coding regions. Finally, we have designed universal primers that yield polymorphic amplicons from all 12 species. Our analysis also suggests that amplicon length polymorphism shows no significant relationship with sequence based phylogeny of SSRs in cpDNA of Brassicaceae family.

Differential response of terpenes and anthraquinones derivatives in Rumex dentatus and Lavandula officinalis to harsh winters across north-western Himalaya

Herbs adapted to diverse climates exhibit distinct variability to fluctuating temperatures and demonstrate various metabolic and physiological adaptations to harsh environments. In this research, Rumex dentatus L. and Lavandula officinalis L. were collected before snowfall in September–November to evaluate variability in major phytoconstituents to diverse seasonal regime. LC-MS was used for simultaneous determination of eight anthraquinone derivatives in R. dentatus, i.e. emodin, physcion, chrysophanol, physcion glucoside, endocrocin, emodin glucoside, chrysophanol glucoside and chromone derivatives and monoterpenes in L. officinalis i.e. (Z)-β-ocimene, (E)-β-ocimene, terpene alcohol, terpin-4-ol, acetate ester-linalyl acetate and bicyclic sesquiterpene (E)-caryophyllene. The correlation analysis confirmed significant variation in anthraquinone glucoside and terpene content within Rumex and Lavender, respectively, and altitude was established as the determinant factor in secondary metabolism of both herbs. The study concludes the propagation of herbs in bioclimatic belts which favour accumulation of major constituents and validate their greater pharmacological activity.

Cloning and expression analysis of chalcone synthase gene from Coleus forskohlii

Flavonoids are an important class of secondary metabolites that play various roles in plants such as mediating defense, floral pigmentation and plant–microbe interaction. Flavonoids are also known to possess antioxidant and antimicrobial activities. Coleus forskohlii (Willd.) Briq. (Lamiaceae) is an important medicinal herb with a diverse metabolic profile, including production of a flavonoid, genkwanin. However, components of the flavonoid pathway have not yet been studied in this plant. Chalcone synthase (CHS) catalyses the first committed step of flavonoid biosynthetic pathway. Full-length cDNA, showing homology with plant CHS gene was isolated from leaves of C. forskohlii and named CfCHS (GenBank accession no. KF643243). Theoretical translation of CfCHS nucleotide sequence shows that it encodes a protein of 391 amino acids with a molecular weight of 42.75 kDa and pI 6.57. Expression analysis of CfCHS in different tissues and elicitor treatments showed that methyl jasmonate (MeJA) strongly induced its expression. Total flavonoids content and antioxidant activity of C. forskohlii also got enhanced in response to MeJA, which correlated with increased CfCHS expression. Induction of CfCHS by MeJA suggest its involvement in production of flavonoids, providing protection from microbes during herbivory or mechanical wounding. Further, our in silico predictions and experimental data suggested that CfCHS may be posttranscriptionally regulated by miR34.

Mannitol Stress Directs Flavonoid Metabolism toward Synthesis of Flavones via Differential Regulation of Two Cytochrome P450 Monooxygenases in Coleus forskohlii.

Cytochrome P450 monooxygenases (CYP450s) are known to play important roles in biosynthesis of all secondary metabolites, including flavonoids. Despite this, few CYP450s have been functionally characterized in model plants and roles of fewer CYP450s are known in non-model, medicinal, and aromatic plants. Our study in Coleus forskohlii indicates that flavone synthase (CYP93B) and flavonoid 3′ monooxygenase (CYP706C) are key enzymes positioned at a metabolic junction, to execute the biosynthesis of different sub-classes of flavonoids (flavones, flavonol, anthocynanin, isoflavones etc.) from a common precursor. Such branch points are favored targets for artificially modulating the metabolic flux toward specific metabolites, through genetic manipulation or use of elicitors that differentially impact the expression of branch point genes. Genkwanin, the only flavone reported from C. forskohlii, is known to possess anti-inflammatory activity. It is biosynthesized from the general flavonoid precursor: naringenin. Two differentially expressed cytochrome P450 genes (CfCYP93B, CfCYP706C), exhibiting maximum expression in leaf tissues, were isolated from C. forskohlii. Mannitol treatment resulted in increased expression of CfCYP93B and decrease in expression of CfCYP706C. Metabolite quantification data showed that genkwanin content increased and anthocyanin levels decreased in response to mannitol treatment. Alignment, phylogenetic analysis, modeling, and molecular docking analysis of protein sequences suggested that CfCYP93B may be involved in conversion of naringenin to flavones (possibly genkwanin via apigenin), while CfCYP706C may act on common precursors of flavonoid metabolism and channel the substrate toward production of flavonols or anthocynanins. Decrease in expression of CfCYP706C and increase in accumulation of genkwanin suggested that mannitol treatment may possibly lead to accumulation of genkwanin via suppression of a competitive branch of flavonoids in C. forskohlii.

RP-thin layer chromatographic method for the quantification of three gingerol homologs of ultrasonic-assisted fresh rhizome extracts of Zingiber officinale collected from North Western Himalayas.

A rapid and sensitive RP high-performance thin-layer chromatographic (RP-HPTLC) methodology was developed and validated for the quantitative estimation of gingerols in methanolic extract of fresh ginger rhizome. The samples were chromatographed on RP-TLC glass plates pre-coated with RP-18 60F(254) as the stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with ternary-solvent system consisting of acetonitrile-water-formic acid (7:2:1 v/v/v) at room temperature (25+/-2 degrees C) and plates were scanned at 500 nm. The system was found to give compact spots for gingerols (R(f) values of 6-gingerol 0.73+/-0.04, 8-gingerol 0.59+/-0.08 and 10-gingerol 0.36+/-0.05). Linearity was found to be in the range of 140-840 ng/spot for 6-gingerol, 168-1008 ng/spot for 8-gingerol and 136-816 ng/spot for 10-gingerol with significantly high value of correlation coefficient. The linear regression analysis data for the calibration plots showed linear relationship (r(2)) and ranged from 0.9992 to 0.9937 for 6-, 8- and 10-gingerol. The method was used for routine analyses and to obtain relative amounts of the gingerols in the fresh rhizomes of the ginger cultivated in different locations of Uttarakhand and Himachal Pradesh of North Western Himalayas (India).

Microclimatic variation in UV perception and related disparity in tropane and quinolizidine alkaloid composition of Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger.

The aim of current research was to evaluate the physiological adjustment in three medicinal herbs viz., Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger to the winter period characterised by intense UV flux in Kashmir valley across the North Western Himalaya. Quinolizidine (QA) and tropane alkaloid (TA) concentrations were analysed in these herbs thriving at two different altitudes via GC-MS and correlated by PCA analysis. This study investigated the hypothesis that UV reflectance and absorbance at low temperatures are directly related to disparity in alkaloid accumulation. Among QAs in L. polyphyllus, ammodendrine and lupanine accumulated at higher concentration and exhibited significant variation of 186.36% and 95.91% in ammodendrine and lupanine respectively in both sites. Tetrahydrohombifoline displayed non-significant variation of about 9.60% irrespective of sites. Among tropane alkaloid (TA), hyoscyamine was recorded as the most abundant constituent irrespective of the plant and site while apotropine accumulated in lesser quantity in A. acuminata than H. niger. However, apotropine demonstrated significant variation of 175% among both sites. The final concentration of quinolizidine (QA) and tropane alkaloid (TA) reflects the interplay between reflectance and absorbance of UV radiation response field. These findings suggest that spectral response of UV light contributes directly to alkaloid biosynthesis.

Ethnobotany, Traditional Knowledge, and Diversity of Wild Edible Plants and Fungi: A Case Study in the Bandipora District of Kashmir Himalaya, India

ABSTRACT This study explored the diversity of traditional knowledge, collected data on utilization, and identified and analyzed the wild edible plants and fungi of Kashmir Himalaya (India). Information was collected by interviews with 113 individuals during 2012–2014 from nine rural and mountainous areas. Data were classified as cooked vegetables, salads, spices, chutneys, herbal teas, home-made alcoholic drinks, soups, raw fruits, and underground snacks. Information was analyzed using use reports (UR), use values (UV), informant consensus (Fic), fidelity level (FL), and cultural importance index (CI) metrics. In total, 111 phytotaxa, distributed into 87 genera and 43 families, were identified as local edible resources. Overall, 94.25% of angiosperms, 3.45% gymnosperms, and 2.30% of cryptogams were documented as being used traditionally. Several taxa were highly cited (Amaranthus caudatus, Angelica archangelica, Asparagus racemosus, Berberis lycium, Fragaria vesca, Hippophae rhamnoides, Oxyria digyna, Juglans regia). Frequently used parts were young leaves (19.82%), fruits (15.32%), young twigs (9.01%), roots (8.11%), and tubers (6.31%). The most usage was mentioned for leafy vegetables (27.93%), followed by raw fruits (22.52%), herbal teas (14.41%), salads (9.91%), alcoholic drinks (6.31%), and snacks (4.50%). The high fidelity scores and informant consensus factors suggest that ethnobotanical knowledge could potentially guide the search for developing new nutraceutical products in the future.