Gentiana Kurroo Research Articles

Hydroethanolic extract of Gentiana kurroo Royle rhizome ameliorates ethanol-induced liver injury by reducing oxidative stress, inflammation and fibrogenesis in rats

Ethnopharmacological relevanceGentiana kurroo Royle is a medicinal plant mentioned as Traymana in Ayurveda. In the folklore, it is used to cure fever, stomach ache, skin diseases and liver disorders. However, limited reports are available on the therapeutic potential of Gentiana kurroo Royle against alcohol-induced liver damage. Aim of the studyTo assess the effectiveness of the hydroethanolic extract of Gentiana kurroo Royle rhizome (GKRE) against alcohol-induced liver injury and explore the mechanism of action. Materials and methodsGKRE was characterized using UHPLC-QTOF-MS/MS. The binding affinity of the identified compound was studied in silico. In vitro studies were performed in the Huh-7 cell line. An acute oral toxicity study (2 g/kg BW) of GKRE was done in rats following OECD 420 guidelines. In the efficacy study, rats were treated with 50% ethanol (5 mL/kg BW, orally) for 4 weeks, followed by a single intraperitoneal dose of CCl4 (30%; 1 mL/kg BW) to induce liver injury. After 4th week, the rats were treated with GKRE at 100, 200 and 400 mg/kg BW doses for the next fifteen days. The biochemical and antioxidant parameters were analyzed using commercial kits and a biochemistry analyzer. Histopathology, gene and protein expressions were studied using qRT PCR and western blotting. ResultsThirteen compounds were detected in GKRE. Few compounds showed a strong interaction with the fibrotic and inflammatory proteins in silico. GKRE reduced (p < 0.05) the ethanol-induced ROS production and inflammation in Huh-7 cells. The acute oral toxicity study revealed no adverse effect of GKRE in rats at 2 g/kg BW. GKRE improved (p < 0.05) the body and liver weights in ethanol-treated rats. GKRE improved (p < 0.05) the mRNA levels of ADH, SREBP1c and mitochondrial biogenesis genes in the liver tissues. GKRE also improved (p < 0.05) the liver damage markers, lipid peroxidation and levels of antioxidant enzymes in the liver. A reduced severity (p < 0.05) of pathological changes, fibrotic tissue deposition and caspase 3/7 activity were observed in the liver tissues of GKRE-treated rats. Further, GKRE downregulated (p < 0.05) the expression of fibrotic (TGFβ, αSMA and SMADs) and inflammatory markers (TNFα, IL6, IL1β and NFκB) in the liver. ConclusionGKRE showed efficacy against alcohol-induced liver damage by inhibiting oxidative stress, apoptosis, inflammation and fibrogenesis in the liver.

Trayamana (Gentiana kurroo Royle): A Substitution or an Adulteration to Katuki (Picrorrhiza kurroa Royle ex Benth.)

ABSTRACT Katuki (Picrorrhiza kurroa Royle ex Benth.) belonging to the family Scrophulariaceae, is one of the most important herbs found in the Himalayan Region at an altitude of 3000–5000 m above the Sea level. It has been used for the management of different diseases such as fever, diabetes, abdominal disorders, and hemorrhoids and most of the formulations such as Arogyavardhini vati, Kumaryasava, and Maha Sudarsana vati also have Katuki. The market demand of Katuki is more and the production is less, so it is either substituted or adulterated with different similar-looking and similar-acting drugs. Mostly, the root of Gentiana kurroo Royle is used in place of Katuki in the market. Katuki and Trayamana rhizomes are subcylindrical to quadrangular and rigid in shape. So, they look similar. Cork cells, tracheids, fibers, etc., are also present in both herbs. Both are bitter in taste and are used in fever, stomachache, dysentery, etc. Hence, an effort has been done to analyse whether Trayamana is actually a substitute or an adulteration depending on rasa, guna, veerya, vipaka, and karma and chemical compounds.

Floristic composition, life history traits and phytogeographic distribution of forest vegetation in the Western Himalaya

IntroductionScientific documentation of the qualitative forest vegetation parameters of a biogeographical area provides baseline information to guide conservation strategies and design policies for biodiversity management regulations.MethodsWe present one of the most comprehensive qualitative vegetation analyses to evaluate the entire structure and function of an ecosystem in the remote northern part of the Kashmir Himalaya, India. Several multivariate ecological community analyses were conducted after determining the presence of plant species in the various habitats using a random sampling technique.ResultsIn total, 155 plant species belonging to 120 genera and 49 families occurred in the area. Asteraceae was the largest family (12% of plant species) followed by Rosaceae (11%). The patterns of species distribution across families were uneven, with 50% of the species belonging to only 7 families, and 23 families being monotypic. In terms of functional groups, the herbaceous growth form dominated. Therophytes were the dominant life form, indicating that the vegetation was disturbed. According to the phytogeographical research, 65% of the species documented in the study area were native, 15% were invasive, 14% naturalized, and 5% being casual. The majority (30%) of exotic species were reported along roadsides. Of all the species found, 39% grew in their natural habitats, such as forests, and 11% were scattered along roadsides. Plant species were grouped in five different clusters based on their floristic similarity. According to the estimated diversity indices, natural forest has the greatest values for Shannon’s and Simpson’s index. We found that the study area serves as the natural habitat for several significant, endangered medicinal plants, including Arnebia benthamii, Bergenia ciliata, Delphinium roylei, Gentiana kurroo, Phytolacca acinosa, Saussurea costus, and Trillium govanianum. Therefore, we recommend that human intervention in natural regeneration efforts be prioritized in these habitats to increase the population of these species.ConclusionExamining species features from the perspective of functional groups contributes to our understanding of the ecological aspects of the flora. It may also be useful in developing management plans to ensure long-term management of forest landscapes in this remote Himalayan region.

Gentiana kurro’s physiology and pharmacological properties: Recent update and future perspective

Gentiana kurroo royle is an endangered bitter medicinal plant of the Indian subcontinent. This medicinal plant mainly grows in Kashmir, Himachal Pradesh, and North-west Himalayas. The medicinal plant's root and rhizome are frequently used by native people for various local remedies. This species has become critically endangered due to its endemic nature and high rate of extraction from its natural environment. In India rhizome and root of this medicinal plant are used as ant periodic, bitter tonics, anti-inflammatory, blood purifier, expectorant, antipsychotic, carminative, and stomachic. Genitiana kurroo can be propagated through micro-filtration of seeds, rhizome cutting, shoot-made segments, and somatic embryogenesis. The roots of Genitiana kurroo are rich in various ingredients such as flavonoids, alkaloids, and terpenoids which are responsible for analgesic, anticancer, and immune-modular effects. Considering these perspectives the present review has intended to provide updated prospects of Gentiana kurroo and provides insights on the nutritional and various pharmacological properties based on updated research literature. This review explores the use of Gentiana kurroo across disciplines for its wide range of prominent-pharmacological properties.

Study on phytochemical, antibacterial and antioxidant properties of Genitiana kurroo Royle

Gentiana kurroo Royle is an endangered bitter medicinal plant of the Indian subcontinent region. This medicinal plant mainly grows in Kashmir, Himachal Pradesh, and North-west Himalayas. Native people frequently use the medicinal plant’s root and rhizome for various local remedies. The leaf and root samples of the plant were studied for their phytochemicals screening and antibacterial and antioxidant properties. The methanolic root extract as a comparison to methanolic leaf extract was detected with high concentration of phytochemicals like alkaloids, flavonoids, saponins, phenol, glycosides whereas, other phytochemicals like carbohydrates, tannins, and terpenoids showed equal concentration in methanolic root and leaf extracts. In antibacterial study, the methanolic root extract was found to exhibit a maximum zone of inhibition (33 ± 1) against E.fecalis and minimum zone (17 ± 2) against E. coli. The methanolic leaf extract showed a maximum zone of inhibition (31 ± 1) against E. fecalis and a minimum zone (18 ± 2) against K. pneumonia. The antioxidant activity of G.kurroo revealed that the methanolic extracts of root as compared to the methanolic extract of leaves showed comparatively high antioxidant activity and this is due to the presence of high phenol and flavonoid content.

Ornamental Potential of &lt;I&gt;Gentiana kurroo&lt;/i&gt; Could be a Boon for its Survival: A Critically Endangered Species

Ornamental Potential of &lt;I&gt;Gentiana kurroo&lt;/i&gt; Could be a Boon for its Survival: A Critically Endangered Species

Development and Validation of a Precise RP- HPLC Method to Determine Gentiopicroside Content in Cultures of Gentiana kurroo Royle

Development and Validation of a Precise RP- HPLC Method to Determine Gentiopicroside Content in Cultures of Gentiana kurroo Royle

Fermentation medium optimization for the 1,4-ß-Endoxylanase production from Bacillus pumilus using agro-industrial waste

A novel cost-effective medium for in vitro cultivation of Gentiana kurroo Royle using sugarcane bagasseShiwani Kaushal, Arushdeep Sidana

Effect of gamma radiation on gentiopicroside production in Gentiana kurroo Royle in vitro cultures

Gentiopicroside (GPD) is one of the vital bioactive molecules present in the rhizomes, roots and aerial parts of Gentiana kurroo Royle. In this study, callus and in vitro shoots cultures of G. kurroo were treated with different doses of gamma radiation to assess its effect on the concentration of GPD. The callus was established on MS media fortified with Indole-3-butyric acid (IBA) and Indole-3-acetic acid (IAA) and exposed to variable radiation doses (0, 10, 20, 30, 40, 50, or 60 Gy 60Co gamma rays). Similarly, in vitro shoots raised in MS media supplemented with 6-Benzylaminopurine (BAP) and IAA were irradiated at (0, 10, 20, 30, 40, 50, 60, or 70 Gy 60Co gamma rays). In vitro cell growth and GPD content were assessed during M1V1, M1V2, M1V3, and M1V4 (Mutation and Vegetative) generations. The lethal dose (LD50) for both callus and shoots were observed at 35 Gy. The results revealed that low radiation doses favored growth and showed increased GPD content in the callus cultures during M1V1 generation, but both parameters were reduced in the M1V2, M1V3 and M1V4 generations. However, shoots irradiated at low doses boosted the growth and GPD level during M1V2, M1V3 and M1V4 generations. This is the first report that used gamma radiation to show the effect of GPD elicitation in the genus Gentiana.

Optimizing Shoot Formation in Gentiana kurroo Royle for Gentiopicroside Production

The roots and shoots of Gentiana kurroo Royle are rich sources of gentiopicroside (GPD). The plant is used traditionally for curing many metabolic diseases. The exploitation of G. kurroo in its native habitat has placed the plant on the critically endangered list of plants in India. One of the ways of creating an alternative source of G. kurroo is through in vitro propagation. Although a number of in vitro propagation methods for G. kurroo exist, there are no studies that have optimized methods for rapid in vitro shoot production and the production of GPD. The objective of this study was to develop an effective in vitro shoot multiplication system of G. kurroo. Furthermore, the influence of solid and liquid induction media were investigated. Shoots were regenerated from embryogenic callus and transferred to solid and liquid Murashige and Skoog (MS) and Gamborg (B5) media fortified with various concentrations of BA containing different auxins. It was observed that the liquid medium produced a higher number of shoots than the solid media. MS supplemented with BA (2 mg/L) and IAA (0.5 mg/L) produced ~ 5.58 shoots per explant on the solid medium, while ~ 16 shoots per explant was obtained in the liquid medium. High-Performance Liquid Chromatography (HPLC) analysis of in vitro shoots grown in the liquid medium produced 9.13 mg/g dry weight (DW) of GPD which is seven-fold higher than that of naturally growing plant shoots. The in vitro protocol for G. kurroo developed in this study may be used for industrial production of GPD.

Optimisation of gentiopicroside production in Gentiana kurroo Royle from adventitious root cultures in a liquid culture system

Gentiana kurroo Royle is a critically endangered medicinal herb of the Indian Himalaya. It has bioactive seco-iridoid glycosides, notably gentiopicroside, in the rhizome and roots. In this study, we report on the in vitro production of gentiopicroside (GPD) from adventitious (AD) roots induced directly from the leaf, nodal, and suspension cell cultures of G. kurroo. Murashige and Skoog (MS) media fortified with Indole-3-butyric acid (IBA) 2 mg L−1 + Naphthaleneacetic acid (NAA) 0.5 mg L−1 and IBA 2 mg L−1 + Indole-3-acetic acid (IAA) 1 mg L−1 produced the maximum number of roots. Suspension cell cultures derived AD roots showed a 1.41-fold higher biomass production than adventitious roots induced from leaf and nodal explants. Among the various concentrations of MS media salts evaluated, half-strength MS suspension media favored the higher biomass and GPD accumulation. The maximum accumulation of GPD (2.58 mg g –l dry weight (DW)) and AD root biomass (18.96 g L−1 DW) was observed on the 48th day. Furthermore, GPD produced from AD root cultures were separated by using TLC, characterized using Nuclear Magnetic Resonance (NMR) and High-Resolution Liquid Chromatograph-Mass Spectrometer (HR LC-MS) and quantified with High-Performance Liquid Chromatography (HPLC).

In-vitro cytotoxicity in relation to chemotypic diversity in diploid and tetraploid populations of Gentiana kurroo Royle

Ethnopharmacological relevanceGentiana kurroo is a multipurpose critically endangered medicinal herb prescribed as medicine in Ayurveda in India and exhibits various pharmacological properties including anti-cancer activity. The species is rich repository of pharmacologically active secondary metabolites together with secoiridoidal glycosides. Aim of the studyThe study aimed to investigate the chemical diversity in different populations/cytotypes prevailing in G. kurroo to identify elite genetic stocks in terms of optimum accumulation/biosynthesis of desired metabolites and having higher in-vitro cytotoxicity potential in relation to chemotypic diversity. Material and methodsThe wild plants of the species were collected from different ranges of altitudes from the Kashmir Himalayas. For cytological evaluation, the standard meiotic analysis was performed. The standard LC-MS/MS technique was employed for phytochemical analysis based on different marker compounds viz. sweroside, swertiamarin, and gentiopicroside. Different tissues such as root-stock, aerial parts, and flowers were used for chemo-profiling. Further, the methanolic extracts of diploid and tetraploid cytotypes were assessed for cytotoxic activity by using MTT assay against four different human cancer cell lines. ResultsThe quantification of major bioactive compounds based on tissue- and location-specific comparison, as well as in-vitro cytotoxic potential among extant cytotypes, was evaluated. The comprehensive cytomorphological studies of the populations from NW Himalayas revealed the occurrence of different chromosomal races viz. n = 13, 26. The tetraploid cytotype was hitherto unreported. The tissue-specific chemo-profiling revealed relative dominance of different phytoconstituents in root-stock. There was a noticeable increase in the quantity of the analyzed compounds in relation to increasing ploidy status along the increasing altitudes. The MTT assay of methanolic extracts of diploid and tetraploid cytotypes displayed significant cytotoxicity potential in tetraploids. The root-stock extracts of tetraploids were highly active extracts with IC50 value ranges from 5.65 to 8.53 μg/mL against HCT-116 colon cancer. ConclusionThe chemical evaluation of major bioactive compounds in diverse cytotypes from different plant parts along different altitudes presented an appreciable variability in sweroside, swertiamarin, and gentiopicroside contents. Additionally, the concentrations of these phytoconstituents varied for cytotoxicity potential among different screened cytotypes. This quantitative difference of active bio-constituents was in correspondence with the growth inhibition percentage of different tested cancer cell lines. Thus, the present investigation strongly alludes towards a prognostic approach for the identification of elite cytotypes/chemotypes with significant pharmacological potential.

Cryopreservation of shoot tips of Gentiana kurroo Royle – a critically endangered medicinal plant of India

Gentiana kurroo Royle, a critically endangered medicinal plant species of India is conserved as in vitro slow growing cultures in the In Vitro Genebank (IVGB) at ICAR-National Bureau of Plant Genetic Resources (NBPGR), New Delhi, India. Shoot tips (about 1 mm in length) excised from 8-weeks-old stock cultures subjected to cold hardening for four weeks at 8 °C (dark) were precultured on Murashige and Skoog (MS) medium supplemented with 5% Dimethylsulfoxide (DMSO) at 8 °C (dark) for two days. Thereafter, shoot tips were dehydrated with PVS2 solution (30 min) at 0 °C and cryopreserved using conventional vitrification (V) and droplet-vitrification (DV) techniques. Average post-thaw regeneration after 4 weeks was ~ 35% by V technique and ~ 60% by DV in all the three accessions tested. Genetic stability assessment on the basis of 40 Inter Simple Sequence Repeats and 30 Expressed Sequence Tagged-Simple Sequence Repeats markers revealed no variation between in vitro-cryopreserved plants and controls, in any of the genotypes. Thus, standardized DV method has the potential for long-term conservation of germplasm of G. kurroo and the three accessions are now safely cryobanked at ICAR-NBPGR. Gentiana kurroo is a critically endangered medicinal plant endemic to North-western Himalayas, India. A protocol to cryopreserve in vitro shoot-tips was developed using droplet vitrification technique with high recovery rates and genetically stable regnerants. The protocol is being applied for cryobanking of Gentiana germplasm at ICAR-NBPGR.

Ecology, distribution mapping and conservation implications of four critically endangered endemic plants of Kashmir Himalaya.

During the last few decades, human-driven activities have led to indiscriminate habitat destruction and exploitation of many plant species in Kashmir Himalaya. As a result, many species have become threatened and are struggling for survival. Of particular concern are the endemic and critically endangered species which have the highest risk of the extinction, hence warranting immediate conservation actions. Therefore the current study was carried out to understand the distribution, ecology and conservation implications of the four critically endangered endemic plants of Kashmir Himalaya. Habitat distribution modelling showed that the suitable potential areas for the species occurred from subalpine to alpine meadowlands with an elevational range of 1500–4600 m asl. The output of the MaxEnt model and field surveys have revealed that their highest potential distribution is in Panchari, Khrew, Ramnagar, Pahalgam, Gurez, Sonamarg, Gulmarg and Kishtwar forest ranges. Based on the field explorations and herbarium records, Saussurea costus (Falc.) Lipsch have 27 distribution areas, Gentiana kurroo Royle 18, Lilium polyphyllum D. Don 12 and Aconitum chasmanthum Stapf have 15. Precipitation of the driest month and annual mean temperature played an important role in the distribution of the studied species. The species started their lifecycle with the onset of the spring season, flowered in summer, fruit in autumn and senesce in the winter season. Under natural conditions, the number of days required for germination ranged from 180 to 210 where cold stratification played a pivotal role. Since last few decades, the populations of these species have been shrinking in their natural habitats due to over-exploitation for medicinal purposes and habitat destruction through amplified humanoid interferences like the expansion of agricultural land, road building, grazing and urbanization. Thus there is an urgent need to come up with positive strategies to save whatever is left and plan long term rescue measures not only to protect these species from extinction but also to reintroduce them along with framing the plans to supply sustained raw materials for medicine.

Conservation and prioritization of threatened plants in Indian Himalayan Region

Assessing the impact of threats, identification, prioritization and predicting conservation pockets are vital for the conservation and management of threatened species. This study investigated the diversity and distribution through inventorization and analysis of International Union for Conservation of Nature (IUCN) categorized threatened plants and their prioritization for conservation. A total of 456 threatened taxa belonging to 233 genera coming under 87 families were recorded as threatened under various threat categories of IUCN. Among lited taxa, 64 species were falls under higher risk of extinction categories of IUCN viz. Critically endangered, Endangered, and Vulnerable with 13, 28 and 23 species respectively. Maximum number of threatened plants was found (84 species) in family Fabaceae followed by Cyperaceae (65 species) and Poaceae (36 species). Richness of threatened plants was found maximum in the Sikkim (203) followed by Himachal Pradesh (190) and Jammu & Kashmir (189). Major causes of depletion of threatened taxa were anthropogenic activities such as habitat loss, overgrazing, deforestation, over-exploitation and unsustainable harvesting of medicinal plants. Therefore, conservation of all the inventorized 64 threatened plants is needful at its earliest. Thus the strategies for the conservation of threatened species must be directed towards those species facing highest risk of extinction, biologically distinct and ecologically valuable for the ecosystem as well having socio-economic significance. Considering this, prioritization of the threatened species was done following existing methodologies. Among the threatened taxa, Angelica glauca, Coptis teeta, Lilium polyphyllum, Nardostachys jatamansi, Aconitum heterophyllum and Gentiana kurroo etc. are ranked at apex, thus needs higher priority for conservation. The study further indicate that six species such as Angelica glauca, Coptis teeta, Gymnocladus assamicus, Illicium griffithii, Piper pedicellatum and Saussurea costus are endemic, having restricted distribution, however highly traded in the IHR. Thus, it is proposed that these species must be taken under special apprehension for conservation planning. Besides this for the conservation of higher risk of extinction species, grid mapping of targeted species was done for identifying specific distribution areas for effective conservation.

Bio-reduction potentials of Gentiana kurroo Royle for single-step bio-genic production of silver nanoparticles

Bio-reduction potentials of Gentiana kurroo Royle for single-step bio-genic production of silver nanoparticles

A novel cost-effective medium for in vitro cultivation of Gentiana kurroo Royle using sugarcane bagasse

Objective: To develop a simple, cost effective and efficient medium by using sugarcane bagasse as a base material to replace the conventional Murashige and Skoog medium. Materials and Methods: Water extracts of sugarcane bagasse alongwith some macronutrients and plant growth regulators were gelled with 0.7% agar-agar powder. Nodal segments of G. kurroo were used as explants and inoculated in the medium and placed in growth chamber under standard conditions of light and temperature. Out of the tested combinations of plant growth regulators, 0.5 mg/l each of KN and BAP showed the excellent shoot multiplication and proliferation rate on the bagasse medium with the same potential as on the MS medium with an average of 5 or - 6 shoots / explant. In vitro rooting was obtained on half strength MS medium supplemented with IBA (0.5 mg/l) with an average length of 7-8 cm and 20-25 roots/ explant. The plants were hardened in a mixture of clay loam and farmyard manure (FYM) in 1:1(w/w) with 70-80% survival rate without any phenotypic abberations. Conclusion: The results from the present investigation indicate that sugarcane bagasse can be used as a cost effective substitute of MS medium for in vitro propagation of G.kurroo.

Somatic embryogenesis and plant regeneration from cell suspension cultures of Gentiana kurroo Royle.

Gentiana kurroo Royle, native to North- Western Himalayas, is one of the critically endangered perennial herbs, which is being overexploited due to its multiple medicinal uses. The goal of this work was to develop a protocol for somatic embryogenesis and efficient plant regeneration through cell suspension culture. Selected ranges of plant growth regulators were experimented for somatic embryogenesis. Light micrographs of the established suspension cell cultures of somatic embryos at different developmental stages were also observed under light microscope. Somatic embryogenesis was induced from the leaf explants and the maximum induction frequency (97.2%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/l each of NAA, BAP and TDZ. Cell suspension cultures were established in MS liquid medium containing IAA (0.5 mg/l) + BAP (1.0 mg/l) with 89-93% viable cells. Transfer of cotyledonary somatic embryos to the MS agar medium containing 2, 4-D (0.4 mg/l) and KN (1.5 mg/l) resulted in somatic embryogenesis at high frequencies (80%) with an average of 28 ± 0.2 embryos/ callus piece. After transfer onto the half strength MS medium without growth regulators approximately 80-90% somatic embryos developed into complete plantlets. The protocol described here could be used for somatic hybridization, genetic transformation, isolation of protoplasts and large-scale propagation of G. kurroo.

Evaluation of antileishmanial potential of Gentiana kurroo Royle by in vitro and in silico methods

Evaluation of antileishmanial potential of Gentiana kurroo Royle by in vitro and in silico methods

Evaluation of antileishmanial potential of Gentiana kurroo Royle by in vitro and in silico methods

Evaluation of antileishmanial potential of Gentiana kurroo Royle by in vitro and in silico methods