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Volume 5 Number 2 2011
CONTENTS AND ABSTRACTS
Anna P. K. Ling, S. L. Ong, H. Sobri (Malaysia) Strategies in Enhancing Secondary Metabolites Production in Plant Cell Cultures (pp 94-101)
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Review: Conventionally, pharmaceutically important secondary metabolites such as flavonoids were extracted directly from whole plants collected from the wild. This conventional method is not cost effective and could even lead to extinction of some endangered plant species. Furthermore, the production of uniform quantity and quality secondary metabolites will be affected as their production is greatly influenced by geographical, seasonal and environmental variations. Biotechnological approaches, specifically, plant tissue culture techniques, have been considered as an attractive solution to the problems of extracting secondary metabolites for industrial applications. Nevertheless, the commercial implementation of pilot scale plant cell suspensions for chemical production is still in the development stage with a few exceptional cases. The major setback is the failure of cell cultures to accumulate significant amounts of secondary metabolites compared to whole plants or organ cultures. Even so, several strategies can be applied in order to substantially increase the yields of secondary metabolites in plant cell cultures. This paper discussed the strategies of nutrient manipulation, precursor feeding and elicitation in enhancing the production of secondary metabolites.
Deepak Ganjewala, Ashwani Kumar Srivastava (India) Recent Progress on Chemical Composition and Bioactivities of Bacopa monnieri (Linn.) a Plant of Ayurveda (pp 102-108)
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Mini-Review: Bacopa monnieri (Linn.) is a highly regarded medicinal plant in Indian traditional Ayurvedic medicine for centuries owing to its broad spectrum bioactivities. The biological effects of B. monnieri are documented in traditional as well as scientific literature. The plant possess many important bioactivities like memory enhancing, anti-oxidant, anti-inflammatory, analgesic, antipyretic, hepatoprotective, sedative, antiepileptic and still many more are constantly being discovered. An ample amount of research on B. monnieri and its major constituents has unraveled its tremendous bioactive potential in the treatment of many serious disorders viz. Alzheimer’s disease, cognitive functions, memory impairment, hepatic carcinoma and cigarette smoking-associated diseases. Since 2005, when the last review on B. monnieri was written, many more new bioactivities have been discovered, although they have not been compiled in one publication. In the present article we describe many new useful bioactivities of B. monnieri and its active chemical constituent bacoside and new chemical constituents that have been isolated and characterized.
Ashutosh Mukherjee (India) Active Compounds from Ginger as Inducers of Mitochondrial Apoptotic Pathway: An in Silico Prediction (pp 109-113)
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Original Research Paper: Knowledge about the mitochondrial apoptotic pathway is very important for chemotherapeutic drug development. The enzyme NADH dehydrogenase can be an important target for natural drugs in this respect. Active compounds of ginger (Zingiber officinale Roscoe) namely gingerol, paradol, shogaol and zingerone were selected as ginger is known to have anti-cancer properties. A three-dimensional model of the human NADH dehydrogenase was developed in silico and docked with gingerol, paradol, shogaol and zingerone. All of them were successfully docked. Among them, two compounds, namely 6-gingerol and 10-gingerol, formed covalent bonds with the enzyme showing that they are the most potent for drug development among the active components of ginger. The successful docking of these natural compounds with the enzyme showed that these compounds are potential inducers of the apoptotic pathway and thus important in chemotherapeutic drug development.
Mohammad J. Saharkhiz, Mohammad Merikhi, Mehdi Zarei (Iran), Jaime A. Teixeira da Silva (Japan) Responses of Ocimum sanctum to Inoculation with Arbuscular Mycorrhizal Fungi and Fertilization with Different Phosphate Sources (pp 114-118)
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Original Research Paper: Arbuscular mycorrhizal fungi (AMF) can change some morphological and physiological characteristics of host plants. A pot experiment was conducted to evaluate the responses of holy basil (Ocimum sanctum) to the inoculation with AMF species (Glomus mosseae and Glomus versiforme) and the use of two different phosphate sources (superphosphate and rock phosphate) as phosphorus fertilizers in a calcareous soil. After a growth period of three months, morphological parameters, root colonization, phosphorus concentration and uptake, chlorophyll content, and the yield of essential oil were measured. AMF significantly (P ≤ 0.05) increased shoot biomass, flower stem length, chlorophyll content, phosphorus concentration and uptake, and root colonization. Phosphorus fertilizers increased all factors except for root colonization compared with the control. Superphosphate was more effective than rock phosphate and control treatments. A synergistic relationship between AMF and phosphorus fertilizers for improving phosphorus uptake and morphological characteristics was observed. Maximum shoot biomass, phosphorus concentration and uptake, as well as essential oil yield were observed when G. versiforme was combined with superphosphate. Rock phosphate combined with inoculation with G. versiforme showed a more positively significant (P ≤ 0.05) effect on all measured traits compared to a single application of rock phosphate. In conclusion, the introduction of AM biofertilizer together with the balanced application of P-fertilizers will be helpful in O. sanctum production.
Artur Manukyan (Germany) Effect of Growing Factors on Productivity and Quality of Lemon Catmint, Lemon Balm and Sage under Soilless Greenhouse Production: I. Drought Stress (pp 119-125)
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Original Research Paper: The formation of bioactive constituents of plants depends on the actual environmental and on growing factors. Controlled environment technology ensures the application of specific stresses, such as drought stress, that can optimize the production of secondary plant metabolites by inducing natural biochemical changes in plants. The impact of drought stress on herbal yield, content and composition of essential oils and polyphenols, as well as on antioxidative capacity of polyphenol-rich extracts of lemon catmint (Nepeta cataria L. f. citriodora), lemon balm (Melissa officinalis L.) and sage (Salvia officinalis L.) was studied under soilless greenhouse cultivation. It was found that the tested substrate moistures (50, 125, 250 hPa) had a considerable effect on herbal yield of lemon catmint, lemon balm and sage with maximum raw medicinal material at 50 hPa. Drought stress influenced the essential oil content of lemon catmint and lemon balm, but not of sage, with 250 hPa resulting higher amount of essential oils. 50 hPa provided high yield of essential oil for all three species. The essential oil composition of studied herbs was mainly affected by tested substrate moistures. The influence of drought stress was also significant for the content of polyphenols in lemon balm and sage, but not in lemon catmint. High drought stress (250 hPa) affected positively polyphenolic content in lemon balm, while in sage polyphenols reached maximum at 125 hPa. HPLC analysis of polyphenols showed differences in sensitivity of main polyphenols in studied herbs to drought stress factor. Polyphenol-rich extracts of lemon catmint and lemon balm showed no differences in antioxidative capacity by the ABTS system, while for sage there were small differences.
Artur Manukyan (Germany) Effect of Growing Factors on Productivity and Quality of Lemon Catmint, Lemon Balm and Sage under Soilless Greenhouse Production: II. Nitrogen Stress (pp 126-132)
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Original Research Paper: Various environmental and growing factors can affect not only the primary, but also the secondary metabolism in herbs. Nitrogen stress considered to be one of the main environmental factors influencing the plant growth and quality. This study investigated the impact of different nitrogen concentrations in nutrient solution (50, 150 and 300 mg/l) on herbal yield, content and composition of essential oils and polyphenols, as well as on antioxidative capacity of polyphenol-rich extracts of lemon catmint (Nepeta cataria L. f. citriodora), lemon balm (Melissa officinalis L.) and sage (Salvia officinalis L.) under soilless greenhouse cultivation. It was found that high nitrogen concentration of 300 mg/l was effective for herbal yield of lemon catmint (52.2 g/plant raw medicinal material). By contrast, low nitrogen concentration of 50 mg/l was positive for herbal productivity of sage (21.1 g/plant raw medicinal material); while for lemon balm both 50 and 150 mg/l concentrations provided maximum results (22.9, 22.1 g/plant raw medicinal material, respectively). Nitrogen stress had an impact on essential oil content of lemon balm and sage, but not of lemon catmint. In case of lemon catmint high nitrogen concentration of 300 mg/l was favourable for maximum yield of essential oil (0.376 ml/plant), while for sage medium concentration of 150 mg/l was the best (0.202 ml/plant) and for lemon balm there was no significant difference between the treatments. The essential oil composition of selected medicinal and aromatic plants was mainly affected by tested nitrogen concentrations. The influence of nitrogen stress was also significant for content and composition of polyphenols in studied herbs, as well as for antioxidative capacity of polyphenol-rich extracts by the ABTS system.
Beemnet Mengesha Kassahun, Solomon Abate Mekonnen, Zinash Teferi Abedena, Haileslassie Gebremeskel Kidanemariam, Beniyam Yalemtesfa, Gizachew Atnafu, Bekri Melka, Wossen Kebede Mengesha (Ethiopia), Jaime A. Teixeira da Silva (Japan) Performance of Lemongrass (Cymbopogon citratus L. (DC) Stapf) Agronomic and Chemical Traits in Different Agro-Ecologies of Ethiopia (pp 133-138)
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Original Research Paper: Lemongrass (Cymbopogon citratus L. (DC) Stapf) is a perennial aromatic and medicinal plant that belongs to the Graminae (Poaceae) family. It has long been introduced and maintained at Wondo Genet, Southern Ethiopia. Despite its long presence, various potential uses and the existence of diverse ecologies, the plant has not been evaluated for the performance of agronomic and chemical traits in different ecologies in Ethiopia. This study was thus designed to contribute to addressing the existing technology gaps and thereby bringing the crop to the level of cultivation in Ethiopia. Data on number of tillers/plant, number of leaves/tiller, number of leaves/plant, longest leaf length, plant height, fresh herbage yield/plant, fresh herbage yield/ha/yr, essential oil (EO) content and EO yield/ha/yr were collected from six locations arranged in a complete randomized block design with four replications. In addition, the overall mean values for citral, myrcene and geraniol contents were quantified and recorded. Growing location demonstrated a highly significant influence (P < 0.01) on the performance of all the parameters considered. With increasing year, the performance of lemongrass was significantly (P < 0.01) influenced in all the studied parameters except for number of leaves/tiller. The respective average fresh herbage yield/ha, EO content and EO yield/ha varied from 7.98-28.01 t, 0.34-0.53% and 42.29-108.16 kg for first year while, during the second year, average annual fresh leaf yield/ha, EO content and EO yield/ha varied from 32.31-75.89 t, 0.45-0.55% and 155.09-342.69 kg, respectively. Increasing age from the first to the second year resulted in an increase of 165.31, 6.66 and 189.91% for fresh leaf yield/ha, EO content and EO yield/ha, respectively. The overall mean performance of lemongrass in terms of citral content over the tested locations varied from 70.81-82.68%.
Ram Swaroop Verma, Amit Chauhan, Laiq ur Rahman, Rajesh Kumar Verma, Anand Singh (India) Aroma Profile of Clary Sage (Salvia sclarea L.): Influence of Harvesting Stage and Post Harvest Storage in Uttarakhand Hills (pp 139-142)
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Original Research Paper: An experiment was conducted to determine the impact of harvesting stage and postharvest storage on essential oil (EO) content and composition of clary sage (Salvia sclarea L.) cv. ‘CIM-Chandni’ in the Kumaon region of western Himalaya. Clary sage inflorescences were collected at five different stages viz., prior to seed setting, all seeds whitish green, half seeds brown, all seeds brown and seed shattering stages. The EO was isolated by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The EO content varied from 0.18 to 0.31% during these stages. The major components of these oils were linalool (25.38-34.32%), linalyl acetate (27.98-48.59%), a-terpineol (4.03-5.25%) and sclareol (1.02-1.59%). Storage of clary sage inflorescences under shade for 96 h prior to distillation significantly reduced the EO recovery (from 0.30 to 0.13%). Linalool increase after storage (28.86 to 37.47%) but linalyl acetate initially increased (from 39.68 to 43.30%) thereafter it decreased after storage.
Mahender Aileni, Venugopal Rao Kokkirala, Rajesh Yarra, Anil Kumar Vemunoori, Kiranmayee Kasula, Pavan Umate, Sadanandam Abbagani (India) In Vitro Regeneration, Flowering and Seed Formation from Leaf Explants of Scoparia dulcis L. (pp 143-146)
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Original Research Paper: This study describes an efficient protocol for the induction of in vitro flowering and fruiting of Scoparia dulcis L. (family - Scrophulariaceae), a multipurpose folk medicinal plant. Murashige and Skoog medium supplemented with kinetin (KIN; 13.93 µM) and indole-3-acetic acid (IAA; 1.14 µM) is optimal for the formation of multiple shoots (20.00 ± 1.67), which induced floral buds (13 ± 0.43). The regenerated plantlets developed flowers and fruits within 40-45 and 50-55 days, respectively. In vitro floral development was asynchronous. The morphology of the in vitro developed floral organs was almost similar to that of the parent plant. The flowers produced in vitro developed ripe fruits with viable seeds. Our finding has important implications in understanding the influence of physiological factors and the molecular basis of floral organ formation in this valuable medicinal plant.
Harish B. Gowdru, Sharath Rajashekarappa, Kumaraswamy H. Malleshappa, Katenahalli R. Maruthi, Venkatarangaiah Krishna (India) In Vitro Regeneration of Inflorescence and Callus Enhancement of Celastrus paniculatus Willd. (pp 147-151)
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Original Research Paper: A protocol for plant regeneration with an intervening callus phase was developed for Celastrus paniculatus Willd.using immature inflorescence segments and flower buds. The floral explants are proliferate into callus mass on Linsmaier and Skoog (LS) medium augmented with 3% fructose as the carbon source and the plant growth regulators kinetin (Kn) and indole-3-butyric acid (IBA) in a range of 6-8 mg L-1 and 0.25 to 0.75 mg L-1, respectively. The interaction of these cytokinins and auxins at a lower level (1-4 mg L-1 Kn and 0.1-0.3 mg L-1 IBA) induced the calli to differentiate into shoot buds. The maximum number of shoots from immature inflorescences (9.3 ± 2.21 per callus) was noticed at 3.0 mg L-1 Kn and 0.2 mg L-1 IBA. Microshoots developed a good root system on half-strength LS medium fortified with 0.3 mg L-1 IBA. The regenerants derived from inflorescence culture exhibit vegetative morphology different to that of the donor plant. Callus from leaf and stem was further enhanced by augmenting the combination of Kn and IBA. There was a significant increase in the callus mass of both leaf and stem. The fresh and dry weight of stem calli (57.36 ± 5.58 g and 1.15 ± 0.12 g) was higher than that of leaf (51.76 ± 1.02 g and 1.04 ± 0.11 g).
Seyed Ali Andi, Vahideh Nazeri, Javad Hadian, Zabihollah Zamani (Iran) Variability of Essential Oil Composition of Origanum vulgare L. ssp. vulgare Populations from Iran (pp 152-155)
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Original Research Paper: In this study, the chemical composition of the essential oils (EOs) extracted from aerial parts of native populations of Origanum vulgare ssp. vulgare from Iran was compared by GC and GC/MS. Plants were collected at bloom stage from their natural habitats located in Kaleybar, Sabalan, Meshkin Shahr and Chalus regions, respectively from East Azerbaijan, Ardabil, Ardabil and Mazandaran provinces. Forty identified compounds accounted for 96.2-99.8% of the EO composition. According to the GC/MS analysis, four chemotypes consisting of carvacrol, sabinene, caryophyllene oxide and linalyl acetate were identified in the populations. Nonetheless, the monoterpenes were included the greatest EO fraction in all the populations. Of the monoterpenes, hydrocarbon compounds accounted for more than the oxygenated ones. This paper represents the first study on the EO composition of Iranian specimens of this plant and can be very useful for future breeding programs and medicinal purposes.
Sunita Sajjekhan, Prashanth Kumar Jha, Shruthi Shirur Dakappa (India) Pharmacognostic Evaluation of Psidium guajava Linn. Leaves (Myrtaceae) (pp 156-159)
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Original Research Paper: The leaves of Psidium guajava (Myrtaceae) (guava) are known to have many medicinal properties. In tropical and sub-tropical countries these leaves are used to treat many disorders such as diarrhoea, cough, gastrointestinal disorders, vomiting, wounds, dysentery, ulcers, toothache and as an antiseptic. In the present study pharmacognostical investigations including morphology, microscopy (both general and quantitative), physico-chemical, preliminary phytochemical and fluorescence studies have been done to establish the specific identity of drugs. Phytochemicals observed with simple qualitative tests were alkaloids, tannins, flavonoids, phytosterols, and glycosides. They could have particular medicinal effects individually or in combined forms on various ailments. The plant has been extensively studied in terms of pharmacological activity of its major components. The findings may provide useful information with regards to its identification and standardization in future.
Sasan Mohsenzadeh, Maryam Gholami (Iran), Jaime A. Teixeira da Silva (Japan) Allelopathic Potential of Ephedra (pp 160-162)
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Short Communication: The ethanolic extract obtained from dried Ephedra pachyclada Boiss. Flowering aerial parts was evaluated in vitro to examine its potential allelopathic effects. The alkaloids ephedrine and pseudoephedrine are the active constituents of the plant. The inhibitory effect of the extract at 0, 2.5, 5, 10 and 20% (i.e., g amounts of original extract in 100 ml of distilled water) on germination and seedling growth of perennial ryegrass (Lolium perenne) and bread wheat (Triticum aestivum) as representative monocotyledonous plants and soybean (Glycine max) and mung bean (Vigna radiata) as representative dicotyledonous plants were tested. All concentrations of the stock Ephedra extract significantly reduced (P ≤ 0.05) germination rate and percentage and seedling growth of the four examined plants. At 20% of the original extract, the germination of perennial ryegrass, soybean and mung bean was completely suppressed. In this study, the allelopathic potential of E. pachyclada extract on seed germination of the two dicotyledonous plants belonging to the Poaceae family was higher than the two monocotyledonous plants belonging to the Fabaceae family.
Wossen K. Mengesha, Daniel M. Bisrat, Solomon M. Abate (Ethiopia) Variability for Antioxidant Activities and Quality Traits of Two Ginger (Zingiber officinale Roscoe) Clones Growing in Ethiopia (pp 163-165)
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Research Note: Ginger rhizomes growing in two different geographical locations, namely Tepi in Southern Regional States and Chilga woreda of Gonder in the North-Western regions of Ethiopia, were extracted by a typical reflux apparatus to see the variability in content of oleoresin, essential oil and gingerol of the two main ginger production areas. In the study, the oleoresin content of the ginger samples from Tepi and Chilga showed 8.0 and 6.5% (w/w), which were statistically different at P < 0.01. Gingerol content within oleoresin was 11% for the Tepi clone and 7.0% for the Chilga clone. The maximum essential oil content (1.0%) was recorded for the Tepi clone and it was only 0.4% for the Chilga clone. Similarly, the antioxidant activities with respect to the free radical 2,2-diphenyl-1-picryl hydrazyl, were also evaluated and showed radical scavenging activity with an IC50 value of 1.81 and 2.84 µg/mL, for Tepi and Chilga clones, respectively.
Kranthi K. Gadidasu, Elagonda N. Murthy, Podishetty Nataraj, Kota Srinivas, Prasad A. Babu (India), Jaime A. Teixeira da Silva (Japan), Vatsavaya S. Raju, Abbagani Sadanandam (India) ISSR Markers Reveal Genetic Polymorphism in Two Morphological Variants of Hyptis suaveolens Invasive to India (pp 166-168)
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Research Note: A white-flowering variant of the invasive Hyptis suaveolens (Lamiaceae) was discovered in India and is reported in this study. Its genetic diversity was measured using 15 ISSR markers. PCR-amplified products were visualized using agarose gel electrophoresis. Traditional morphological traits such as leaf morphology, length of calyx bristles, colour of the petals, among others, were useful in unveiling infraspecific variation in H. suaveolens. Polymorphism was 35% between the white-flowering and typical blue forms. Maximum polymorphism was detected by primer Hy6 in 9 alleles while only one allelic variation was found for each of primers Hy10, Hy13, Hy14 and Hy15. ISSR molecular markers are useful by disclosing genetic differences underscoring morphological variations in H. suaveolens. Minor differences, however prevents a taxonomic status to be assigned to the white-flowering variant.
Muluh E. Khan, Benard Mela (Nigeria), Jaime A. Teixeira da Silva (Japan), Joseph O. Amupitan, Dimas Kubmarawa, Rejoyce Atiko (Nigeria) Phytochemical and Antimicrobial Evaluation of the Potency of Nepata cateria Leaves against Some Pathogens (pp 169-172)
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Research Note: A phytochemical and antimicrobial investigation was undertaken to determine the efficacy of the methanolic extract of the leaves of Nepata cateria, locally used for the treatment of diarrhea and dysentery in Gombe State, North Eastern Nigeria. Phytochemical screening confirmed the presence of bioactive secondary metabolites, namely phenols, tannins, terpenoids, glycosides, resins, phlobatannins, alkaloids, saponnins and volatile oils. Carbohydrates, flavonoids and sterols were absent. The leaf extract showed significant antimicrobial activity against Escherichia coli, Shigella dysenterica, Streptococcus pyogenes but had no effect on Salmonella typhii and Staphylococcus aureus. This correlates with the ethnomedicinal claims, thus providing for the first time, a pharmacological basis for the folkloric use of leaves of N. cateria as a cure for diarrhea and dysentery in Gombe State.
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