T. S. Rana

Genetic diversity in inter-simple sequence repeat profiles across natural populations of Indian pomegranate (Punica granatum L.).

Pomegranate (Punica granatum L.), in the monogeneric family Punicaceae, is found in Iran, Afghanistan, India and Mediterranean countries. Iran is considered to be its primary centre of origin. In India, pomegranate occurs naturally only in the Western Himalayan regions of Jammu and Kashmir, Himachal Pradesh and Uttarakhand States. However, there is no information about genetic variation in wild pomegranate at population level. In this paper, we describe genetic diversity across natural populations of Indian pomegranate based on inter-simple sequence repeat (ISSR) markers. Forty-nine accessions representing eight populations from two regions were analysed using ISSR. Seventeen ISSR primers resulted in 268 polymorphic bands, with 87.01% polymorphism throughout the accessions. Pair-wise population genetic distances ranged from 0.05 to 0.45, with a mean of 0.25 between populations. amova and Neis genetic diversity analyses revealed higher genetic variation within populations than among populations. A higher genetic differentiation (G(ST)) was observed between the spatially distant populations, indicating a low level of genetic exchange (Nm) among these populations. However, clustering of populations was not in accordance with their geographical affiliations in the tree. The results indicate that the ISSR method is sufficiently informative and powerful to assess genetic variability in pomegranate, and that patterns of genetic variability observed among populations of wild pomegranate from the Western Himalaya differ. Estimation of genetic variation reported here provides a significant insight for in situ conservation and exploitation of genetic resources for this economically important species as potential breeding material.

SPAR profiles and genetic diversity amongst pomegranate (Punica granatum L.) genotypes.

We are interested in studying the distribution and range of diversity amongst the pomegranates in India. Single Primer Amplification Reaction (SPAR) profiling using Random Amplified Polymorphic DNA (RAPD) and Directed Amplification of Minisatellite DNA (DAMD) methods enabled the determination of the genetic diversity amongst a total of 64 Indian pomegranate genotypes including 15 wild, 34 semi-wild and 14 cultivated types. SPAR profile data were scored for the computation of pairwise distances as well as a Neighbour Joining (NJ) tree of all the genotypes. Eight RAPD and four DAMD primers showed discrete polymorphic patterns amongst these genotypes. From the profiles obtained with all the 12 primers considered together, 259 bands were scored. The NJ tree generated after a 1000 bootstrap test using Jaccard coefficient showed separation of Lagerstroemia speciosa used as the out-group taxon, while the pomegranate genotypes were resolved into distinct genetic lineages such that all the cultivated (except CBd70), and wild genotypes (except W101) clearly separated from other genotypes in distinct sub clusters while the semi-wild genotypes were resolved into three sub-clusters. The greatest and least distances detected between genotypes were 0.94 and 0.12, 0.97 and 0.24 and 0.95 and 0.38, amongst the cultivated, semi-wild and the wild genotypes respectively. The results indicate the high levels of genetic diversity present amongst the genotypes. Significantly, the wild genotypes also have a reasonably good range of diversity. A good germplasm collection, especially including the wild genotypes will enable a better pomegranate improvement program. Both SPAR methods, RAPD and DAMD, are found to be useful for studying the genetic diversity of pomegranate.

Seed protein electrophoresis of some cultivated and wild species of Chenopodium

Seed protein profiles of 40 cultivated and wild taxa of Chenopodium have been compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The relative similarity between various taxa, estimated by Jaccard’s similarity index and clustered in UPGMA dendrogram, is generally in accordance with taxonomic position, crossability relationships and other biochemical characters. Eight accessions of C. quinoa studied are clustered together and show genetic similarity with closely related C. bushianum and C. berlandieri subsp. nuttalliae. The taxa included under C. album complex are clustered in two groups which show that these taxa are a heterogenous assemblage and their taxonomic affinities need a reassessment. Other wild species studied are placed in the dendrogram more or less according to their taxonomic position.

Molecular differentiation of Chenopodium album complex and some related species using ISSR profiles and ITS sequences.

The present study was undertaken to understand the genetic differentiation and relationships in various components of C. album complex, C. giganteum and some related species using inter simple sequence repeats (ISSR) profiles and internal transcribed spacer (ITS) sequences. The relationships based on UPGMA dendrograms have shown the heterogenous nature of C. album complex. The 2x taxa while showing close relation among themselves are sharply segregated from 4x and 6x taxa belonging to C. album and C. giganteum. Among the three cytotypes from North Indian plains the 4x shows greater similarity to 6x than to 2x which is corroborated by the karyotypic studies. Furthermore, the 6x C. album and C. giganteum accessions of American and European origin are clearly segregated from those of Indian origin which may show their separate origin. Other related species show relationships according to their taxonomic position. The present study based on ISSR profiles and ITS sequences has therefore been very useful in explaining the relationships between various components of C. album complex and related species. However, more work needs to be done using different CpDNA loci to define correct species boundary of the taxa under C. album complex from Himalayas and North Indian Plains.

Genetic Relationships Among Wild and Cultivated Accessions of Curry Leaf Plant ( Murraya koenigii (L.) Spreng.), as Revealed by DNA Fingerprinting Methods

Murraya koenigii (L.) Spreng. (Rutaceae), is an aromatic plant and much valued for its flavor, nutritive and medicinal properties. In this study, three DNA fingerprinting methods viz., random amplification of polymorphic DNA (RAPD), directed amplification of minisatellite DNA (DAMD), and inter-simple sequence repeat (ISSR), were used to unravel the genetic variability and relationships across 92 wild and cultivated M. koenigii accessions. A total of 310, 102, and 184, DNA fragments were amplified using 20 RAPD, 5 DAMD, and 13 ISSR primers, revealing 95.80, 96.07, and 96.73% polymorphism, respectively, across all accessions. The average polymorphic information content value obtained with RAPD, DAMD, and ISSR markers was 0.244, 0.250, and 0.281, respectively. The UPGMA tree, based on Jaccard’s similarity coefficient generated from the cumulative (RAPD, DAMD, and ISSR) band data showed two distinct clusters, clearly separating wild and cultivated accessions in the dendrogram. Percentage polymorphism, gene diversity (H), and Shannon information index (I) estimates were higher in cultivated accessions compared to wild accessions. The overall high level of polymorphism and varied range of genetic distances revealed a wide genetic base in M. koenigii accessions. The study suggests that RAPD, DAMD, and ISSR markers are highly useful to unravel the genetic variability in wild and cultivated accessions of M. koenigii.

Isolation, Characterization and Anticancer Potential of Cytotoxic Triterpenes from Betula utilis Bark.

Betula utilis, also known as Himalayan silver birch has been used as a traditional medicine for many health ailments like inflammatation, HIV, renal and bladder disorders as well as many cancers from ages. Here, we performed bio-guided fractionation of Betula utilis Bark (BUB), in which it was extracted in methanol and fractionated with hexane, ethyl acetate, chloroform, n-butanol and water. All six fractions were evaluated for their in-vitro anticancer activity in nine different cancer cell lines and ethyl acetate fraction was found to be one of the most potent fractions in terms of inducing cytotoxic activity against various cancer cell lines. By utilizing column chromatography, six triterpenes namely betulin, betulinic acid, lupeol, ursolic acid (UA), oleanolic acid and β-amyrin have been isolated from the ethyl acetate extract of BUB and structures of these compounds were unraveled by spectroscopic methods. β-amyrin and UA were isolated for the first time from Betula utilis. Isolated triterpenes were tested for in-vitro cytotoxic activity against six different cancer cell lines where UA was found to be selective for breast cancer cells over non-tumorigenic breast epithelial cells (MCF 10A). Tumor cell selective apoptotic action of UA was mainly attributed due to the activation of extrinsic apoptosis pathway via up regulation of DR4, DR5 and PARP cleavage in MCF-7 cells over non-tumorigenic MCF-10A cells. Moreover, UA mediated intracellular ROS generation and mitochondrial membrane potential disruption also play a key role for its anti cancer effect. UA also inhibits breast cancer migration. Altogether, we discovered novel source of UA having potent tumor cell specific cytotoxic property, indicating its therapeutic potential against breast cancer.

Composition and in vitro cytotoxic activities of essential oil of Hedychium spicatum from different geographical regions of western Himalaya by principal components analysis.

The rhizome of Hedychium spicatum has been widely used in traditional medicines. The present study deals with the evaluation of the cytotoxic potential of rhizome essential oils from four different regions of the Western Himalaya (India) along with comparative correlation analysis to characterise the bioactive cytotoxic component. The essential oils were coded as MHS-1, MHS-2, MHS-3 and MHS-4, and characterised using GC-FID and GC–MS. The main volatile compounds identified were 1,8-cineol, eudesmol, cubenol, spathulenol and α-cadinol. In vitro cytotoxic activities were assessed against human cancer cell lines such as, the lung (A549), colon (DLD-1, SW 620), breast (MCF-7, MDA-MB-231), head and neck (FaDu), and cervix (HeLa). MHS-4 is significantly active in comparison to other samples against all cancer cell lines. Sample MHS-4 has major proportion of monoterpene alcohol mainly 1,8-cineol. Principal components analysis was performed for the experimental results and all four samples were clustered according to their percentage inhibition at different doses.

Chemical Constituents and Antimicrobial Potential of Essential Oil from Betula utilis Growing in High Altitude of Himalaya (India)

Abstract Compounds having antimicrobial activity obtained from plant sources are useful because they are new sources against antibiotic resistance. The present study is based on the evaluation of antimicrobial activity of the essential oil of the fresh bark of Betula utilis and to identify the chemical constituents responsible for antimicrobial potential, grown in high altitude of Himalaya, India. The chemical constituents of essential oil were identified by gas chromatography-mass spectrometry (GC-MS). The essential oil was evaluated for the antimicrobial potential against human pathogenic bacteria and fungi by using a micro-dilution assay. GC-MS analysis revealed that the major constituents of the essential oil were geranic acid (11.38 %), β-seleneol (10.98 %), β-linalool (10.91 %), terragon (10.61 %), β-sesquiphellendrene (8.02 %), champacol (6.33 %) and 1,8-cineol (5.49 %). Result showed that B. utilis essential oil has a strong antimicrobial activity against the fungus Candida albicans and Gram (+) and Gram (-) human pathogenic bacteria (MIC: 60.5-240 µg/ml).

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

Abstract 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.

ETHNOBOTANICAL OBSERVATION AMONG JAUNSARIS OF JAUNSAR-BAWAR, DEHRA DUN(U.P.), INDIA

AbstractThis paper highlights the ethnobotanical uses of some plants by the Jaunsari tribes of Chakrata Tehsil in the Dehra Dun district. The Jaunsari tribe of Jaunsar-Bawar is a primitive group of people who have their own culture, custom and traditions. The region is quite rich in plant wealth. Living close to nature the tribals use a number of wild plants such as Aesculus indica, Asparagus filicinus, Berberis chitria, Bidens biternata, Cannabis sativa, Cinnamomum tamala, Cissampelos pareira, Crotalaria spectabilis, Cryptolepis buchananii, Datura stramonium, Oxalis corniculata, Rumex nepalensis, Solanum ferox, Thalictrum foliolosum, Viola canescens, Zanthoxylum armatum etc., to fill their day to day requirements. Local names, -habits and flowering periods are provided for each species. The family names (in parentheses) are followed by detailed uses. Finally the authors emphasize the urgent need to document and develop databases on the ethnobotanical uses of the areas plants so that the local inhabitant...