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Kaempferia galanga Linn. is an endangered rhizomatous medicinal plant belonging to the Zingiberaceae family. It has evolved as an emerging industrial crop and dominates pharma as well as aroma sector. Though the extracts of this species have been extensively used in herbal medicine across the globe for the treatment of numerous diseases, but still the composition of the extract is not characterized properly. Thus, methanol extracts of K. galanga leaves and rhizomes were subjected to phytochemical screening, total phenolic content (TPC), total flavonoid content (TFC), and Gas Chromatography-Mass Spectrometry (GC-MS) to analyze the phytoconstituents. Leaf extract contained more TPC and TFC values as compared to rhizome extract. A total of eight and ten compounds were identified in the leaf and rhizome extract accounting for 61.44% and 96.97% of the total peak area respectively. Ethyl p-methoxycinnamate was found as the main constituent in rhizome extract covering 80.39% of the total area. Other important compounds like ethyl cinnamate (9.61±0.45%), pentadecane (3.12±0.2%) were also found in the rhizome extract, whereas leaf extract contained 2-(3,4-dimethoxyphenyl)-7-hydroxy-3-methoxy-4H-chromen-4-one (18.26%), 2-(3-hydroxy-4- methoxyphenyl)-3,7-dimethoxy-4H-chromen-4-one (14.01%), octamethylcyclotetrasiloxane (11.79%). The study indicated that K. galanga is a good source of phytoconstituents which can be used at the industrial level to produce pharmaceuticals, perfumes and flavouring agents.

Andrographis paniculata is an Indian medicinal plant with tremendous therapeutic values due to the presence of active diterpenoids in its aerial parts. However, high domestic and export demand has led to overexploitation of wild populations of this species. With a view to bringing A. paniculata into cultivation and to reduce the pressure on wild populations, the present study was undertaken to identify elite germplasm from different locations of eastern India by analysing intraspecific variation in the content of four major active diterpenoids. A total of 166 wild accessions of A. paniculata analysed through high-performance liquid chromatography (HPLC) revealed remarkable variation in the sum of four active diterpenoids in the aerial parts, ranging from 0.41 to 8.55% on a dry weight basis. Three elite accessions (AP-6, AP-8, AP-46) having respectively 8.02, 8.36 & 8.55% of the sum of four major active diterpenoids were identified. These germplasm could be used for commercial cultivation and genetic improvement of A. paniculata.

In the present study, the extracted volatiles from dried leaf and fruit of Piper longum were analysed by gas chromatography-mass spectrometry (GC-MS) and each detected 53 constituents having 92.41% and 96.31% of the total volatiles respectively. E-nerolidol (19.56%), β-pinene (17.07%) and α-pinene (6.8%) were main constituents in leaf volatiles whereas the fruit volatiles dominated by germacrene-D (23.38%), 8-heptadecene (8.95%) and β-caryophyllene (8.20%). Antioxidant potential of the volatiles were assessed by DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) assays. The fruit volatiles revealed higher radical scavenging activities as compared to leaf. The samples were also evaluated against multidrug resistant (MDR) isolates including one non MDR fungal strain. The fruit volatiles showed a very strong activity against Acinetobacter baumannii than others whereas leaf volatiles possessed strong activity against Klebsiella pneumoniae as compared to other strains. Thus, the dried fruits can be exploited for drug development towards therapeutic purpose.

Turmeric is an important medicinal plant possessing valuable secondary metabolites. The present study was conducted for evaluating the effect of environmental parameters on quality of essential oil from leaf and rhizome of ‘Lakadong’ turmeric cultivar. Five different agroclimatic regions of Odisha were selected for this study. The GC–MS analysis revealed significant differences in the abundance of constituents by means of area percentage as well as in essential oil yield. The area percentage of alpha phellandrene in leaf varied from 31.78 to 39.89%, while the ar-turmerone content in rhizome varied from 36.18 to 44.36% across the agroclimatic regions. However, no study has been done about the sensitive environmental factors responsible for these variations with respect to location. Thus, in the present study, relative sensitivity analysis has been carried out which revealed that temperature, humidity and altitude are most important factors for enhancing the quality and yield of leaf and rhizome essential oil. The present investigation will help in selection of suitable areas of cultivation for increasing the essential oil yield with quality in Lakadong cultivar.

Alpinia galanga is commonly known as ‘greater galangal’ is extensively present across India and Southeast Asia. The essential oil of this zingiberaceae species has high demand in the global market ...

Protection and utilization of plant genomic resources are greatly perceived these days. The possibility of their dwindling genetic diversity along with the augmented demands for the resources has necessitated immediate concern for their conservation and sustainable plant utilization. The genus Curcuma (family Zingiberaceae) is well known for its valuable multipurpose products for medicine, spices, food, fragrance, flavoring agents, condiments as well as ornamentals. Because of the resurgence of curiosity in the commercial development and conservation of Curcuma, it has become essential to precisely characterize the existing genetic diversity at the species level. In the present study, 109 EST-SSR markers showed cross-transferability among 9 Curcuma species, among which 33 showed polymorphism. These 33 polymorphic EST-SSRs were utilized to assess genetic resemblance and distance among Curcuma species. The results of cluster analysis and principal coordinate analysis highly correspond to each other. AMOVA showed higher genetic variation among Curcuma species. The population genetic study revealed the genetic differentiation of species. The mean values of Nei's gene diversity (H), Shannon's information index (I), polymorphic information content (PIC), percentage of polymorphic bands (PPB), and resolving Power (RP) were found to be 0.3314, 0.49, 0.66, 88.57% and 7.24, respectively. Values of Nei's gene diversity in C. longa indicated the need for its large-scale cultivation and conservation prioritization of wild taxa of C. caesia, C. aromatica and C. amada. These SSR markers could also be utilized to identify morphologically similar Curcuma species.

Curcuma zedoaria (Christm.) Roscoe (white turmeric) is a perennial herbaceous plant of family Zingiberaceae and mainly found in the wild areas of tropical and subtropical regions worldwide. The cytochrome proteins in plants play important roles in promoting their growth and development, as well as protecting them from stresses and diseases. Cytochrome proteins like psbF, psbE, petB, petD, petN, petG, and ccsA play important roles in degradation of mis-folded proteins, ATP formation, cyclic electron flow and biogenesis of c-type cytochrome of C. zedoaria. However, due to lack of structural availability of these C. zedoaria cytochrome proteins in structural databases, the physiochemical parameters of sequences were estimated using Expasy ProtParam web tool. Self-Optimized Prediction Method with Alignment (SOPMA) server and MODELLER version 9.23 were used for modelling along with Qualitative Model Energy Analysis (QMEAN) and Protein Structure Analysis (ProSA) servers were implemented for validating the secondary and tertiary structures of these proteins. The obtained QMEAN4 values of the modelled cytochrome proteins were -2.04, -1.20, -3.01, -1.57, -2.11, -1.74 and -12.87. The Z-scores obtained from ProSA server were 0.5, -0.83, -1.5, -0.58, -0.02, 0.14 and -3.73. All seven modelled structures have been submitted to protein model database (PMDB). The derived results will be helpful in further investigations towards determining the crystal structure of the hypothetical proteins, structural motifs, physiochemical properties, and also protein-protein interaction studies of various cytochrome proteins.

Rhizome essential oil of Curcuma caesia has myriad of therapeutic activities which is being practiced since ages as folklore remedy but commercially remained underutilized till date. Though previous reports had characterized the phytoconstituents of C. caesia essential oil, a comparative assessment will lead to identify a good germplasm for commercialization. Quality germplasm should have high yield and quality constituents. Hence, the current study deals with intraspecific variability of C. caesia rhizome essential oil associated with two geographical regions, i.e., Odisha and West Bengal. The Principal Component Analysis and Agglomerative Hierarchical Clustering analysis distinguished two groups of essential oils on the basis of the phytoconstituents. The highest oil yield was found in Mayurbhanj (0.64 ± 0.015% w/w) population, whereas the lowest oil yield was in South 24-Parganas population (0.39 ± 0.01% w/w). The chemical constituents of C. caesia were analyzed by Gas Chromatography–Mass Spectrometry which detected a total of 34 compounds accounting to 90.08–97.94% of the total oil. Oxygenated monoterpenes constituted the major fraction followed by oxygenated sesquiterpenes. West Bengal accessions were dominated by oxygenated monoterpenes (34.8–45.41%) followed by oxygenated sesquiterpenes (26.63–38.06%). However, Odisha accessions were rich in oxygenated monoterpenes (37.6–54.36%) followed by sesquiterpene hydrocarbons (21.9–33.8%). In this study, phytochemical variability in C. caesia associated with geographical distribution patterns has been successfully studied in respect of essential oil yield and quality.

Eclipta prostrata (Linn.) (Family: Asteraceae) is an important Indian medicinal herb having myriad therapeutic potential. In the present study, the essential oil was extracted from the aerial parts...

Zingerone (4-(4-Hydroxy-3-methoxyphenyl)-2-butanone) is one of the non-volatile and nontoxic compounds of ginger. It is also called vanillylacetone with a crystalline solid form which is sparingly soluble in water and more soluble in ether. The contribution of this compound in ginger is about 9.25%. The chemical structure is made of a phenolic ring with methoxy group attached to benzene ring. Gingerol can be heated to form zingerone by retroaldol reaction. It has been reported that zingerone has multiple pharmacological activities. It is effective against diarrhoea causing enterotoxigenic bacteria that leads to infant death. It is also used against intestinal gastric, oxidative stress, weak immunity, obesity. During its activity against cancer, it governs the expression of different cell cycle protein and TGF-?1 expression. Antioxidant response is controlled by inducing the activity of ROS neutralising enzymes like superoxide dismutase, catalase and glutathione reductase. It can also reduce various inflammations by restricting the activity of interleukins. This review summarizes the multiple pharmacology activities of zingerone against various important diseases like cancers, tumors, inflammations, oxidative conditions, microbial infections, biofilm formations, thrombosis and other diseases. In addition, the molecular regulation of these pharmacological responses by zingerone is also critically discussed.

The essential oil of

Turmeric contains a yellow color pigment that is curcumin, based on which different cultivars of turmeric have been released so far. Limited molecular studies on different pigment synthesis genes and lack of cultivar specific SSRs and SNPs in turmeric have confined the scope of identification of turmeric and establishment of their genetic makeup. In this study de novo transcriptome assembly of four turmeric cultivars i.e. Suroma, Roma, Lakadong and Megha was done. From the transcriptome analysis, cultivar specific SSRs, SNPs and transcripts encoding pigment synthesis genes were derived. A total of 113,862 contigs for Suroma, 125,762 for Roma, 113,021 for Lakadong and 116,711 for Megha were obtained. Multiple databases were used to acquire a complete annotation and the transcripts were functionally categorized using GO, KOG, KEGG and PlantCyc database. While presenting a large repertoire of cultivar specific quality SNPs and variations in SSRs, we have also demonstrated the functional basis of differentially expressed pigment synthesis genes among the four cultivars. The results of this study have clearly indicated their different genetic makeup. This report would enable authentic identification of these morphologically similar elite cultivars through use of cultivar specific SSRs and SNPs and to understand the molecular mechanisms behind differential expression of specific pigment synthesis genes in turmeric while providing scope for overall improvement of the taxa.

Hedychium coronarium (Zingiberaceae) is a perennial rhizomatous herb widely used for its aromatic, cosmetic and medicinal value. In the present research, the morphological, phytochemical and molecular diversity was assessed for 50 H. coronarium accessions collected from different states of Eastern India. The accessions were morphologically diverse with leaf number, leaf length, leaf width, rhizome diameter and rhizome dry weight characters showing the highest variation. A total of 44 constituents were identified in the essential oils with β-pinene (11.87–35.16%), eucalyptol (13.12–39.82%), linalool (1.58–41.33%) and coronarin-E (1.14–33.87%) as the predominant constituents. Using 10 Inter-simple sequence repeats (ISSR) and 5 Random amplified polymorphic DNA (RAPD) primers, a total of 71 and 34 bands were amplified, respectively. The average polymorphism, polymorphic information content, marker index were found to be 86.30%, 0.57, 3.26 and 87.58%, 0.55, 3.01 for ISSR and RAPD markers, respectively. Cluster analysis based on phytochemical and molecular characterization exhibited a clear relationship between diversity pattern and geographical origin. The percentage of variance explained using principal coordinate analysis (PCoA) was 70.31% and 71.91% for ISSR and RAPD markers, respectively. Analysis of molecular variance (AMOVA) analysis revealed high genetic variability occurred among the population than within populations. Bayesian analytical model and cluster analysis showed similar grouping patterns. The findings of the present research revealed a high degree of molecular and phytochemical variation among H. coronarium populations in relation to geographic locations. The results of the present research can be exploited for conservation and breeding programmes of H. coronarium for its genetic improvement.

Objective: The present study was undertaken to evaluate the total phenolic and flavonoid content and the antioxidant property of two important seagrass species namely, Halophila ovalis and Halophila beccarii occurring in Chilika lagoon, Odisha, India. Methods: Total Phenolic Content (TPC) of the extracts of Halophila species was determined by Folin-Ciocalteu method with little modifications and the total flavonoid content (TFC) was measured by aluminum chloride colorimetric assay. The antioxidant activity of different extracts was investigated by DPPH and ABTS radical scavenging activity. IC50 values were calculated for the DPPH and ABTS methods. Result: The study revealed that the methanol extract of H. ovalis has greater antioxidant activity than H. beccarii. Methanol extract of both the species (H. ovalis and H. beccarii) was found to possess high phenolic content at value of 70.25 mg GAE/g of extract and 48.53 mg GAE/g of extract respectively. Similarly flavonoid contents was found highest in methanol extract for both H. ovalis (76.82 mg quercetin equivalent/ g of extract) and H. beccarii (64.28 mg quercetin equivalent/ g of extract). The antioxidant activity of different extracts of these two species were evaluated using DPPH and ABTS radical assay. The methanol extract of both H. ovalis and H. beccarii showed high radical scavenging activity with IC50 values of 37.77 μg/ml and 52.25 μg/ml for DPPH and 25.62 μg/ml and 45.45 μg/ml for ABTS respectively. Conclusion: The study revealed the potential of the Halophila species as natural sources of antioxidants having considerable commercial importance.

Curcuma longa L. (turmeric), family Zingiberaceae, is an industrially valued plant having multipurpose use in cosmetics, dye, food and pharmaceutical industries. In the present study, the essential oil content and composition of 8 Indian turmeric cultivars has been analysed under similar environmental condition. The essential oil content of the fresh rhizomes varied from 0.5-0.9 % (v/w) and oil yield per plant varied from 1.6-3.49 ml. The GC/MS analysis has shown the presence of about 24-32 compounds representing 78.6-90.2 % of the total essential oil among 8 cultivars. The major compounds identified were Ar-Turmerone (39.5-45.5 %), Curlone (9.8-11.7 %), α-Phellandrene (5.5-7.7 %), Eucalyptol (3.2-5.5 %), β-Himachalene (1.6-5.5 %) and α-Copen-11-ol (2.3-5.4 %). Sesquiterpene hydrocarbons were found to be the main constituents in all eight cultivars analysed. Thus the result revealed that the cultivars Suroma and Lakadong having significantly high essential oil yield and quality could be used for commercial production of essential oil for their stable supply to national and international market.

Essential oil of twenty-five germplasms of Zingiber zerumbet from different accessions of Eastern India were analysed using Gas Chromatography-Mass Spectroscopy (GC/MS). A total of 29 constituents ...

Background Hedychium coronarium Koen. (Zingiberaceae) is traditionally used as medicine in countries such as India, China, and Vietnam for treatment of various ailments including cancer. However, in spite of its implied significance in cancer treatment regimes, there are no reports so far involving the anticancerous attributes of H, coronarium ethanol extract (HCEE) on cancer cells and a more comprehensive study on its mechanism is still lacking. Materials and methods The cytotoxicity of HCEE was evaluated by MTT and clonogenic survival assay. Annexin V/propidium iodide (PI), Hoechst 33342 staining, and TUNEL assay were performed to detect apoptosis. Cell cycle analysis was performed using PI staining. JC-1 and 2',7'-dichlorodihydrofluorescein diacetate assay were used to check the levels of MMP and ROS, respectively. Western blot analysis was carried out to measure the expression levels of proteins. Migration and invasion activity were assessed by wound healing and Transwell membrane assay, respectively. Results Antiproliferative effect of HCEE was investigated in various cancerous and normal cell lines. Among these, HCEE significantly inhibited the survival of HeLa cells without affecting the viability of normal human umbilical vein endothelial cells. Annexin V/PI, Hoechst staining, and TUNEL assay showed HCEE induced apoptosis in HeLa cells in a dose-dependent manner. HCEE promoted cell cycle arrest at G1 phase in HeLa cells by upregulating the levels of p53 and p21 and downregulating the levels of cyclin D1, CDK-4, and CDK-6. Moreover, HCEE treatment upregulated the expression of Bax and downregulated the expression of Bcl-2. Additionally, HCEE activated the caspase cascade by increasing the activities of caspase-9, caspase-8, and caspase-3. The expression levels of Fas ligand and Fas were also upregulated. Further, HCEE inhibited the migratory potential of HeLa cells by downregulating MMP-2 and MMP-9 expression levels. Conclusion Our results indicate H. coronarium exerts antiproliferative and apoptotic effects against HeLa cells, and therefore may be used for treatment against cervical cancer.

The Hedychium genus (Zingiberaceae) produces a plethora of secondary metabolites that include terpenoids, some of which are known for their aromatic properties. The volatile compositions of several members of this genus have not been chemically defined, as many species are endemic to remote ecosystems. In the present research, the essential oil composition of ten Hedychium species (H. gardnerianum, H. flavescens, H. thyrisiforme, H. flavum, H. ellipticum, H. aurantiacum, H. gracile, H. greenii, H. spicatum, and H. coronarium) was studied by gas chromatography flame ionization detector (GC-FID) and gas chromatography mass spectrometry (GC MS). The essential oil yield obtained by hydrodistillation varied from 0.05 (%v/w) in H. greenii to 0.75 (%v/w) in H. ellipticum and H. thyrsiforme, respectively. A total of 56 constituents representing 75.93–99.66% of volatile constituents were characterized in the essential oil of 10 Hedychium species. The predominant components were β-pinene in H. gardnerianum, H. gracile, H. thyrsiforme, H. flavum and H. flavescens; bornyl acetate in H. greenii; humulene epoxide I in H. aurantiacum; 1,8-cineole in H. spicatum and H. coronarium. Agglomerative Hierarchical Clustering (AHC) and Principal component analysis (PCA) based on essential oil compositions grouped Hedychium species into three different clusters. In view of the ambiguity existing among different Hedychium species, the developed chemical fingerprint in the present study would assume great significance in identification and classification of Hedychium species.

The present investigation was carried out to establish an efficient and reproducible micropropagation protocol for the production of morphologically, genetically and chemically uniform plants of Curcuma zedoaria. Axillary bud explants of C. zedoaria were inoculated into MS basal medium supplemented with various combinations and concentrations of 6-benzyladenine (2.2–22.2 µM, BA), kinetin (2.3–23.2 µM, Kin), indole-3-acetic acid (2.9–11.4 µM, IAA), α-naphthalene acetic acid (2.7–10.2 µM, NAA) and adenine sulphate (33.9–203.6 µM, Ads). Almost 95% of rhizome buds sprouted on MS medium supplemented with 13.3 μM BA, 5.7 μM IAA and 63.9 μM Ads giving rise to an average of 12.89 ± 0.02 shoots within 6 weeks. However, the maximum number of roots (25.8 ± 0.07 roots per explant) was obtained on half strength MS medium supplemented with 7.4 µM of IBA after 4 weeks of inoculation. Morphological characteristics were similar in both conventionally propagated and micropropagated plants. Additionally, genetic homogeneity of in vitro plants was further confirmed through ISSR and flow cytometry analysis. A total of 27 ISSR primers were screened, out of which 13 ISSR primers generated 58 monomorphic and reproducible bands thereby confirming the genetic uniformity of obtained plants. The mean 2C DNA content of the mother plant (2.96 pg) was similar to that of in vitro derived plants (3.07 pg). Gas chromatography-mass spectrometry (GC–MS) analysis showed similarity in the qualitative profile of chemical constituents of essential oil and high-performance liquid chromatography analysis revealed no significant differences in curcumin content in the tissue culture regenerants and mother plants of C. zedoaria. Therefore, the present micropropagation protocol could be effectively employed to generate true to type plantlets of C. zedoaria.

A high-frequency clonal propagation protocol was developed for Curcuma angustifolia Roxb., a high valued traditional medicinal plant. Axillary bud explants of C. angustifolia were explanted on Murashige and Skoog (MS) medium fortified with 4.4–22.2 µM 6-benzyladenine (BA), 2.9–5.7 µM indole-3-acetic acid (IAA), 2.3–23.2 µM kinetin (Kin), 2.7–5.4 µM naphthalene acetic acid (NAA) and 67.8-271.5 µM adenine sulphate (Ads) in different combinations. The maximum number of shoots per explants (14.1 ± 0.55) and roots per shoot (7.6 ± 0.47) was achieved on media containing 13.3 µM BA, 5.7 µM IAA and 135.7 µM Ads. Stability in phytomedicinal yield potential of micropropagated plants was assessed through GC–MS and HPTLC. Gas chromatogram of essential oil of conventional and micropropagated plants of C. angustifolia had similar essential oil profile. HPTLC analysis of rhizome extracts of in vitro and field grown plants revealed no significant differences in the fingerprint pattern and in curcumin content. Genetic integrity of in vitro and field grown derived plants were evaluated with inter-simple sequence repeat (ISSR) primers and flow cytometry using Glycine max as an internal standard. A total of 1260 well resolved bands were generated by 12 ISSR primers showing monomorphic banding patterns across all plants analyzed. The mean 2C DNA content of conventionally and micropropagated plant was estimated to be 2.26 pg and 2.31 pg, respectively. As no somaclonal variations were detected in tissue culture plantlets, the present micropropagation protocol could be applied for in vitro conservation and large-scale production of C. angustifolia.

The anticancerous and immunomodulatory activities of Alpinia nigra essential oils and extracts were evaluated in the present study. Besides, volatile constituents of essential oils were also analys...

Hedychium coronarium Koen. is a perennial herb of family Zingiberaceae that is valued for its high quality essential oil. In the present study, the variability in chemical constituents, antioxidant and antimicrobial property of essential oils of Hedychium coronarium collected from different locations of Eastern India were evaluated. Essential oil hydrodistilled from rhizome parts of H. coronarium varied from 0.05 to 0.47% (v/w). A total of 44 components accounting for 86.18–99.94% of H. coronarium essential oil were identified by gas chromatography flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC–MS). The major components are β-pinene (11.07–42.74%), eucalyptol (11.48–40.59%), linalool (1.56–45.11%), coronarin-E (1.01–39.57%), α-pinene (3.80–16.60%), p-cymene (1.05–8.89%), γ-terpinene (1.73–5.82%) and 10-epi-γ-eudesmol (1.11–4.86%). According to agglomerative hierarchical clustering (AHC) and principal component analysis (PCA), the essential oils of H. coronarium populations were grouped into four different chemotypes. They were (I) eucalyptol, (II) linalool, (III) coronarin-E, and (IV) β-pinene. Furthermore, essential oils exhibited considerable levels of reducing power activity (IC50, 0.39–1.66 mg/ml), 2,2-diphenyl-1-picrylhydrazyl (DPPH) (IC50, 0.57–2.19 mg/ml), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (IC50, 0.12–0.67 mg/ml) radical-scavenging activities. Results of antimicrobial activity concluded that essential oil has good inhibitory activity against Candida albicans and Fusarium oxysporum. The results showed that geographic origin greatly influenced the chemical composition of essential oil of H. coronarium and their associated bioactivities. Four chemotypes identified in the present study could be developed as promising bioresource in perfumery, food and pharmaceutical industry.

The antioxidant and antimicrobial activities of the essential oil and extract of Alpinia nigra (Gaertn.) Burtt (Zingiberaceae) were assessed in vitro. Besides total phenolic and flavonoid content o...

Rhizome extracts of Hedychium coronarium are widely used as phytotherapeutics. As of date, there is no documented study on the standardization of H. coronarium extract, and the following research is an effort in this direction. Coronarin D is an important bioactive compound present in H. coronarium which shows chemopreventive activity against cancer. H. coronarium extracts were assessed for coronarin D content for the first time. The extraction was checked using different solvents: n-hexane, acetone, and methanol. Coronarin D was separated on silica gel 60F254 high-performance thin-layer chromatography (HPTLC) plates by isocratic gradient method using n-hexane-ethyl acetate (80:20 v/v) as mobile phase. Densitometric quantification was performed at 231 nm in absorption mode. This method gave a well-defined peak at Rf 0.20 corresponding to coronarin D. The method was validated using International Conference on Harmonization (ICH) guidelines in terms of precision, repeatability, and accuracy. Linearity range...

Hedychium coronarium J. koenig (Zingiberaceae) is well known for to its medicinal and aromatic properties because of it terpenic components. The paper aims to characterize the terpenic composition present in the essential oil of Hedychium coronarium J. koenig by two-dimensional gas chromatography with time-of-flight mass spectrometry (GCxGC-TOF-MS). A total of 101 terpenic constituents were identified, 53 of which were reported for the first time. On the basis of their chemical structure, the constituents were grouped into four classes: hydrocarbon monoterpenes, oxygenated monoterpenes, sesquiterpene hydrocarbons and oxygenated sesquiterpenes. The identified compounds include 35 alcohols, 34 hydrocarbons, 12 ketones, 7 aldehydes, 7 oxides, 3 ethers and 3 esters. The predominant compounds identified by GCxGC-TOF-MS were eucalyptol, p -cymene and p -menth-1-en-8-ol. A database containing retention indices of terpenoids was created for the bi-dimensional column, thus proving to be a remarkable step for analysis of terpenoids using a two dimensional gas chromatography (GCxGC) system. GCxGC enhanced the separation efficiency of constituents as well as resolved a number of co-eluting components that were unresolved on a one dimensional gas chromatography column. The current research enabled in comprehensive profiling of terpenic composition present in H. coronarium oil, an important initiative that could add to the valuation and bioprospecting potential of H. coronarium as a source of terpenic constituents.

Kewda (Pandanus fascicularis Lam.) is a well known medicinal and aromatic plant. The paper aims to precisely characterize volatile constituents present in Kewda flower oil using comprehensive two-dimensional gas chromatography coupled to time of flight mass spectrometry (GCxGC-TOFMS). A total of 159 components were identified due to enhanced chromatographic separation and mass spectral deconvolution of GCxGC-TOFMS. On the basis of its chemical structure, the identified compounds were grouped into hydrocarbons, alcohols, ethers, ketones, esters, nitrogen compounds, aldehydes, acids, lactones, halides and sulfur containing compounds. Ethers were the major components. The predominant compounds identified by GCxGC-TOFMS were kewda ether, ortho-cymene and terpinen-4-ol. A database containing retention indices of compounds was created for the bi-dimensional column, thus proving to be a remarkable step for analysis of constituents using a GCxGC system. GCxGC-TOFMS separated a number of co-eluting components which were unresolved on a single GC column.

The present investigation was carried out to establish an efficient and reproducible micropropagation protocol for the production of morphologically, genetically and chemically uniform plants of Curcuma zedoaria. Axillary bud explants of C. zedoaria were inoculated into MS basal medium supplemented with various combinations and concentrations of 6-benzyladenine (2.2–22.2 µM, BA), kinetin (2.3–23.2 µM, Kin), indole-3-acetic acid (2.9–11.4 µM, IAA), α-naphthalene acetic acid (2.7–10.2 µM, NAA) and adenine sulphate (33.9–203.6 µM, Ads). Almost 95% of rhizome buds sprouted on MS medium supplemented with 13.3 μM BA, 5.7 μM IAA and 63.9 μM Ads giving rise to an average of 12.89 ± 0.02 shoots within 6 weeks. However, the maximum number of roots (25.8 ± 0.07 roots per explant) was obtained on half strength MS medium supplemented with 7.4 µM of IBA after 4 weeks of inoculation. Morphological characteristics were similar in both conventionally propagated and micropropagated plants. Additionally, genetic homogeneity of in vitro plants was further confirmed through ISSR and flow cytometry analysis. A total of 27 ISSR primers were screened, out of which 13 ISSR primers generated 58 monomorphic and reproducible bands thereby confirming the genetic uniformity of obtained plants. The mean 2C DNA content of the mother plant (2.96 pg) was similar to that of in vitro derived plants (3.07 pg). Gas chromatography-mass spectrometry (GC–MS) analysis showed similarity in the qualitative profile of chemical constituents of essential oil and high-performance liquid chromatography analysis revealed no significant differences in curcumin content in the tissue culture regenerants and mother plants of C. zedoaria. Therefore, the present micropropagation protocol could be effectively employed to generate true to type plantlets of C. zedoaria. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-019-2009-9) contains supplementary material, which is available to authorized users.

Alpinia galanga Wild.(L.) is well known for its aromatic constituents. Though the aromatic composition is already known, but lots of constituents which contributing overall aroma of the oil are still unknown due to the co-eluting factor of single column in GC-MS. Thus the current study aims to characterise maximum volatile constituents present in the essential oil of A. galanga using thermal desorption modulator of two-dimensional gas chromatography and time-of-flight mass spectrometry. The 102 compounds with good match and high probability value were identified out of which 42 were identified for the first time. The total identified compounds include 47 hydrocarbons 25 alcohols, 7 ketones, 7 esters, 3 aldehyde, 4 ethers and 9 other classified aromatic compounds. It was further categorised into Monoterpene Hydrocarbons, Oxygenated Monoterpenes, Sesquiterpene Hydrocarbons and Oxygenated Sesquiterpenes. The major constituent also varies with respect to area percentage. The in-depth characterisation will help in its qualitative analysis.

Betelvine (Piper betle L., Family: Piperaceae) is one of the important cultivated cash crop and medicinal plant of India, lacking sufficient molecular resources in various databases for exploration of biosynthetic pathways involved in terpenoid synthesis. Molecular markers like SSRs (Simple Sequence Repeats) and SNPs (Single Nucleotide Polymorphisms) are generally used for genetic diversity study. However, limited number of nucleotides and absence of SSRs as well as SNPs restrict molecular study of betelvine landraces. These problems also minimise molecular characterization of novel genes that are involved in terpenoid biosynthesis. To elucidate the biochemical pathways of terpenoid biosynthesis, transcriptome profiling of RNA sample was performed from leaves of betelvine. The pair end transcriptome profiling generated 6.7 G bases (4.4Gb) of data containing 33,235,667 raw reads for each end of RNA Seq. After removing contaminants, 30,104,811 clean reads were subjected to de novo transcriptome assembling, which subsequently produced 46,656 clean transcripts. The clean transcripts were annotated and their functions were identified using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Eukaryotic Orthologous Groups (KOG), PlantCyc, Swissprot, and TrEMBL database. These annotations resulted mining of various trait specific genes including important terpenoid biosynthesis genes. Presence of 43 terpenoid compounds in essential oil of four betelvine landraces has also been confirmed through GC MS. Further, gene expression analysis of five terpenoid biosynthesis genes were carried out by real time polymerase chain reaction (RT PCR). The expression of monoterpene synthase genes (limonene synthase, linalool synthase, and α-terpeneol synthase) and sesquiterpene synthase genes (germacrene synthase and sesquiterpene) were validated using four landraces of betelvine. Landraces specific high quality 4181 SSRs and 350,676 SNPs were identified for further genetic diversity study. In this study, terpenoid biosynthesis specific transcription factors and 22 miRNAs having 33 target sites were also predicted. Overall, this study will enrich the molecular repository of betelvine for further improvement of the taxa and to develop new genotype of betelvine crop with quality traits.

Clematis chinensis (family: Ranunculaceae) is a deciduous climbing plant having multipurpose use in traditional Chinese medicine, but the use of molecular markers in Clematis chinensis is yet to gain momentum. Recently EST-SSRs are the free by-product of the currently growing EST databases. SSRs (microsatellites) have been extensively used as molecular marker in several genetic studies. The present study reports the frequency of SSRs and appropriate primers for the C. chinensis EST sequences. The EST sequences of C. chinensis were obtained from NCBI database and screened for the presence of SSRs. MISA tool was used to predict the microsatellites and Primer3 plus software was used for designing the primers. By using the MISA tool a total of 155 SSRs were identified in contigs and 275 SSRs were identified in singletones. Occurrence of trinucleotide repeats were most abundant both in contigs (52.3%) and singletones (41.5%) followed by dinucleotides. Maximum trinucleotide motifs code for Leucine (CTT) followed by glutamic acid (GAA) and Serine (TCT) i.e., whereas in singleton sequences glutamic acid (GAA) is dominated followed by glutamine (CAA) and asparagine (AAC). AG/CT was the most recurrent repeat of dinucleotide. We also - designed 60 primers flanking the SSR repeats. The identified SSRs and their related markers can be used in future studies of linkage mapping, comparative genomics study, genetic diversity analysis and different breeding programs in Clematis chinensis.

Curcuma angustifolia Roxb. (Family: Zingiberaceae) is an important rhizomatous medicinal herb, grows wild in India. The leaf essential oil of C. angustifolia was extracted by hydro-distillation and...

Curcuma longa L. belonging to the family Zingiberaceae is an important spice and medicinal plant. Despite an ever growing interest and commercial importance of curcumin and Curcuma essential oil, it is still not clear as to the nature and mode of influence the different soil and environmental factors have on the production and quality of curcumin, leaf and rhizome essential oil of this species. The present report deals with ‘Surama’, a high yielding turmeric cultivar that was cultivated at nine agroclimatic zones with an objective to analyse the effects the different soil and environmental parameters have on product optimization as well the quality of curcumin and essential oil contents. Variation in curcumin (1.5–5%), leaf (0.37–0.8%) and rhizome oil (0.45–0.7%) were recorded across all the zones. Gas chromatography mass spectrometry (GCMS) analysis of leaf and rhizome essential oil for different experimental zones showed wide variation in their quantity and quality. It was found that for curcumin content, most sensitive factors were; the altitude, soil pH, nitrogen and Potassium content. In contrast, the temperature and phosphorous content were found critical for leaf essential oil while, nitrogen, phosphorous and potassium contents were the key quantitative and qualitative determinants for rhizome essential oil. This study provides essential experimental data that could find effective use in yield optimization and managing varying environmental parameters of this important species for high and quality yield of curcumin, rhizome and leaf essential oil. Identification and critical analysis of sensitive/determinant agro-climatic factors are significant for commercial exploitation as well value addition of turmeric products especially with high alpha phellandrene and tumerone content.

Alpinia galanga is a wild medicinal plant having tremendous industrial importance. However, lack of quality planting material in natural resources restricted its wide exposure in industrial sector. Thus, a standardised protocol for stable supply of quality planting materials with genetical and phytochemical stability is need of the hour to meet commercial requirements. The current study reports an efficient protocol to regenerate micropropagated plantlets with long term stability in molecular as well as phytochemical level. Murashige and Skoog (MS) media augmented with 3 mg/l 6-Benzyladenine (BA), 3 mg/l Kinetin (Kn) and 1 mg/l Napthalene Acetic acid (NAA) was found optimum for regeneration and multiplication of plantlets. In order to establish genetic stability molecular marker-based profiling of micropropagated plants were done at every 6 months intervals up to 6 years. Monomorphic banding pattern were found that are identical to the mother plants. After six years of in vitro conservation, the regenerated plants were transplanted to the field and its drug yielding potential were evaluated through phytoconstituent analysis along with bioactivity studies. A comparative Gas chromatography (GC) and mass spectroscopy (MS) analysis of essential oils showed no significant differences in phytoconstituents between the conventional and in vitro propagated plants. The bioactivities like antioxidant, antimicrobial and anticancerous study along with total phenolic and total flavonoid content of in vitro plants were compared with mother plants and found stable with negligible variation. Thus, the present study has enough significance towards commercial exploitation of A. galanga with phytochemical and molecular stability.

Pandanus odorifer (Forssk.) Kuntze is an economically important aromatic plant. The essential oil from male flowers is widely used in aromatherapy, cosmetics and as food flavouring agent. Phenylethyl methyl ether (PEME), the major constituent of essential oil, gives the chief characteristic fragrance to the oil. In the present study, 180 samples from 12 different regions were collected and hydrodistilled for essential oil isolation. The oil was then subjected to GC and GC-MS analysis to find out the percentage of the constituents. The results revealed PEME as the major constituent ranging from 58.03 to 81.86% and terpinen-4-ol, the second major constituent ranging from 7.81 to 21.46%. Soil nitrogen was found to be the most influential factor for oil yield and PEME content. The flowers containing high essential oil yield and PEME content could be used as elite chemotypes with enough potential for large-scale commercial cultivation to meet the demand of kewda industries.