Archana Mathur

Saponin production in callus and cell suspension cultures of Panax quinquefolium

Abstract Callus and cell suspension cultures of American ginseng (Panax quinquefolium) were compared for growth and in vitro ginsenoside production over a 35-day culture cycle on modified Murashige and Skoogs medium. A time course study at five day intervals revealed that biomass yield in suspension and callus cultures was maximal on the 25th and 30th day of growth, respectively. Both types of cultures were able to produce ginsenosides in amounts and quality comparable to the cultivated plants. TLC-densitometry and HPLC analyses of the crude ginsenosides revealed that yield and relative distribution of different fractions belonging to the Rb and Rg groups of ginsenosides were greatly influenced by culture age. For the Rb group components, 25-day-old callus or suspension cultures were the best source of these compounds, while for the Rg group fractions 30–35-day-old cell cultures gave the maximum yield. Appreciable amounts of ginsenosides, particularly Rg1, were found to leach out in the culture medium of 30–35-day-old suspension cultures.

Screening and evaluation of agronomically useful somaclonal variations in Japanese mint (Mentha arvensis L.)

SummaryA procedure has been standardized for high frequency plant regeneration response from nodal explant cultures of Mentha arvensis Linn. var. piperascens Holmes. Murashige and Skoogs medium supplemented with IAA or NAA (0.5–2.0 mgl−1) alone, supported axillary shoot elongation while BAP (2.0–3.0 mgl−1) alongwith IAA (1.0 mgl−1) supported multiple shoot production. In vitro-derived shoots readily developed roots when cultured on NAA (1.0 mgl−1) fortified MS medium. Regenerated plantlets were successfully transferred to glasshouse (90–95% survival rate) and ultimately to the field. Among 280 plants transferred to the field a wide range of variation was observed for various agronomic traits i.e. plant height (32.0–92.0 cm), leaf-stem weight ratio (0.53–2.32), herb yield (105.0–870.0 g), oil content (0.32–1.10%) and oil yield (0.66–5.22 ml/plant). In addition, variations were also recorded for four major constituents of the essential oil i.e. menthol (65.2–94.77%), menthone (1.40–20.89%), isomenthone (0.96–5.14%) and menthyl acetate (0.75–8.52%). A positive correlation is found for oil yield with plant height and herb yield, whereas a negative correlation exists between herb yield and oil content. Based on the initial agronomic assessments on individual plant basis, 27 somaclones were selected and further evaluated in a replicated plant to row trial with parent plant CIMAP/Hy-77 as standard check. Somaclones Sc 59 and Sc 179, selected on the basis of higher herb yield in the initial screening, recorded 55.8% and 64.3% increase in oil yield over the control, respectively. Somaclones Sc 93, Sc 114, Sc 121 and Sc 124 that were selected for their better oil content exhibited 47.2%, 50.6%, 57.5% and 48.2% increase in oil yield over the parent variety, respectively. The performance of these clones in evaluation trials is discussed in relation to the possibility of genetic improvement of mints through somaclonal breeding.

Differential morphogenetic responses, ginsenoside metabolism and RAPD patterns of three Panax species

Genetic and metabolomic demarcations between two Indian and one American congeners of Genus Panax have been discerned. Genomic DNA was isolated from the root derived callus cultures of these species and amplified by AP-PCR. RAPD analyses of the DNA with six most responding arbitrary oligonucleotide decamers provided a total of 70 reproducible bands for computation of the similarity matrix amongst the Panax species. Only 18 of these were monomorphic giving an estimate of about 74% polymorphism among the test species examined. The similarity coefficient values based on the amplification pattern support an equidistant position of the three test species. The molecular demarcations between the species are also manifested in terms of their characteristic cultural requirements, in vitro growth kinetics, regeneration competence and ginsenoside complement of their calli. The Indian congeners i.e. P. sikkimensis and P. pseudoginseng were distinguishable by higher proportions of ginsenoside Rf and Ro (40% and 20%, respectively) in the crude saponin fractions. Furthermore P. quinquefolium calli mainly accumulated ginsenoside Rb2 and Rg1, whilst P. sikkimensis callus was rich in Rd fraction. P. quinquefolium showed high similarity with P. sikkimensis with respect to plasticity and totipotency for somatic embryogeny whereas P. pseudoginseng callus was highly recalcitrant and lacked regenerative capacity. The chemical and genetic fingerprints alongwith morphogenetic responses of the three species under controlled in vitro environment strongly advance the case of P. sikkimensis as an independent species, rather than a conglomerate of location specific variety or sub-species of P. pseudoginseng. The findings are also of relevance to formulations and evaluation of ginseng-based health foods.

In Vitro Conservation of Twenty-Three Overexploited Medicinal Plants Belonging to the Indian Sub Continent

Twenty-three pharmaceutically important plants, namely, Elaeocarpus spharicus, Rheum emodi, Indigofera tinctoria, Picrorrhiza kurroa, Bergenia ciliata, Lavandula officinalis, Valeriana wallichii, Coleus forskohlii, Gentiana kurroo, Saussurea lappa, Stevia rebaudiana, Acorus calamus, Pyrethrum cinerariaefolium, Aloe vera, Bacopa monnieri, Salvia sclarea, Glycyrrhiza glabra, Swertia cordata, Psoralea corylifolia, Jurinea mollis, Ocimum sanctum, Paris polyphylla, and Papaver somniferum, which are at the verge of being endangered due to their overexploitation and collection from the wild, were successfully established in vitro. Collections were made from the different biodiversity zones of India including Western Himalaya, Northeast Himalaya, Gangetic plain, Western Ghats, Semiarid Zone, and Central Highlands. Aseptic cultures were raised at the morphogenic level of callus, suspension, axillary shoot, multiple shoot, and rooted plants. Synseeds were also produced from highly proliferating shoot cultures of Bacopa monnieri, Glycyrrhiza glabra, Stevia rebaudiana, Valeriana wallichii, Gentiana kurroo, Lavandula officinalis, and Papaver somniferum. In vitro flowering was observed in Papaver somniferum, Psoralea corylifolia, and Ocimum sanctum shoots cultures. Out of 23 plants, 18 plants were successfully hardened under glasshouse conditions.

Establishment and multiplication of colchi-autotetraploids of Rauvolfia serpentina L. Benth. ex Kurz. through tissue culture

A tissue culture procedure was developed for the establishment and propagation of a colchi-autotetraploid of Rauvolfia serpentina for possible commercial exploitation. Multiplication of autotetraploid shoots was obtained either through axillary bud elongation on Murashige and Skoog [1] medium (MS) containing 2.65 μM (0.5 mgl−1) α-naphthaleneacetic acid and 0.33 μM (0.05 mgl−1) kinetin, or via multiple shoot formation on MS medium supplemented with 4.44 μM (1.0 mgl−1) 6-benzylaminopurine and 0.53 μM (0.1 mgl−1) α-naphthaleneacetic acid. Rooting could be induced by transferring the shoots to MS medium containing 7.95 μM (1.5 mgl−1) α-naphthaleneacetic acid alone. The plantlets, thus formed, were tetraploid in nature by cytological observations of the root tips. They exhibited 80–90% success in establishment under glass house and field conditions.

Biotransformation of artemisinin derivatives by Glycyrrhiza glabra , Lavandula officinalis , and Panax quinquefolium

Biotransformation of α-artemether and dihydroartemisinin (DHA) by Glycyrrhiza glabra (Linn.), Lavandula officinalis (L.), and Panax quinquefolium was investigated. Two metabolites: tetrahydrofuran derivative (3) and a 13-carbon ring-rearranged product (4) were produced from α-artemether (1). DHA (2) provided metabolite 4. The structure of the metabolites were characterized by proton (1H) and carbon (13C) nuclear magnetic resonance (NMR) imaging, fourier transform infrared spectroscopy, and mass spectroscopy. This is the first report that G. glabra and L. officinalis have the capability to biotransform α-artemether, and P. quinquefolium to biotransform DHA. Metabolite 3 is a new compound and metabolite 4 is reported here for the first time from artemisinin derivatives 1 and 2. The presence of acetate function in the derivative 3 and hydroxyl and C-12 deoxo groups in 4 obtained in our study make them interesting synthones for further modification into new clinically potent molecules.

Solvent-based extraction optimisation for efficient ultrasonication-assisted ginsenoside recovery from Panax quinquefolius and P. sikkimensis cell suspension lines

The present study aims at developing an extraction protocol for efficient ginsenoside recovery from cell suspensions of Panax quinquefolius and P. sikkimensis. Methanol (100%, 70% and 30%), water (40°C, 90°C), water-saturated butanol and butanol-saturated water were compared for their ultrasonication-assisted ginsenoside retrieval efficacy. HPLC and HP-TLC analysis revealed 100% methanol as the best solvent for maximum retrieval of Rb (diol) and Rg (triol) ginsenosides (P. quinquefolius: Rb: 0.189, Rg: 3.163 mg/g DW; P. sikkimensis: Rb: 0.245, Rg: 4.073 mg/g DW), followed by water (90°C). Methanolic solutions, especially 70%, proved to be significant retrievers of Rg1 (1.812 and 1.327 mg/g DW in P. quinquefolius and P. sikkimensis), with poor Re recovery (0.328 and 0.342 mg/g DW). Water-saturated butanol also led to significant ginsenoside extraction (72.4% of content extracted by methanol), selectively in P. quinquefolius, with a less than 50% of total content extracted by methanol, in P. sikkimensis.

A literature update elucidating production of Panax ginsenosides with a special focus on strategies enriching the anti-neoplastic minor ginsenosides in ginseng preparations

Ginseng, an oriental gift to the world of healthcare and preventive medicine, is among the top ten medicinal herbs globally. The constitutive triterpene saponins, ginsenosides, or panaxosides are attributed to ginseng’s miraculous efficacy towards anti-aging, rejuvenating, and immune-potentiating benefits. The major ginsenosides such as Rb1, Rb2, Rc, Rd., Re, and Rg1, formed after extensive glycosylations of the aglycone “dammaranediol,” dominate the chemical profile of this genus in vivo and in vitro. Elicitations have successfully led to appreciable enhancements in the production of these major ginsenosides. However, current research on ginseng biotechnology has been focusing on the enrichment or production of the minor ginsenosides (the less glycosylated precursors of the major ginsenosides) in ginseng preparations, which are either absent or are produced in very low amounts in nature or via cell cultures. The minor ginsenosides under current scientific scrutiny include diol ginsenosides such as Rg3, Rh2, compound K, and triol ginsenosides Rg2 and Rh1, which are being touted as the next “anti-neoplastic pharmacophores,” with better bioavailability and potency as compared to the major ginsenosides. This review aims at describing the strategies for ginsenoside production with special attention towards production of the minor ginsenosides from the major ginsenosides via microbial biotransformation, elicitations, and from heterologous expression systems.

Saponin Production by Cell/Callus Cultures of Panax Species

Roots of ginseng (“Elixir of Life”) have long been recognised as a miraculous herbal medicine to combat ageing and nervous stresses in the Orient [1]. Ginseng is characterised by the presence of a group of triterpene glycosidic saponins — the ginsenosides [2]. Owing to the strong immuno-modulatory, adaptogenic and aphrodisiac actions of these saponins, ginseng roots are the fourth largest selling health care product in the international market today [3]. Priced at around 1500 US

Emerging trends in research on spatial and temporal organization of terpenoid indole alkaloid pathway in Catharanthus roseus: a literature update.

per kg, the annual demand of Panax roots is estimated to be about 40 thousand tons. Indian pharmaceutical companies alone import 400–500 tons of ginseng roots annually. The bulk of this market demand is met by the sale of 4–7 year old roots of P. ginseng (Korean ginseng) and P. quinquefolium (American ginseng). The agricultural production of ginseng is extremely slow and poses great demands on labour, soil and climate [1, 4]. This, with the backdrop of their increasing industrial demand and soaring price, have made Panax species a forerunner for applying modern biotechnological tools to enhance ginsenoside production. Therefore, employment of cell and tissue cultures for the in vitro synthesis of ginsenosides has been actively pursued in many laboratories [2–10].