Volume 1 Number 2 2007

CONTENTS AND ABSTRACTS

M.N.V. Prasad, K. Padmalatha, K. Jayaram, N.L. Raju (India), Jaime A. Teixeira da Silva (Japan) Medicinal Plants from the Deccan Ecoregion, India: Traditional Knowledge, Ethnophamacology, Cultivation, Utilization, Biotechnology and Conservation - Opportunities and Impediments (pp 155-208)

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Review: An annotated synopsis of prominent Medicinal and Aromatic plants from the Deccan ecoregion of India viz., Aegle marmelos, Andrographis paniculata, Argyreia nervosa, Aristolochia indica, Asparagus racemosus, Azadirachta indica, Bacopa monnieri, Bixa orellana, Boswellia serrata, Butea monosperma, B. superba, Centella asiatica, Celastrus paniculatus, Chlorophytum arundinaceum, C. tuberosum, Commiphora wightii, Costus speciosus, Curcuma pseudomontana, Cycas circinalis, Decalepis hamiltonii, Drosera burmanii, D. indica, Embelia ribes, Entada pursaetha, Fagonia cretica, Gardenia gummifera, Gloriosa superba, Gymnema sylvestre, Ocimum sanctum, Oroxylum indicum, Piper longum, P. nigrum, Plumbago rosea, Pterocarpus marsupium, P. santalinus, Pueraria tuberosa, Rauvolfia serpentina, R. tetraphylla, Santalum album, Saraca asoka, Schleichera oleosa, Sterculia urens, Strychnos nux-vomica, S. potatorum, Terminalia arjuna, T. chebula, T. pallida, Tinospora cordifolia, Urginea nagarjunae, Vanda tessellate, Vitex trifolia, V. negundo, Withania somnifera, and Zanthoxylum alatum, ethnopharmacological utilization and conservation are presented in this paper together with the issues and challenges to achieve this goal. Ex situ conservation and cultivation of selected Redlisted plants of the Deccan ecoregion was initiated in 2000 at the University of Hyderabad and are being maintained in a field gene bank and seed bank. A special feature of the medicinal flora in the Deccan ecoregion is the preponderance of plants that provide raw material for addressing a wide range of medical disorders and pharmaceutical requirements.

Neeta Shrivastava, Tejas Patel (India) Clerodendrum and Healthcare: An Overview - Part II. Phytochemistry and Biotechnology (pp 209-223)

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Invited Mini-Review: The genus Clerodendrum is very widely distributed throughout the world and has more than five hundred species. Many species of this genus have been described in various indigenous systems of medicine and are used in preparation of folklore medicines for the treatment of various life-threatening diseases. From the genus few species are very well studied for their chemical constituents and biological activities, the latter having been covered in our previous review. This review mainly focuses on phytochemistry i.e. isolation, identification and characterization of chemical constituents and biotechnological prospects of the Clerodendrum genus. Some of the species described in the review are Clerodendrum trichotomum, C. bungei, C. chinense, C. colebrookianum, C. inerme, C. phlomidis, C. petasites, C. grayi, C. indicum, C. serratum, C. campbellii, C. calamitosum and C. cyrtophyllum. The major chemical constituents present in this genus were identified as phenolics, flavonoids, terpenes, steroids and oils. Biotechnological aspects have also been discussed in the review.

T.K. Behera (USA/India), J.E. Staub, Snigdha Behera, Philipp W. Simon (USA) Bitter Gourd and Human Health (pp 224-226)

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Invited Research Note: Bitter gourd (Momordica charantia L.; Family: Cucurbitaceae) is a vegetable with tropical and subtropical distribution. Bitter gourd fruit is a rich source of nutrients and ranks first among cucurbits for its nutritive value, being a good source of carbohydrates, proteins, vitamins, and minerals. Natural antioxidants in bitter gourd are primarily plant phenolics and polyphenolic compounds derived from fruits and seeds, and thus are alternatives to replace synthetic antioxidants to enhance food quality. Fruit contains as many as 14 carotenoids (five at the immature stage, and six and 14 in the mature-green and ripe stages, respectively) and cryptoxanthin, which is the principal chloroplast- and chromoplast-based pigment in ripe fruit.

A. Gramza-Michalowska (Poland) Antiradical Methods in Evaluating Antioxidant Potential of Preparations of Plant Origin (pp 227-233)

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Invited Mini-Review: The formula for antioxidants is very difficult to define, although they had been used in food for centuries. Antioxidants are compounds used to delay the accumulation of free radicals and strengthen the oxidative stability of food. According to the activity mechanism of antioxidants, they can act by means of many different processes. The properties of antioxidants are also the result of many factors, like mixing ability and activity in different reaction environment systems, stability in processing time, as well as research methods used. However, there exist limitations concerning the use of these substances. Research reflects much interest on the influence of radicals generated by different environmental factors on human health and food stability, but there is still a need to find the substances that have wide antiradical properties. There are many methods for evaluating antiradical activity. This manuscript discusses the most popular and adequate methods. Widely used parameters of free radical scavenging ability as well as the evaluation of antioxidant potential are reactions with ABTS^+・ and DPPH^・ radicals, the luminol chemiluminescence method, ORAC, FRAP, TRAP and many other assays. The aim of the comparison of these methods and this review is to qualify the effectiveness of different natural substances as scavengers possessing antiradical activity.

Thayamini H. Seran (Sri Lanka) Anther Culture in Tea Improvement (pp 234-239)

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Invited Mini-Review: The important economic species in the genus Camellia is C. sinensis, which is a high priority crop in Sri Lanka and is cultivated commercially for its tender leaves. Therefore, breeding methods in tea should be directed towards leaf improvement. Improvement of tea depends on the presence of adequate genetic variability. Anther culture plays an important role in crop improvement programs to overcome barriers experienced in conventional breeding methods. Thus this technique enhances plant breeding and speeds up the release of superior cultivars within a short time in large quantities. Several workers have attempted to produce haploids that could be used for future improvement of woody plants, i.e. tea, rubber etc. In this paper, the work carried out by various workers in relation to tea improvement and potential of in vitro anther culture technique are discussed.

Edison J. Mavundza, Thilivhali E. Tshikalange, Fhatuwani N. Mudau, Jacobus J.M. Meyer (South Africa) Evaluation of Antibacterial and Antioxidant Activities in Ethanol Extract of Wild Bush Tea (Athrixia phylicoides (DC.)) (pp 240-242)

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Short Communication: South Africa is well known for its indigenous herbal tea production such as honeybush and rooibos tea. Bush tea, like any other herbal tea, has been used for many years as a herbal or medicinal tea by traditional African people. To validate its medicinal potential, an ethanol extract from wild bush tea leaves was used in vitro for antibacterial and antioxidant activity determination. The antibacterial assay showed inhibitory activity against five bacterial strains evaluated with minimum inhibitory concentration (MIC) values between 3.13 and 6.12 mg×ml^-1. The extract demonstrated the potent antioxidant activity by effectively scavenging DPPH, a free radical, with an EC₅₀ value of 28.12 µg×ml^-1.

Hintsa T. Araya, Puffy Soundy, Elsa S. du Toit, Fhatuwani N. Mudau (South Africa) Influence of Cutting Position, Medium, Hormone and Season on Rooting of Bush Tea (Athrixia phylicoides DC.) Stem Cuttings (pp 243-252)

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Original Research Paper: The objective of this investigation was to study the effect of cutting position, rooting medium and hormone on the rooting of bush tea stem cuttings. The experiment was carried out in four consecutive seasons from 2002 to 2003 at the Hatfield Experimental Farm of the University of Pretoria, Pretoria, South Africa. The parameters recorded were cutting position (apical vs. basal), rooting medium (composted pine bark vs. sand), rooting hormone (with Seradix^® No. 2 vs. no hormone), sampling days (15, 20, 25 and 30 days) and season (summer, autumn, winter and spring). Cutting position had a highly significant effect (P<0.001) on rooting and transplanting survival of bush tea with better rooting percentage, root length, root number and survival percentage from apical than from basal cuttings. Composted pine bark improved the number of roots developed but had no effect on rooting percentage, survival percentage nor on root length of the cuttings. The application of rooting hormone (Seradix^® No. 2) during the propagation period increased root number and further increased shoot length after transplanting but not rooting percentage, survival percentage or root length. Season also showed highly significant differences (P<0.001) on rooting percentage, root number and root length. Rooting of cuttings was improved when propagated in autumn (longer roots) and spring (number of roots) but not in summer or winter. The results of this study suggest that vegetative propagation of bush tea could be achieved by apical cuttings propagated in composted pine bark with Seradix^® No. 2 hormone in spring for 30 days since root number was the most important factor for the successful establishment of the cuttings.

Cüneyt Çirak (Turkey), Jolita Radusiene (Lithuania) Hypericins in Hypericum montbretii: Variation among Plant Parts and Phenological Stages (pp 253-256)

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Original Research Paper: In the present study, morphogenetic and phenological variations of hypericin and pseudohypericin were investigated in Hypericum montbreti, a perennial herbaceous plant from Turkish flora for the first time. Wild growing plants were harvested at the vegetative, floral budding, full flowering, fresh fruiting and mature fruiting stages and dissected into stem, leaf and reproductive tissues and assayed for hypericin and pseudohypericin by HPLC. Phenological fluctuation in hypericin and pseudohypericin content of plant material including whole shoots, stems, leaves and reproductive parts was found to be significant (P<0.01). Hypericin and pseudohypericin content in whole shoots, leaves and reproductive parts increased during the course of ontogenesis. The highest level of both compounds was reached at full flowering. In contrast, hypericin and pseudohypericin content in stems decreased with an advancement of plant development and stems from newly emerged shoots at the vegetative stage produced the highest level of both compounds. Among different plant tissues, reproductive parts were found to be superior than leaves and stems with regard to both hypericin and pseudohypericin accumulation.

Manisha Dhasmana (India), Luke Simon (UK/India), P. Narayanaswamy, R.K.S. Rathore, B.S. Sreeramu (India) Characterization of Solanum nigrum L. Genotypes by Morphological and RAPD Markers (pp 257-262)

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Original Research Paper: Randomly amplified polymorphic DNA markers were used to assess genetic diversity among 12 cultivated genotypes of Solanum nigrum L. obtained from diverse locations of India and were compared with their morphological data obtained for vegetative and reproductive characters. DNA was isolated using CTAB method and 113 amplified fragments were obtained from 15 random 10-mer primers. Intense, clear and repeatable bands ranging from 250 bp to 6 Kb were used for RAPD analysis. Dendrograms were constructed for both morphological and RAPD data using Wards method, which grouped the genotypes into two clusters ‘A’ and ‘B’ at 5.56 and 4.0 linkage distances, respectively. Cluster ‘A’ consisted of eight genotypes, which were tall plants and required less time to flower. Cluster ‘B’ consisted of four genotypes showing short and widely spread plants with high pollen viability and had more number of seeds per berry. The genetic dissimilarity matrix calculated for the RAPD markers was based on Squared Euclidean Distance, showed a variation from 16 to 53% among the cultivars. Thus, RAPD markers combined with morphological analysis proved to be a quick, simple and significant testing method to assess genetic diversity among S. nigrum L. genotypes.

K. Padmalatha, M.N.V. Prasad (India) Morphological and Molecular Diversity in Pterocarpus santalinus L.f - an Endemic and Endangered Medicinal Plant (pp 263-273)

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Original Research Paper: The present study is the first report of morphological and molecular variations in accessions of Pterocarpus santalinus L.f (endemic, endangered, commercially and medicinally very important) collected from Kerala, Karnataka and mostly from Andhra Pradesh, India. It addresses the determination of genetic variation among accessions using few morphological parameters and RAPD markers. A total of 27 accessions were collected out of which morphological variations were characterized for 14, 15 and 16 and molecular variations for 15 of them. Morphological data opened up lot of variations among the accessions. Molecular investigation revealed that, out of the 40 primers screened, 26 primers selected for the data analysis generated a total of 217 scorable markers, all of which were polymorphic. This high proportion of polymorphism i.e., almost 100%, was found with 53 unique markers. Cluster analysis based on Dice’s coefficient showed two major groups indicating that in cross-pollinated plants, high levels of differentiation among accessions exist. The grouping of these accessions was independent of the geographical distance. The significant molecular variations in the accession collected from Tirupathi when compared to other accessions need to be further investigated. Hence the results of the present study can be viewed 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 conservational management practices, rootstock breeding and hybridization programmes.

Kalpana N. Reddy, A. Manoj Kumar, B. Abdul Rahiman (India) Rapid in Vitro Protocol for High Multiple Shoot Induction, Rooting and Flowering in Asparagus racemosus L. (pp 274-277)

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Original Research Paper: A high frequency shoot multiplication protocol for Asparagus racemosus L. was developed by culturing shoot tips and nodal bud explants. Murashige and Skoog (MS) medium with 3 mg l^-1 and 0.1 mg l^-1α-naphthalene acetic acid (NAA), respectively formed green morphogenic calli which produced more than 95 multiple shoots in the same medium after 35 days of culture. The stunted shoots were transferred to shoot elongation medium with 2 mg l^-1 6-benzylamino purine (BAP) and 0.1 mg l^-1 NAA. Rooting was obtained on half MS with 7 mg l^-1 BAP after 45 days of culture. The shoots inoculated on lower concentrations of NAA did not show any change. Some shoots produced floral buds which bloomed to produce normal flowers in vitro when left undisturbed on NAA medium (3 mg l^-1) for 60 days and the plants bearing flowers wilted after 3 days. The plantlets with a morphologically normal appearance produced from adventitious shoots were transferred to soil and acclimatized in the growth chamber with a 62% survival rate.

Jin-ichi Sasaki, Chao Lu, Einosuke Machiya, Mami Tanahashi, Katsunori Hamada (Japan) Processed Black Garlic (Allium sativum) Extracts Enhance Anti-Tumor Potency against Mouse Tumors (pp 278-281)

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Short Communication: Black garlic (Allium sativum) was produced from ordinary white garlic (Allium sativum) clove by processing it in a temperature (70°C)- and humidity (75%)-controlled room for a month. The final product by this procedure was soft and sweet with a less irritating odor and fruity taste. The heat-extracts of black garlic were chemically and bio-functionally analyzed and compared with those of ordinary fresh garlic extracts. The aged black garlic contained an increasing amount of amino acids, and organo-sulfur substance, S-allyl-L-cysteine (SAC), which probably contributed much to the enhancement of anti-tumor potency. The tumor cure rate by black garlic extracts attained 50% against Meth A fibrosarcoma of BALB/c mouse by intra-tumor injection of 1 mg extracts, three times every other day. By contrast, fresh garlic extracts used as a reference failed to induce tumor-free animals, even though they reduced tumor size to 60% to compare with the tumor mass in non-treated control mouse. Additionally, the black garlic extracts showed anti-bacterial activity against medically important bacteria such as MRSA (methicillin-resistant Staphylococcus aureus), enterohemorrhagic Escherichia coli O157:H7, Pseudomonas aeruginosa, and Candida albicans, however its potency was less than that of fresh garlic extracts.

Iram Siddique, M. Anis (India) High Frequency Multiple Shoot Regeneration and Plantlet Formation in Cassia angustifolia (Vahl.) Using Thidiazuron (pp 282-284)

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Short Communication: An in vitro propagation system for Cassia angustifolia Vahl. has been developed. De novo shoot organogenesis was induced by culturing cotyledonary node explants excised from seven days-old aseptic seedlings. Of the different concentrations (0.1,0.3, 0.5, 0.8, 1.0, 2.5, 5.0, 7.5 and 10.0 µM) of thidiazuron (TDZ) evaluated as supplements to Murashige and Skoog (MS) medium, TDZ at an optimal concentration of 1.0 µM was effective in inducing multiple shoots. The number of shoots increased significantly when the responding explants were transferred to a medium devoid of TDZ. The highest number of shoots and shoot length were recorded at the end of fourth subculture. Ex vitro rooting was achieved when the basal cut end of regenerated shoots were dipped in 200 µM indole-3-butyric acid (IBA) for half an hour followed by their transplantation in plastic pots filled with sterile Soilrite^TM where 80% plantlets grew and all exhibited normal development.

P. Ujwal, M.P.M. Pramod Kumar, H. Raja Naika, B.B. Hosetti (India) Antimicrobial Activity of Different Extracts of Pongamia pinnata (pp 285-287)

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Short Communication: Plant material was collected from Western Ghats, India, shade-dried and extracted successively with different polarity solvents of petroleum ether and ethyl acetate. The extracts were tested for their antimicrobial activity against Gram-positive and Gram-negative bacteria and against a yeast species using the disc-diffusion method. The antimicrobial activity of extracts of Pongamia pinnata was tested against bacterial strains such as Bacillus subtilis (NCIM 2117), Escherichia coli (NCIM 2079), Pseudomonas aeruginosa (NCIM 2036), Staphylococcus aureus (NCIM 2079) and a fngal species Candida albicans (NCIM 3100). In both extracts of petroleum ether and ethyl acetate, the extracts from leaf, bark and seeds showed an inhibition zone on B. subtilis and S. aureus cultures. Extract from the seeds of petroleum ether and ethyl acetate showed maximum inhibition zone on B. subtilis, while the leaf extract of petroleum ether and ethyl acetate showed comparable activity against S. aureus. E. coli and C. albicans were unaffected by any of the extracts of P. pinnata, except for the bark extract of petroleum ether. These findings also support the use of this plant in traditional medicine for the treatment of bacterial and fungal infections.