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Volume 4 Special Issue 1 2010
Medicinal Plants of the Himalayas: Advances and Insights
How to reference: Bantawa P, Ghosh SK, Bhandari P, Singh B, Ghosh PD, Ahuja PS, Mondal TK (2010) Micropropagation of an Elite Line of Picrorhiza scrophulariiflora, Pennell, an Endangered High Valued Medicinal Plant of the Indo-China Himalayan Region. In: Husaini AM (Ed) Medicinal Plants of the Himalayas: Advances and Insights. Medicinal and Aromatic Plant Science and Biotechnology 4 (Special Issue 1), 1-7
Guest Editor
Amjad M. Husaini
Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, India
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http://skuastkashmir.ac.in/
CONTENTS AND ABSTRACTS
Pranay Bantawa, Swapan Kumar Ghosh, Pamita Bhandari, Bikram Singh, Partha Deb Ghosh, Paramvir Singh Ahuja, Tapan Kumar Mondal (India) Micropropagation of an Elite Line of Picrorhiza scrophulariiflora,Pennell, an Endangered High Valued Medicinal Plant of the Indo-China Himalayan Region (pp 1-7)
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ABSTRACT
Original Research Paper: An elite genotype of Picrorhiza scrophulariiflora Pennell was multiplied in vitro for its conservation. Rhizomes of mature plants collected from various locations of the eastern Himalayan region of Indo-China border were characterized morphologically and analyzed by HPLC to determine the content of marker compounds, namely picroside I and II. Amidst the genotypes, one from Ha, Bhutan was found to contain the highest amount of total picroside (7.33% dw). Subsequently, a rapid and highly reproducible method of micropropagation from rhizome or shoot tips was developed. While 100% bud break from rhizomes was achieved on Woody Plant Medium (WPM) containing 0.44 mM BAP (6 benzyl amino purine), 40-fold multiplication was achieved on WPM fortified with 2.3 mM Kn (kinetin) within 12 weeks. The multiplied shoots were elongated on WPM supplemented with 0.44 mM BAP. Around 90% of in vitro shoots were rooted without basal callus formation on WPM supplemented with 5.3 mM NAA (α-naphthalene acetic acid) within 4 weeks. Following this protocol, 1100 micropropagated plantlets of an elite line (Ha, Bhutan) were hardened in their natural habitat. The present study illustrates the usefulness of additives for mass propagation and germplasm conservation and is, to the best of our knowledge, the first report of in vitro propagation of P. scrophulariiflora.
Manoj Kumar Goel, Shilpa Goel, Suchitra Banerjee, Karuna Shanker, Arun Kumar Kukreja (India) Agrobacterium rhizogenes-Mediated Transformed Roots of Rauwolfia serpentina for Reserpine Biosynthesis (pp 8-14)
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ABSTRACT
Original Research Paper: Root extracts of Rauwolfia serpentina have been used for centuries in Ayurvedic medicine as a panacea for a wide variety of physical as well as mental disorders. The potential of Agrobacterium rhizogenes-mediated genetic transformation for the synthesis of phytomolecules of high pharmaceutical value is now well established and documented. Transgenic roots were induced from R. serpentina leaf explants in response to A. rhizogenes A4 strain on semi-solid ½-strength MS medium. Amongst 200 hairy root clones developed, 27 showing persistent and incessant growth over several generations were selected. Transformed roots grew vigorously and branched profusely on hormone-free liquid B5 medium with 3% sucrose with higher biomass yields compared to the control and showed two stable and distinct morphotypes. Medium devoid of any carbon source served as the control. The transformed nature of the roots was confirmed by PCR amplification with rolA primers. Growth kinetic studies exhibited the highest growth index (58.57 ± 1.92) at the 10th week followed by slow growth in the subsequent period up to 14 weeks. Reserpine content increased with root growth and was highest in 10-weeks-old cultures. Hairy root clones showed a wide array of variation in relative reserpine content, varying from 0.0064 to 0.0858% dry weight (DW). On the basis of relative reserpine content, these hairy root clones were classified into 5 different groups. SM12 clone had the highest reserpine level (0.0858% DW) producing 2-3 times more than the content of field-grown roots harvested after 18-24 months. A distinct relationship between root morphology and reserpine content was observed. The present study is the first report of reserpine production in quantifiable amounts from the hairy roots of any Rauwolfia species.
Amita Misra, Ashutosh K. Shukla, Ajit K. Shasany, V. Sundaresan, Shital P. Jain, Subhash C. Singh, Guru D. Bagchi, Suman P. S. Khanuja (India) AFLP Markers for Identification of Aconitum Species (pp 15-19)
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ABSTRACT
Original Research Paper: The genus Aconitum is highly complex and its taxonomy has been traditionally difficult due to the high level of variation among the various species. The Aconitum species are known for their highly toxic diterpenoid alkaloids but have been described in traditional medicine systems as high-value medicine after proper and prescribed detoxification. In India, A. heterophyllum, A. balfourii and A. violaceum are found mainly in the North-Western Himalayas whereas A. ferox is found in the North-Eastern Himalayan region. Among these species, A. heterophyllum is the most significant in terms of therapeutic importance and herbal drug market value. It has become critically endangered due to high demand of the herb and indiscriminate overexploitation. There is an existing demand in the bulk herbal drug industry to have an authentic identification system for the Aconitum species in order to enable their commercial use as genuine phytoceuticals. In the present study we have used Amplified Fragment Length Polymorphism (AFLP) for developing DNA fingerprints for 4 Aconitum species. A total of 10 accessions (4 of A. heterophyllum, 3 of A. violaceum, 2 of A. balfourii and 1 of A. ferox) from the 4 species were used in the study, which employed 64 AFLP selective primer pairs. Only 26 selective primer pairs were found to respond with all the accessions and generated a total of 4112 fragments. A number of species-specific markers were identified for all the 4 Aconitum species (16 for A. heterophyllum, 125 for A. violaceum, 79 for A. balfourii,and226 for A. ferox). These AFLP fingerprints of the Aconitum species could be used in future for authentication of the drug and checking the adulteration-related problems faced by the commercial users of the herb.
Rafia Rasool, Bashir Ahmad Ganai, Azra Nahaid Kamili, Seema Akbar, Akbar Masood (India) Antioxidant and Antibacterial Activities of Extracts from Wild and in Vitro-Raised Cultures of Prunella vulgaris L. (pp 20-27)
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Original Research Paper: MeOH, EtOH, CHCl3 and aqueous extracts from the whole plant of wild Prunella vulgaris, a Kashmir Himalayan perennial medicinal herb,as well as from in vitro-regenerated plants were evaluated and compared for their antioxidant and antimicrobial properties. Antioxidant activity was screened by using various in vitro models: scavenging of the free radicals using DPPH, riboflavin photo oxidation, DNA damage, inhibition of lipid oxidation via PMS, FTC and TBA assay. The MeOH and CHCl3 extract from wild and in vitro-regenerated plants possessed an almost equal radical scavenging effect. In vitro and wild grown plant extracts in different solvent systems were also screened for antimicrobial activity against medically important bacterial strains by the agar well diffusion method. The MeOH extract of both (wildand in vitro) plants extracts were almost equally effective against Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and Kleibsella pneumonae. Both in vitro and wild dried plant extracts showed an almost similar concentration-dependent antioxidant and antimicrobial inhibition. Therefore, the commercial manufacture of active constituents from these improved elite lines would be useful and profitable. The present study provides first evidence that in vitro grown P. vulgaris has antioxidant and antibacterial activities, suggesting the potential of the tissue culture technique to substitute wild P. vulgaris in the pharmaceutical industry.
Ripu M. Kunwar (Nepal/USA), Chundamani Burlakoti (USA), Chhote L. Chowdhary (Nepal), Rainer W. Bussmann (USA) Medicinal Plants in Farwest Nepal: Indigenous Uses and Pharmacological Validity (pp 28-42)
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ABSTRACT
Original Research Paper: Medicinal plants have been used indigenously since ancient past as medicines for the treatment of various ailments. However, the knowledge of indigenous therapies have been distorting to these days due to changing perception, acculturation, commercialization and socio-economic transformations. The present study compares indigenous knowledge of therapies of 48 medicinal plants with the latest common pharmacological findings. Traditional indigenous plant knowledge and phytomedicine are consistently gaining acceptance in global society. The present study found that over two-thirds of traditionally used plants in the region show clear pharmacological efficacy. Total 23 species possessed strong resemblances and the species Euphorbia royleana, Ricinus communis, Plantago major, Chenopodium album, Cordyceps sinensis, etc. contributed the most. The complementarity of indigenous therapies and pharmacological uses is obvious and it is base of the modern therapeutic medicine. The increasing use of indigenous therapies demands more scientifically sound evidence, therefore further investigation and phytochemical screening of ethnopharmacologically used plants and assessment of the validity to the indigenous uses is worthwhile.
Mukesh Joshi, Munesh Kumar (India), Rainer W. Bussmann (USA) Ethnomedicinal Uses of Plant Resources of the Haigad Watershed in Kumaun Himalaya, India (pp 43-46)
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ABSTRACT
Research Note: The present study was carried out in the Haigad watershed of Kumaun Himalaya. A total of 32 medicinal plant species belonging to 26 families were recorded. A major proportion of species were in forested landscape (62%) and the rest in cultural landscape (38%) of the watershed. The plants used for medicinal purposes in the local health traditions are gradually becoming extinct due to developmental activities, population explosion and other anthropogenic reasons. To avoid overexploitation and promote sustainable use, rapid conservation efforts are needed. Farmers should be involved in the cultivation of medicinal plants emphasizing suitable production methods, especially on barren and fallow land.
Parveen Kumar Sharma, N. S. Chauhan, Brij Lal, Amjad M. Husaini (India), Jaime A. Teixeira da Silva (Japan), Punam (India) Conservation of Phyto-diversity of Parvati Valley in Northwestern Himalayas of Himachal Pradesh, India (pp 47-63)
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Research Note: This study provides information about the traditional indigenous uses of plants by the inhabitants of the Parvati Valley of Kullu district in the western Himalayas of India. Since no published literature from the past 10 years exists, an ethnobotanical survey was conducted among the ethnic groups of the Parvati valley and first hand information on these plant species was recorded. A total of 266 species belonging to 180 genera and 71 families (including 44 species as recorded for the first time in the area) were collected. Out of these, 223 species within 152 genera of 61 families belong to dicots; 31 species and 22 genera under 7 families belong to monocots and 10 species with 6 genera in 3 families belong to gymnosperms.
Venkat Kishore Ryakala (India), Shahin Sharif Ali (Ireland/India), Hallihosur Sharanabasava, Naushaba Hasin, Pragya Sharma, Utpal Bora (India) Ethnobotany of Plants Used to Cure Diabetes by the People of North East India (pp 64-68)
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Research Note: Northeast India is considered as an ecological hot spot and has a wide variety of flora and fauna. Diverse ethnic communities inhabit the area, each having their own traditional medical cures for different diseases. During the course of present studies it was found that 52 species of plants belonging to 36 families are used as antidiabetic agents in folk medicinal practice. Leaves and bark were found to be the two major plant parts used for making hypoglycemic herbal preparations. Around 26 treatments involve administration of decoction to the diabetic patient. These decoctions are either prepared from leaves, bark, fruit, root, seeds or from whole plants. Out of the 52 plants 12 are also reported to have antidiabetic properties in the Diabetes Medicinal Plant Database. The remaining plants could be a potential source of new and efficient cures for diabetes.
Ram Swaroop Verma, Rajendra Chandra Padalia, Amit Chauhan, Ajai Kumar Yadav (India) Chemical Composition of Leaf and Flower Essential Oils of Two Thymus spp. from Western Himalaya (pp 69-72)
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Original Research Paper: Thymus species (Lamiaceae) are considered to be very beneficial whether used as food or as a medicament. Essential oils (EOs) derived from leaves and flowers of Thymus serpyllum and Thymus linearis grown in northern India were analyzed by GC and GC-MS. A total of 37 components forming 94.8-98.4% of EO composition were identified. The EOs of both species were rich in thymol, p-cymene and g-terpinene. Thymol was higher in the EO of T. linearis (74.6-75.8%) compared to T. serpyllum (51.9-70.1%). The amount of thymol methyl ether, p-cymene, 1-octen-3-ol, camphor and borneol was relatively higher in T. serpyllum EO. Further, phenolic monoterpenes were higher in flower EOs of both species than in leaf EOs.
Ram Swaroop Verma, Rajendra Chandra Padalia, Amit Chauhan (India) Chemical Profiling of Mentha spicata L. var. ‘viridis’ and Mentha citrata L. Cultivars at Different Stages from the Kumaon Region of Western Himalaya (pp 73-76)
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Research Note: Two spearmint (Mentha spicata L. var. ‘viridis’)cultivars viz. ‘Neerkalka’ and ‘Supriya’ and one bergamot mint (Mentha citrata L.) cultivar ‘Kiran’ cultivated in the Kumaon region of northern India were investigated for their essential oil content and composition at different stages of crop growth. Essential oil content and composition were both affected by crop age in all cultivars. All the cultivars accumulated maximum essential oil at 150 days after transplanting. The percentage carvone in ‘Neerkalka’ was higher at 90 days (67.0%) followed by the 150-days-old crop (61.68%), while in ‘Supriya’, carvone concentration increased at 150 days (72.47%). In ‘Kiran’, linalool and linalyl acetate were highest in 150- and 180-days-old crops, respectively.
Amit Chauhan, Ram Swaroop Verma (India) Cultivation Potential of Three Rose-scented Geranium (Pelargonium graveolens) Cultivars in the Kumaon Region of Western Himalayas (pp 77-79)
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Research Note: A field experiment was conducted to evaluate the production potential of three cultivars of rose-scented geranium viz., ‘Bourbon’, ‘CIM-Pawan’ and ‘Kelkar’ in the temperate region of Uttarakhand. ‘CIM-Pawan’ had the highest essential oil yield (103.87 g plot-1) followed by ‘Kelkar’ (79.93 g plot-1) and ‘Bourbon’ (72.01 g plot-1). The essential oil profile of ‘Bourbon’ was rich (relative percentages) in citronellol (29.05), geraniol (24.36), citronellyl formate (5.94), isomenthone (5.82); the oil of ‘CIM-Pawan’ was rich in citronellol (32.60), geraniol (21.38), 10-epi-γ-eudesmol (6.83), citronellyl formate (6.29) while the essential oil of ‘Kelkar’ showed a different profile with citonellol (61.48) and isomenthone (10.56) being almost twice that of other cultivars.
Javid Ahmad Parray, Azra N. Kamilli, Raies Qadri, Rehana Hamid (India), Jaime A. Teixeira da Silva (Japan) Evaluation of Antibacterial Activity of Euryale ferox Salisb., a Threatened Aquatic Plant of Kashmir Himalaya (pp 80-83)
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Research Note: The antibacterial activity of methanolic extract of seeds and leaves of Euryale ferox was tested against nine clinically isolated bacterial strains (Staphylococus aureus, Escherichia coli, Pseudomonas aureoginosa, Citrobacter freundi, Shigella flexneri, Kliebsella pneumoniae, Proteus vulgaris, Salmonella typhi and Salmonella typhimurium)and subsequently was also tested for minimum inhibitory concentration (MIC) values which ranged from 0.25 to 500 mg/l against six ATCC bacterial strains using micro broth dilution method. The broad spectrum activity displayed by the seed and leaf extracts appears to provide a scientific basis for the use of E. ferox in kidney problems and urinary tract infections in ethno medicines.
Rehana Hamid, Azra N. Kamili, Mahmood uz Zaffar (India), Jaime A. Teixeira da Silva (Japan), A. Mujib, Javid Ahmad Parray (India) Callus-Mediated Shoot Organogenesis from Shoot Tips of Cichorium intybus (pp 84-86)
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Research Note: Cichorium intybus L. is a medicinally important plant with anti-cancerous and anti-hepatotoxic properties. An efficient method for totipotent callus formation has been developed in C. intybus from the basal portion of shoot tip explants on MS medium supplemented with different concentrations of plant growth regulators (PGRs) like 6-benzylamino purine (BAP) and kinetin (Kn) with an auxin, indole-3-butyric acid (IBA). Cultures growing under the influence of BAP+IBA produced considerably more callus than cytokinins used alone. Re-differentiation of such callus led to multiple shoot formation on the same medium after 3 weeks. Isolated shoots were individually rooted in the presence of different concentrations of IBA. Plantlets obtained were transplanted into small pots containing peat, vermiculite, sand and soil mixture (1:1:1:1), 60% of which survived.
Rehana Hamid, Mahmood uz Zaffar (India), Jaime A. Teixeira da Silva (Japan), Azra N. Kamili, Javid Ahmad Parray (India) Impact of Chromium on the Oxidative Defense System of Brassica juncea L. cv. ‘Pusa Jai Kissan’ under Hydroponic Culture (pp 87-89)
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Research Note: Brassica juncea is a medicinally important plant and is commonly used as a diuretic, stimulant and to treat arthritis. Seed are used for the treatment of tumors and stomach disorders. B. juncea can hyperaccumulate cadmium and many other soil trace elements like selenium, chromium, iron and zinc food supplements. Chromium (Cr)-induced oxidative damage and changes in the contents of proline and glutathione in leaves of B. juncea. L. cv. ‘Pusa Jai Kissan’ were investigated after 3 and 5 days of treatment under hydroponic culture. Cr was supplied as K2Cr2O7. The main response was an increase in superoxide dismutase activity and proline content which subsequently reduced the activity of catalase and glutathione content in plants.
Janifer Raj, Ballabh Basanth, Pal M. Murugan (India), Jaime A. Teixeira da Silva (Japan), Kumar Saurav, Om P. Chaurasia, Shashi Bala Singh (India) Screening Phytochemical Constituents of 21 Medicinal Plants of Trans-Himalayan Region (pp 90-93)
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Research Note: Alkaloids, tannins, flavonoids, saponins, steroids, and cardiac glycoside distribution in 1 high altitude medicinal plants belonging to different families (Apiaceae, Asteraceae, Crassulaceae, Lamiaceae, Rosaceae, Rubiaceae, Urticaceae, and Zygophyllaceae) were assessed and compared. The plants investigated were Achillea millefolium, Artemesia dracunculus, Bidens pilosa, Carum carvi, Dracocephalum heterophyllum, Ferula jaeskiana, Gallium pauciflorum, Heracleum pinnatum, Hippophae rhamnoides, Inula racemosa, Mentha longifolia, Nepeta podostachys, Origanum vulgare, Peganum harmala, Rhodiola imbricata, Rhodiola heterodenta, Rosa webbiana, Rosa macrophylla, Rubia cordifolia, Tanacetum gracile,and Utrica hyperborea,which have been widely used for time immemorial in the traditional Amchi system of medicine in the Ladakh region of India. Phytochemicals were qualitatively detected using aqueous extracts and solvent fractions of plants using various biochemical tests. These plants are a potential source of useful drugs. Future studies will isolate, identify, characterize and elucidate the structure of novel bioactive compounds. The significance of these plants in traditional medicine and the importance of the distribution of their chemical constituents are discussed in the context of the role of these plants in ethnomedicine in Ladakh. |
