Medicinal and Aromatic Plant Science and Biotechnology

Medicinal and Aromatic Plant Science and Biotechnology
Abbreviation: Medicinal Aromatic Plant Sci. Biotech.
Print: ISSN 1752-3389
Frequency and Peer status: Biannual, Peer reviewed
Scope and target readership: Medicinal and Aromatic Plant Science and Biotechnology will provide a complete analysis and
understanding of all aspects of medicinal and aromatic plant science and biotechnology.
Medicinal and Aromatic Plant Science and Biotechnology primarily wishes to examine:
1) Cultural practices (greenhouse growth, hydroponics, aeroponics, organic farming);
2) In vitro propagation (micropropagation, somatic embryogenesis, tissue culture, bioreactor system production);
3) Mycorrhizal symbioses (and effects on plant physiology, productivity, reproduction and disease resistance);
4) Novel techniques for analysis (genetic, biochemical, biophysical);
5) Physiology, genetics, molecular biology, structural botany (integrated, pure and applied);
6) Pathology;
7) Production of secondary metabolites, organic and inorganic biochemistry, and phytochemistry; pharmacological properties;
8) Storage of valuable genetic material (cold-storage or cryopreservation).
For publication in Medicinal and Aromatic Plant Science and Biotechnology the research must provide a highly significant new
contribution to our understanding of medicinal and aromatic plants (from any climactic or geographic origin) and must generally be
supported by a combination of either: physiological, biochemical, genetic or molecular analyses. All areas of study are welcome and the
experimental approaches used can be wide-ranging. Results that simply provide a description without an integrated multi-disciplinary
approach might not be considered, as might descriptive or overly-localized studies and reports on conventional propagation without a
wide impact. Manuscripts pertaining to breeding, post-harvest technology, nutritional aspects or influences on human health of medicinal
and aromatic plants will also be considered.
Editor-in-Chief
Jaime A. Teixeira da Silva, Kagawa University, Japan
Technical Editor
Kasumi Shima, Japan
Statistics Advisor
Marcin Kozak, Warsaw University of Life Sciences, Poland
Editorial Board and Advisory Panels (Listed alphabetically)
Khalid Ali Khalid Ahmed, National Research Centre, Egypt
Noureddine Benkeblia, The University of the West Indies,
Jamaica
Debprasad Chattopadhyay, Indian Council of Medical Research,
India
Sashi Kanta Dash, Orissa University of Agriculture & Technology,
India
Alberto Dias, Universidade do Minho, Portugal
Patricia Dias Fernandes, Federal University of Rio de Janeiro,
Brazil
Manuel Fernandes-Ferreira, Universidade do Minho, Portugal
Harzallah-Skhiri Fethia, Higher Biotechnology Institute of
Monastir, University of Monastir, Tunisia
Ashoke Kumar Ghosh, Bengal Institute of Pharmaceutical
Sciences, India
C. Gopi, K. S. Rangasamy College of Technology, India
Mazuru Gundidza, University of Zimbabwe/University of Fort
Hare, Zimbabwe/South Africa
Nirmal Joshee, Fort Valley State University, USA
Natchimuthu Karmegam, VMKV Engineering College/Vinayaka
Missions University, India
Ara Kirakosyan, The University of Michigan, USA
Shao-ping Li, University of Macau, China
Pavol Mártonfi, P. J. Šafárik University, Slovakia
Ákos Máthé, Szent István University, Hungary
Paraskev T. Nedialkov, Medical University-Sofia, Bulgaria
Majeti Narashima Vara Prasad, University of Hyderabad, India
Chandi Charan Rath, North Orissa University, India
Alberto Ritieni, University of Naples Federico II, Italy
Mohammad Jamal Saharkhiz, Shiraz University, Iran
Sabuj Sahoo, Utkal University, India
Ivan Salamon, Presov University, Slovak Republic
Sanjai Saxena, Thapar University, India
Mohd. Shahid, Aligarh Muslim University, India
Junjie Shan, Beijing Institute of Pharmacology & Toxicology,
China
Chunmeng Shi, Third Military Medical Univeristy, China
Neeta Shrivastava, B. V. Patel Pharmaceutical Education and
Research Development (PERD) Centre, India
Perumal Siddhuraju, Bharathiar University, India
Angelina Subotić, Institute for Biological Research "Siniša
Stanković", Serbia
Sivaramakrishnan Sudhakaran, AIMST University, Malaysia
Klára Szentmihályi, Hungarian Academy of Sciences, Hungary
Ilias S. Travlos, Benaki Phytopathological Institute, Greece
Rong Tsao, Agriculture & Agri-Food Canada, Canada
Supayang P. Voravuthikunchai, Prince of Songkla University,
Thailand
Anand K. Yadav, Fort Valley State University, USA

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Guest Editor
Dr. Amjad Masood Husaini
Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, India
Cover photos/figures: Left plate: Musical instrument of local deity Jamlu, Malana village (top), Angelica glauca (center
left), Hedychium spicatum (center right), Udithach (largest alpine pasture in the valley) (bottom) (Sharma et al., pp 47-63).
Top right row: Some Himalayan medicinal plants. (Left) Rhus parviflora. Fruits are indigenously used for diarrhea and
dysentery. (Center) Urtica dioica. Stem juice is valued for sprain and fractures. (Right) Euphorbia royleana. Plant is kept in
roof of house for protecting from evil. (Kunwar et al., pp 28-42). Center plate and gel: Agrobacterium rhizogenes-mediated
genetic transformation in Rauwolfia serpentina with rolA (top gel) and virD1 (top gel) PCR detection (Goel et al., pp 8-14).
Center right: Mature rhizomes of different genotypes of Picrorhiza scrophulariiflora. (Top) Bhutan, (Center) North Sikkim,
(Bottom) East Sikkim (Bantawa et al., pp 1-7). Bottom right: In vitro propagation and acclimatization of Cichorium intybus
through indirect callus culture on MS + 10 µM IBA (Hamid et al., pp 84-86).
Disclaimers: All comments, conclusions, opinions, and recommendations are those of the author(s), and do not necessarily
reflect the views of the publisher, or the Editor(s). GSB does not specifically endorse any product mentioned in any
manuscript, and accepts product descriptions and details to be an integral part of the scientific content.
Printed in Japan on acid-free paper.
Published: December, 2010.

The Guest Editor
Dr. Amjad Masood Husaini
Dr. Amjad Masood Husaini, a young Scientist working as Assistant Professor in Sher-e-Kashmir University of
Agricultural Sciences & Technology of Kashmir (India) holds a Ph.D. in Biotechnology and PG Diploma in Bioinformatics
(Jamia Hamdard, New Delhi), besides certificates in Intellectual Property Rights (Indian Law Institute, New Delhi) and
Remote Sensing Applications in Agriculture (Indian Agricultural Research Institute-Indian Space Research Organization).
Recipient of Young Scientist Award-2009 in Agriculture (Jammu & Kashmir State Council for Science & Technology,
Government of J&K), Jawahar Lal Nehru Award for Agricultural Research-2008 (Indian Council of Agricultural Research,
Government of India), Junior Scientist of the Year Award-2007 (National Environmental Sciences Academy, New Delhi), he
is listed among Top 100 Scientists of 2010 by the International Biographical Centre (IBC, Cambridge), and his biography
included in 27 th edition of Marquis Who’s Who in the World. With an illustrious academic career Dr. Husaini holds the
distinction of being top position holder in National Eligibility Tests for Life Sciences and Agricultural Biotechnology in
India. His publications include book entitled ‘Strawberry- Transgenics for stresses’ and more than two dozen research/
review papers in National and International journals of repute, discussing different aspects of agricultural research and
technology.
Dr. Husaini serves as member of professional associations like World Association of Young Scientists, New York
Academy of Sciences, The Indian Science Congress Association, Biotechnology Society of India, National Environmental
Science Academy (India), Young Professionals’ Platform for Agricultural Research for Development, Scientists Without
Borders, International Association of Computer Science and Information Technology, Royal Society of Crop Science,
International Society for Biosafety Research, and serves in the capacity of editor/ associate editor etc. in editorial boards of
various International journals of repute.

Foreword
Amjad Masood Husaini
Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, J&K 191121, India
E-mail: dr.amjadhusaini@hotmail.com
“Man, ever desirous of knowledge, has already explored many things, but more and greater still remains concealed;
perhaps reserved for far distant generations, who shall prosecute the examination of their Creator’s work in remote countries,
and make many discoveries for the pleasure and convenience of life” (Linnaeus, 1754). One such vast unexplored region
and a biodiversity hot spot, lies between two great ancient civilizations of India and China and is famous as “The Great
Himalayan Region”. The main Himalaya range runs west to east, from the Indus river valley to the Brahmaputra river valley,
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
eastern Tibet-Arunachal Pradesh region. The range consists of three coextensive sub-ranges, with the northernmost, and
highest, known as the Great or Inner Himalayas. The ancient religious scripture of Hindus, Atharvaveda is the earliest
celebrated treatise mentioning the use of medicinal plants of the region. Atharvaveda contains 114 hymns or formulations
for the treatment of diseases. Ayurveda, a system of traditional medicine native to the Indian subcontinent, originated in and
developed from these hymns. The Suśruta Saṃhitā and the Charaka Saṃhitā are two important works on this traditional
system of medicine. In addition there is a famous reference in Valmiki’s Ramayana, a religious scripture of Hindus, about
the existence of rare medicinal plant Sanjivani (Selaginella bryopteris) in Himalayas, which saved the life of Lakshmana
(brother of the Hindu god Lord Rama).
Over the centuries people have depended on these medicinal plants for treating daily ailments like cough, colds,
indigestion, ulcers, sore eyes etc. In fact Sir Lawrence, a British Settlement Commissioner in his book, ‘The Valley of
Kashmir’ (1895) refers to this point as, “when I have made inquiries as to various herbs which I have seen in the valley and
on hillsides, I am always told that they are hot and good for cold humours, cold and good for hot humours, dry and
beneficial to damp humours, damp and beneficial to dry humours.”
In this Special Issue (SI) on Himalayan MAPS, an attempt has been made to present various issues pertaining to
conservation, documentation, biotechnological applications and medicinal uses of plants of Himalayan region. The SI
comprises of 13 research articles related to different areas of plant biotechnology. In the first paper Bantawa et al. take-up
an important highly valued endangered medicinal plant of Indo-China Himalayas viz. Picrorhiza scrophulariiflora Pennell
and describe in detail its micropropagation. This study is first such report on this plant and illustrates the usefulness of
additives for mass propagation and germplasm conservation. In a similar study Hamid et al. describe a method for in vitro
shoot organogenesis of Cichorium intybus using shoot tips as explants. Cichorium intybus is known for its anti-cancerous
and anti-hepatotoxic properties and their successful transfer to pots with 60% survival percentage is a step forward towards
its ex situ conservation. The potential of Agrobacterium rhizogenes-mediated genetic transformation for the synthesis of
phytomolecules of high pharmaceutical value is well established. Goel et al. present the first report of reserpine production
in quantifiable amounts from the Agrobacterium rhizogenes-generated transgenic hairy roots of Rauwolfia serpentine,
whose root-extracts have been used for centuries in Ayurvedic medicine. In one clone the reserpine level was found to be 2-
3 times that of field grown roots, which is quite encouraging.
Supply of authentic medicinal plants to herbal drug industry is an important requisite for enabling their commercial use
in production of genuine phytoceuticals. An authentic identification system based on amplified fragment length
polymorphism (AFLP) for Aconitum heterophyllum, A. violaceum, A. balfourii and A. ferox has been reported in an original
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 and antimicrobial properties of wild and in vitro-regenerated plants of a
Kashmir Himalayan perennial medicinal herb, Prunella vulgaris. Their study is probably the first report giving evidence
that in vitro grown P. vulgaris has antioxidant and antibacterial activities similar to that of wild, suggesting the substitution
of wild P. vulgaris with tissue culture raised plants for use in pharmaceutical industry. In another study on antibacterial
activity, the potential of methanolic extract of seeds and leaves of Euryale ferox was tested against nine clinically isolated
bacterial strains by Parray et al. The broad spectrum activity displayed by these extracts appears to provide logic for the use
of E. ferox as ethno-medicine in urinary tract infections. The issues related to ethno-medicinal uses and overexploitation of
medicinal plants of Haigad watershed of Kumaun Himalaya have been discussed by Joshi et al. They argue that for
sustainable use, in addition to rapid conservation efforts, farmers should be involved in the cultivation of medicinal plants.
An exhaustive ethno-botanical survey on phyto-diversity, spanning over more than 250 species, of Parvati Valley in
Northwestern Himalayas described by Sharma et al. is highly informative. They stress the involvement of local inhabitants
for conservation of indigenous knowledge and traditional practices. In a similar study on medicinal plants of west Nepal,
Kunwar et al. compare indigenous knowledge of therapies of 48 medicinal plants with the latest common pharmacological
findings, suggesting complementarities and thus forming base for use in modern therapeutic medicine. Similar correlation
was reported by Ryakala et al. while studying the ethnobotany of 52 plant species used to cure diabetes by the inhabitants of
north eastern India. Raj et al. have screened phytochemical constituents of 21 medicinal plants used in traditional Amchi
system of medicine in the Ladakh region of India. The significance of these plants is discussed in the context of their role in
ethnomedicine All these studies have generated the possibilities of using the unexplored plants as potential sources of future
drugs.
Verma et al. have contributed an informative paper describing the chemical composition of leaf and flower essential oils
of Thymus serpyllum and T. linearis from Western Himalaya, while Hamid et al. discuss the impact of chromium on the
oxidative defense system of Brassica juncea L., a medicinally important plant commonly used as a diuretic and stimulant.
I hope that the scientists working on medicinal plants will find this Special Issue helpful in moving forward in their
important quest of contributing in the area of medicine, drug discovery, and conservation of medicinal plants etc. I would
like to thank Dr. Jaime A. Teixeira da Silva and Ms. Kasumi Shima at Global Science Books Ltd., UK for their cooperation
and helpful suggestions; and my family for their understanding and support during the prolonged and time-consuming work
on this volume.
December, 2010

Muhammad Iqbal, PhD, FNASc
Professor
Department of Botany
JAMIA HAMDARD
(Declared as Deemed-to-be University under Section 3 of
the UGC Act, 1956 vide Notification No. F.9-18/85-U.3
dated 10.5.1989 of the Government of India)
December 24, 2010
The tradition of herbal treatment for curing the human ailments is pretty old in India. Formal
accounts of medicinal plants appeared as early as during the Vedic period in the Vedico-
Brahminic treatises like Atharvaveda (2000 BC), Sushruta Samhita (1300 BC) and Charaka
Samhita (300 BC). Of the tradional Indian systems of medicine, the Ayurveda (science of life)
and the Unani (Greeco-Arabian) systems are based largely on medicinal plants, whereas
Siddha depends mainly on minerals. Over the centuries, the traditional practitioners have
developed a number of herbal formulations for the treatment of various ailments with special
emphasis on sexual debility, liver disorders and kidney problems. As the popular alternative
medicine, these preparations now constitute an important segment of the integrated health
management all over the world.
The Himalayas, often called "The Roof of the World", encompass a number of biodiversity hot
spots and repositories of medicinal plants. The whole Himalayan range is envisaged as a
trove of medicinal herbs, offering refuge to a variety of rare plants in its varied mountain
ecosystems. The research work carried out in the recent past has accumulated enough
scientific information on a variety of medicinal plants inhabiting the various zones of the
Himalayan range with diverse climatic conditions. Given the above, the document in hand is a
commendable effort that duly elucidates the various aspects of the medicinal-plant research
that have a potential promise for a safe herbal medication without the much talked about
adverse after-effects. The information covered by this issue is comprehensive and most of the
plants mentioned are well known for their therapeutic efficacy. Information on the traditional
knowledge, describing the ethno-medicinal uses of plants is also included.
I heartily appreciate Dr. Amjad Masood Husaini of the Sher-e-Kashmir University of
Agricultural Sciences & Technology of Kashmir, India, for editing this useful volume that
focuses on the medicinal plants of the Himalayas, and also the galaxy of distinguished
scientists and researchers who have contributed for this special issue of the Medicinal and
Aromatic Plant Science and Biotechnology, an emerging research journal of the Global
Science Books (GSB), UK. This document must prove a useful guide to botanists, cultivators
and collectors of medicinal plants and a pride possession of all those who are keen on the
Himalayan vegetation.
(Muhammad Iqbal)
Hamdard Nagar, New Delhi – 110 062, INDIA
Phone: +91-11-2605 9688, Extn.: 5530 (O), 5531 (R); Fax: +91-11-2605 9663
E-mail: iqbalg5@yahoo.co.in

SPECIAL ISSUE: CONTENTS
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
Manoj Kumar Goel, Shilpa Goel, Suchitra Banerjee, Karuna Shanker, Arun Kumar Kukreja (India) Agrobacterium
rhizogenes-Mediated Transformed Roots of Rauwolfia serpentina for Reserpine Biosynthesis
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
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.
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
Mukesh Joshi, Munesh Kumar (India), Rainer W. Bussmann (USA) Ethnomedicinal Uses of Plant Resources of the Haigad
Watershed in Kumaun Himalaya, India
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
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
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
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
Amit Chauhan, Ram Swaroop Verma (India) Cultivation Potential of Three Rose-scented Geranium (Pelargonium
graveolens) Cultivars in the Kumaon Region of Western Himalayas
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
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
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
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
1
8
15
20
28
43
47
64
69
73
77
80
84
87
90

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)
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 μM BAP (6 benzyl amino purine),
40-fold multiplication was achieved on WPM fortified with 2.3 μM Kn (kinetin) within 12 weeks. The multiplied shoots were
elongated on WPM supplemented with 0.44 μM BAP. Around 90% of in vitro shoots were rooted without basal callus formation
on WPM supplemented with 5.3 μM 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)
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 B 5 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 10 th 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. SM 12 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)
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, and 226 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)
ABSTRACT
Original Research Paper: MeOH, EtOH, CHCl 3 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 CHCl 3 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 (wild and 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)
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)
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)
ABSTRACT

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)
ABSTRACT
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)
ABSTRACT
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 γ-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)
ABSTRACT
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)
ABSTRACT
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)
ABSTRACT
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)
ABSTRACT
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)
ABSTRACT
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 K 2Cr 2O 7. 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)
ABSTRACT
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.