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Peppermint (Mentha × piperita L) is a perennial, glabrous and strongly scented herb belongs to the family Lamiaceae. It is cultivated in a temperate region of Europe, Asia, United States, India and Mediterranean countries due to their commercial value and distinct aroma. In addition to traditional food flavouring uses, M. × piperita is well recognized for their traditional use to treat fever, cold, digestive, anti-viral, anti-fungal and oral mucosa and throat inflammation. The scientific studies provide awareness on the use of M. × piperita for biological effects such as anti-oxidant, anti-microbial, anti-viral, anti-inflammatory, biopesticidal, larvicidal, anticancer, radioprotective effect, genotoxicity and anti-diabetic activity have been ascribed. A wide spectrum of bioactive phytochemicals such as flavonoids, phenolics lignans and stilbenes and essential oils are expected to be responsible for the aroma effects. In this sense, this present review provides an extensive overview of the traditional medicinal, phytochemical and multiple biological activities of this "Peppermint." [ABSTRACT FROM AUTHOR]

In traditional, herbal medicine, and aromatherapy, use of essential oils and their aroma compounds have been known since long, for the management of various human diseases. The essential oil is a mixture of highly complex, naturally occurring volatile aroma compounds synthesized by medicinal and aromatic plants as secondary metabolites. Essential oils widely used in pharmaceutical, cosmetic, sanitary, food industry and agriculture for their antibacterial, antiviral, antifungal, antiparasitic, insecticidal, anticancer, neuroprotective, psychophysiological, and anti-aging activities. Moreover, volatile aroma compounds comprise a chemically diverse class of low molecular weight organic compounds with significant vapor pressure. However, aroma compounds produced by plants, mainly attract pollinators, seed dispersers and provide defense against pests or pathogens. However, in humans, about 300 active olfactory receptor genes are involved to detect thousands of different aroma compounds and modulates expression of different metabolic genes regulating human psychophysiological activity, brain function, pharmacological signaling, and therapeutic potential. Keeping in mind this importance, present database, namely, AromaDb (http://bioinfo.cimap.res.in/aromadb/) covers information of plant varieties/chemotypes, essential oils, chemical constituents, GC-MS profile, yield variations due to agromorphological parameters, trade data, aroma compounds, fragrance type, and bioactivity details. The database includes 1,321 aroma chemical structures, bioactivities of essential oil/aroma compounds, 357 fragrance type, 166 commercially used plants, and their high yielding 148 varieties/chemotypes. Also includes calculated cheminformatics properties related to identification, physico-chemical properties, pharmacokinetics, toxicological, and ecological information. Also comprises interacted human genes affecting various diseases related cell signaling pathways correlating the use of aromatherapy. This database could be a useful resource to the plant's growers/producers, an aroma/fragrance industrialist, health professionals, and researchers exploring the potential of essential oils and aroma compounds in the development of novel formulations against human diseases. [ABSTRACT FROM AUTHOR]

The quinolinate phosphoribosyl transferase (QPT) is a key enzyme that converts quinolinic acid into nicotinic acid mononucleotide. The QPT gene plays an essential role in the pyridine nucleotide cycle as well as in the biosynthetic pathway of the alkaloid nicotine. However, a clear role for QPT is yet to be characterized to validate the actual function of this gene in planta. In this study, an RNA interference (RNAi) approach was used to reveal the functional role of QPT. Transformation and analysis of the hairy roots (HRs) of the Nicotiana leaf explants was used, followed by plant regeneration and analysis. High-performance liquid chromatography (HPLC) analysis of the HRs and of the regenerated plants both revealed altered alkaloid biosynthetic cycle, with a substantially reduced content of nicotine and anabasine. The transgenic plants exhibited a significantly altered phenotype and growth pattern. Also, silencing of QPT led to a decrease in chlorophyll content, maximum quantum efficiency of PSII, net CO2 assimilation and starch content. Results clearly demonstrated that QPT was not only involved in the biosynthetic pathway of the alkaloids but also affected plant growth and development. Our results provide information to be considered when trying to engineer the secondary metabolite quality and quantity. [ABSTRACT FROM AUTHOR]

Background: Pyrethrins are monoterpenoids and consist of either a chrysanthemic acid or pyrethric acid with a rethrolone moiety. Natural pyrethrins are safe and eco-friendly while possessing strong insecticidal properties. Despite such advantages of commercial value coming with the eco-friendly tag, most enzymes/genes involved in the pyrethrin biosynthesis pathway remain unidentified and uncharacterized. Since the flowers of Tanacetum cinerariifolium are rich in major pyrethrins, next generation transcriptome sequencing was undertaken to compare the flowers and the leaves of the plant de novo to identify differentially expressed transcripts and ascertain which among them might be involved in and responsible for the differential accumulation of pyrethrins in T. cinerariifolium flowers. Results: In this first tissue specific transcriptome analysis of the non-model plant T. cinerariifolium, a total of 23,200,000 and 28,500,110 high quality Illumina next generation sequence reads, with a length of 101 bp, were generated for the flower and leaf tissue respectively. After functional enrichment analysis and GO based annotation using public protein databases such as UniRef, PFAM, SMART, KEGG and NR, 4443 and 8901 unigenes were identified in the flower and leaf tissue respectively. These could be assigned to 13344 KEGG pathways and the pyrethrin biosynthesis contextualized. The 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway was involved in the biosynthesis of acid moiety of pyrethrin and this pathway predominated in the flowers as compared to the leaves. However, enzymes related to oxylipin biosynthesis were found predominantly in the leaf tissue, which suggested that major steps of pyrethrin biosynthesis occurred in the flowers. Conclusions: Transcriptome comparison between the flower and leaf tissue of T. cinerariifolium provided an elaborate list of tissue specific transcripts that was useful in elucidating the differences in the expression of the biosynthetic pathways leading to differential presence of pyrethrin in the flowers. The information generated on genes, pathways and markers related to pyrethrin biosynthesis in this study will be helpful in enhancing the production of these useful compounds for value added breeding programs. Related proteome comparison to overlay our transcriptome comparison can generate more relevant information to better understand flower specific accumulation of secondary metabolites in general and pyrethrin accumulation in particular. [ABSTRACT FROM AUTHOR]

A field experiment was conducted to study the influence of combined use of farm yard manure and inorganic fertilizers on biomass and essential oil yields, chemical and microbial biomass carbon and nitrogen of soil grown with peppermint (Mentha piperita) in a hilly region, Bageshwar, India. Combined application of both farm yard manure and inorganic fertilizers increased the biomass and oil yields of peppermint compared to sole inorganic fertilizers treatments. The treatments receiving both farm yard manure and inorganic fertilizers improved the soil organic carbon and total N from initial value of 3.9 to 7.0 g kg-1soil and 0.27 to 0.58 g kg-1soil, respectively. A significant improvement has been noticed in soil respiration, soil microbial biomass carbon and nitrogen compared to inorganic fertilizer alone. Moreover, different proportions of inorganic fertilizers with farm yard manure had significant variation on available major soil nutrients. A significant correlation coefficient (r= 0.41* to 0.98**) was observed between yields and soil properties. The quality of essential oil was good and acceptable for the market. [ABSTRACT FROM PUBLISHER]

Vermicompost (VC) produced from distillation waste of geranium (Pelargonium graveolens), farmyard manure (FYM) produced from animal excreta mixed with pine needle (Pinussp.), and biofertilzer (Azotobacter) were utilized for this experiment. The plant growth attributes, biomass, and oil yield of geranium were significantly increased with integrated nutrient supply, and maximum increase was found in T8treatments (N100P60K60+ 5t VC). Soil organic carbon (Corg) significantly increased by 4.2% to 81.8% in T4and T8treatments, respectively, over the control. Data obtained on total nitrogen (Nt) and available N, phosphorus (P), and potassium (K) clearly showed that the integrated nutrient supply considerably improved the soil health and sustainability. The soil respiration and microbial biomass C (Cmic) and N (Nmic) were increased by the manures according to the application rate. The Cmicaccounted for 1.8 to 2.7% of the soil Corgcontent and microbial N accounted for 3.9 to 5.8 % of Ntunder different treatment combinations. [ABSTRACT FROM AUTHOR]

To enhance the production of terpenoid indole alkaloids in Rauwolfia serpentina, Catharanthus tryptophan decarboxylase ( Crtdc) gene was over-expressed in transgenic hairy root cultures using Agrobacterium rhizogenes-mediated transformation. Among six transgenic hairy root lines, line RT4 accumulated the highest alkaloid content, with 0.1202 % dry weight (DW) reserpine and 0.0064 % DW ajmalicine, after 10 weeks of culture. Whereas, wild-type roots accumulated 0.0596 ± 0.003 % DW reserpine and 0.0011 ± 0.001 % DW ajmalicine. Transgenic hairy root line RT7 produced the lowest alkaloid content (reserpine: 0.0896 ± 0.002 % DW; ajmalicine: 0.002 ± 0.0 % DW). On the basis of alkaloid content the six hairy root lines were grouped as RT4/RT2 > RT3/RT5 > RT7/RT8. Analysis of gene expression profile indicated that Crtdc was expressed at a higher level in transgenic lines, which could be correlated with enhanced metabolite accumulation in roots. This study confirms that over-expression of Crtdc is a superlative method to improve the biosynthetic potential of Rauwolfia hairy root cultures. Enhanced reserpine and ajmalicine production can serve as an alternative choice to provide resources for relative pharmaceutical industries. [ABSTRACT FROM AUTHOR]

To determine the seasonal influence on essential oil yield and composition of three rose-scented geranium cultivars, namely Bourbon type, CIM-Pawan and Kelkar, an experiment was carried out in western Himalayan region, India. The essential oil yield varied from 0.05% to 0.20%, from 0.10% to 0.25% and from 0.03% to 0.12% in fresh biomass of the cv. Bourbon type, CIM-Pawan and Kelkar, respectively. Gas chromatography/flame ionization detector (GC/FID) and /mass spectrometry (GC/MS) analyses revealed significant variations in the essential oil composition of rose-scented geranium due to cultivar and season of harvesting. The major components in the essential oil of cv. Bourbon type were geraniol (14.1–34.6%), citronellol (15.2–31.3%), linalool (2.9–9.2%), citronellyl formate (4.4–9.2%), isomenthone (4.5–6.6%), 10-epi-γ-eudesmol (4.7–6.7%) and geranyl formate (3.8–6.2%). The dominant constituents of the cv. CIM-Pawan essential oil were geraniol (11.9–31.9%), citronellol (16.1–30.2%), citronellyl formate (5.2–8.9%), linalool (3.7–6.4%), isomenthone (4.0–6.3%), 10-epi-γ-eudesmol (4.4–5.2%) and geranyl formate (4.3–5.0%). However, the chemical composition and odor of cv. Kelkar was quite different from the other two cultivars and the major components found in this oil were citronellol (51.0–63.4%) and isomenthone (9.8–17.8%). [ABSTRACT FROM PUBLISHER]

Abstract: As part of a study to explore the potential for new or modified bio-product formation, Beta vulgaris (sugar beet) has been genetically modified to express in root-organ culture a bacterial gene of phenylpropanoid catabolism. The HCHL gene, encoding p-hydroxycinnamoyl-CoA hydratase/lyase, was introduced into B. vulgaris under the control of a CaMV 35S promoter, using Agrobacterium rhizogenes LBA 9402. Hairy root clones expressing the HCHL gene, together with non-expressing clones, were analysed and revealed that one expression-positive clone accumulated the glucose ester of p-hydroxybenzoic acid (pHBA) at about 14% on a dry weight basis. This is the best yield achieved in plant systems so far. Determination of cell-wall components liberated by alkaline hydrolysis confirmed that the ratio of pHBA to ferulic acid was considerably higher in the HCHL-expressing clones, whereas only ferulic acid was detected in a non-expressing clone. The change in cell-wall components also resulted in a decrease in tensile strength in the HCHL-expressing clones. [Copyright &y& Elsevier]

Agrobacterium rhizogenes-mediated 'hairy root' cultures were established in Atropa acuminata. The chemical profiling of the hairy roots was carried out by a new mass spectrometric technique, direct analysis in real time (DART). The intact hairy roots were directly analyzed by holding them in the gap between the DART ion source and mass spectrometer. Two alkaloids, atropine and scopolamine, were characterized. The structural confirmation of the two alkaloids was made through their accurate molecular formula determinations. This is the first report of establishing hairy roots in A. acuminata as well as application of the DART technique for the chemical profiling of its hairy roots. Copyright © 2008 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abstract: A protocol has been established for the induction of disease resistance in callus cultures of rose-scented geranium, Pelargonium graveolens cv. Hemanti against leaf blight disease caused by the fungal pathogen Alternaria alternata. The callus cultures were raised and maintained on semi-solid Murashige and Skoog''s (MS) medium supplemented with 10mgl−1 Kn and 1.0mgl−1 NAA. The calli were subjected to various concentrations of culture filtrate (0%, 4%, 8%, 12%, 16% and 20%) obtained from A. alternata. Resistant calli were selected and placed on regeneration medium (MS supplemented with 0.5mgl−1 BAP and 0.1mgl−1 NAA). The regenerants were confirmed for A. alternata resistance by exposing their leaves to the same concentrations of culture filtrate as used previously. While the parental wild type demonstrated typical susceptibility, the leaves of putative resistant clones remained green and viable in the presence of toxin and regenerated shoots directly on the toxin-free regeneration medium (MS supplemented with 5mgl−1 Kn and lmgl−1 NAA). The above experiment demonstrated the induction of disease resistance in rose-scented geranium plants at the cellular level. This approach could be successfully exploited in raising new disease-resistant cultivars in geranium against various fungal pathogens. [Copyright &y& Elsevier]

In the present investigation, pollen morphology and viability of four Ocimum L. species viz. O. sanctum (CIM-Angana), O. basilicum (CIM-Saumya), O. gratissimum, and O. kilimandscharicum were studied. Morphological characters were observed using scanning electron and light microscopy. Pollen of all the four studied Ocimum species have hexacolpate apertures with varied aperture length and width. The exine ornamentation was reticulate and bi-reticulate. O. basilicum has large size (53.9 µm) pollen grains and rest three have medium sized pollen. The viability of pollen was assessed through staining dyes such as 2,5-diphenyl tetrazolium bromide (MTT) and 2,3,5-triphenyl tetrazolium chloride (TTC) and in vitro pollen germination method. O. basilicum possessed maximum viability (92.4%) whereas least viability was found in O. sanctum (38.2%). The maximum pollen germination percentage and tube growth were observed in the semisolid medium gelled with phytagel supplemented with 15% sucrose, 15% PEG, 6.4 mM boric acid and 7.3 mM calcium nitrate. O. basilicum showed the highest germination percentage (90.3%) followed by O. kilimandscharicum (57.1%), O. gratissimum (42.5%) and O. sanctum (25.5%). Pollen of the four Ocimum species were stored at different temperatures viz. 37 °C, + 4 °C, 0 °C, − 20 °C, and − 80 °C for 75 days. The highest pollen germinability of all the four Ocimum species was observed at − 20 °C. This study can be helpful in taxonomic identification of Ocimum species and further utilized in conservation and improvement of different Ocimum species by cross-hybridization breeding programmes. • Pollen morphology and viability of four Ocimum species were studied. • All the four studied Ocimum species have hexacolpate apertures. • Pollen viability was done by staining and in vitro pollen germination method. • O. basilicum have largest size with maximum viability and germination percentage. • Pollen grains of four Ocimum species were stored at different temperatures. [ABSTRACT FROM AUTHOR]

Abstract: Agrobacterium rhizogenes induced hairy root cultures are entering into a new juncture of functional research in generating pharmaceutical lead compounds by bringing about chemical transformations aided through its inherent enzyme resources. Rational utilization of hairy root cultures as highly effective biotransformation systems has come into existence in the last twenty years involving a wide range of plant systems as well as exogenous substrates and diverse chemical reactions. To date, hairy root cultures are preferred over plant cell/callus and suspension cultures as biocatalyst due to their genetic/biochemical stability, hormone-autotrophy, multi-enzyme biosynthetic potential mimicking that of the parent plants and relatively low-cost cultural requirements. The resultant biotransformed molecules, that are difficult to make by synthetic organic chemistry, can unearth notable practical efficacies by acquiring improved physico-chemical properties, bioavailability, lower toxicity and broader therapeutic properties. The present review summarizes the overall reported advances made in the area of hairy root mediated biotransformation of exogenous substrates with regard to their reaction types, plant systems associated, bacterial strains/molecules involved and final product recovery. [Copyright &y& Elsevier]

Abstract: Hyoscyamus albus hairy roots with/without an exogenous gene (11 clones) were established by inoculation of Agrobacterium rhizogenes. All clones cultured under iron-deficient condition secreted riboflavin from the root tips into the culture medium and the productivity depended on the number and size of root tips among the clones. A decline of pH was observed before riboflavin production and root development. By studying effects of proton-pump inhibitors, medium acidification with external organic acid, and riboflavin addition upon pH change and riboflavin productivity, we indicate that riboflavin efflux is not directly connected to active pH reduction, and more significantly active riboflavin secretion occurs as a response to an internal requirement in H. albus hairy roots under iron deficiency. [Copyright &y& Elsevier]

Vanillin is a naturally occurring phenolic compound, which has broad spectrum importance in food, cosmetics, flavouring agent and bioprservative. Aditionally, it also imparts many therapeutic benfits in the form of antimicrobial, antimutagenic and anticarcinogenic agent. Present study demonstrated the production of vanillin from two transgenic hairy root (HR) lines of Beta vulgaris bearing HCHL gene from Pseudomonas fluorescens , encoding p -hydroxycinnamoyl-CoA hydratase/lyase under the control of CaMV 35S promoter. These HCHL expressing HR clones viz . Bv 42 and Bv 18, showed conversion of inherently available phenypropanoid precursor (ferulic acid) into vanillin. Optimization of central parameters, such as media formulation, concentration of sucrose (1–5%), pH of the media (17), culture duration in combination with accessibility to higher amount of precursor by feeding with ferulic acid (2–10 mM), was carried out in detail, which have noticeably influenced the growth as well as vanillin synthesis potential of the roots. The HR clone Bv 42 showed maximum growth (GI = 1426 ± 22.71) in MS medium with 3% sucrose at pH 5.8 within 45 days, which was two times and five times higher than that with pH 7 and pH 3 respectively. Evidently, although the maximum synthesis of vanillin (0.052% ± 0.00013) could be obtained at pH 3 without any additional precursor feeding, its overall yield was better at 5.8 pH, which may be due to retardation in biomass yield as observed at pH 3. Conspicuously, addition of 10 mM of ferulic acid as the precursor enhanced the vanillin yield upto four fold (0.209% ± 0.0021), relative to that in the cultures devoid of any added precursor. The presently established transgenic HR mediated vanillin production technology adds much significance in realising the industrial goal of meeting the global demand of this biomolecule. [ABSTRACT FROM AUTHOR]

Ocimum gratissimum is renowned for its immense role in traditional, ayurvedic and unani system of medicines. The essential oil has a plethora of active ingredient like triterpenes, polyphenols and phenylpropanoids. The active ingredients claim most of the properties like anticancerous, anti HIV, antipyretic, antinociceptive, antiurolithiatic etc. Screening of biologically active ingredients from this valuable plant may provide us the basis to produce novel pharmaceutically important drugs and medicines. Considering the importance of this plant, it is the need to have a well developed regeneration and transformation protocol. The present report focuses on the development of an efficient protocol for direct regeneration and Agrobacterium mediated genetic transformation of O. gratissimum . CN; when inoculated on MS + 1.5 mg/L + 0.5 mg/L + 0.5 mg/L KN medium has shown the best response with a regeneration frequency of 98% ± 0.4. A. tumefaciens LBA4404 containing binary vector pBI121, harboring gus - A reporter gene with intron under the transcriptional control of Cauliflower Mosaic Virus (CaMV) 35S promoter and neomycin phosphotransferase ( npt-II) as selection marker genes, to achieve maximum transformation frequency (20% ± 0.7). To carry out transformation, parameters like O.D., infection duration, concentration of AS, surfactant, (O.D 600 -0.6, 15 min, 300 μM, 0.01%) were standardized . The putative transformants were screened on SMM having 50 mg/L kanamycin and subsequently rooted on ½ MS. The confirmation was done through PCR. The established regeneration and transformation protocol could be used as a tool to elucidate the functional genomics of important/novel compound which can be used in the preparation of therapeutic formulations for future. [ABSTRACT FROM AUTHOR]

Swertia chirayita (Roxb.) H. Karst. is a medicinal herb used in traditional medicines. Scientific authentications of its multiple health benefits against several diseases have incessantly increased its commercial demand in the domestic and international market. As a result, alternative strategies for conventional cultivation/collection are necessary to eliminate the huge gap between demand and supply. Therefore, the goal of this work was to design an efficient hairy root induction protocol and to explore the effects of methyl jasmonate (MJ), sodium nitroprusside (SNP), and salicylic acid (SA) on the improvement of swerchirin (1,8-dihydroxy-3,5-dimethoxyxanthone) and 1,2,5,6-tetrahydroxyxanthone in S. chirayita hairy roots. To produce hairy roots, in vitro -cultured leaf explants were infected Rhizobium rhizogenes LBA 9402 with different cell density, infection time, and acetosyringone concentrations. The highest root induction frequency (87.50 ± 4.94%) and better hairy roots were observed at bacterial cell density (OD 600 = 0.8) + 20 min infection time along with 200 µM of acetosyringone. The highest hairy root biomass (222.00 ± 8.33 g/L fresh weight (FW) and 21.64 ± 0.83 g/L dry weight (DW) were observed after 60 days. The highest content of swerchirin (0.710 ± 0.13 mg/g DW, 6th days after elicitation) and 1,2,5,6-tetrahydroxyxanthone (5.501 ± 0.73 mg/g, 6th days after elicitation) was displayed in 100 μM MJ experiment which was 1.80 and 6.0-fold increase than the control. Hence, an efficient developed hairy root protocol of S. chirayita is cost-effective and could be efficiently used for large-scale industrial manufacture of bioactive xanthones. [Display omitted] • Establishment of an efficient hairy root induction protocol for Swertia chirayita. • The bacterial density, infection time and acetosyringone influenced the hairy root induction. • Swerchirin and 1,2,5,6-tetrahydroxyxanthone were enhanced by methyl jasmonate. • 50 µM of salicylic acid increased 4.8-fold 1,2,5,6-tetrahydroxyxanthone content. [ABSTRACT FROM AUTHOR]

Swertia chirayita (Roxb.) H. Karst. is a well-documented traditional plant used to cure liver diseases, malaria fever, jaundice, hepatitis and diabetes. Xanthones are produced by the plant and provide a variety of health benefits. The goal of this research is to develop an efficient and reliable approach for hairy root induction and to explore the biosynthesis of xanthones in hairy root lines. Infection of S. chirayita leaves with Rhizobium rhizogenes strain (LBA 9402) resulted in distinct hairy root lines. The rolB gene integration in the root was validated by polymerase chain reaction (PCR) in four hairy root lines. Hairy root line SA produced the maximum biomass of 13.32 ± 0.50 g/flask FW (fresh weight) and 1.29 ± 0.05 g/flask DW (dry weight) after 60 days. A novel 1,2,5,6-tetrahydroxyxanthone (2) was isolated and characterized from SA hairy root line using flash chromatography. Swerchirin (1) (0.382 ± 0.089 mg/g DW) and 1,2,5,6-tetrahydroxyxanthone (2) (0.935 ± 0.02 mg/g DW) along with swertianin (0.0195 ± 0.007 mg/g DW) were accumulated higher amount in SA line. 1,2,5,6-tetrahydroxyxanthone (2) exhibited strong antioxidant activity in DPPH (IC 50 = 1.60 ± 0.004 µg/mL) and FRAP (4.83 ± 0.02 Eq. to FeSO 4) assay. Further, swerchirin (1) exhibited anti-proliferative activity by suppressing the growth of MDA-MB-231 cell line with IC 50 of 18.82 μg/mL (65.83% at 50 μg/mL) and 1,2,5,6-tetrahydroxyxanthone (2) inhibited the growth of K562 cell line with IC 50 of 22.83 μg/mL (67.82% at 50 μg/mL). The characterized compounds with promising biological activity could be taken up for its commercial production. [Display omitted] • An efficient hairy root induction protocol of Swertia chirayita was developed. • Hairy root SA line produced the maximum biomass after 60 days. • A novel xanthone was characterized from hairy root SA line by flash column chromatography. • Higher production of swerchirin, 1,2,5,6-tetrahydroxyxanthone and swertianin in hairy root SA line. • Swerchirin and 1,2,5,6-tetrahydroxyxanthone showed strong antioxidant and anti-proliferative activity. [ABSTRACT FROM AUTHOR]

Abstract: Curcuma amada Roxb. (family Zingiberaceae), is gaining global attention as potential source of new drug molecule(s) as it is credited with compounds having remarkable pharmacological properties. Recognizing the risk of loss of genetic variability of C. amada in response to imprudent exploitation of the natural resources coupled with unsystematic cultivation practices, preservation of its genetic resources has attracted the current research attention. The present study exploited the multifarious properties of Luffa sponge (i.e. sound physical strength, stability in texture/shape over wide pH range and repeated autoclaving, cost effective availability, etc.) as a novel matrix for synthetic seed conservation. In this study, we have evaluated the consequences of the presence of two fungicides, namely – bavistin and rosebengal, in the gel matrix as well as in the Luffa sponge for conservation of contamination free germplasm of C. amada. A maximum of 50% and 75% regrowth could be recorded amongst rosebengal and bavistin containing synthetic seeds respectively after 4 weeks of plating on plant growth regulator-free MS medium. Further growth of the encapsulated microshoots was significantly impeded by the presence of rosebengal within the synthetic seeds. On the contrary, the bavistin containing synthetic seeds demonstrated five times better regeneration ability to contamination-free shoots. A total of 78% survival was achieved upon transfer of these healthy plantlets to glass-house. RAPD fingerprinting revealed 84.62% genetic similarity between randomly selected synthetic seed derived plantlets. This report strengthened the vital conservation approach of C. amada using inexpensive Luffa sponge as storage matrix and bavistin for eradication of contaminations. [Copyright &y& Elsevier]

• Enhanced production of gymnemic acids through cell suspension culture of Gymnema sylvestre. • Callus and suspension culture established at 3.0 mg/ L (2,4-dichlorophenoxy acetic) + 1.0 mg / L (Kinetin). • Sodium nitroprusside treatments enhanced cell biomass production in suspension culture. • Highest content of deacylgymnemic acid and gymnemic acid XVII increased by sodium nitroprusside treatments. • 50 μM salicylic acid enhanced 4.12-fold gymnemagenin content. Gymnemic Acids (GAs) belong to the class of triterpenoid saponins present majorly in the leaves of Gymnema sylvestre. These bioactive compounds are responsible for hypoglycemic activity. Therefore, it is expansively used in Ayurveda treatment for diabetes. This study is aimed to develop an effective method for suspension culture and to investigate the effect of sodium nitroprusside (SNP), salicylic acid (SA) and methyl jasmonate (MeJA) on the enhancement of deacylgymnemic acid, gymnemagenin, gymnemic acid IV and gymnemic acid XVII in G. sylvestre cell suspension culture. To induce callus, in vitro -derived leaf explants were cultivated on Murashige and Skoog's (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or α-naphthalene-acetic acid (NAA) either alone or in combination with 6-benzylaminopurine (BAP) or kinetin (Kin) at 1.0 to 5.0 mg/L. The highest callus initiation frequency (95.30 ± 1.40%) and better friable callus observed on MS medium supplemented with 3.0 mg/L (2,4-D) + 1.0 mg/L (Kin). Optimized callus induction medium was used to develop cell suspension culture, growth kinetic study, elicitors treatment and quantify the production of major bioactive compounds. At 40 days, the highest fresh cell suspension culture biomass was 339.08 g/L fresh weight (FW). Biomass production, 366.68 g/L FW and 30.30 g/L DW were the highest in the treatment of 20 μM SNP among the tested elicitors. The highest accumulation of deacylgymnemic acid (2936.90 μg/g DW, 96 h treatment) and gymnemic acid XVII (2414.20 μg/g dry weight (DW), 48 h treatment) were obtained in 20 μM SNP treatments which were 6.1 and 5.2 fold higher than the control. Furthermore, the highest values of gymnemagenin (1179.77 μg/g DW at 50 μM treatment) and gymnemic acid IV (731.04 μg/g DW at 10 μM treatment) were obtained in SA treatment which was 4.12 and 5.11 fold greater than the control was studied through our newly developed HPLC method. The findings of the present study showed that the SNP could be a good strategy for large-scale industrial production of triterpenoid saponins in G. sylvestre cell suspension cultures. [ABSTRACT FROM AUTHOR]