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CHAPTER
Introduction
1
Nikhil Malhotra, Mohar Singh
ICAR-National Bureau of Plant Genetic Resources Regional Station, Shimla, Himachal Pradesh,
India
The Himalayan center of plant diversity is a narrow band of biodiversity lying on the
southern margin of the Himalayas, the world’s highest mountain range with elevations exceeding 8000 m (Barthlott et al., 2005). The Himalayan region, likewise
other biomes of the world, is known since centuries for harboring a rich wealth of
extremely valuable medicinal plants (Kala, 2005, 2010). The Indian Himalayas
are home to more than 8000 species of vascular plants of which 1748 possess
medicinal properties (Singh and Hajra, 1996; Samant et al., 1998; Joshi et al.,
2016). At present, the trade of these plants from the Himalayas to the other parts
of the world is speeding up due to increase in their demand, which subsequently
affect the traditional collection practices of medicinal plants (Olsen, 1998; Bhat
et al., 2013). Earlier, traditional healers mainly practiced the harvesting of medicinal
plants but with the high demand at regional to international markets, many of the
untrained collectors begin to participate in collection (Sharma and Kala, 2016),
which has also resulted in adulteration of plant material (Sagar, 2014). Numerous
wild and cultivated medicinal plants possessing bioactive com pounds known as
secondary metabolites have been utilized as curative agents since ancient times
and medicinal plants have gained importance recently, not only as herbal medicines
but also as natural ingredients for the cosmetic industries. As a result, a significant
number of Himalayan plants now figure in the Red Data Book of rare, endangered,
and threatened medicinal plants and require urgent conservation efforts (Rana and
Samant, 2010; Goraya, 2011). Some of the important medicinal plants species of
the Indian Himalayas include Aconitum heterophyllum (Atis), Dactylorhiza
hatagirea (Salampanja), Picrorhiza kurroa (Kutki), Podophyllum hexandrum (Bankakdi), Rauwolfia serpentina (Sarpagandha), etc.
Secondary metabolites are a unique group of compounds produced by plants to
protect against various biotic and abiotic factors (Kennedy and Wightman, 2011).
These compounds, however, do not influence the primary metabolic activities
such as growth and reproduction of plants (Arbona et al., 2013). The major classes
of secondary metabolites include phenolics, alkaloids, tannins, saponins, lignins,
glycosides, and terpenoids. Some of these compounds have become an integral
part of plantemicrobe interactions toward adapting to environmental irregularities.
They regulate symbiosis, induce seed germination, and show allelopathic effect, i.e.,
Himalayan Medicinal Plants. https://doi.org/10.1016/B978-0-12-823151-7.00004-0
Copyright © 2021 Elsevier Inc. All rights reserved.
1
2 CHAPTER 1 Introduction
inhibit other competing plant species in their environment. Moreover, these compounds induce adverse physiological activities such as reduced digestive efficiency,
reproductive failure, neurological problems, and gangrene and also possess high
toxicity. The discovery of such unique compounds in the majority of Himalayan
medicinal plants has inspired many scientific communities to explore their potential
applications in various industries. The use of natural bioactive compounds and their
products is thereby considered the most suitable source of alternative medicine.
Thus, there is an unprecedented task to meet the increasing demand for plant secondary metabolites from flavor and fragrance, food, and pharmac eutical industries.
However, their supply has become a major constraint since their large-scale cultivation is very limited. Moreover, it is difficult to obtain a constant quantity of
compounds from cultivated plan ts as their yield fluctuates due to several factors
including genotypic variations, geography, and edaphic conditions along with
harvesting and processing methods.
Further, in situ and ex situ conservations are the most efficient methods to
conserve genetic diversity of plants, including medicinal herbs. Traditionally,
in situ conservation efforts have utilized the delineation of protected areas, whereas
ex situ conservation efforts have included in vitro approaches and gene banks.
However, conservation efforts have taken new dimensions with the advent of new
technologies in recent years. As per the present scenario, these new approaches
have integrated with traditional well-developed methods of conservation. The recent
technological developments in high-throughput next-generation sequencing and
other molecular biology techniques have provided greater opportunities to identify
and characterize a large number of genes involved in important metabolite pathways. However, linking genotype to phenotype, predicting gene regulations, and
ascertaining mutations require the utilization of vast genomic information and
encompass the incorporation of intraspecific and environmental variability.
Unfortunately, there remains a paucity of information relating biological activities of essential metabolites with the ethnobotanical uses of the plants. In many
cases, this may be due to the activity residing in nonvolatile components. Additionally, many researchers have neglected bioactivity screening related to ethnopharmacological uses. Thus, detailed work should be carried out to identify
phytochemicals associated with biological activities, which support traditional
uses of medicinal plants. Moreover, an integrated approach including conventional
as well as emerging technologies should be utilized for the effective conservation
of Himalayan herbs (Sharma and Kala, 2018). The latest ecological ana lysis
methods coupled with whole-genome and transcriptome sequencing, metabolic
engineering, and big data analytics should become an integral part of programs
for the conservation and genetic improvement of the Himalayan plant wealth for
future generations. Further, increased interdisciplinary collaboration and multiinstitutional focus for a resolute effort to this effect is urgently required. We
encourage the pre ser vation of trad itio na l knowledge and uses o f Himalayan
medicinal plants and hope that additional steps should be undertaken to protect
and maintain the Himalayan ecology.
References 3
References
Arbona, V., Manzi, M., de Ollas, C., Go´mez-Cadenas, A., 2013. Metabolomics as a tool to
investigate abiotic stress tolerance in plants. Int. J. Mol. Sci. 14, 4885e4911.
Barthlott, W., Mutke, J., Rafiqpoor, D., Kier, G., Kreft, H., 2005. Global centers of vascular
plant diversity. Nova Acta Leopold. 92, 61e83.
Bhat, J., Kumar, M., Bussmann, R.W., 2013. Ecological status and traditional knowledge of
medicinal plants in Kedarnath Wildlife Sanctuary of Garhwal Himalaya, India.
J. Ethnobiol. Ethnomed. 9, 1e18.
Goraya, G., 2011. Conservation concerns for medicinal plants of Himachal Pradesh. ENVIS
News Lett. Med. Plants 3, 15.
Joshi, R.K., Satyal, P., Setzer, W.N., 2016. Himalayan aromatic medicinal plants: a review of
their ethnopharmacology, volatile phytochemistry, and biological activities. Medicines 3, 6.
Kala, C.P., 2005. Indigenous uses, population density, and conservation of threatened medic-
inal plants in protected areas of the Indian Himalayas. Conserv. Biol. 19, 368e378.
Kala, C.P., 2010. Medicinal Plants of Uttarakhand: Diversity Livelihood and Conservation.
Biotech Books, Delhi, India, p. 188.
Kennedy, D.O., Wightman, E.L., 2011. Herbal extracts and phytochemicals: plant secondary
metabolites and the enhancement of human brain function. Adv. Nutr. 2, 32e50.
Olsen, C.S., 1998. The trade in medicinal and aromatic plants from central Nepal to Northern
India. Econ. Bot. 52, 279e292.
Rana, M.S., Samant, S.S., 2010. Threat categorisation and conservation prioritisation of
floristic diversity in the Indian Himalayan region: a state of art approach from Manali
wildlife sanctuary. J. Nat. Conserv. 18, 159e168.
Sagar, P.K., 2014. Adulteration and substitution in endangered, ASU herbal medicinal plants
of India, their legal status, scientific screening of active phytochemical constituents. Int. J.
Pharmaceut. Sci. Res. 5, 4023e4039.
Samant, S.S., Dhar, U., Palni, L.M.S., 1998. Medicinal Plants of Indian Himalaya: Diversity
Distribution Potential Values. G.B. Pant Institute of Himalayan Environment and Development, Almora, India.
Sharma, N., Kala, C.P., 2016. Utilization pattern, population density and supply chain of
Rhododendron arboreum and Rhododendron campanulatum in Dhauladhar mountain
range of Himachal Pradesh, India. Appl. Ecol. Environ. Sci. 4, 102e107.
Sharma, N., Kala, C.P., 2018. Harvesting and management of medicinal and aromatic plants in
the Himalaya. J. Appl. Med. Aromat. Plants 8, 1e9.
Singh, D.K., Hajra, P.K., 1996. Floristic diversity. In: Gujral, G.S., Sharma, V. (Eds.), Chang-
ing Perspective of Biodiversity Status in the Himalaya. British Council Division, British
High Commission Publication, Wildlife Youth Services, New Delhi, India, pp. 23e38.
Aconitum heterophyllum
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CHAPTER
2
1
ICAR-National Bureau of Plant Genetic Resources Regional Station, Shimla, Himachal Pradesh, India;
2
Himalayan Forest Research Institute (HFRI), Conifer Campus, Shimla, Himachal Pradesh, India
Nikhil Malhotra1, Shivani Sharma
2.1 Introduction
Out of many important medicinal plants cultivated in present times, Aconitum species finds a key position for their conservation and cultivation. The genus Aconitum
belongs to the family Ranunculaceae. There are w400 species of Aconitum occurring worldwide (Lane, 2004; Yin et al., 2019). In the northwest Himalayas, it is represented by 10 species and 2 varieties. Some of the important species of Aconitum
are Aconitum balfourii, Aconitum bisma, Aconitum carmichaeli, Aconitum chasman-
thum, Aconitum deinorrhizum, Aconitum ferox, Aconitum japonicum, Aconitum
napellus, and Aconitum violaceum along with A. heterophyllumdthe only nontoxic
species of this genus (Chauhan, 2006; Buddhadev and Buddhadev, 2017). These herbaceous biennial plants are primarily natives of the mountainous parts of the Northern Hemisphere, growing in moisture retentive but well-drained soils on the
mountain meadows (Tamura, 1995). These plants are tall, with erect stem being
crowned by racemes of large and eye-catching blue, purple, white, yellow, or pink
zygomorphic flowers with numerous stamens. The root is best harvested in the
autumn as soon as the plant dies down and is dried for later use. In recent years,
the demand for medicinal and aromatic plants has grown rapidly because of accelerated local, national, and international interest. Aconitum genus is the center of
attraction in the field of herbal medicines because of its property of curing a wide
range of diseases and, hence, the pressure on its natural habitat has increased.
This is one of the most prized plant genuses which has been enlisted in the Red
Data Book and is widely considered as a mystifying group due to fatal as well as
therapeutic behavior (Tai et al., 2015). The pharmacological analysis of Aconitum
species and their compounds have shown various therapeutic effects pertaining to
cardiovascular and central nervous system (Dzhakhangirov et al., 1997;
Friese et al., 1997; Ameri 1998; Polyakov et al., 2005) alongside anticancer (Solyanik et al., 2004), antimicrobial, and cytotoxic activities (Gavı´n et al., 2004; Gonza´lez
et al., 2005). In recent years, a large number of studies have investigated the toxico-
logical characteristics of Aconitum, its main alkaloids, and their derivatives (Xie
et al., 2005; Fujita et al., 2007; Jaiswal et al., 2013, 2014). It has been observed
that the whole plant of Aconitum is highly toxic with the concentration of toxic
2
Himalayan Medicinal Plants. https://doi.org/10.1016/B978-0-12-823151-7.00015-5
Copyright © 2021 Elsevier Inc. All rights reserved.
5
6 CHAPTER 2 Aconitum heterophyllum
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compounds higher in roots and flowers than in leaves and stems (Ding et al., 1993).
The symptoms of toxicity affect mainly the central nervous system and the heart,
with concomitant gastrointestinal signs. The cause of death is the development of
ventricular tachyarrhythmia and heart arrest. No specific therapy exists for Aconitum
poisoning, although cardiovascular supportive treatment is usually applied (Lin
et al., 2004). The toxicity of Aconitum is mainly due to the diester diterpene alka-
loids and monoester diterpene alkaloids such as deoxyaconitine, benzoylmesaconitine, jesaconitine, benzoylhypaconine, and benzoylaconine (Chinese
Pharmacopoeia Commission, 2005; Srivastava et al., 2010; Nyirimigabo et al.,
2015). Through various physical and chemical methods of treatment, highly toxic
Aconitum alkaloids could be transformed into less toxic derivatives.
A. heterophyllum Wall, commonly known as “atis,” is a rare diploid (2n ¼ 16)
Himalayan plant species found between 2400 and 3600 m amsl (Fig. 2.1). Ayurveda
classical texts of 15the16th century introduced “Abhava-Pratinidhi Dravya”
concept, wherein it was categorized as an “abhava dravya” (unavailable drug). Its
roots are ovoid-conical, tapering downward to a print, 2.0e7.5 cm long,
0.4e1.6 cm or more thick at its upper extremity, gradually decreasing in thickness
toward tapering end, externally light ash-gray, white or gray-brown, while internally
FIGURE 2.1
Mature Aconitum heterophyllum plant.
2.1 Introduction 7
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starch white, external surface wrinkled marked with scars of fallen rootlet, and with
a rosette of scaly rudimentary leaves on top. It is a cross-pollinated plant which
flowers in the second year. The flowers are helmet shaped, bright blue or greenish
blue in color and have a purple vein. For med icinal use, the roots from plants bearing
fully developed tubers are collected (Kumar et al., 2016; Rajakrishnan et al., 2016).
The tubers sometimes occur as a pair of mother and daughter tubers. Tuberization in
A. heterophyllum is a distinctive process from young rootlet to fully mature storage
roots which are committed to the storage of primary as well as secondary metabolites (Pal et al., 2015). A. heterophyllum has been listed as “critically endangered medicinal herb” by the International Union for Conservation of Nature and Natural
Resources (IUCN, 1993; Nautiyal et al., 2002; CAMP, 2003; Srivastava et al.,
2011), which has thereby prohibited the export of its plants, plant portions and their
derivatives, and extracts obtained from the wild (Shah, 2005; Chinese Pharmaco-
poeia Commission, 2015). Owing to the huge cost for dried tuberous roots of
A. heterophyllum (w₹10,000 per kg), and an ever-rising demand of raw material
(>20 tons per year) (Aneesh et al., 2009; NMPB, 2015), overharvesting of its tubers
has been facilitated over the years. This reckless collection has led to reduction in its
population in natural habitat. Although efforts have been done to maintain its population in farm fields by conventional breeding and propagation methods (Fig. 2.2),
nothing has been significantly achieved in R&D programs globally (Rawat et al.,
2016). Nontoxic active components like atisine, hetisine, and heteratisine, collec-
tively termed as “aconites,” accumulating in tuberous roots of A. heterophyllum
have wide pharmacological effects on immune, digestive, and nervous systems
(Murti and Khorana, 1968; Pelletier et al., 1968 ; Mori et al., 1989; Rastogi and Meh-
rotra, 1991; Zhaohong et al., 2006; Nisar et al., 2009; Malhotra et al., 2014; Malhotra, 2017).
FIGURE 2.2
Field plantation of Aconitum heterophyllum at HFRI Farm, Shillaru, Himachal Pradesh,
India.
8 CHAPTER 2 Aconitum heterophyllum
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Biotechnological interventions have substantially contributed in terms of higher
aconites production and conservation in various Aconitum species, but the contemporary breakthroughs are still lacking in A. heterophyllum, besides a few research
interventions made in the recent past. Although this plant has been circumspectly
studied for its cultivation (Nautiyal et al., 2006; Srivastava et al., 2011), conservation
and sustainable utilization (Pandey et al., 2005; Seethapathy et al., 2014; Kumar
et al., 2016), cytology (Siddique et al., 1998; Rani et al., 2011; Jeelani et al.,
2015), ecology (Nautiyal et al., 2002; Bhat et al., 2014; Jeelani et al., 2015), medic-
inal uses (Ukani et al., 1996; Nyirimigabo et al., 2015), phytochemical constituents
(Gajalakshmi et al., 2011; Jaiswal et al., 2013; Jaiswal et al., 2014; Malhotra et al.,
2014; Nagarajan et al., 2015a,b; Nyirimigabo et al., 2015; Kumar et al., 2016),
reproductive biology (Siddique et al., 1998) along with reports on plant tissue culture (Giri et al., 1993, 1997; Jabeen et al., 2006; Solanki and Siwach, 2012), and
OMICS-assisted approaches (Malhotra et al., 2014, 2016; Pal et al., 2015; Kumar
et al., 2016), the comprehensive coverage of botany, production, and research ad-
vancements in A. heterophyllum have not been attempted till date. Thus, this chapter
becomes very unique and important for the researchers and readers across the globe
working on this high-value plant species.
2.2 Origin and distribution
Classification of the genus Aconitum has been extremely difficult because aconites
are morphologically highly variable (Yang, 1990; Tamura, 1995; Luo, 2003).
Numerous categorizations in this genus have been proposed (de Candolle, 1824;
Nakai, 1953; Wang, 1965; Tamura, 1995), but due to the difference in explanation
of features considered, these are still in great dispute. The major centers of Aconitum
diversity are northwest and east Himalayas, southwest China, and Japan. Although
the chloroplast DNA, nuclear ribosomal DNA (nrDNA), and nuclear internal transcribed spacer (ITS) sequence data have been used to study the phylogenetic relationships within Aconitum subgenus Aconitum (Kita et al., 1995; Kita and Ito,
2000; Luo et al., 2005) along with a study on chromosomal and molecular patterns
(Mitika et al., 2007), significant information on adequate understanding of its phylogeny is still lacking. Moreover, separate studies for tracing the evolutionary history
of each Aconitum species have not been done; therefore, no records are available for
justifying the origin of A. heterophyllum also.
In India, A. heterophyllum is found and cultivated in the Himalayan states of
Jammu and Kashmir, Ladakh, Himachal Pradesh, and Uttarakhand in the northwest
along with Sikkim and Arunachal Pradesh in the east. It also occurs in Nepal,
Bhutan, and parts of southwest China.
2.3 Medicinal properties 9
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2.3 Medicinal properties
From ancient times, A. heterophyllum has been used in different formulations in the
Indian Ayurvedic System for curing various diseases. Balachaturbhadra Churna,
Caspa Drops, Chandraprabha Vati, Chaturbhadraka Vati, Chitrakadi vati, Kutajghan
Vati, Livex, Panchatikta Guggulu Ghrita, Rasnerandadi Kwatha, Satyadi Yoga,
Shaddharana churna, and Sudarshan Churna are some of the popular multidrug herbal formulations in which A. heterophyllum is used as one of the main ingredients
(Lather et al., 2010; Nariya et al., 2011; Ajanal et al., 2012; Sojitra et al., 2013; Joshi
et al. 2014, 2016; Kumar et al., 2014; Selvaraj et al., 2014; Chaudhary et al., 2015;
Gupta et al., 2015; Dhamankar and Jadhav, 2016; Baishya et al., 2020). These drugs
find common use in the treatment of diarrhea, fever, indigestion, inflammation, helminthiasis, hyperlipidemia, and other ailment s. Some of the important medicinal
properties of A. heterophyllum are listed in Table 2.1.
2.3.1 Antibacterial activity
Ahmad et al. (2008) isolated the new aconitine type norditerpenoid alkaloids, 6-
dehydroacetylsepaconitine, and 13-hydroxylappaconitine from the tubers of
A. heterophyllum along with the known alkaloids lycoctonine, delphatine, and lappaconitine, which were screened for antibacterial activity against different bacterial
strains. They showed antibacterial activity against diarrhea causing gram-negative
bacteria Escherichia coli, Shigella flexneri, Pseudomonas aeruginosa, and Salmo-
nella typhi. This report strengthens the use of A. heterophyllum as an antimicrobial
Table 2.1 Medicinal properties of Aconitum heterophyllum.
Use References
Abdominal distension Imtiyaz et al. (2013)
Anti-Alzheimer’s disease Ahmad et al. (2017)
Antibacterial Srivastava et al. (2011), Sinam et al. (2014)
Antidiabetic Prasad et al. (2014), Nirja and Sharma (2016)
Antidiarrheal Prasad et al. (2014), Paramanick et al. (2017)
Antihelminthic Pattewar et al. (2012), Rungsung et al. (2013)
Antiinflammatory Verma et al. (2010), Paramanick et al. (2017)
Antileucorrhea Rana et al. (2013)
Antioxidant Prasad et al. (2012), Rah et al. (2016)
Antiulcer Rajakrishnan et al. (2020)
Aphrodisiac Imtiyaz et al. (2013), Sojitra et al. (2013)
Arthritis Lone and Bhardwaj (2013)
Hypolipidemic Subash and Augustine (2012)
Immunomodulatory Nagarajan et al. (2015b) , Joshi et al. (2016)
Nephroprotective Konda et al. (2016)
10 CHAPTER 2 Aconitum heterophyllum
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and/or antihelminthic agent. In another study by Sinam et al. (2014), the root alkaloid extract of A. heterophyllum showed antibacterial activity against Bacillus sub-
tilis, Bordetella bronchiseptica, Pseudomonas putida, Staphylococcus aureus, and
Xanthomonas campestris.
2.3.2 Antidiarrheal activity
The antidiarrheal activity of roots of A. heterophyllum may be attributed to an antisecretory and antienteropooling type effect as a result of reactivation of Na
ATPase activity mediated through nitric oxide pathway. They cause either a decrease
in mucosal secretion or increase in mucosal absorption, which allows the feces to
become desiccated, thus retarding its movement through the colon (Prasad et al.,
2014).
2.3.3 Antihelminthic activity
Aqueous and alcoholic extracts of tubers of A. heterophyllum gave encouraging results when evaluated against Pheretima posthuma, using piperazine citrate as standard. Time required for initial three paralytic attacks and deaths was used as
parameters to evaluate the drug (Pattewar et al., 2012). It was revealed that a dose
of 100% aqueous root extract was responsible for anthelmintic activity.
þ
and K
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2.3.4 Antihyperlipidemic activity
The methanolic extract of tubers of A. heterophyllum had a hypolipidemic effect on
diet-induced obese rats. It was observed that the pharmacological effect was due to
the inhibition of hydroxymethylglutarate-Coenzyme A reductase and activation of
lecithin-cholesterol acyltransferase. This resulted in lowering apolipoprotein B, total
cholesterol, low-density lipoprotein cholesterol, and triglycerides in the blood serum
along with the decrease in intestinal fat absorption and increase in apolipoprotein A
with high-density lipoprotein cholesterol. These results supported the use of
A. heterophyllum as an antihyperlipidemic agent (Subash and Augustine, 2012).
2.3.5 Antiinflammatory and antipyretic activity
For the assessment of antiinflammatory activity of A. heterophyllum, cottonpelleteinduced granuloma method was used. It was found that ethanolic extract
of A. heterophyllum tuber had significant antiinflammatory activity, thereby
providing scientific evidence for a traditional use as an antiinflammatory agent.
Further, the antipyretic effects of roots of A. heterophyllum in the form of aqueous,
chloroform, and hexane extracts were examined using the method of yeast-induced
pyrexia, with aspirin as a standard antipyretic agent for comparison. These studies
showed that the extracts were nontoxic with nonsignificant antipyretic activity
(Verma et al., 2010).
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