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Volume 2 Number 2 2008
CONTENTS AND ABSTRACTS
Fei-fei Qin (Japan/China), Hui-lian Xu (Japan) Active Compounds in Gingers and Their Therapeutic Use in Complimentary Medication (pp 72-78)
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Invited Mini-Review: The rhizome of ginger is used widely both as a seasoning and herbal medicine. The active compounds contained in ginger are divided into two groups: volatile essential oils and fragrant or harsh phenol compounds. Two types of gingers, fresh and dried, are used in the herbal medicine. There exist differences in the chemical composition between fresh and dried gingers that might result in differences in medicinal functions of the herbs. The therapeutic functions, which include lowering cholesterol, relieving allergies and asthma, arthritis, colds, and nausea, protecting the digestive tract and liver against toxins and parasites, and avoiding gallstone, cerebral vascular and cardiovascular diseases, are reviewed and the toxicity and safety issues are also discussed in this article.
Salim Khan, Amjad M. Husaini, Usha Kiran, Kamaluddin, Mauji Ram, M. Z. Abdin (India) SCAR Markers for Authentication of Herbal Drugs (pp 79-85)
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Mini-Review: Correct identification and quality assurance is indispensable to ensure reproducible medicinal quality of herbal drugs. Authentication is especially useful in case of those medicinal herbs that are frequently substituted or adulterated with other species or varieties, morphologically and phytochemically indistinguishable. Morphological as well as biochemical markers used in authentication of herbal drugs have many limitations due to the impact of environmental conditions. Molecular markers therefore, are an important tool in quality assurance and preservation of germplasm of medicinal plant species in the plant kingdom. Randomly Amplified Polymorphic DNA (RAPD) is an easy and simple molecular marker, but lack of reproducibility makes it a lesser reliable authentication method for herbal drugs. Besides RAPD, other popular PCR and non-PCR based markers like AFLP, ISSR, SSR and RFLP are also used for authentication. However, these also have disadvantages like use of radioactive isotopes, costly and absolute requirement of sequence information and hence it is a better option to improve the reproducibility of RAPD by converting RAPD amplicons into Sequence Characterized Amplified Region (SCAR) markers. SCAR markers are easy, reliable and reproducible thus, have an advantage over RAPD markers for authentication of medicinal herbs used in the preparation of traditional medicines. These markers however, have been developed for only a few medicinal herbs. This review is an attempt to evaluate critically the role of SCAR markers in authentication of medicinal herbs used in traditional formulations.
Neelu Joshi, Gurinder J. Randhawa, Prashant K. Firke, Sunil D. Purohit (India) RAPD Analysis of Diversity in ‘Safed Musli’ (Chlorophytum borivilianum Sant. et Fernand.), a Rare Indian Medicinal Herb (pp 86-89)
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Original Research Paper: RAPD markers were used to assess genetic diversity in seven accessions of a rare Indian medicinal herb Chlorophytum borivilianum collected from different geographical regions of India. A total of 290 amplified bands were scored from 33 random decamer primers out of which 242 (83.44%) were found to be polymorphic. The average number of polymorphic bands per primer was 7.33. The Jaccard’s similarity coefficient ranged from 0.108 to 0.533 with a mean of 0.338. A dendrogram generated by UPGMA analysis grouped the accessions into four clusters which were not based on their geographical distribution.
K. Padmalatha, M. N. V. Prasad (India) Genetic Diversity in Centella asiatica (L.) Urb., a Memory-Enhancing Neutraceutical Herb, using RAPD Markers (pp 90-95)
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Original Research Paper: The present study is the first report of genetic diversity analysis of Centella asiatica (L.) Urb, a medicinal and aromatic plant collected from various locations of Andhra Pradesh, India. Though there are several reports on diversity analysis of C. asiatica using RAPD analysis the data presented in this article is the first report on diversity studied in accessions collected from Andhra Pradesh State. It addresses the determination of genetic variations using a few visual morphological parameters and by RAPD markers. We collected sixteen accessions which demonstrated clear morphological variations and used nine to test for molecular variation. Wide phenotypic variations were observed. Molecular analyses revealed that out of the 30 primers screened, the 16 that were selected for data analysis generated a total of 137 scorable polymorphic markers out of 156 total number of markers. An 87% polymorphism was observed. Cluster analysis based on Dice’s coefficient showed two major groups indicating that in cross-pollinated plants, high levels of differentiation among accessions exists. The grouping of these accessions was independent of the geographical distance. Hence the results of the present study can be seen as a starting point for future research on the population and evolutionary genetics of this species and understanding such variation would facilitate their use in various conservation management practices, and for generating an elite variety from which the production of secondary metabolites can be enhanced.
Shailendra Vyas, Manohar Singh Rao, Rajesh Kumar Suthar, Sunil Dutta Purohit (India) Liquid Culture System Stimulates in Vitro Growth and Shoot Multiplication in Four Medicinally Important Plants (pp 96-100)
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Original Research Paper: Agar is the most widely used gelling agent, and accounts for 10-20% of the cost of the culture medium. Besides this, agar can contain impurities leading to inconsistent responses. We show, in this study, that the in vitro propagation in liquid medium is an effective means for the establishment of cultures of some medicinally important plants of Aravallis in Rajasthan, so as to achieve stimulated growth and shoot multiplication. The shoots of Chlorophytum borivilianum grown in liquid culture medium supported by glass beads showed a much higher rate of multiplication (4.75-fold) than the control (solid medium). In the case of Celastrus paniculatus, a 7-fold rate of shoot multiplication was achieved on liquid medium compared to a 4.5-fold rate on 0.8 % agar-gelled solid medium. We could stimulate shoot growth with good leaf expansion in Terminalia bellerica and double the rate of shoot multiplication in Boswellia serrata as compared to control. In all plant species, the growth of shoots was improved on liquid medium more than on solid medium. Glass beads provided mechanical support to the multiplying shoots. No adverse effects such as hyperhydricity were observed in any of the cases under investigation and plants could be acclimatized easily.
Manohar Singh Rao, Sunil Dutta Purohit (India) In Vitro Growth and Multiplication of Celastrus paniculatus Willd. under Carbon Dioxide Enriched Environment (pp 101-104)
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Original Research Paper: Celastrus paniculatus shoots were grown in vitro on MS (Murashige and Skoog 1962) medium containing 0.5 mg l-1 BAP (6-benzylaminopurine) under sucrose-free (SFSM) or sucrose-containing (30.0 g l-1) (SCSM) medium with varying CO2 concentrations (0.0, 0.6, 10.0, 40.0 g m-3) provided in small acrylic chambers using different concentrations and combinations of NaHCO3 (sodium bicarbonate), Na2CO3 (sodium carbonate), KHCO3 (potassium bicarbonate) and K2CO3 (potassium carbonate). The complete absence of any carbon source resulted in the death of shoots within 20 days. The shoots of C. paniculatus were grown fully photoautotrophically on a medium without sucrose (SFSM). Compared to the SCSM cultures grown under ambient air of the growth room (control), SFSM cultures grown under 10.0 g m-3 CO2 showed an equal rate of shoot multiplication (3.0-fold) with increased leaf area. The best response, however, was obtained when SCSM cultures were grown under 10.0 g m-3 CO2. At this concentration, the rate of shoot multiplication was nearly double the standard rate obtained in the SCSM control. Total fresh and dry weight, average shoot length, leaf number and leaf area per cluster also showed best response under this condition.
Mahmoud A. Sharaf-Eldin, Abeer Y. Ibrahim, Hassan M. Korkar (Egypt) Effect of Gibberellic Acid and Dry Yeast on Growth, Yield, and Essential Oil of Lemon Balm (Melissa officinalis L.) (pp 105-109)
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Original Research Paper: In a field experiment during two successive seasons (2005-2006 and 2006-2007), the effect of gibberellic acid (GA3) and active dry yeast on growth, yield, and essential oil (EO) of lemon balm plants was investigated. Application of GA3 and/or active dry yeast increased vegetative characters (i.e. plant height, number of branches, and herb fresh and dry weight per plant) compared to control (sprayed with water only). The maximum mean values of growth characters were obtained as a result of spraying with 6 g l-1 yeast + 300 ppm GA3. The lowest fresh and dry weight of plants were observed with the treatment of 2 g l-1 yeast + 0 ppm GA3 in the first harvest. EO content in the lemon balm herb increased due to the application of GA3 and/or active dry yeast compared to control. The highest EO yield per plant was observed with the treatment of 6 g l-1 yeast + 300 ppm GA3. The lowest amount of EO yield was obtained with the control treatment. The highest geranial in lemon balm EO occurred with the treatment of 6 g l-1 yeast + 300 ppm GA3.
Raoufa Abd El-Rahman, Hussein Taha, Mohamed El-Bahr (Egypt) Harmine in Peganum harmala L. in Vitro Cultures (pp 110-113)
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Original Research Paper: Explant type and choice of plant growth regulators significantly affected callus growth, shoot regeneration and harmine production in Peganum harmala callus cultures produced from excised hypocotyl, leaf and root explants of seedlings germinated in vitro. From a range of auxins and cytokinins tested, Murashige and Skoog (MS) medium supplemented with 5 mg/l Kinetin and 1 mg/l 1-naphthaleneacetic acid (NAA) resulted in the greatest callus mass and best growth parameters from all three explant types while MS medium supplemented with 5 mg/l N6-benzylaminopurine (BAP) and 0.1 mg/l of zeatin and NAA, resulted in greatest shoot production. MS medium supplemented with 3 mg/l BAP and 1 mg/l 2,4-D, however, resulted in higher harmine accumulation (0.962 mg/g dry weight) from root-derived calli than from callus derived from leaves or hypocotyls. Maximum levels of harmine (1.45 mg/g dry weight) were produced from shoots regenerated from hypocotyl-derived calli.
Puffy Soundy, Kwena W. Mpati, Elsa S. du Toit, Fhatuwani N. Mudau, Hintsa T. Araya (South Africa) Influence of Cutting Position, Medium, Hormone and Season on Rooting of Fever Tea (Lippia javanica L.) Stem Cuttings (pp 114-116)
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Original Research Paper: Cutting position, rooting medium and rooting hormone are critical factors that affect rooting development of stem cuttings. In this study, the objectives were to determine the best cutting position, ideal propagation medium and the effect of hormone on rooting of fever tea (Lippia javanica L.) stem cuttings. Results of this study demonstrated that apical cuttings rooted earlier than basal cuttings, but at 15 to 20 days after establishment both cuttings rooted similarly. Composted pine bark growing medium, compared to sand, resulted in improved root length of the cuttings. Basal cuttings had thicker stem circumferences and more number of leaves as compared to apical cuttings. Seradix® No. 2 hormone (0.3% IBA) increased fresh mass, stem circumference, number of roots and number of leaves for both apical and basal cuttings. Therefore, the results of this study suggest that for the establishment of fever tea stem cuttings, both apical and basal cuttings can be used but composted pine bark is the ideal propagation medium. Furthermore, fever tea stem cuttings can be ready for transplanting at 15 to 20 days after establishment and Seradix® No. 2 hormone is recommended to promote rooting.
Fei-fei Qin (Japan/China), Hui-lian Xu, G. Ma (Japan) Garlic Sprouts Grown Indoors at Kitchen Sites (pp 117-122)
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Original Research Paper: Recently, garlic shoots have been used as vegetable and functional food in additional to the bulbs. In the present research, garlic sprouts were produced from garlic cloves with different sizes. The cloves were grown onto paper towels sprayed with tap water every two days at kitchen sites under room temperature (fluctuated from 6°C in the night to 16°C around midday) or in a growth chamber (25±1/20±1°C, day/night). The final fresh and dry biomass indicated that the tiny cloves could also produce edible sprouts though they were not as strong as those produced by large and medium cloves. Fresh biomass was measured each day until harvest and the data were used for analysis of the dynamic growth of these garlic sprouts by a sigmoid model, gG = gT{1+(1-βt)e[-α(t-τ)]}^(-1)+g0(1-βt). One incidence named as “Biomass Downcast Phenomenon” occurred as shoot fresh biomass sharply declined when the nutrients in the clove were used up at the later growth stages of garlic sprouts. The curve phase of the sharp declining was analyzed using an exponential model, gD= g’T+(gmax-g’T)e[-γ(t-14)], and the biomass downcast was quantified using definite integral formula, g(δ) = *1. Leaf photosynthesis was low due to the weak light in kitchen site conditions. It quickly reached light saturation point (LSP) as the photosynthetic phonon flux (PPF) increased to 250 μmol m-2 s-1. Photoinhibition occurred once PPF suddenly increased above LSP and it was quantified by definite integral formula, P(i)= *2. In conclusion, garlic cloves, even in small size of no marketing value, can be used to produce fresh sprouts with high edible value at kitchen sites. The sprouts should be harvested before the nutrition in the cloves is used up when the so-called “Biomass Downcast Phenomenon” occurs. The mathematical approach adopted in this research for garlic sprout growth is of high reference value to plant scientists.
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Anshu Srivastava, Neeta Shrivastava (India) Phytochemical and Preclinical Screening of Aseptically Produced Herbal Raw Material: Bacopa monnieri (pp 123-127)
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Original Research Paper: The demand for herbal raw material has increased tremendously due to renewed interest in plant-based medicines, especially in developed countries. This has led to the indiscriminate cutting and collection of medicinal plants from natural resources resulting in depletion of natural resources and in some cases extinction of species. In the recent past, aseptically grown raw material and cell cultures have received recognition as an alternative source for the production of herbal raw material and chemical constituents. A low concentration of therapeutically important chemical constituents, which is directly related to the efficacy of the plant, was the major limitation in the commercial application of aseptically grown raw material and cell cultures. Due to this limitation, the acceptance of aseptically regenerated plants in the world of herbal medicines and Ayurvedic products was not very encouraging. In the present investigation, we have not only established the protocol for aseptic culture of Bacopa monnieri (L.) Pennell, but also checked the aseptically grown material for its chemical constituents and efficacy through established bioassay models. Assessment of the presence of major phytochemicals and in vitro preclinical bioassays indicated that aseptically grown shoots can be a better source of raw material. This can stop further depletion of the species from nature and also provide a consistent quality raw material. Aseptically grown improved quality shoots can also be good quality planting material for standardized cultivation practice.
Muluh E. Khan, Stephen S. Hati, Kabiru B. Abdu, Aliyu Babale, Micah I. Achi (Nigeria) Chemical Analysis and Antibacterial Activity of Acacia nilotica and Tapinanthus dodoneifolius Growing in Nigeria (pp 128-130)
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Original Research Paper: Pods of Acacia nilotica and leaves of Tapinanthus dodoneifolius were spectrophotometrically analysed for their mineral constituents (Ca, Cu, Fe, K, Mg, Mn, Na, P and Zn). Phytochemical analysis was conducted on successive Soxhlet extracts of ethanolic, n-hexane and ethyl acetate of the pods and leaves, respectively. Furthermore, the antibacterial activities of the extracts were investigated. Results showed high Ca and K with low Cu and Zn concentrations in both plants. The concentration of P in the pods of A. nolitica was highest. Phytochemical analysis revealed the presence of steroids, tannins and saponins in both ethanol and ethyl acetate extracts respectively, for both A. nolitica and T. dodoneifolius. Flavonoids were present only in the ethanol extract of A. nolitica. The ethanol extract generally had more phytochemical agents in both plants studied. All solvent extracts of T. dodoneifolius showed activity against Escherichia coli, Staphylococcus aureus, Klebsiella aerogenes and Proteus mirabilis showed varying activities only for A. nolitica solvent extracts, and only ethanol extract recorded activity against S. aureus, while the n-hexane extract showed no activity against any pathogen. The ethyl acetate extract of A. nolitica showed significant activity against K. aerogenes and P. mirabilis. The results in this work tend to agree with the ethno-medical claims.
Adelodun L. Kolapo, Mudashiru B. Okunade, Jacob A. Adejumobi, Mathew O. Ogundiya (Nigeria) In Vitro Antimicrobial Activity and Phytochemical Composition of Dichrostachys cinerea (pp 131-133)
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Original Research Paper: Studies on phytochemical composition and antimicrobial activity of aqueous and ethanol extracts of roots and stems of Dichrostachys cinerea against clinical isolates of Candida albicans, Streptococcus mutans and Staphylococcus saprophyticus werecarried out. The main phytochemicals present in the stem and roots included alkaloids, saponins and tannins, with roots containing the greater share. Steroids and cyanoglycoside were present in the stem. Both ethanol and aqueous extracts of the tested chewing stick inhibited the growth of all three tested microorganisms. There was no significant difference (P>0.05) between the inhibitory effect of the aqueous and ethanolic extracts of the roots of D. cinerea on C. albicans. However, the ethanolic extract of the stem exhibited a significantly higher (P<0.05) bioactivity than that exhibited by the ethanolic extract of the root. The pattern of inhibition of S. mutans and S. saprophyticus by the extracts were similar. Solvent used in extraction did not produce any significant effect (P>0.05), but the stem extracts exhibited a significant inhibition (P<0.05) compared to the root extract. Our results clearly show that D. cinerea is a potential candidate plant that could be used in the development of a dentifrice.
B. Muthukumar, D. Natarajan, N. Nagamurugan (India) Antimicrobial Assay of Zizyphus oenoplia (L.) Mill. (pp 134-136)
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Original Research Paper: The antimicrobial activity of acetone and methanolic leaf extracts from Zizyphus oenoplia was tested against three Gram-negative (Pseudomonas putida, Vibrio cholerae, Shigella flexneri)andtwo Gram-positive bacteria(Staphylococcus aureus and Bacillus sp.)and two fungal strains (Candida albicans and Cryptococcus neoformans) by conducting a well-in-agar method. P. putida and V. cholerae exhibited better resistance against the plant extracts followed by Shigella flexneri and Bacillus sp. Other pathogens were ineffective against the plant extracts.
Rotimi Ayodele Oderinde, Ibironke Adetolu Ajayi, Adewale Adewuyi (Nigeria) Nutritional Elements, Antibacterial Activity and Cytotoxicity of the Leaf, Root and Stem Bark of Blighia unijugata Baker(Sapindaceae) (pp 137-140)
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Original Research Paper: A comparative study was carried out on the mineral nutrient, cytotoxicity and antibacterial activity of the ethanol extracts of the leaf (BUL), root (BUR) and stem bark (BUB) of Blighia unijugata Baker (Sapindaceae). The phytochemical test showed the presence of steroid, saponin and tannin in all the extracts. A total of ten metals, six trace metals (Fe, Mn, Cu, Pb, Cd and Zn) and four macro nutrients (Na, K, Mg and Ca) were determined using Atomic Absorption Spectrophotometry. The concentration of the macronutrients was the highest with Mg being the highest (21.01 ± 1.21 ppm) in the BUR and the Na being the lowest (517.01 ± 0.50 ppm) in BUB. The concentration of the trace metal also differs with Mn being the highest (104.35 ± 0.11 ppm) in BUR and Pb being the lowest (0.36 ± 0.01 ppm) in the BUB. Mg also had the highest transfer factor (0.9501) in BUL. The antibacterial activity of these extracts against pathogenic bacterial showed significant inhibitory activity. BUL was active against all the tested bacterial strains.Both BUR and BUB did not show any activity against the growth of Klebsiella pneumoniae. BUR has no activity against Enterococcus faecalis as BUB was not also active against Salmonella typhi and Pseudomonas aeruginosa. All the extracts had high sensitivity to Staphylococcus aureus. The ethanol extracts of BUF and BUR showed potent cytotoxicity with LC50 of 196.50 and 269.05µg/ml, respectively when tested against Brine shrimp larvae, which supports the ethnomedical claims for the plant.
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