Medicinal and Aromatic Plant Science and Biotechnology
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Medicinal and Aromatic Plant Science and Biotechnology
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<strong>Medicinal</strong> <strong>and</strong> <strong>Aromatic</strong> <strong>Plant</strong> <strong>Science</strong> <strong>and</strong> <strong>Biotechnology</strong><br />
Abbreviation: <strong>Medicinal</strong> <strong>Aromatic</strong> <strong>Plant</strong> Sci. Biotech.<br />
Print: ISSN 1752-3389<br />
Frequency <strong>and</strong> Peer status: Biannual, Peer reviewed<br />
Scope <strong>and</strong> target readership: <strong>Medicinal</strong> <strong>and</strong> <strong>Aromatic</strong> <strong>Plant</strong> <strong>Science</strong> <strong>and</strong> <strong>Biotechnology</strong> will provide a complete analysis <strong>and</strong><br />
underst<strong>and</strong>ing of all aspects of medicinal <strong>and</strong> aromatic plant science <strong>and</strong> biotechnology.<br />
<strong>Medicinal</strong> <strong>and</strong> <strong>Aromatic</strong> <strong>Plant</strong> <strong>Science</strong> <strong>and</strong> <strong>Biotechnology</strong> primarily wishes to examine:<br />
1) Cultural practices (greenhouse growth, hydroponics, aeroponics, organic farming);<br />
2) In vitro propagation (micropropagation, somatic embryogenesis, tissue culture, bioreactor system production);<br />
3) Mycorrhizal symbioses (<strong>and</strong> effects on plant physiology, productivity, reproduction <strong>and</strong> disease resistance);<br />
4) Novel techniques for analysis (genetic, biochemical, biophysical);<br />
5) Physiology, genetics, molecular biology, structural botany (integrated, pure <strong>and</strong> applied);<br />
6) Pathology;<br />
7) Production of secondary metabolites, organic <strong>and</strong> inorganic biochemistry, <strong>and</strong> phytochemistry; pharmacological properties;<br />
8) Storage of valuable genetic material (cold-storage or cryopreservation).<br />
For publication in <strong>Medicinal</strong> <strong>and</strong> <strong>Aromatic</strong> <strong>Plant</strong> <strong>Science</strong> <strong>and</strong> <strong>Biotechnology</strong> the research must provide a highly significant new<br />
contribution to our underst<strong>and</strong>ing of medicinal <strong>and</strong> aromatic plants (from any climactic or geographic origin) <strong>and</strong> must generally be<br />
supported by a combination of either: physiological, biochemical, genetic or molecular analyses. All areas of study are welcome <strong>and</strong> the<br />
experimental approaches used can be wide-ranging. Results that simply provide a description without an integrated multi-disciplinary<br />
approach might not be considered, as might descriptive or overly-localized studies <strong>and</strong> reports on conventional propagation without a<br />
wide impact. Manuscripts pertaining to breeding, post-harvest technology, nutritional aspects or influences on human health of medicinal<br />
<strong>and</strong> aromatic plants will also be considered.<br />
Editor-in-Chief<br />
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Ch<strong>and</strong>i Charan Rath, North Orissa University, India<br />
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Mohammad Jamal Saharkhiz, Shiraz University, Iran<br />
Sabuj Sahoo, Utkal University, India<br />
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Mohd. Shahid, Aligarh Muslim University, India<br />
Junjie Shan, Beijing Institute of Pharmacology & Toxicology,<br />
China<br />
Chunmeng Shi, Third Military Medical Univeristy, China<br />
Neeta Shrivastava, B. V. Patel Pharmaceutical Education <strong>and</strong><br />
Research Development (PERD) Centre, India<br />
Perumal Siddhuraju, Bharathiar University, India<br />
Angelina Subotić, Institute for Biological Research "Siniša<br />
Stanković", Serbia<br />
Sivaramakrishnan Sudhakaran, AIMST University, Malaysia<br />
Klára Szentmihályi, Hungarian Academy of <strong>Science</strong>s, Hungary<br />
Ilias S. Travlos, Benaki Phytopathological Institute, Greece<br />
Rong Tsao, Agriculture & Agri-Food Canada, Canada<br />
Supayang P. Voravuthikunchai, Prince of Songkla University,<br />
Thail<strong>and</strong><br />
An<strong>and</strong> K. Yadav, Fort Valley State University, USA
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<strong>Medicinal</strong> <strong>and</strong> <strong>Aromatic</strong> <strong>Plant</strong> <strong>Science</strong> <strong>and</strong> <strong>Biotechnology</strong> ©2010 Global <strong>Science</strong> Books, Ltd.<br />
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Guest Editor<br />
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Sher-e-Kashmir University of Agricultural <strong>Science</strong>s & Technology of Kashmir, India<br />
Cover photos/figures: Left plate: Musical instrument of local deity Jamlu, Malana village (top), Angelica glauca (center<br />
left), Hedychium spicatum (center right), Udithach (largest alpine pasture in the valley) (bottom) (Sharma et al., pp 47-63).<br />
Top right row: Some Himalayan medicinal plants. (Left) Rhus parviflora. Fruits are indigenously used for diarrhea <strong>and</strong><br />
dysentery. (Center) Urtica dioica. Stem juice is valued for sprain <strong>and</strong> fractures. (Right) Euphorbia royleana. <strong>Plant</strong> is kept in<br />
roof of house for protecting from evil. (Kunwar et al., pp 28-42). Center plate <strong>and</strong> gel: Agrobacterium rhizogenes-mediated<br />
genetic transformation in Rauwolfia serpentina with rolA (top gel) <strong>and</strong> virD1 (top gel) PCR detection (Goel et al., pp 8-14).<br />
Center right: Mature rhizomes of different genotypes of Picrorhiza scrophulariiflora. (Top) Bhutan, (Center) North Sikkim,<br />
(Bottom) East Sikkim (Bantawa et al., pp 1-7). Bottom right: In vitro propagation <strong>and</strong> acclimatization of Cichorium intybus<br />
through indirect callus culture on MS + 10 µM IBA (Hamid et al., pp 84-86).<br />
Disclaimers: All comments, conclusions, opinions, <strong>and</strong> recommendations are those of the author(s), <strong>and</strong> do not necessarily<br />
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manuscript, <strong>and</strong> accepts product descriptions <strong>and</strong> details to be an integral part of the scientific content.<br />
Printed in Japan on acid-free paper.<br />
Published: December, 2010.
The Guest Editor<br />
Dr. Amjad Masood Husaini<br />
Dr. Amjad Masood Husaini, a young Scientist working as Assistant Professor in Sher-e-Kashmir University of<br />
Agricultural <strong>Science</strong>s & Technology of Kashmir (India) holds a Ph.D. in <strong>Biotechnology</strong> <strong>and</strong> PG Diploma in Bioinformatics<br />
(Jamia Hamdard, New Delhi), besides certificates in Intellectual Property Rights (Indian Law Institute, New Delhi) <strong>and</strong><br />
Remote Sensing Applications in Agriculture (Indian Agricultural Research Institute-Indian Space Research Organization).<br />
Recipient of Young Scientist Award-2009 in Agriculture (Jammu & Kashmir State Council for <strong>Science</strong> & Technology,<br />
Government of J&K), Jawahar Lal Nehru Award for Agricultural Research-2008 (Indian Council of Agricultural Research,<br />
Government of India), Junior Scientist of the Year Award-2007 (National Environmental <strong>Science</strong>s Academy, New Delhi), he<br />
is listed among Top 100 Scientists of 2010 by the International Biographical Centre (IBC, Cambridge), <strong>and</strong> his biography<br />
included in 27 th edition of Marquis Who’s Who in the World. With an illustrious academic career Dr. Husaini holds the<br />
distinction of being top position holder in National Eligibility Tests for Life <strong>Science</strong>s <strong>and</strong> Agricultural <strong>Biotechnology</strong> in<br />
India. His publications include book entitled ‘Strawberry- Transgenics for stresses’ <strong>and</strong> more than two dozen research/<br />
review papers in National <strong>and</strong> International journals of repute, discussing different aspects of agricultural research <strong>and</strong><br />
technology.<br />
Dr. Husaini serves as member of professional associations like World Association of Young Scientists, New York<br />
Academy of <strong>Science</strong>s, The Indian <strong>Science</strong> Congress Association, <strong>Biotechnology</strong> Society of India, National Environmental<br />
<strong>Science</strong> Academy (India), Young Professionals’ Platform for Agricultural Research for Development, Scientists Without<br />
Borders, International Association of Computer <strong>Science</strong> <strong>and</strong> Information Technology, Royal Society of Crop <strong>Science</strong>,<br />
International Society for Biosafety Research, <strong>and</strong> serves in the capacity of editor/ associate editor etc. in editorial boards of<br />
various International journals of repute.
Foreword<br />
Amjad Masood Husaini<br />
Sher-e-Kashmir University of Agricultural <strong>Science</strong>s & Technology of Kashmir, J&K 191121, India<br />
E-mail: dr.amjadhusaini@hotmail.com<br />
“Man, ever desirous of knowledge, has already explored many things, but more <strong>and</strong> greater still remains concealed;<br />
perhaps reserved for far distant generations, who shall prosecute the examination of their Creator’s work in remote countries,<br />
<strong>and</strong> make many discoveries for the pleasure <strong>and</strong> convenience of life” (Linnaeus, 1754). One such vast unexplored region<br />
<strong>and</strong> a biodiversity hot spot, lies between two great ancient civilizations of India <strong>and</strong> China <strong>and</strong> is famous as “The Great<br />
Himalayan Region”. The main Himalaya range runs west to east, from the Indus river valley to the Brahmaputra river valley,<br />
forming an arc 2,400 km long, which varies in width from 400 km in the western Kashmir-Xinjiang region to 150 km in the<br />
eastern Tibet-Arunachal Pradesh region. The range consists of three coextensive sub-ranges, with the northernmost, <strong>and</strong><br />
highest, known as the Great or Inner Himalayas. The ancient religious scripture of Hindus, Atharvaveda is the earliest<br />
celebrated treatise mentioning the use of medicinal plants of the region. Atharvaveda contains 114 hymns or formulations<br />
for the treatment of diseases. Ayurveda, a system of traditional medicine native to the Indian subcontinent, originated in <strong>and</strong><br />
developed from these hymns. The Suśruta Saṃhitā <strong>and</strong> the Charaka Saṃhitā are two important works on this traditional<br />
system of medicine. In addition there is a famous reference in Valmiki’s Ramayana, a religious scripture of Hindus, about<br />
the existence of rare medicinal plant Sanjivani (Selaginella bryopteris) in Himalayas, which saved the life of Lakshmana<br />
(brother of the Hindu god Lord Rama).<br />
Over the centuries people have depended on these medicinal plants for treating daily ailments like cough, colds,<br />
indigestion, ulcers, sore eyes etc. In fact Sir Lawrence, a British Settlement Commissioner in his book, ‘The Valley of<br />
Kashmir’ (1895) refers to this point as, “when I have made inquiries as to various herbs which I have seen in the valley <strong>and</strong><br />
on hillsides, I am always told that they are hot <strong>and</strong> good for cold humours, cold <strong>and</strong> good for hot humours, dry <strong>and</strong><br />
beneficial to damp humours, damp <strong>and</strong> beneficial to dry humours.”<br />
In this Special Issue (SI) on Himalayan MAPS, an attempt has been made to present various issues pertaining to<br />
conservation, documentation, biotechnological applications <strong>and</strong> medicinal uses of plants of Himalayan region. The SI<br />
comprises of 13 research articles related to different areas of plant biotechnology. In the first paper Bantawa et al. take-up<br />
an important highly valued endangered medicinal plant of Indo-China Himalayas viz. Picrorhiza scrophulariiflora Pennell<br />
<strong>and</strong> describe in detail its micropropagation. This study is first such report on this plant <strong>and</strong> illustrates the usefulness of<br />
additives for mass propagation <strong>and</strong> germplasm conservation. In a similar study Hamid et al. describe a method for in vitro<br />
shoot organogenesis of Cichorium intybus using shoot tips as explants. Cichorium intybus is known for its anti-cancerous<br />
<strong>and</strong> anti-hepatotoxic properties <strong>and</strong> their successful transfer to pots with 60% survival percentage is a step forward towards<br />
its ex situ conservation. The potential of Agrobacterium rhizogenes-mediated genetic transformation for the synthesis of<br />
phytomolecules of high pharmaceutical value is well established. Goel et al. present the first report of reserpine production<br />
in quantifiable amounts from the Agrobacterium rhizogenes-generated transgenic hairy roots of Rauwolfia serpentine,<br />
whose root-extracts have been used for centuries in Ayurvedic medicine. In one clone the reserpine level was found to be 2-<br />
3 times that of field grown roots, which is quite encouraging.<br />
Supply of authentic medicinal plants to herbal drug industry is an important requisite for enabling their commercial use<br />
in production of genuine phytoceuticals. An authentic identification system based on amplified fragment length<br />
polymorphism (AFLP) for Aconitum heterophyllum, A. violaceum, A. balfourii <strong>and</strong> A. ferox has been reported in an original<br />
research paper by Misra et al., which could be used for checking adulteration-related problems faced by commercial users
of the herb. Rasool et al. compared antioxidant <strong>and</strong> antimicrobial properties of wild <strong>and</strong> in vitro-regenerated plants of a<br />
Kashmir Himalayan perennial medicinal herb, Prunella vulgaris. Their study is probably the first report giving evidence<br />
that in vitro grown P. vulgaris has antioxidant <strong>and</strong> antibacterial activities similar to that of wild, suggesting the substitution<br />
of wild P. vulgaris with tissue culture raised plants for use in pharmaceutical industry. In another study on antibacterial<br />
activity, the potential of methanolic extract of seeds <strong>and</strong> leaves of Euryale ferox was tested against nine clinically isolated<br />
bacterial strains by Parray et al. The broad spectrum activity displayed by these extracts appears to provide logic for the use<br />
of E. ferox as ethno-medicine in urinary tract infections. The issues related to ethno-medicinal uses <strong>and</strong> overexploitation of<br />
medicinal plants of Haigad watershed of Kumaun Himalaya have been discussed by Joshi et al. They argue that for<br />
sustainable use, in addition to rapid conservation efforts, farmers should be involved in the cultivation of medicinal plants.<br />
An exhaustive ethno-botanical survey on phyto-diversity, spanning over more than 250 species, of Parvati Valley in<br />
Northwestern Himalayas described by Sharma et al. is highly informative. They stress the involvement of local inhabitants<br />
for conservation of indigenous knowledge <strong>and</strong> traditional practices. In a similar study on medicinal plants of west Nepal,<br />
Kunwar et al. compare indigenous knowledge of therapies of 48 medicinal plants with the latest common pharmacological<br />
findings, suggesting complementarities <strong>and</strong> thus forming base for use in modern therapeutic medicine. Similar correlation<br />
was reported by Ryakala et al. while studying the ethnobotany of 52 plant species used to cure diabetes by the inhabitants of<br />
north eastern India. Raj et al. have screened phytochemical constituents of 21 medicinal plants used in traditional Amchi<br />
system of medicine in the Ladakh region of India. The significance of these plants is discussed in the context of their role in<br />
ethnomedicine All these studies have generated the possibilities of using the unexplored plants as potential sources of future<br />
drugs.<br />
Verma et al. have contributed an informative paper describing the chemical composition of leaf <strong>and</strong> flower essential oils<br />
of Thymus serpyllum <strong>and</strong> T. linearis from Western Himalaya, while Hamid et al. discuss the impact of chromium on the<br />
oxidative defense system of Brassica juncea L., a medicinally important plant commonly used as a diuretic <strong>and</strong> stimulant.<br />
I hope that the scientists working on medicinal plants will find this Special Issue helpful in moving forward in their<br />
important quest of contributing in the area of medicine, drug discovery, <strong>and</strong> conservation of medicinal plants etc. I would<br />
like to thank Dr. Jaime A. Teixeira da Silva <strong>and</strong> Ms. Kasumi Shima at Global <strong>Science</strong> Books Ltd., UK for their cooperation<br />
<strong>and</strong> helpful suggestions; <strong>and</strong> my family for their underst<strong>and</strong>ing <strong>and</strong> support during the prolonged <strong>and</strong> time-consuming work<br />
on this volume.<br />
December, 2010
Muhammad Iqbal, PhD, FNASc<br />
Professor<br />
Department of Botany<br />
JAMIA HAMDARD<br />
(Declared as Deemed-to-be University under Section 3 of<br />
the UGC Act, 1956 vide Notification No. F.9-18/85-U.3<br />
dated 10.5.1989 of the Government of India)<br />
December 24, 2010<br />
The tradition of herbal treatment for curing the human ailments is pretty old in India. Formal<br />
accounts of medicinal plants appeared as early as during the Vedic period in the Vedico-<br />
Brahminic treatises like Atharvaveda (2000 BC), Sushruta Samhita (1300 BC) <strong>and</strong> Charaka<br />
Samhita (300 BC). Of the tradional Indian systems of medicine, the Ayurveda (science of life)<br />
<strong>and</strong> the Unani (Greeco-Arabian) systems are based largely on medicinal plants, whereas<br />
Siddha depends mainly on minerals. Over the centuries, the traditional practitioners have<br />
developed a number of herbal formulations for the treatment of various ailments with special<br />
emphasis on sexual debility, liver disorders <strong>and</strong> kidney problems. As the popular alternative<br />
medicine, these preparations now constitute an important segment of the integrated health<br />
management all over the world.<br />
The Himalayas, often called "The Roof of the World", encompass a number of biodiversity hot<br />
spots <strong>and</strong> repositories of medicinal plants. The whole Himalayan range is envisaged as a<br />
trove of medicinal herbs, offering refuge to a variety of rare plants in its varied mountain<br />
ecosystems. The research work carried out in the recent past has accumulated enough<br />
scientific information on a variety of medicinal plants inhabiting the various zones of the<br />
Himalayan range with diverse climatic conditions. Given the above, the document in h<strong>and</strong> is a<br />
commendable effort that duly elucidates the various aspects of the medicinal-plant research<br />
that have a potential promise for a safe herbal medication without the much talked about<br />
adverse after-effects. The information covered by this issue is comprehensive <strong>and</strong> most of the<br />
plants mentioned are well known for their therapeutic efficacy. Information on the traditional<br />
knowledge, describing the ethno-medicinal uses of plants is also included.<br />
I heartily appreciate Dr. Amjad Masood Husaini of the Sher-e-Kashmir University of<br />
Agricultural <strong>Science</strong>s & Technology of Kashmir, India, for editing this useful volume that<br />
focuses on the medicinal plants of the Himalayas, <strong>and</strong> also the galaxy of distinguished<br />
scientists <strong>and</strong> researchers who have contributed for this special issue of the <strong>Medicinal</strong> <strong>and</strong><br />
<strong>Aromatic</strong> <strong>Plant</strong> <strong>Science</strong> <strong>and</strong> <strong>Biotechnology</strong>, an emerging research journal of the Global<br />
<strong>Science</strong> Books (GSB), UK. This document must prove a useful guide to botanists, cultivators<br />
<strong>and</strong> collectors of medicinal plants <strong>and</strong> a pride possession of all those who are keen on the<br />
Himalayan vegetation.<br />
(Muhammad Iqbal)<br />
Hamdard Nagar, New Delhi – 110 062, INDIA<br />
Phone: +91-11-2605 9688, Extn.: 5530 (O), 5531 (R); Fax: +91-11-2605 9663<br />
E-mail: iqbalg5@yahoo.co.in
SPECIAL ISSUE: CONTENTS<br />
Pranay Bantawa, Swapan Kumar Ghosh, Pamita Bh<strong>and</strong>ari, Bikram Singh, Partha Deb Ghosh, Paramvir Singh Ahuja,<br />
Tapan Kumar Mondal (India) Micropropagation of an Elite Line of Picrorhiza scrophulariiflora, Pennell, an Endangered<br />
High Valued <strong>Medicinal</strong> <strong>Plant</strong> of the Indo-China Himalayan Region<br />
Manoj Kumar Goel, Shilpa Goel, Suchitra Banerjee, Karuna Shanker, Arun Kumar Kukreja (India) Agrobacterium<br />
rhizogenes-Mediated Transformed Roots of Rauwolfia serpentina for Reserpine Biosynthesis<br />
Amita Misra, Ashutosh K. Shukla, Ajit K. Shasany, V. Sundaresan, Shital P. Jain, Subhash C. Singh, Guru D. Bagchi,<br />
Suman P. S. Khanuja (India) AFLP Markers for Identification of Aconitum Species<br />
Rafia Rasool, Bashir Ahmad Ganai, Azra Nahaid Kamili, Seema Akbar, Akbar Masood (India) Antioxidant <strong>and</strong><br />
Antibacterial Activities of Extracts from Wild <strong>and</strong> in Vitro-Raised Cultures of Prunella vulgaris L.<br />
Ripu M. Kunwar (Nepal/USA), Chundamani Burlakoti (USA), Chhote L. Chowdhary (Nepal), Rainer W. Bussmann<br />
(USA) <strong>Medicinal</strong> <strong>Plant</strong>s in Farwest Nepal: Indigenous Uses <strong>and</strong> Pharmacological Validity<br />
Mukesh Joshi, Munesh Kumar (India), Rainer W. Bussmann (USA) Ethnomedicinal Uses of <strong>Plant</strong> Resources of the Haigad<br />
Watershed in Kumaun Himalaya, India<br />
Parveen Kumar Sharma, N. S. Chauhan, Brij Lal, Amjad M. Husaini (India), Jaime A. Teixeira da Silva (Japan), Punam<br />
(India) Conservation of Phyto-diversity of Parvati Valley in Northwestern Himalayas of Himachal Pradesh, India<br />
Venkat Kishore Ryakala (India), Shahin Sharif Ali (Irel<strong>and</strong>/India), Hallihosur Sharanabasava, Naushaba Hasin, Pragya<br />
Sharma, Utpal Bora (India) Ethnobotany of <strong>Plant</strong>s Used to Cure Diabetes by the People of North East India<br />
Ram Swaroop Verma, Rajendra Ch<strong>and</strong>ra Padalia, Amit Chauhan, Ajai Kumar Yadav (India) Chemical Composition of<br />
Leaf <strong>and</strong> Flower Essential Oils of Two Thymus spp. from Western Himalaya<br />
Ram Swaroop Verma, Rajendra Ch<strong>and</strong>ra Padalia, Amit Chauhan (India) Chemical Profiling of Mentha spicata L. var.<br />
‘viridis’ <strong>and</strong> Mentha citrata L. Cultivars at Different Stages from the Kumaon Region of Western Himalaya<br />
Amit Chauhan, Ram Swaroop Verma (India) Cultivation Potential of Three Rose-scented Geranium (Pelargonium<br />
graveolens) Cultivars in the Kumaon Region of Western Himalayas<br />
Javid Ahmad Parray, Azra N. Kamilli, Raies Qadri, Rehana Hamid (India), Jaime A. Teixeira da Silva (Japan)<br />
Evaluation of Antibacterial Activity of Euryale ferox Salisb., a Threatened Aquatic <strong>Plant</strong> of Kashmir Himalaya<br />
Rehana Hamid, Azra N. Kamili, Mahmood uz Zaffar (India), Jaime A. Teixeira da Silva (Japan), A. Mujib, Javid Ahmad<br />
Parray (India) Callus-Mediated Shoot Organogenesis from Shoot Tips of Cichorium intybus<br />
Rehana Hamid, Mahmood uz Zaffar (India), Jaime A. Teixeira da Silva (Japan), Azra N. Kamili, Javid Ahmad Parray<br />
(India) Impact of Chromium on the Oxidative Defense System of Brassica juncea L. cv. ‘Pusa Jai Kissan’ under Hydroponic<br />
Culture<br />
Janifer Raj, Ballabh Basanth, Pal M. Murugan (India), Jaime A. Teixeira da Silva (Japan), Kumar Saurav, Om P.<br />
Chaurasia, Shashi Bala Singh (India) Screening Phytochemical Constituents of 21 <strong>Medicinal</strong> <strong>Plant</strong>s of Trans-Himalayan<br />
Region<br />
1<br />
8<br />
15<br />
20<br />
28<br />
43<br />
47<br />
64<br />
69<br />
73<br />
77<br />
80<br />
84<br />
87<br />
90
Pranay Bantawa, Swapan Kumar Ghosh, Pamita Bh<strong>and</strong>ari, Bikram Singh, Partha Deb Ghosh, Paramvir Singh Ahuja,<br />
Tapan Kumar Mondal (India) Micropropagation of an Elite Line of Picrorhiza scrophulariiflora, Pennell, an Endangered High<br />
Valued <strong>Medicinal</strong> <strong>Plant</strong> of the Indo-China Himalayan Region (pp 1-7)<br />
ABSTRACT<br />
Original Research Paper: An elite genotype of Picrorhiza scrophulariiflora Pennell was multiplied in vitro for its conservation.<br />
Rhizomes of mature plants collected from various locations of the eastern Himalayan region of Indo-China border were<br />
characterized morphologically <strong>and</strong> analyzed by HPLC to determine the content of marker compounds, namely picroside I <strong>and</strong> II.<br />
Amidst the genotypes, one from Ha, Bhutan was found to contain the highest amount of total picroside (7.33% dw).<br />
Subsequently, a rapid <strong>and</strong> highly reproducible method of micropropagation from rhizome or shoot tips was developed. While<br />
100% bud break from rhizomes was achieved on Woody <strong>Plant</strong> Medium (WPM) containing 0.44 μM BAP (6 benzyl amino purine),<br />
40-fold multiplication was achieved on WPM fortified with 2.3 μM Kn (kinetin) within 12 weeks. The multiplied shoots were<br />
elongated on WPM supplemented with 0.44 μM BAP. Around 90% of in vitro shoots were rooted without basal callus formation<br />
on WPM supplemented with 5.3 μM NAA (α-naphthalene acetic acid) within 4 weeks. Following this protocol, 1100<br />
micropropagated plantlets of an elite line (Ha, Bhutan) were hardened in their natural habitat. The present study illustrates the<br />
usefulness of additives for mass propagation <strong>and</strong> germplasm conservation <strong>and</strong> is, to the best of our knowledge, the first report<br />
of in vitro propagation of P. scrophulariiflora.<br />
Manoj Kumar Goel, Shilpa Goel, Suchitra Banerjee, Karuna Shanker, Arun Kumar Kukreja (India) Agrobacterium<br />
rhizogenes-Mediated Transformed Roots of Rauwolfia serpentina for Reserpine Biosynthesis (pp 8-14)<br />
ABSTRACT<br />
Original Research Paper: Root extracts of Rauwolfia serpentina have been used for centuries in Ayurvedic medicine as a<br />
panacea for a wide variety of physical as well as mental disorders. The potential of Agrobacterium rhizogenes-mediated genetic<br />
transformation for the synthesis of phytomolecules of high pharmaceutical value is now well established <strong>and</strong> documented.<br />
Transgenic roots were induced from R. serpentina leaf explants in response to A. rhizogenes A4 strain on semi-solid ½-strength<br />
MS medium. Amongst 200 hairy root clones developed, 27 showing persistent <strong>and</strong> incessant growth over several generations<br />
were selected. Transformed roots grew vigorously <strong>and</strong> branched profusely on hormone-free liquid B 5 medium with 3% sucrose<br />
with higher biomass yields compared to the control <strong>and</strong> showed two stable <strong>and</strong> distinct morphotypes. Medium devoid of any<br />
carbon source served as the control. The transformed nature of the roots was confirmed by PCR amplification with rolA primers.<br />
Growth kinetic studies exhibited the highest growth index (58.57 ± 1.92) at the 10 th week followed by slow growth in the<br />
subsequent period up to 14 weeks. Reserpine content increased with root growth <strong>and</strong> was highest in 10-weeks-old cultures.<br />
Hairy root clones showed a wide array of variation in relative reserpine content, varying from 0.0064 to 0.0858% dry weight<br />
(DW). On the basis of relative reserpine content, these hairy root clones were classified into 5 different groups. SM 12 clone had<br />
the highest reserpine level (0.0858% DW) producing 2-3 times more than the content of field-grown roots harvested after 18-24<br />
months. A distinct relationship between root morphology <strong>and</strong> reserpine content was observed. The present study is the first<br />
report of reserpine production in quantifiable amounts from the hairy roots of any Rauwolfia species.<br />
Amita Misra, Ashutosh K. Shukla, Ajit K. Shasany, V. Sundaresan, Shital P. Jain, Subhash C. Singh, Guru D. Bagchi,<br />
Suman P. S. Khanuja (India) AFLP Markers for Identification of Aconitum Species (pp 15-19)<br />
ABSTRACT<br />
Original Research Paper: The genus Aconitum is highly complex <strong>and</strong> its taxonomy has been traditionally difficult due to the<br />
high level of variation among the various species. The Aconitum species are known for their highly toxic diterpenoid alkaloids<br />
but have been described in traditional medicine systems as high-value medicine after proper <strong>and</strong> prescribed detoxification. In<br />
India, A. heterophyllum, A. balfourii <strong>and</strong> A. violaceum are found mainly in the North-Western Himalayas whereas A. ferox is<br />
found in the North-Eastern Himalayan region. Among these species, A. heterophyllum is the most significant in terms of<br />
therapeutic importance <strong>and</strong> herbal drug market value. It has become critically endangered due to high dem<strong>and</strong> of the herb <strong>and</strong><br />
indiscriminate overexploitation. There is an existing dem<strong>and</strong> in the bulk herbal drug industry to have an authentic identification<br />
system for the Aconitum species in order to enable their commercial use as genuine phytoceuticals. In the present study we<br />
have used Amplified Fragment Length Polymorphism (AFLP) for developing DNA fingerprints for 4 Aconitum species. A total of<br />
10 accessions (4 of A. heterophyllum, 3 of A. violaceum, 2 of A. balfourii <strong>and</strong> 1 of A. ferox) from the 4 species were used in the<br />
study, which employed 64 AFLP selective primer pairs. Only 26 selective primer pairs were found to respond with all the
accessions <strong>and</strong> generated a total of 4112 fragments. A number of species-specific markers were identified for all the 4 Aconitum<br />
species (16 for A. heterophyllum, 125 for A. violaceum, 79 for A. balfourii, <strong>and</strong> 226 for A. ferox). These AFLP fingerprints of the<br />
Aconitum species could be used in future for authentication of the drug <strong>and</strong> checking the adulteration-related problems faced by<br />
the commercial users of the herb.<br />
Rafia Rasool, Bashir Ahmad Ganai, Azra Nahaid Kamili, Seema Akbar, Akbar Masood (India) Antioxidant <strong>and</strong><br />
Antibacterial Activities of Extracts from Wild <strong>and</strong> in Vitro-Raised Cultures of Prunella vulgaris L. (pp 20-27)<br />
ABSTRACT<br />
Original Research Paper: MeOH, EtOH, CHCl 3 <strong>and</strong> aqueous extracts from the whole plant of wild Prunella vulgaris, a Kashmir<br />
Himalayan perennial medicinal herb, as well as from in vitro-regenerated plants were evaluated <strong>and</strong> compared for their<br />
antioxidant <strong>and</strong> antimicrobial properties. Antioxidant activity was screened by using various in vitro models: scavenging of the<br />
free radicals using DPPH, riboflavin photo oxidation, DNA damage, inhibition of lipid oxidation via PMS, FTC <strong>and</strong> TBA assay.<br />
The MeOH <strong>and</strong> CHCl 3 extract from wild <strong>and</strong> in vitro-regenerated plants possessed an almost equal radical scavenging effect. In<br />
vitro <strong>and</strong> wild grown plant extracts in different solvent systems were also screened for antimicrobial activity against medically<br />
important bacterial strains by the agar well diffusion method. The MeOH extract of both (wild <strong>and</strong> in vitro) plants extracts were<br />
almost equally effective against Escherichia coli, Staphylococcus aureus, Salmonella typhimurium <strong>and</strong> Kleibsella pneumonae.<br />
Both in vitro <strong>and</strong> wild dried plant extracts showed an almost similar concentration-dependent antioxidant <strong>and</strong> antimicrobial<br />
inhibition. Therefore, the commercial manufacture of active constituents from these improved elite lines would be useful <strong>and</strong><br />
profitable. The present study provides first evidence that in vitro grown P. vulgaris has antioxidant <strong>and</strong> antibacterial activities,<br />
suggesting the potential of the tissue culture technique to substitute wild P. vulgaris in the pharmaceutical industry.<br />
Ripu M. Kunwar (Nepal/USA), Chundamani Burlakoti (USA), Chhote L. Chowdhary (Nepal), Rainer W. Bussmann (USA)<br />
<strong>Medicinal</strong> <strong>Plant</strong>s in Farwest Nepal: Indigenous Uses <strong>and</strong> Pharmacological Validity (pp 28-42)<br />
ABSTRACT<br />
Original Research Paper: <strong>Medicinal</strong> plants have been used indigenously since ancient past as medicines for the treatment of<br />
various ailments. However, the knowledge of indigenous therapies have been distorting to these days due to changing<br />
perception, acculturation, commercialization <strong>and</strong> socio-economic transformations. The present study compares indigenous<br />
knowledge of therapies of 48 medicinal plants with the latest common pharmacological findings. Traditional indigenous plant<br />
knowledge <strong>and</strong> phytomedicine are consistently gaining acceptance in global society. The present study found that over<br />
two-thirds of traditionally used plants in the region show clear pharmacological efficacy. Total 23 species possessed strong<br />
resemblances <strong>and</strong> the species Euphorbia royleana, Ricinus communis, <strong>Plant</strong>ago major, Chenopodium album, Cordyceps<br />
sinensis, etc. contributed the most. The complementarity of indigenous therapies <strong>and</strong> pharmacological uses is obvious <strong>and</strong> it is<br />
base of the modern therapeutic medicine. The increasing use of indigenous therapies dem<strong>and</strong>s more scientifically sound<br />
evidence, therefore further investigation <strong>and</strong> phytochemical screening of ethnopharmacologically used plants <strong>and</strong> assessment<br />
of the validity to the indigenous uses is worthwhile.<br />
Mukesh Joshi, Munesh Kumar (India), Rainer W. Bussmann (USA) Ethnomedicinal Uses of <strong>Plant</strong> Resources of the Haigad<br />
Watershed in Kumaun Himalaya, India (pp 43-46)<br />
ABSTRACT<br />
Research Note: The present study was carried out in the Haigad watershed of Kumaun Himalaya. A total of 32 medicinal plant<br />
species belonging to 26 families were recorded. A major proportion of species were in forested l<strong>and</strong>scape (62%) <strong>and</strong> the rest in<br />
cultural l<strong>and</strong>scape (38%) of the watershed. The plants used for medicinal purposes in the local health traditions are gradually<br />
becoming extinct due to developmental activities, population explosion <strong>and</strong> other anthropogenic reasons. To avoid<br />
overexploitation <strong>and</strong> promote sustainable use, rapid conservation efforts are needed. Farmers should be involved in the<br />
cultivation of medicinal plants emphasizing suitable production methods, especially on barren <strong>and</strong> fallow l<strong>and</strong>.<br />
Parveen Kumar Sharma, N. S. Chauhan, Brij Lal, Amjad M. Husaini (India), Jaime A. Teixeira da Silva (Japan), Punam<br />
(India) Conservation of Phyto-diversity of Parvati Valley in Northwestern Himalayas of Himachal Pradesh, India (pp 47-63)<br />
ABSTRACT
Research Note: This study provides information about the traditional indigenous uses of plants by the inhabitants of the Parvati<br />
Valley of Kullu district in the western Himalayas of India. Since no published literature from the past 10 years exists, an<br />
ethnobotanical survey was conducted among the ethnic groups of the Parvati valley <strong>and</strong> first h<strong>and</strong> information on these plant<br />
species was recorded. A total of 266 species belonging to 180 genera <strong>and</strong> 71 families (including 44 species as recorded for the<br />
first time in the area) were collected. Out of these, 223 species within 152 genera of 61 families belong to dicots; 31 species <strong>and</strong><br />
22 genera under 7 families belong to monocots <strong>and</strong> 10 species with 6 genera in 3 families belong to gymnosperms.<br />
Venkat Kishore Ryakala (India), Shahin Sharif Ali (Irel<strong>and</strong>/India), Hallihosur Sharanabasava, Naushaba Hasin, Pragya<br />
Sharma, Utpal Bora (India) Ethnobotany of <strong>Plant</strong>s Used to Cure Diabetes by the People of North East India (pp 64-68)<br />
ABSTRACT<br />
Research Note: Northeast India is considered as an ecological hot spot <strong>and</strong> has a wide variety of flora <strong>and</strong> fauna. Diverse<br />
ethnic communities inhabit the area, each having their own traditional medical cures for different diseases. During the course of<br />
present studies it was found that 52 species of plants belonging to 36 families are used as antidiabetic agents in folk medicinal<br />
practice. Leaves <strong>and</strong> bark were found to be the two major plant parts used for making hypoglycemic herbal preparations.<br />
Around 26 treatments involve administration of decoction to the diabetic patient. These decoctions are either prepared from<br />
leaves, bark, fruit, root, seeds or from whole plants. Out of the 52 plants 12 are also reported to have antidiabetic properties in<br />
the Diabetes <strong>Medicinal</strong> <strong>Plant</strong> Database. The remaining plants could be a potential source of new <strong>and</strong> efficient cures for<br />
diabetes.<br />
Ram Swaroop Verma, Rajendra Ch<strong>and</strong>ra Padalia, Amit Chauhan, Ajai Kumar Yadav (India) Chemical Composition of Leaf<br />
<strong>and</strong> Flower Essential Oils of Two Thymus spp. from Western Himalaya (pp 69-72)<br />
ABSTRACT<br />
Original Research Paper: Thymus species (Lamiaceae) are considered to be very beneficial whether used as food or as a<br />
medicament. Essential oils (EOs) derived from leaves <strong>and</strong> flowers of Thymus serpyllum <strong>and</strong> Thymus linearis grown in northern<br />
India were analyzed by GC <strong>and</strong> GC-MS. A total of 37 components forming 94.8-98.4% of EO composition were identified. The<br />
EOs of both species were rich in thymol, p-cymene <strong>and</strong> γ-terpinene. Thymol was higher in the EO of T. linearis (74.6-75.8%)<br />
compared to T. serpyllum (51.9-70.1%). The amount of thymol methyl ether, p-cymene, 1-octen-3-ol, camphor <strong>and</strong> borneol was<br />
relatively higher in T. serpyllum EO. Further, phenolic monoterpenes were higher in flower EOs of both species than in leaf EOs.<br />
Ram Swaroop Verma, Rajendra Ch<strong>and</strong>ra Padalia, Amit Chauhan (India) Chemical Profiling of Mentha spicata L. var. ‘viridis’<br />
<strong>and</strong> Mentha citrata L. Cultivars at Different Stages from the Kumaon Region of Western Himalaya (pp 73-76)<br />
ABSTRACT<br />
Research Note: Two spearmint (Mentha spicata L. var. ‘viridis’) cultivars viz. ‘Neerkalka’ <strong>and</strong> ‘Supriya’ <strong>and</strong> one bergamot mint<br />
(Mentha citrata L.) cultivar ‘Kiran’ cultivated in the Kumaon region of northern India were investigated for their essential oil<br />
content <strong>and</strong> composition at different stages of crop growth. Essential oil content <strong>and</strong> composition were both affected by crop<br />
age in all cultivars. All the cultivars accumulated maximum essential oil at 150 days after transplanting. The percentage carvone<br />
in ‘Neerkalka’ was higher at 90 days (67.0%) followed by the 150-days-old crop (61.68%), while in ‘Supriya’, carvone<br />
concentration increased at 150 days (72.47%). In ‘Kiran’, linalool <strong>and</strong> linalyl acetate were highest in 150- <strong>and</strong> 180-days-old<br />
crops, respectively.<br />
Amit Chauhan, Ram Swaroop Verma (India) Cultivation Potential of Three Rose-scented Geranium (Pelargonium<br />
graveolens) Cultivars in the Kumaon Region of Western Himalayas (pp 77-79)<br />
ABSTRACT<br />
Research Note: A field experiment was conducted to evaluate the production potential of three cultivars of rose-scented<br />
geranium viz., ‘Bourbon’, ‘CIM-Pawan’ <strong>and</strong> ‘Kelkar’ in the temperate region of Uttarakh<strong>and</strong>. ‘CIM-Pawan’ had the highest<br />
essential oil yield (103.87 g plot -1 ) followed by ‘Kelkar’ (79.93 g plot -1 ) <strong>and</strong> ‘Bourbon’ (72.01 g plot -1 ). The essential oil profile of<br />
‘Bourbon’ was rich (relative percentages) in citronellol (29.05), geraniol (24.36), citronellyl formate (5.94), isomenthone (5.82);<br />
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<br />
the essential oil of ‘Kelkar’ showed a different profile with citonellol (61.48) <strong>and</strong> isomenthone (10.56) being almost twice that of
other cultivars.<br />
Javid Ahmad Parray, Azra N. Kamilli, Raies Qadri, Rehana Hamid (India), Jaime A. Teixeira da Silva (Japan) Evaluation<br />
of Antibacterial Activity of Euryale ferox Salisb., a Threatened Aquatic <strong>Plant</strong> of Kashmir Himalaya (pp 80-83)<br />
ABSTRACT<br />
Research Note: The antibacterial activity of methanolic extract of seeds <strong>and</strong> leaves of Euryale ferox was tested against nine<br />
clinically isolated bacterial strains (Staphylococus aureus, Escherichia coli, Pseudomonas aureoginosa, Citrobacter freundi,<br />
Shigella flexneri, Kliebsella pneumoniae, Proteus vulgaris, Salmonella typhi <strong>and</strong> Salmonella typhimurium) <strong>and</strong> subsequently<br />
was also tested for minimum inhibitory concentration (MIC) values which ranged from 0.25 to 500 mg/l against six ATCC<br />
bacterial strains using micro broth dilution method. The broad spectrum activity displayed by the seed <strong>and</strong> leaf extracts appears<br />
to provide a scientific basis for the use of E. ferox in kidney problems <strong>and</strong> urinary tract infections in ethno medicines.<br />
Rehana Hamid, Azra N. Kamili, Mahmood uz Zaffar (India), Jaime A. Teixeira da Silva (Japan), A. Mujib, Javid Ahmad<br />
Parray (India) Callus-Mediated Shoot Organogenesis from Shoot Tips of Cichorium intybus (pp 84-86)<br />
ABSTRACT<br />
Research Note: Cichorium intybus L. is a medicinally important plant with anti-cancerous <strong>and</strong> anti-hepatotoxic properties. An<br />
efficient method for totipotent callus formation has been developed in C. intybus from the basal portion of shoot tip explants on<br />
MS medium supplemented with different concentrations of plant growth regulators (PGRs) like 6-benzylamino purine (BAP) <strong>and</strong><br />
kinetin (Kn) with an auxin, indole-3-butyric acid (IBA). Cultures growing under the influence of BAP+IBA produced considerably<br />
more callus than cytokinins used alone. Re-differentiation of such callus led to multiple shoot formation on the same medium<br />
after 3 weeks. Isolated shoots were individually rooted in the presence of different concentrations of IBA. <strong>Plant</strong>lets obtained<br />
were transplanted into small pots containing peat, vermiculite, s<strong>and</strong> <strong>and</strong> soil mixture (1:1:1:1), 60% of which survived.<br />
Rehana Hamid, Mahmood uz Zaffar (India), Jaime A. Teixeira da Silva (Japan), Azra N. Kamili, Javid Ahmad Parray<br />
(India) Impact of Chromium on the Oxidative Defense System of Brassica juncea L. cv. ‘Pusa Jai Kissan’ under Hydroponic<br />
Culture (pp 87-89)<br />
ABSTRACT<br />
Research Note: Brassica juncea is a medicinally important plant <strong>and</strong> is commonly used as a diuretic, stimulant <strong>and</strong> to treat<br />
arthritis. Seed are used for the treatment of tumors <strong>and</strong> stomach disorders. B. juncea can hyperaccumulate cadmium <strong>and</strong> many<br />
other soil trace elements like selenium, chromium, iron <strong>and</strong> zinc food supplements. Chromium (Cr)-induced oxidative damage<br />
<strong>and</strong> changes in the contents of proline <strong>and</strong> glutathione in leaves of B. juncea. L. cv. ‘Pusa Jai Kissan’ were investigated after 3<br />
<strong>and</strong> 5 days of treatment under hydroponic culture. Cr was supplied as K 2Cr 2O 7. The main response was an increase in<br />
superoxide dismutase activity <strong>and</strong> proline content which subsequently reduced the activity of catalase <strong>and</strong> glutathione content<br />
in plants.<br />
Janifer Raj, Ballabh Basanth, Pal M. Murugan (India), Jaime A. Teixeira da Silva (Japan), Kumar Saurav, Om P.<br />
Chaurasia, Shashi Bala Singh (India) Screening Phytochemical Constituents of 21 <strong>Medicinal</strong> <strong>Plant</strong>s of Trans-Himalayan<br />
Region (pp 90-93)<br />
ABSTRACT<br />
Research Note: Alkaloids, tannins, flavonoids, saponins, steroids, <strong>and</strong> cardiac glycoside distribution in 1 high altitude medicinal<br />
plants belonging to different families (Apiaceae, Asteraceae, Crassulaceae, Lamiaceae, Rosaceae, Rubiaceae, Urticaceae,<br />
<strong>and</strong> Zygophyllaceae) were assessed <strong>and</strong> compared. The plants investigated were Achillea millefolium, Artemesia dracunculus,<br />
Bidens pilosa, Carum carvi, Dracocephalum heterophyllum, Ferula jaeskiana, Gallium pauciflorum, Heracleum pinnatum,<br />
Hippophae rhamnoides, Inula racemosa, Mentha longifolia, Nepeta podostachys, Origanum vulgare, Peganum harmala,<br />
Rhodiola imbricata, Rhodiola heterodenta, Rosa webbiana, Rosa macrophylla, Rubia cordifolia, Tanacetum gracile, <strong>and</strong> Utrica<br />
hyperborea, which have been widely used for time immemorial in the traditional Amchi system of medicine in the Ladakh region<br />
of India. Phytochemicals were qualitatively detected using aqueous extracts <strong>and</strong> solvent fractions of plants using various<br />
biochemical tests. These plants are a potential source of useful drugs. Future studies will isolate, identify, characterize <strong>and</strong><br />
elucidate the structure of novel bioactive compounds. The significance of these plants in traditional medicine <strong>and</strong> the
importance of the distribution of their chemical constituents are discussed in the context of the role of these plants in<br />
ethnomedicine in Ladakh.