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Assessment, extraction pattern and restoration through propagation of Angelica glauca, a critically endangered Himalayan medicinal herb

Assessment, extraction pattern and restoration through propagation of Angelica glauca, a... International Journal of Biodiversity Science and Management 2 (2006) 105–113 Assessment, extraction pattern and restoration through propagation of Angelica glauca, a critically endangered Himalayan medicinal herb Anil Kumar Bisht, Arvind Bhatt, R. S. Rawal and Uppeandra Dhar G.B. Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora, India Key words: Conservation, assessment, Angelica glauca, ethnobotanical survey, germination, seedling performance SUMMARY Himalayan medicinal plants are at risk of over-exploitation. Many medicinal plants of the region are being exploited unsustainably to meet local as well as global demands. Hence, conservation initiatives must be formulated after gathering information on availability and the type and extent of pressure on a particular species and these must be supported by simple propagation methods. The present paper analyses the status of Angelica glauca Edgew. in two centres of Western Himalaya. The availability of the species in the wild is low and it is subject to severe threat largely from overgrazing and anthropogenic pressure. Because of the influence of traders from outside, local inhabitants are losing interest in traditional conservation mechanisms. The existing scenario shows that natural popula- tions of A. glauca need to be preserved. One option for promoting this is seed propagation to offset existing trends and encourage cultivation. Addition of indole-acetic acid (IAA) was found to be effective in improving germination. Seedling survival and performance was significantly better at higher elevations. The study thus assesses the status and threats A. glauca and suggests an easy propagation method. INTRODUCTION Steady depletion of biodiversity is a matter of about four billion people or nearly 80% of the growing concern (WCMC 1992). Flowering plants population rely on herbal medicine (Sheldon et al. are important resources because of their medici- 1997) and 25% of prescription drugs are plant- nal, phytochemical and pharmacological value derived chemicals (Rai et al. 2000). (Harnischfeger 2000). Medicinal plant species and The increasing popularity of herbal drugs, their parts are used singly or in combination to cure accompanied by depletion of habitats, has various human ailments. As such, the need for revi- increased pressure on natural populations of many talization of the Indian system of medicine, based valuable medicinal plant species. Thus, research on primarily on the herbal drugs, has been recognized conservation and sustainable use of medicinal all over the world. According to a WHO estimate, plants of the Indian Himalayan Region (IHR) is of Correspondence: Arvind Bhatt, G.B. Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora 263 643 (UA), India. Email: arvin_bhatt@rediffmail.com 105 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. considerable interest. The IHR is a rich repository of plant diversity and supports over 1748 (23.2% of India) plant species (1685 angiosperms, 51 gymno- sperms, 12 pteridophytes) of medicinal importance (Samant et al. 1998). A recent study indicates that about 175 species of the IHR (most of which are under threat) are used for the preparation of over 316 drug formulations (Dhar et al. 2000), resulting in depletion of natural populations of many species. Thus, conservation of rare plant popula- tions of Himalayan medicinal plants has emerged as an important priority. In order to develop strate- gies for conservation of such populations, it is important to assess distribution patterns and iden- tify preferred habitats. Further, identification of important populations of such plants provides a first step for perpetuation of stock through various propagation methods. Angelica glauca Edgew. (Apiaceae), a critically endangered Himalayan perennial herb (Mamgain et al. 1998), has been studied because of its aromatic and medicinal value and is in the top three priori- tized medicinal plants of Western Himalaya (Sastry and Chatterjee 2000). The species is distributed across the Indian Himalaya (northwest to east Himalaya), between 1,800–3,700 m (Figure 1). The root is used as a spice by indigenous communities Figure 1 Location map of study sites in Uttaranchal (Collett 1980) and considered cardioactive. It is and Himachal Pradesh used in constipation (Anonymous 1985), in bron- chitis (Gaur 1999), for rheumatism and urinary disorders (Shah 1983), to cure stomach troubles, (3) develop suitable propagation protocols via bilious complaints, menorrhagia, infantile atrophy, seeds and to monitor seedling performance at rinderpest and also as a stimulant (Chopra et al. different altitudes. 1956; Anonymous 1985). In spite of its multiple uses and local/global demand, the extent of its distribution in the wild has not been studied and METHODS knowledge of extraction from the wild is com- Phytosociological study pletely lacking. Methods of conventional and/or non-conventional propagation have also not been A phytosociological study was undertaken in developed. two distant centres of species occurrence in The present state of distribution of A.glauca Western Himalaya, namely the Kumaon region of requires assessment of representative populations Uttaranchal and Kullu and Parvati valley of using phytosociological methods. The species is Himachal Pradesh (Figure 1). Eight representative thought to be declining in the wild (Mamgain et al. sites, Sunderdunga (SD), Phurkiya (PH), Oonch 1998). Factors responsible for the decline need to (O), Naumour (N), Solang Nala (SN), Dohra (D), be identified and addressed. Developing simple Fojal (F) and Pulga (P), were identified for the methods of propagation will serve to offset mount- detailed study. Criteria for site selection were based ing pressure on the species. Considering the above on: (1) secondary information obtained from factors, the present study attempts to: (1) analyze floras, (2) a reconnaissance survey, and (3) accessi- distribution patterns and availability, (2) identify bility of populations. Specific locations (i.e. lati- factors responsible for its decline in the wild, and tude, longitude, altitude) of the sites were recorded 106 International Journal of Biodiversity Science and Management Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. with a hand-held Global Positioning System and interviewed individually. Respondents from differ- aspect was recorded with a compass (Table 1). ent villages were selected at random, half of the Studies were conducted during the flowering respondents were > 50 years old and half were < 50 period (July/August, 2001). The data were col- years old (Figure 2). Thus, there were four catego- lected from 20 randomly placed quadrats (1 × 1 m) ries: (1) women < 50; (2) women > 50; (3) men within an identified stand (20 × 20 m) in each site. < 50; and (4) men > 50. Small population size (spread) was the major con- Extent of extraction was assessed on the basis of straint for stand replication. Data on occurrence the quantity of rhizomes collected (dry weight) per and availability were recorded following the season, giving three categories; (A) < 10 kg (B) standard phytosociological methods (Smith 1974; 10–25 kg (C) > 25 kg. Muller-Dombois and Ellenberg 1974; Krebs 1989). Data on species use were calculated and catego- Individuals of A. glauca and associated species were rized as domestic use, trade, domestic use and counted in each quadrat and density, frequency, trade, domestic use and barter. Data were analyzed abundance and A/F ratio was analyzed using stan- and calculated on a percentage basis for each dard phytosociological formulae (Krebs 1989). village. Survey Seed germination experiments A survey was carried out in 13 villages of Seeds were collected during October 2000 from six West Himalaya (Kullu and Parvati valley of sites (Solang Nala, SN; Naumor, N; Oonch, O; Himachal Pradesh, Kumaon and Garhwal region of Fojal, F; Sunderdhunga, SD; Phurkia, PH). Insuffi- Uttaranchal) and a part of Trans Himalaya (Lahaul cient seed numbers prevented collection from valley). The adjacent villages were selected for Pulga and Dohra, and these sites were not consid- data collection on use category and extraction ered for the seed germination study. Immediately pattern, using a structured, closed questionnaire. after collection, seeds were dried at room tempera- From each village 15 men and 15 women were ture for one week and stored separately in brown Table 1 Population data for A. glauca Latitude Longitude Altitude Centres/population (N) (E) (m asl) Aspect Habitat Uttaranchal Sunderdunga (SD) 30° 12′ 79° 54′ 3740 SE Moist, steep slope at timberline zone among Rhododendron spp. Phurkiya (PH) 30° 13′ 79° 59′ 3500 SW Steep slope in moist Rhododendron forest Himachal Pradesh Oonch (O) 32° 01′ 77° 23′ 2940 SW Moist, grassy steep slope in pure Picea forests Naumor (N) 32° 19′ 77° 12′ 2798 NE Gentle slope in mixed forests (Platanus spp., Picea spp. and Quercus spp.) Solang Nala (SN) 32° 20′ 77° 08′ 2760 NE Moist, gentle slope in mixed forest (Platanus spp. and Picea spp.) Dohra (D) 32° 09′ 77° 15′ 2700 NE Moist slope along stream in mixed forest canopy (Platanus spp and Abies spp) Fojal (F) 32° 12′ 77° 04′ 2660 NW Mixed forest (Platanus spp, Abies spp and wild cherry) Pulga (P) 32° 03′ 77° 19′ 2500 NW Very moist, grassy patch in forest canopy (Picea spp, Abies spp, and Acer spp,) International Journal of Biodiversity Science and Management 107 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. paper bags. Seed viability was determined using the chamber at 25 ± 2°C. Experiments were laid in ran- 2,3,5-triphenyl tetrazolium chloride (TTZ) test domized complete block design. Germination was (Hartmann and Kester 1989), after removing the monitored daily from the date of sowing until first wings. Fifty seeds from each site were immersed in germination and thereafter germinated seeds were 1% tetrazolium chloride solution and incubated in counted at weekly intervals. The seeds were con- the dark for 24 h. Viable embryos stained dark red. sidered to have germinated when the radical was For germination experiments, seeds that 2 mm long. These germinated seeds were removed collapsed when pinched gently with forceps and and used for seedling performance experiments. those showing fungal infection were discarded. The whole experiment was monitored for approx. Several methods were tested in an attempt to maxi- 70 days. mise germination of A. glauca (Table 2). For each treatment, three replicates of 15 seeds from each Seedling performance site were used. Seeds were washed thoroughly and soaked in test solutions for 24 h. Thereafter they Performance and survival of A. glauca seedlings were washed again with tap water and placed on were determined at the GB Pant Institute nursery at Petri dishes (95 × 17 mm) containing Whatman Katarmal (1240 m) and Lata village in Nanda Devi (No. 44) filter paper and moistened with distilled Biosphere Reserve (NDBR, 2220 m). One-month- water twice a week. The dishes were kept in a growth old seedlings in three replicates of 24 seedlings each were planted at these sites in May 2001 and watered twice a week. Percentage survival and growth performance were recorded in October Statistical analysis Data on extraction category, quantity of rhizomes extracted per growing season, seed germination and seedling performance was statistically tested using SYSTAT (Wilkinson 1986). Significant differences between means were separated by the least significant difference (LSD) (p < 0.05) test (Snedechor and Cochran 1968). RESULTS Assessment of availability and threats to the species Assessment of the species in selected natural sites showed that it occurred on north-facing (five Figure 2 Flow chart of the methodology adopted for phytosociological study and survey sites) moist slopes between 2500 to 3900 m. Plant Table 2 Details of treatments used for improving seed germination in the present study Concentration Treatment SN Treatments (ppm) duration (h) Remarks 1. Gibberelic acid (GA ) 100, 500, 1000 24 Seeds thoroughly washed with tap water 2. Thiourea 100, 200, 300 24 Seeds thoroughly washed with tap water 3. Potassium nitrate (KNO ) 100, 500, 1000 24 Seeds thoroughly washed with tap water 4. Indole acetic acid (IAA) 800, 1000, 1200 24 Seeds thoroughly washed with tap water 108 International Journal of Biodiversity Science and Management Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. density was low in all the sites, ranging from 0.95 (p < 0.05) improvement was noticed in germina- 2 2 individuals/m (SD, PH, D) to 1.8 individuals/m tion percentage in SN (73.3%) and N (53.3%) with (SN, F). A. glauca contributed 1.8% (SN) to 7.1% potassium nitrate (500 ppm) treatment, as com- (D) to total plant density, decreasing towards pared to control (SN 37.8%; N 8.9%). However, higher elevations (r = −0.510; p > 0.05). In spite germination at higher altitude sites improved of the low density, A. glauca maintained a high significantly. Thiourea also enhanced germination frequency of occurrence (70–75%) in all sites, com- for all the sites, particularly at SN and N and at parable with other dominant taxa of these sites higher altitude sites. IAA (800 ppm) was most (Table 3). The ratio of abundance to frequency effective in improving percentage germination (A/F) was used to interpret the distribution pattern across the sites, with 91.1% at SD, which was signifi- at different scales, i.e. regular, < 0.025; random, cantly (p < 0.05) higher as compared to the control 0.025–0.05 and contagious, > 0.05 distribution (64.4%). (Curtis and Cottam 1956). A. glauca was randomly distributed in SD, O, N, SN and F sites and regularly Seedling performance distributed in PH, D and P sites. Survival of seedlings planted at the higher altitudes (Lata village) was significantly (p < 0.05) higher (61.4%) as compared to those growing at Kosi Extraction (48.8%). Similarly, growth (plant height, stem dia- The extraction was exclusively by local inhabitants meter and leaf number) was significantly (p < 0.05) from wild populations (Table 4a) and for domestic higher at Lata (Table 6). use (84.1%), with few respondents involved in trade (13.3%). There was no significant gender differ- ence for extraction categories. Significantly more DISCUSSION men (p < 0.05) extracted the species for trade, Angelica glauca is a high-value medicinal plant of the while older people (> 50 years) mainly collected Himalaya, which has attained endangered status as plants for domestic use. Domestic use and barter a result of several pressures on its populations and did not differ significantly among age classes, but habitat. Overgrazing, destructive and unscientific significantly more of the younger age group harvesting are gradually altering the habitat of the (14.3%) were non-users. None of the older people species. There is a clear relationship between plant were identified as the non-users. Extraction of part harvested and the degree of disturbance to the rhizomes per growing season in the study area sites. The removal of whole plants and roots has differed little between men (60.5%) and women a more immediate and damaging effect than the (71.3%). However, significantly more (p < 0.05) harvesting of leaves and fruits (Cunningham 1998). men (14.9%) were found to collect plants under As the root is the part used in medicine, the plant category C (Table 4b), while 9.8% of younger is susceptible to destructive harvesting and plant people (< 50 years) and 5.4% of older people (> 50 productivity and sustainable utilization are contin- years) collected plants under category C. gent to the severity of harvesting practices. Distribution of the species in natural sites sug- gests that A. glauca prefers north-facing slopes in Propagation moist-shady subalpine and timberline habitats. Seeds of A. glauca from different sites had different Although the species is randomly distributed and viability: 98.3% for N followed by 95.0% for PH, has higher frequency of occurrence in such habi- 93.0% for SN, 91.7% for O, 86.9% for F and 83.3% tats, anthropogenic pressures might be responsible for SD. for the low plant density. The field survey revealed Germination differed between sites and treat- that local people are generally not concerned ments. Untreated seeds showed a marked variation about conservation, despite knowing the usefulness in percentage germination, ranging from 64.4% of the species. The Convention on Biological Diver- (SD) to 6.7% (PH) (Table 5). Soaking seeds in sity (CBD) states that indigenous people play a vital different concentrations of gibberelic acid did role in environmental management and develop- not affect percentage germination. Significant ment through their traditional practices (Glowka International Journal of Biodiversity Science and Management 109 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. Table 3 Phytosociological status of sites of A. glauca with associate species Population Species Frequency Density Indiv./m Relative density (%) A/F ratio Sunderdunga (SD) A. glauca 75 0.95 5.09 0.025 1. Rheum moorcroftiana 80 5.05 27.08 0.081 2. Polygonum polystachyum 75 2.75 14.75 0.053 3. Fragaria nubicola 85 2.60 13.94 0.047 Others 7.30 39.14 Phurkiya (PH) A. glauca 75 0.95 3.51 0.020 1. Senecio sp. 90 8.30 30.68 0.114 2. Polygonum polystachyum 100 6.90 25.51 0.076 3. Fragaria nubicola 60 2.25 8.32 0.065 Others 8.65 31.98 Oonch (O) A. glauca 75 1.20 5.96 0.028 1. Impatiens sp. 40 2.30 11.66 0.143 2. Dryopteris sp. 55 1.90 9.84 0.065 3. Fragaria nubicola 60 1.60 8.29 0.047 Others 12.40 64.21 Naumor (N) A. glauca 75 1.35 2.15 0.026 1. Impatiens sp. 100 24.30 38.81 0.246 2. Polygonum amplexicaule 100 9.55 15.25 0.108 3. Ligularia sp. 65 4.15 6.62 0.106 Others 23.25 37.08 Solang (S) A. glauca 70 1.80 1.97 0.026 1. Viola sp. 100 28.65 49.14 0.295 2. Dryopteris sp. 60 9.25 15.87 0.266 3. Rosa sp. 95 8.10 13.89 0.098 Others 10.50 19.09 Dohra (D) A. glauca 75 0.95 7.14 0.022 1. Viola sp. 80 3.85 28.94 0.065 2. Polygonum amplexicaule 65 2.10 15.78 0.053 3. Impatiens sp. 45 2.05 15.41 0.103 Others 4.35 32.67 Fojal (F) A. glauca 70 1.80 3.23 0.040 1. Fragaria sp. 100 12.95 23.20 0.131 2. Coniogramme intermedia 90 8.20 14.70 0.105 3. Viola sp. 60 4.45 7.97 0.123 Others 28.40 50.87 Pulga (P) A. glauca 75 1.15 2.59 0.022 1. Dryopteris sp. 65 13.70 30.86 0.323 2. Impatiens sp. 90 13.10 29.50 0.162 3. Ligularia sp. 80 7.45 16.78 0.129 Others 9.00 20.23 A/F = Abundance/Frequency 1997), however, cash incentives encourage local Despite a comparatively lower percentage of people to ignore traditional harvesting practices. people in the high extraction category C, a large In the present study, greater awareness of the quantity of plant material is also being extracted potential of the plant by men could be because they from the wild under categories A and B. The higher are involved in collection and trade. percentage of people (men and women) in the 110 International Journal of Biodiversity Science and Management Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. Table 4(a) Categories of extraction of A. glauca Gender (%) Age class (%) Male Female Use (n=15 ¥ 13) (n=15 ¥ 13) LSD F < 50 yr > 50 yr LSD F DU 80.5 87.7 13.6 1.3 79.7 88.5 6.1 10.2 TR 18.9 7.7 11.6 4.5 18.7 5.8 9.1 9.5 DU + BT 0.5 4.6 6.1 2.2 0.6 4.3 4.7 3.0 NU 2.1 9.2 8.6 1.7 14.3 0.0 11.5 7.2 DU domestic use, TR trade, BT barter, NU not used; (n = 15 people × 13 = village) Table 4(b) Amount of rhizome extracted by villagers in a growing season Gender % Age Class % Class Male Female LSD F < 50 yr > 50 yr LSD F A 60.5 71.3 18.2 1.2 62.3 73.8 8.8 8.1 B 24.6 28.7 15.1 0.3 29.5 23.3 7.9 2.91 C 14.9 0.0 11.4 8.1 9.8 5.4 4.9 3.8 A < 10 kg dry weight, B 10–25 kg dry weight, C > 25 kg dry weight Table 5 Seed germination responses of A. glauca with various treatments Population Treatment SN N O F SD PH LSD Control 37.8 8.9 31.1 13.3 64.4 6.7 13.9 IAA 800 55.6 24.4 37.8 37.8 91.1 20.0 16.6 1000 42.2 55.6 40.0 37.8 80.0 15.6 18.3 1200 53.3 66.7 55.7 13.3 80.0 17.8 33.1 LSD 20.6 33.9 18.9 18.5 20.3 16.3 KNO 100 60.0 37.8 35.6 13.3 75.6 8.9 12.2 500 73.3 53.3 31.1 33.3 73.3 6.7 23.4 1000 35.6 33.3 44.5 13.3 71.2 8.9 17.7 LSD 22.4 16.3 22.9 16.6 18.9 10.3 Thiourea 100 71.1 24.4 40.0 13.3 37.9 22.2 16.3 200 42.2 26.7 33.3 11.1 71.1 24.5 26.7 300 44.4 22.2 35.6 13.3 73.3 11.1 23.4 LSD 31.4 9.6 24.1 23.5 22.9 12.6 GA 100 40.0 20.0 26.7 20.0 66.7 6.7 13.9 500 40.0 11.1 31.1 13.3 46.7 8.9 10.8 1000 40.0 11.1 31.1 13.3 24. 5 8.9 16.3 LSD 16.2 8.9 23.3 7.3 14.9 11.5 high extraction category in villages with more sites inaccessible areas, involving travel for long dis- containing the plants probably reflects the influ- tances on tough terrain, which might not be ence of traders in areas of high availability. Signifi- possible for older people and women. It also shows cantly more young men are in the high extraction that younger men are important players in the category C because plants are generally found in marketing chain. International Journal of Biodiversity Science and Management 111 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. Table 6 Seedling performance of A. glauca at different altitudes Altitude Total planted Survival Plant height Stem diameter Locality (m asl) seedlings (%) (cm) (cm) Leaf number Kosi 1240 72 48.6 43.9 3.1 2.6 Lata 2220 72 61.4 53.9 3.6 3.2 LSD (p < 0.05) – – 12.9 2.5 0.6 0.6 In the present scenario, where collection of utilization of high-value medicinal plants of the plant material is adversely affecting plant regenera- Himalaya. The plan must focus on the following tion, it is important that a simple and effective strategies: regeneration protocol via seeds is developed so that 1. An aggressive awareness and education pro- local inhabitants can initiate farming of A. glauca gramme should be initiated among local and reduce their dependency on wild populations. people to encourage conservation; In this context, the results of germination experi- ment showed differences between sites, treatments 2. Legislative measures should be enacted for and concentrations of growth promoters. Such regulating the extraction of medicinal plants variations in germination have been reported in from the wild; other species (Baskin and Baskin 1998). IAA 3. Propagation protocols must be developed for produced the largest improvement in germination rapid and mass multiplication of the species; of A. glauca seeds, while Chauhan et al. (1998) and found that lower concentrations of IAA promoted germination of Saussurea costus. 4. Demonstration and dissemination of propaga- Significantly better survival and growth of tion packages among end users should be ini- seedlings of A. glauca at Lata (2220 m) might be tiated to promote cultivation and to reduce due to in situ environmental conditions of tempera- pressure on natural populations. ture and moisture, suggesting that maximum returns could be achieved by establishing nurseries close to natural populations. Successful cultivation ACKNOWLEDGEMENTS of Podophyllum hexandrum, Aconitum heterophyllum and Aconitum balfouroii at comparable altitudes has Authors wish to thank ID Bhatt, S Airi and col- been reported (Prasad 2000; Bahuguna et al. 2000). leagues from the CBD group for their support and help. Financial assistance from the Department of Biotechnology, Government of India (BT/ RECOMMENDATIONS PR1118/PB/17/050/98) and National Medicinal An area-specific action plan needs to be devel- Plant Board, New Delhi (No. 135/2002) is grate- oped for the conservation and sustainable fully acknowledged. REFERENCES Anonymous. The Wealth of India, A Dictionary of Indian Baskin CC and Baskin JM. Seeds: Ecology, Biogeography Raw Materials and Industrial Products, Vol. 1. New and Evolution of Dormancy and Germination. San Delhi: Publication and Information Directorate, Diego: Academic Press; 1998 CSIR; 1985 Bisht AK, Bhatt A, Rawal RS and Dhar U. Prioritiza- Bahuguna R, Purohit MC, Rawat MSM and Purohit tion and conservation of Himalayan medicinal AN. Qualitative and quantitative variations in alka- plants: Angelica glauca Edgew. as a case study. loids of Aconitum species from Garhwal Himalaya. 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Journal Talk April 1998;29–31 of Herbs, Spices and Medicinal Plants 2000;5:43–50 Smith RL. Ecology and Field Biology. New York: Harper Hartmann HT and Kester DE. Plant propagation: and Row; 1974 Principals and practices. New Delhi: Prentice Hall; Snedechor GW and Cochran WG. Statistical Methods. 1989 New Delhi: Oxfords and IBH Publishing; 1968 Krebs CJ. Ecological methodology. Cambridge: Harper WCMC. Global biodiversity: Status of earth’s living and Row; 1989 resources. London: Chapman and Hall; 1992 Mamgain SK, Goel AK and Sharma SC. Conservation Wilkinson J. SYSTAT: A System for statistics. Evaston IL: assessment of some important medicinal plants of Systat Inc.; 1986 International Journal of Biodiversity Science and Management 113 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png International Journal of Biodiversity Science & Management Taylor & Francis

Assessment, extraction pattern and restoration through propagation of Angelica glauca, a critically endangered Himalayan medicinal herb

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Abstract

International Journal of Biodiversity Science and Management 2 (2006) 105–113 Assessment, extraction pattern and restoration through propagation of Angelica glauca, a critically endangered Himalayan medicinal herb Anil Kumar Bisht, Arvind Bhatt, R. S. Rawal and Uppeandra Dhar G.B. Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora, India Key words: Conservation, assessment, Angelica glauca, ethnobotanical survey, germination, seedling performance SUMMARY Himalayan medicinal plants are at risk of over-exploitation. Many medicinal plants of the region are being exploited unsustainably to meet local as well as global demands. Hence, conservation initiatives must be formulated after gathering information on availability and the type and extent of pressure on a particular species and these must be supported by simple propagation methods. The present paper analyses the status of Angelica glauca Edgew. in two centres of Western Himalaya. The availability of the species in the wild is low and it is subject to severe threat largely from overgrazing and anthropogenic pressure. Because of the influence of traders from outside, local inhabitants are losing interest in traditional conservation mechanisms. The existing scenario shows that natural popula- tions of A. glauca need to be preserved. One option for promoting this is seed propagation to offset existing trends and encourage cultivation. Addition of indole-acetic acid (IAA) was found to be effective in improving germination. Seedling survival and performance was significantly better at higher elevations. The study thus assesses the status and threats A. glauca and suggests an easy propagation method. INTRODUCTION Steady depletion of biodiversity is a matter of about four billion people or nearly 80% of the growing concern (WCMC 1992). Flowering plants population rely on herbal medicine (Sheldon et al. are important resources because of their medici- 1997) and 25% of prescription drugs are plant- nal, phytochemical and pharmacological value derived chemicals (Rai et al. 2000). (Harnischfeger 2000). Medicinal plant species and The increasing popularity of herbal drugs, their parts are used singly or in combination to cure accompanied by depletion of habitats, has various human ailments. As such, the need for revi- increased pressure on natural populations of many talization of the Indian system of medicine, based valuable medicinal plant species. Thus, research on primarily on the herbal drugs, has been recognized conservation and sustainable use of medicinal all over the world. According to a WHO estimate, plants of the Indian Himalayan Region (IHR) is of Correspondence: Arvind Bhatt, G.B. Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora 263 643 (UA), India. Email: arvin_bhatt@rediffmail.com 105 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. considerable interest. The IHR is a rich repository of plant diversity and supports over 1748 (23.2% of India) plant species (1685 angiosperms, 51 gymno- sperms, 12 pteridophytes) of medicinal importance (Samant et al. 1998). A recent study indicates that about 175 species of the IHR (most of which are under threat) are used for the preparation of over 316 drug formulations (Dhar et al. 2000), resulting in depletion of natural populations of many species. Thus, conservation of rare plant popula- tions of Himalayan medicinal plants has emerged as an important priority. In order to develop strate- gies for conservation of such populations, it is important to assess distribution patterns and iden- tify preferred habitats. Further, identification of important populations of such plants provides a first step for perpetuation of stock through various propagation methods. Angelica glauca Edgew. (Apiaceae), a critically endangered Himalayan perennial herb (Mamgain et al. 1998), has been studied because of its aromatic and medicinal value and is in the top three priori- tized medicinal plants of Western Himalaya (Sastry and Chatterjee 2000). The species is distributed across the Indian Himalaya (northwest to east Himalaya), between 1,800–3,700 m (Figure 1). The root is used as a spice by indigenous communities Figure 1 Location map of study sites in Uttaranchal (Collett 1980) and considered cardioactive. It is and Himachal Pradesh used in constipation (Anonymous 1985), in bron- chitis (Gaur 1999), for rheumatism and urinary disorders (Shah 1983), to cure stomach troubles, (3) develop suitable propagation protocols via bilious complaints, menorrhagia, infantile atrophy, seeds and to monitor seedling performance at rinderpest and also as a stimulant (Chopra et al. different altitudes. 1956; Anonymous 1985). In spite of its multiple uses and local/global demand, the extent of its distribution in the wild has not been studied and METHODS knowledge of extraction from the wild is com- Phytosociological study pletely lacking. Methods of conventional and/or non-conventional propagation have also not been A phytosociological study was undertaken in developed. two distant centres of species occurrence in The present state of distribution of A.glauca Western Himalaya, namely the Kumaon region of requires assessment of representative populations Uttaranchal and Kullu and Parvati valley of using phytosociological methods. The species is Himachal Pradesh (Figure 1). Eight representative thought to be declining in the wild (Mamgain et al. sites, Sunderdunga (SD), Phurkiya (PH), Oonch 1998). Factors responsible for the decline need to (O), Naumour (N), Solang Nala (SN), Dohra (D), be identified and addressed. Developing simple Fojal (F) and Pulga (P), were identified for the methods of propagation will serve to offset mount- detailed study. Criteria for site selection were based ing pressure on the species. Considering the above on: (1) secondary information obtained from factors, the present study attempts to: (1) analyze floras, (2) a reconnaissance survey, and (3) accessi- distribution patterns and availability, (2) identify bility of populations. Specific locations (i.e. lati- factors responsible for its decline in the wild, and tude, longitude, altitude) of the sites were recorded 106 International Journal of Biodiversity Science and Management Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. with a hand-held Global Positioning System and interviewed individually. Respondents from differ- aspect was recorded with a compass (Table 1). ent villages were selected at random, half of the Studies were conducted during the flowering respondents were > 50 years old and half were < 50 period (July/August, 2001). The data were col- years old (Figure 2). Thus, there were four catego- lected from 20 randomly placed quadrats (1 × 1 m) ries: (1) women < 50; (2) women > 50; (3) men within an identified stand (20 × 20 m) in each site. < 50; and (4) men > 50. Small population size (spread) was the major con- Extent of extraction was assessed on the basis of straint for stand replication. Data on occurrence the quantity of rhizomes collected (dry weight) per and availability were recorded following the season, giving three categories; (A) < 10 kg (B) standard phytosociological methods (Smith 1974; 10–25 kg (C) > 25 kg. Muller-Dombois and Ellenberg 1974; Krebs 1989). Data on species use were calculated and catego- Individuals of A. glauca and associated species were rized as domestic use, trade, domestic use and counted in each quadrat and density, frequency, trade, domestic use and barter. Data were analyzed abundance and A/F ratio was analyzed using stan- and calculated on a percentage basis for each dard phytosociological formulae (Krebs 1989). village. Survey Seed germination experiments A survey was carried out in 13 villages of Seeds were collected during October 2000 from six West Himalaya (Kullu and Parvati valley of sites (Solang Nala, SN; Naumor, N; Oonch, O; Himachal Pradesh, Kumaon and Garhwal region of Fojal, F; Sunderdhunga, SD; Phurkia, PH). Insuffi- Uttaranchal) and a part of Trans Himalaya (Lahaul cient seed numbers prevented collection from valley). The adjacent villages were selected for Pulga and Dohra, and these sites were not consid- data collection on use category and extraction ered for the seed germination study. Immediately pattern, using a structured, closed questionnaire. after collection, seeds were dried at room tempera- From each village 15 men and 15 women were ture for one week and stored separately in brown Table 1 Population data for A. glauca Latitude Longitude Altitude Centres/population (N) (E) (m asl) Aspect Habitat Uttaranchal Sunderdunga (SD) 30° 12′ 79° 54′ 3740 SE Moist, steep slope at timberline zone among Rhododendron spp. Phurkiya (PH) 30° 13′ 79° 59′ 3500 SW Steep slope in moist Rhododendron forest Himachal Pradesh Oonch (O) 32° 01′ 77° 23′ 2940 SW Moist, grassy steep slope in pure Picea forests Naumor (N) 32° 19′ 77° 12′ 2798 NE Gentle slope in mixed forests (Platanus spp., Picea spp. and Quercus spp.) Solang Nala (SN) 32° 20′ 77° 08′ 2760 NE Moist, gentle slope in mixed forest (Platanus spp. and Picea spp.) Dohra (D) 32° 09′ 77° 15′ 2700 NE Moist slope along stream in mixed forest canopy (Platanus spp and Abies spp) Fojal (F) 32° 12′ 77° 04′ 2660 NW Mixed forest (Platanus spp, Abies spp and wild cherry) Pulga (P) 32° 03′ 77° 19′ 2500 NW Very moist, grassy patch in forest canopy (Picea spp, Abies spp, and Acer spp,) International Journal of Biodiversity Science and Management 107 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. paper bags. Seed viability was determined using the chamber at 25 ± 2°C. Experiments were laid in ran- 2,3,5-triphenyl tetrazolium chloride (TTZ) test domized complete block design. Germination was (Hartmann and Kester 1989), after removing the monitored daily from the date of sowing until first wings. Fifty seeds from each site were immersed in germination and thereafter germinated seeds were 1% tetrazolium chloride solution and incubated in counted at weekly intervals. The seeds were con- the dark for 24 h. Viable embryos stained dark red. sidered to have germinated when the radical was For germination experiments, seeds that 2 mm long. These germinated seeds were removed collapsed when pinched gently with forceps and and used for seedling performance experiments. those showing fungal infection were discarded. The whole experiment was monitored for approx. Several methods were tested in an attempt to maxi- 70 days. mise germination of A. glauca (Table 2). For each treatment, three replicates of 15 seeds from each Seedling performance site were used. Seeds were washed thoroughly and soaked in test solutions for 24 h. Thereafter they Performance and survival of A. glauca seedlings were washed again with tap water and placed on were determined at the GB Pant Institute nursery at Petri dishes (95 × 17 mm) containing Whatman Katarmal (1240 m) and Lata village in Nanda Devi (No. 44) filter paper and moistened with distilled Biosphere Reserve (NDBR, 2220 m). One-month- water twice a week. The dishes were kept in a growth old seedlings in three replicates of 24 seedlings each were planted at these sites in May 2001 and watered twice a week. Percentage survival and growth performance were recorded in October Statistical analysis Data on extraction category, quantity of rhizomes extracted per growing season, seed germination and seedling performance was statistically tested using SYSTAT (Wilkinson 1986). Significant differences between means were separated by the least significant difference (LSD) (p < 0.05) test (Snedechor and Cochran 1968). RESULTS Assessment of availability and threats to the species Assessment of the species in selected natural sites showed that it occurred on north-facing (five Figure 2 Flow chart of the methodology adopted for phytosociological study and survey sites) moist slopes between 2500 to 3900 m. Plant Table 2 Details of treatments used for improving seed germination in the present study Concentration Treatment SN Treatments (ppm) duration (h) Remarks 1. Gibberelic acid (GA ) 100, 500, 1000 24 Seeds thoroughly washed with tap water 2. Thiourea 100, 200, 300 24 Seeds thoroughly washed with tap water 3. Potassium nitrate (KNO ) 100, 500, 1000 24 Seeds thoroughly washed with tap water 4. Indole acetic acid (IAA) 800, 1000, 1200 24 Seeds thoroughly washed with tap water 108 International Journal of Biodiversity Science and Management Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. density was low in all the sites, ranging from 0.95 (p < 0.05) improvement was noticed in germina- 2 2 individuals/m (SD, PH, D) to 1.8 individuals/m tion percentage in SN (73.3%) and N (53.3%) with (SN, F). A. glauca contributed 1.8% (SN) to 7.1% potassium nitrate (500 ppm) treatment, as com- (D) to total plant density, decreasing towards pared to control (SN 37.8%; N 8.9%). However, higher elevations (r = −0.510; p > 0.05). In spite germination at higher altitude sites improved of the low density, A. glauca maintained a high significantly. Thiourea also enhanced germination frequency of occurrence (70–75%) in all sites, com- for all the sites, particularly at SN and N and at parable with other dominant taxa of these sites higher altitude sites. IAA (800 ppm) was most (Table 3). The ratio of abundance to frequency effective in improving percentage germination (A/F) was used to interpret the distribution pattern across the sites, with 91.1% at SD, which was signifi- at different scales, i.e. regular, < 0.025; random, cantly (p < 0.05) higher as compared to the control 0.025–0.05 and contagious, > 0.05 distribution (64.4%). (Curtis and Cottam 1956). A. glauca was randomly distributed in SD, O, N, SN and F sites and regularly Seedling performance distributed in PH, D and P sites. Survival of seedlings planted at the higher altitudes (Lata village) was significantly (p < 0.05) higher (61.4%) as compared to those growing at Kosi Extraction (48.8%). Similarly, growth (plant height, stem dia- The extraction was exclusively by local inhabitants meter and leaf number) was significantly (p < 0.05) from wild populations (Table 4a) and for domestic higher at Lata (Table 6). use (84.1%), with few respondents involved in trade (13.3%). There was no significant gender differ- ence for extraction categories. Significantly more DISCUSSION men (p < 0.05) extracted the species for trade, Angelica glauca is a high-value medicinal plant of the while older people (> 50 years) mainly collected Himalaya, which has attained endangered status as plants for domestic use. Domestic use and barter a result of several pressures on its populations and did not differ significantly among age classes, but habitat. Overgrazing, destructive and unscientific significantly more of the younger age group harvesting are gradually altering the habitat of the (14.3%) were non-users. None of the older people species. There is a clear relationship between plant were identified as the non-users. Extraction of part harvested and the degree of disturbance to the rhizomes per growing season in the study area sites. The removal of whole plants and roots has differed little between men (60.5%) and women a more immediate and damaging effect than the (71.3%). However, significantly more (p < 0.05) harvesting of leaves and fruits (Cunningham 1998). men (14.9%) were found to collect plants under As the root is the part used in medicine, the plant category C (Table 4b), while 9.8% of younger is susceptible to destructive harvesting and plant people (< 50 years) and 5.4% of older people (> 50 productivity and sustainable utilization are contin- years) collected plants under category C. gent to the severity of harvesting practices. Distribution of the species in natural sites sug- gests that A. glauca prefers north-facing slopes in Propagation moist-shady subalpine and timberline habitats. Seeds of A. glauca from different sites had different Although the species is randomly distributed and viability: 98.3% for N followed by 95.0% for PH, has higher frequency of occurrence in such habi- 93.0% for SN, 91.7% for O, 86.9% for F and 83.3% tats, anthropogenic pressures might be responsible for SD. for the low plant density. The field survey revealed Germination differed between sites and treat- that local people are generally not concerned ments. Untreated seeds showed a marked variation about conservation, despite knowing the usefulness in percentage germination, ranging from 64.4% of the species. The Convention on Biological Diver- (SD) to 6.7% (PH) (Table 5). Soaking seeds in sity (CBD) states that indigenous people play a vital different concentrations of gibberelic acid did role in environmental management and develop- not affect percentage germination. Significant ment through their traditional practices (Glowka International Journal of Biodiversity Science and Management 109 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. Table 3 Phytosociological status of sites of A. glauca with associate species Population Species Frequency Density Indiv./m Relative density (%) A/F ratio Sunderdunga (SD) A. glauca 75 0.95 5.09 0.025 1. Rheum moorcroftiana 80 5.05 27.08 0.081 2. Polygonum polystachyum 75 2.75 14.75 0.053 3. Fragaria nubicola 85 2.60 13.94 0.047 Others 7.30 39.14 Phurkiya (PH) A. glauca 75 0.95 3.51 0.020 1. Senecio sp. 90 8.30 30.68 0.114 2. Polygonum polystachyum 100 6.90 25.51 0.076 3. Fragaria nubicola 60 2.25 8.32 0.065 Others 8.65 31.98 Oonch (O) A. glauca 75 1.20 5.96 0.028 1. Impatiens sp. 40 2.30 11.66 0.143 2. Dryopteris sp. 55 1.90 9.84 0.065 3. Fragaria nubicola 60 1.60 8.29 0.047 Others 12.40 64.21 Naumor (N) A. glauca 75 1.35 2.15 0.026 1. Impatiens sp. 100 24.30 38.81 0.246 2. Polygonum amplexicaule 100 9.55 15.25 0.108 3. Ligularia sp. 65 4.15 6.62 0.106 Others 23.25 37.08 Solang (S) A. glauca 70 1.80 1.97 0.026 1. Viola sp. 100 28.65 49.14 0.295 2. Dryopteris sp. 60 9.25 15.87 0.266 3. Rosa sp. 95 8.10 13.89 0.098 Others 10.50 19.09 Dohra (D) A. glauca 75 0.95 7.14 0.022 1. Viola sp. 80 3.85 28.94 0.065 2. Polygonum amplexicaule 65 2.10 15.78 0.053 3. Impatiens sp. 45 2.05 15.41 0.103 Others 4.35 32.67 Fojal (F) A. glauca 70 1.80 3.23 0.040 1. Fragaria sp. 100 12.95 23.20 0.131 2. Coniogramme intermedia 90 8.20 14.70 0.105 3. Viola sp. 60 4.45 7.97 0.123 Others 28.40 50.87 Pulga (P) A. glauca 75 1.15 2.59 0.022 1. Dryopteris sp. 65 13.70 30.86 0.323 2. Impatiens sp. 90 13.10 29.50 0.162 3. Ligularia sp. 80 7.45 16.78 0.129 Others 9.00 20.23 A/F = Abundance/Frequency 1997), however, cash incentives encourage local Despite a comparatively lower percentage of people to ignore traditional harvesting practices. people in the high extraction category C, a large In the present study, greater awareness of the quantity of plant material is also being extracted potential of the plant by men could be because they from the wild under categories A and B. The higher are involved in collection and trade. percentage of people (men and women) in the 110 International Journal of Biodiversity Science and Management Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. Table 4(a) Categories of extraction of A. glauca Gender (%) Age class (%) Male Female Use (n=15 ¥ 13) (n=15 ¥ 13) LSD F < 50 yr > 50 yr LSD F DU 80.5 87.7 13.6 1.3 79.7 88.5 6.1 10.2 TR 18.9 7.7 11.6 4.5 18.7 5.8 9.1 9.5 DU + BT 0.5 4.6 6.1 2.2 0.6 4.3 4.7 3.0 NU 2.1 9.2 8.6 1.7 14.3 0.0 11.5 7.2 DU domestic use, TR trade, BT barter, NU not used; (n = 15 people × 13 = village) Table 4(b) Amount of rhizome extracted by villagers in a growing season Gender % Age Class % Class Male Female LSD F < 50 yr > 50 yr LSD F A 60.5 71.3 18.2 1.2 62.3 73.8 8.8 8.1 B 24.6 28.7 15.1 0.3 29.5 23.3 7.9 2.91 C 14.9 0.0 11.4 8.1 9.8 5.4 4.9 3.8 A < 10 kg dry weight, B 10–25 kg dry weight, C > 25 kg dry weight Table 5 Seed germination responses of A. glauca with various treatments Population Treatment SN N O F SD PH LSD Control 37.8 8.9 31.1 13.3 64.4 6.7 13.9 IAA 800 55.6 24.4 37.8 37.8 91.1 20.0 16.6 1000 42.2 55.6 40.0 37.8 80.0 15.6 18.3 1200 53.3 66.7 55.7 13.3 80.0 17.8 33.1 LSD 20.6 33.9 18.9 18.5 20.3 16.3 KNO 100 60.0 37.8 35.6 13.3 75.6 8.9 12.2 500 73.3 53.3 31.1 33.3 73.3 6.7 23.4 1000 35.6 33.3 44.5 13.3 71.2 8.9 17.7 LSD 22.4 16.3 22.9 16.6 18.9 10.3 Thiourea 100 71.1 24.4 40.0 13.3 37.9 22.2 16.3 200 42.2 26.7 33.3 11.1 71.1 24.5 26.7 300 44.4 22.2 35.6 13.3 73.3 11.1 23.4 LSD 31.4 9.6 24.1 23.5 22.9 12.6 GA 100 40.0 20.0 26.7 20.0 66.7 6.7 13.9 500 40.0 11.1 31.1 13.3 46.7 8.9 10.8 1000 40.0 11.1 31.1 13.3 24. 5 8.9 16.3 LSD 16.2 8.9 23.3 7.3 14.9 11.5 high extraction category in villages with more sites inaccessible areas, involving travel for long dis- containing the plants probably reflects the influ- tances on tough terrain, which might not be ence of traders in areas of high availability. Signifi- possible for older people and women. It also shows cantly more young men are in the high extraction that younger men are important players in the category C because plants are generally found in marketing chain. International Journal of Biodiversity Science and Management 111 Angelica glauca, a critically endangered Himalayan medicinal herb Bisht et al. Table 6 Seedling performance of A. glauca at different altitudes Altitude Total planted Survival Plant height Stem diameter Locality (m asl) seedlings (%) (cm) (cm) Leaf number Kosi 1240 72 48.6 43.9 3.1 2.6 Lata 2220 72 61.4 53.9 3.6 3.2 LSD (p < 0.05) – – 12.9 2.5 0.6 0.6 In the present scenario, where collection of utilization of high-value medicinal plants of the plant material is adversely affecting plant regenera- Himalaya. The plan must focus on the following tion, it is important that a simple and effective strategies: regeneration protocol via seeds is developed so that 1. An aggressive awareness and education pro- local inhabitants can initiate farming of A. glauca gramme should be initiated among local and reduce their dependency on wild populations. people to encourage conservation; In this context, the results of germination experi- ment showed differences between sites, treatments 2. Legislative measures should be enacted for and concentrations of growth promoters. Such regulating the extraction of medicinal plants variations in germination have been reported in from the wild; other species (Baskin and Baskin 1998). IAA 3. Propagation protocols must be developed for produced the largest improvement in germination rapid and mass multiplication of the species; of A. glauca seeds, while Chauhan et al. (1998) and found that lower concentrations of IAA promoted germination of Saussurea costus. 4. Demonstration and dissemination of propaga- Significantly better survival and growth of tion packages among end users should be ini- seedlings of A. glauca at Lata (2220 m) might be tiated to promote cultivation and to reduce due to in situ environmental conditions of tempera- pressure on natural populations. ture and moisture, suggesting that maximum returns could be achieved by establishing nurseries close to natural populations. Successful cultivation ACKNOWLEDGEMENTS of Podophyllum hexandrum, Aconitum heterophyllum and Aconitum balfouroii at comparable altitudes has Authors wish to thank ID Bhatt, S Airi and col- been reported (Prasad 2000; Bahuguna et al. 2000). leagues from the CBD group for their support and help. Financial assistance from the Department of Biotechnology, Government of India (BT/ RECOMMENDATIONS PR1118/PB/17/050/98) and National Medicinal An area-specific action plan needs to be devel- Plant Board, New Delhi (No. 135/2002) is grate- oped for the conservation and sustainable fully acknowledged. REFERENCES Anonymous. The Wealth of India, A Dictionary of Indian Baskin CC and Baskin JM. Seeds: Ecology, Biogeography Raw Materials and Industrial Products, Vol. 1. New and Evolution of Dormancy and Germination. San Delhi: Publication and Information Directorate, Diego: Academic Press; 1998 CSIR; 1985 Bisht AK, Bhatt A, Rawal RS and Dhar U. Prioritiza- Bahuguna R, Purohit MC, Rawat MSM and Purohit tion and conservation of Himalayan medicinal AN. Qualitative and quantitative variations in alka- plants: Angelica glauca Edgew. as a case study. loids of Aconitum species from Garhwal Himalaya. 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Journal

International Journal of Biodiversity Science & Management – Taylor & Francis

Published: Jun 1, 2006

Keywords: CONSERVATION; ASSESSMENT; ANGELICA GLAUCA; ETHNOBOTANICAL SURVEY; GERMINATION; SEEDLING PERFORMANCE

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