Reena Parida

In vitro propagation of Hedychium coronarium Koen. through axillary bud proliferation

This report describes an efficient and reproducible protocol for large-scale multiplication of Hedychium coronarium plantlets. Axillary bud explants were cultured on Murashige and Skoog medium supplemented with 3 mg/l benzylaminopurine, 3 mg/l kinetin (KIN), and 0.2 mg/l thidiazuron, yielding a maximum of 13.2 ± 0.3 number of shoots. Sub-culturing of shoots every 4 weeks on fresh multiplication medium yielded a consistent proliferation rate. Shoot clusters containing three to five shoots were successfully rooted in KIN (3 mg/l) and indole acetic acid (0.5 mg/l), yielding a maximum of 6.3 ± 0.5 number of roots. Plantlets grown in vitro were acclimatized and subsequently transferred to the field for phenotypic evaluation. Random amplified polymorphic DNA and inter-simple sequence repeat analysis has confirmed the genetic uniformity of in vitro plantlets up to 2 years. After 2 years, these plantlets were transplanted to field, and evaluation of phenotypic characteristics was done. This study is of high significance as these could be commercially utilized for large-scale production of true-to-type plantlets.

Chemical Composition of Essential Oil from Leaf and Rhizome of Micropropagated and Conventionally Grown Hedychium coronarium Koen. from Eastern India

Abstract The essential oil from leaves and rhizomes of micropropagated and conventionally grown Hedychium coronarium Koen. of family Zingiberaceae, was extracted through hydro-distillation process and its chemical composition was analyzed by GC-MS. A total of 30 (95.23%) and 7 (98.12%) compounds were identified from the leaf and rhizome of conventionally grown plants where as 16 (95.96%) and 3 (99.65%) compounds were identified from leaf and rhizome samples of micropropagated plants. The major compound identified in leaf oil of micropropagated and conventional plants was β-pinene (26.87% and 15.02%) and in rhizome oil was eucalyptol 47.97% and 33.71% respectively. As compared to the conventionally propagated plants the micropropagated plants were more uniform hence could be used for commercial purposes.

Evaluation of yield, quality and antioxidant activity of essential oil of in vitro propagated Kaempferia galanga Linn.

Abstract Objective To determine chemical constituents and antioxidant properties of essential oil from rhizome of the medicinal plant, Kaempferia galanga (K. galanga) Linn. (Zingiberaceae) in conventionally propagated (CP) and in vitro propagated (IVP) plants. Methods In vitro (micro) propagation of K. galanga was done by inoculating explants on to Murashige and Skoog agar medium, supplemented with suitable combinations of phytohormones; the regenerants were transferred to soil for further growth. Essential oil preparations of both CP and IVP rhizomes grown in soil, obtained by the hydro-distillation method were analyzed by gas chromatography-mass spectrometry. Antioxidant activities of essential oil samples were monitored. Results Maximum numbers of regenerated shoots were found in the medium supplemented with 1 mg/L benzyl adenine and 0.5 mg/L indole-3-acetic acid. A total of 6 compounds were identified from rhizomes from CP and IVP plants that yielded 96.9% and 97.81% of the total oil contents, respectively. The major compound of rhizome oil identified from CP and IVP rhizomes was ethyl p-methoxy cinnamate in quantities, 82.01% and 71.77%, respectively, without any compositional variation. Antioxidant properties of essential oil preparations were assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydrogen peroxide radical scavenging assays. Moreover, antioxidant activities of rhizome-oil from IVP plants were better than that of CP oil samples. Conclusions As IVP rhizomes had better oil yield, those could be used for a large scale commercial propagation for sustainable use of essential oil. The principal chemical in the essential oil, ethyl p-methoxy cinnamate could help apothecary, for several ailments.

Chemical composition and antioxidant activity of some important betel vine landraces

Abstract The essential oil composition and in vitro antioxidant activity of eight important betel vine landraces, such as Balia, Chandrakala, Dandabalunga, Desibangla, Godibangla, Karpada local, Maghai and Nahua were evaluated in this research. GC and GC-MS analysis of the essential oil resulted in identification of 50 compounds representing 95.97% of the oil. Major compounds were eugenol, terpinolene, terpinene, cadinene, caryophyllene, 3-carene, etc. The plant essential oil was also subjected to screening for the evaluation of their antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl assay. The highest antioxidant activity was shown by Balia landraces, whereas the lowest activity by Godibangla landraces. Essential oil showed a great variety of compounds in their chemical composition and antioxidant activity. This might be due to the affect of different environmental factors and soil nutrients of betel vine cultivating zones. The GC-MS chromatogram with antioxidant activity of the above betel vine landraces would not only help to select the elite chemotypes, but also to enhance their future industrial applications.

Variation in Volatile Constituents and Eugenol Content of Five Important Betelvine (Piper betle L.) Landraces Exported from Eastern India

Abstract Piper betle L. commonly known as betel vine has been consumed as mouth freshener and traditionally used for medicinal purposes in many countries around the world. The volatile constituents of Piper betle landraces from eastern India were analysed through GC FID and GC-MS analysis. Thirty eight compounds were identified accounting to a total of 98.96% in Paradeep, 98.07% in Birikuli, 95.46% in Matiali, 99.94% in Sanchi and 95.51% in Kali Bangla respectively. Isoeugenol was found to be the major constituent in Paradeep (34.56%), Matiali (48.2%) and in Sanchi (59.2%) where as in Birikuli, trans-isoeugenol (28.45%) and in Kali Bangla, acetyl eugenol (54.76%) were the major constituent. β-ocemene was the unique molecule present in Paradeep where as Birikuli was marked by the presence of eugenol, γ-cadinene , cubebene and β-caryophyllene. Isoeugenol acetate, α-phellandrene and methyl eugenol were only present in Matiali. Sanchi can be distinguished from other landraces by the presence of β-elemene. B-cardinene, chavicol and chavicolacetate were unique to Kalibangla. The volatile constituents can be used for authentic identification of landraces and also enhance their export potential by means of important phytoconstituents and bioactivity studies.

Retention of drug yielding potential of micropropagated Hedychium coronarium

Abstract The aim of the present study was to analyze the chemical constituents present in the essential oil obtained from both conventional and micropropagated Hedychium coronarium rhizomes using gas chromatography - mass spectroscopy analysis. A total number of 7 compounds were identified from the rhizomes of conventional plants, whereas 3 compounds in case of the micropropagated plants. The major compounds identified in micropropagated rhizome oil were eucalyptol (47.97%), β-pinene (39.23%) and linalool (12.45%). The antioxidant activity of H. coronarium essential oil exhibited a moderate result using 2, 2-diphenyl-1-picrylhydrazyl free-radical scavenging assay. In spite of its high medicinal value, this endangered wild species is going to extinct for which micropropagation is necessary for retention of its drug yield potential present in its essential medicinal oils. All above analysis showed that the micropropagated H. coronarium could be used for a large scale commercial propagation bearing its medicinal constituents.

Coronarin-D Production from Tissue Cultured Plantlets of Hedychium coronarium

Abstract Hedychium coronarium known as white ginger lily, an endangered medicinal plant species belonging to family Zingiberaceae. Coronarin D, a labdane diterpene isolated from rhizomes of H.coronarium inhibits NF-kB activation induced by different inflammatory stimuli and carcinogenesis. In the international market the price of coronarin D is USD 260/5 mg. Callus cultures were initiated, plantlets were regenerated and experimental study of in vitro cultures of H. coronarium was done to estimate the highest presence of coronarin D in lines using different growth regulators as compared to that of the mother plant and its genetic study. Explants were inoculated on Murashige and Skoog media and cultures were maintained. The enhanced coronarin D content from matured rhizomes of established cultures were quantified using High Performance Thin Layer Chromatography with the standard. Inter simple sequence repeats study was performed to characterize the plants genetically. The maximum presence of coronarin D content was found to be 3.51 % whereas in control (mother plant) was 2.35 %. The molecular characterization showed no polymorphism in banding pattern as compared to the control. The present protocol reports somaclone with high coronarin D content which could be further analyzed to be used in large scale for commercial exploitation.

Assessment of genetic diversity of Curcuma aromatica from eastern India using ISSR and RAPD markers

Molecular characterization of Curcuma aromatica from 4 different populations of Odisha using inter simple sequence repeats and random amplified polymorphic DNA markers to indicate the closeness of species and hybrids quickly and efficiently. A dendrogram was constructed through sequential agglomerative hierarchial and nested (SAHN) clustering and unweighted pair group method with arithmetic mean (UPGMA) analysis using Jaccard’s similarity coefficient of combined markers using this particular species. Two major clusters were found i.e., cluster-I (Koraput-1, Koraput-2, Koraput-3, G.Udaigiri-1, G.Udaigiri2, G.Udaigiri-3 and Phulabani-1, Phulabani-2, Phulabani-3) and cluster-II (Raikia-1, Raikia-2 and Raikia-3). The clustering pattern also revealed moreover the extent of genetic similarity between germplasms collected from four different regions population. The potential of this technique would be further realized to fullest extent for the identification and tagging of important novel gene in different taxa, unexplored yet, thus facilitating the improvement of desired taxa of Zingiberaceae. The findings would be of immense enough significance for complementing the strategies of conservation and characterization of these important taxa of Zingibearceae following modern biotechnological approach.

Edible plant-derived essential oils synergistically enhance the Th1, Th2 and anti-inflammatory cytokines in neonatal cord blood monocytic cell line

ABSTRACT Edible medicinal plants Curcuma longa, Kaempferia galanga and Zingiber officinale are considered to be suitable for pregnant mother and neonates. Their essential oils are claimed to have therapeutic effects, yet its immunomodulatory activities need to be investigated. To validate the immunomodulatory effect, cord blood monocyte cells (CBMCs) treated with different concentrations of essential oils ranging from 0 to 40 µg/ml. K. galanga oil at 40 µg/ml concentrations upregulated IFN-γ at 49.23 pg/ml, while C. longa oil downregulated the IL-10 at 10.45 pg/ml showing immunostimulant activity on CBMCs. However, Z. officinale oil showed no significant result at any concentration. K. galanga and C. longa oils also showed an increase in lymphocyte proliferation in a dose-dependent manner. The present study was aimed to evaluate the immunomodulatory potentialities of essential oils which will attract the interest of researchers and pharmaceutical industries for clinical studies and other applications in the therapy of diseases related to neonatal infections. GRAPHICAL ABSTRACT