Medicinal and Aromatic Plant Science and Biotechnology

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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
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