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Assessment of availability and habitat preference of Jatamansi a critically endangered medicinal plant of west Himalaya
S. Airi, R. S. Rawal, U. Dhar and A. N. Purohit*
G. B. Pant Institute of Himalayan Environment and Development (GBPIHED), Kosi-Katarmal, Almora 263 643, India *High Altitude Plant Physiology Research Centre (HAPPRC), H.N.B. Garhwal University, Srinagar 246 174, India

Nardostachys jatamansi (D. Don) DC., a critically endangered rhizome-bearing medicinal plant, is restricted to specialized habitats in high altitudes of the Himalaya, ranging from 3000 to 5000 m asl. The plant is collected from natural habitats for local consumption and trade. The existing status of the species and variations in its performance in different habitats were studied in selected sites in Kumaun, west Himalaya. Dripping moss-laden rocks (frequency 40.7%, density 15.9 individual/m2) and moist boulders (frequency 25.9% and density 16.8 individual/m2) are the most preferred habitats of this plant. Generally, density and frequency had significant (P < 0.05) positive relationship with altitude. The mean density in two contrasting slopes differed significantly (P < 0.05), showing relatively higher density on west-facing slopes. Several biological and environmental features of the individual plants contributing towards wholesome below-ground biomass were identified. For example, among biological parameters, plant density (P < 0.01), plant height (P < 0.01) and above ground biomass (P < 0.01) were positively correlated. So were soil nitrogen (P < 0.05) and moisture content (P < 0.01) with below ground biomass. T HE Indian Himalayan region is a rich repository of medicinal plants with a total of 1,748 species. Increasing attention is being paid to medicinal plants value both due to their economic and conservation concerns1. Seventeen species are listed in the Red Data Book of Indian Plants2. The list includes Nardostachys jatamansi (D. Don) DC. (Hindi: Jatamansi; English: Spikenard), considered vulnerable3 at high altitudes (Figure 1 ac). A recent Conservation Assessment and Management Plan (CAMP) review identifies N. jatamansi as a critically endangered species of north West Himalaya4. Over-exploitation of the rhizome (Figure 1 d, e) for medicinal use and consequent degradation of natural habitats are reported to be the major threats to this plant3. On account of narrow distribution range, small population size and high use value, the species figure among top priority taxa of Askot Wildlife Sanctuary of
For

correspondence. (e-mail: joshidn@vsnl.com)

Kumaun Himalaya5. In spite of these attributes, the existing information of the species is inadequate, especially with regard to (i) frequency of its occurrence in natural habitat, (ii) quantitative data on its availability, and (iii) performance under different conditions. Nardostachys (Family Valerianaceae) is a small herbaceous Himalayan genus, represented by two broad range endemic species, N. grandiflora DC. and N. jatamansi (D. Don) DC. in India6. Until recently, the two species were considered synonymous7. However, critical taxonomic evaluation has separated them6. N. grandiflora has been reported to be scarcely occurring in certain localities (above 4000 m) of Kumaun and Sikkim Himalaya, whereas N. jatamansi is relatively common in Garhwal, Kumaun and Sikkim Himalaya between 3000 and 5000 m asl. The underground part of N. jatamansi is used as a substitute for valerian and the extracts find use in over 26 Ayurvedic preparations6. The root is also used for treatment of heart disease, high blood pressure and insomnia3,811. The root and rhizome contain active compounds with carminative, sedative, antispasmodic and tranquilizing properties12. Information on taxonomy, general distribution, uses, conventional and in vitro methods of propagation of N. jatamansi are available2,6,8,9,1315. Also, the phytochemical properties have been investigated in detail11,12,1621. Considering the increasing demand for herbal drugs in general and Himalayan medicinal plants1,2 in particular, and consequent depletion of several species including N. jatamansi, it is imperative to initiate urgent steps for conservation. The present communication attempts to assess the status of occurrence and quantum of availability of N. jatamansi in the natural habitats of Kumaun Himalaya, essential for developing a conservation strategy. The study area (30639N lat. and 705580E long.), located in Kumaun (west Himalaya), extends between 3100 and 4000 m asl and consists of three river catchments, viz. Pindari, Sunderdhunga and Kaphani. Usually the site remains snow bound from late October through April. The soil of the study area, a residue from crystalline rocks, is dark grey to dark brown and black and silty loam to loam in constitution. Keeping in view the altitudinal range of its occurrence, the sampling was restricted to altitudes above 3000 m asl. The study area was divided into three arbitrary altitude zones < 3400 m (S1); 34013600 m (S2) and > 3600 m (S3). In each zone of a catchment, two sites (one each in east and west facing slopes) were identified. A total of 18 sample sites were identified. However, considering localized occurrence of the species, the sampling sites were established in areas of its availability. Therefore, all the estimates will be applicable for such sites where it occurs. The estimates cannot be extrapolated for entire area above 3000 m asl.
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Figure 1. a, Jatamansi habitat; b and c, habit and d, collection in the high altitude zone of Kumaun Himalaya, e, sun drying of the rhizomes.

Vegetation sampling was conducted through vertical belt transects (one in each site 20 m wide and 150 to 200 m long) involving a total area of 0.30.4 ha. Three (20 20 m) plots (stands) were marked at three positions of each transect (viz. base, mid and top). Habitat characteristics of each plot were recorded. The detailed information on species performance was obtained from ten (1 1 m) randomly-placed quadrats in each plot. The data was analysed for frequency and abundance22. The frequency of occurrence and abundance of the species in a particular habitat was determined as an indicator of its preference. Data on morphological features (i.e. plant height, number of leaves, flowers, rhizome length, diameter, etc.) were collected from randomly-selected mature individuals (n = 10) of each site of a transect in July August, 1998. Selected individuals (n = 5) were re1468

moved (whole) from the site and brought to the laboratory for biomass studies22. The substrate of N. jatamansi varied from dry rocky crevices to moss laden rock surfaces. The substrate samples (500700 g) collected from different stands of a transect23 were dried in the sun for one week. Dried soil samples were packed in perforated polythene bags and stored at room temperature. Moisture content, pH and organic carbon were determined24. Total nitrogen was determined by Kjeldhal technique, using Kjelplus Pelicon unit for distillation. Relationships between studied parameters were statistically tested using SYSTAT25. Significant differences among means were separated using LSD mean separation test (P < 0.05)26. The density patterns of N. jatamansi among different altitude zones exhibited a significant (P < 0.05) differCURRENT SCIENCE, VOL. 79, NO. 10, 25 NOVEMBER 2000

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ence in mean density of two sites (on opposite slopes) in a particular altitude zone (Table1). The mean density (25.6 individual/m2) at > 3600 m altitude zone on west facing slope (site S3W) was significantly high compared to all other sites. This suggests favourableness of the altitude (> 3600 m asl) and aspect (west) for species proliferation. The density of species showed significant increase (r = 0.462, P < 0.05) with altitude. This is in general agreement with the statement that altitude, to a large extent determines the vegetation pattern in the mountains23,27,28. However, while comparing the distribution on two contrasting hill slopes (east and west), significantly higher increase in density (14.325.6 individual/m2) with altitude was revealed (r = 0.638, P < 0.05) on west-facing slope. Since the north and west-facing slopes in alpine zone of this region are broadly considered as relatively shady areas with low light and high moisture29, the results suggest that the species prefers such conditions. All the investigated (54) stands of N. jatamansi can be grouped under seven arbitrary habitat types (Figure 2). Considering frequency of occurrence in stands and relatively high mean density of individuals, dripping moss-laden rock (frequency 40.74%; density 15.9 individual/m2) followed by moist boulders (frequency 25.93% and density 16.75 individual/m2) are the most preferred habitats. The substrate of N. jatamansi was invariably acidic (pH 5.696.02), with high mean organic carbon (6.21 9.46%) and nitrogen (0.560.78%) (Table 2) and comparable to the values reported for alpine soils of the region22,30. In general, the higher elevation stands exhibited significant increase in nitrogen (r = 0.599,
Table 2. Soil characteristics of Nardostachys jatamansi in three different sites (values represent means of all catchments) Site S1 S1 S2 S2 S3 S3 E W E W E W Moisture content (%) 35.43 37.67 34.83 39.81 36.50 39.89 3.95 pH 5.69 5.97 5.88 6.02 5.72 5.70 0.98 Organic carbon (%) 6.21 6.80 7.58 7.97 9.28 9.46 1.31 Nitrogen (%) 0.56 0.61 0.57 0.62 0.67 0.78 0.20

LSD (<0.05)

Site details as in Table 1.

Table 1. Distribution pattern of Nardostachys jatamansi in three different catchments Density (individuals/m 2) Site S1 E S1 W S2 E S2 W S3 E S3 W Catchment mean Pindari 9.17 (2.14) 16.17 (3.62) 9.17 (3.20) 18.50 (7.45) 17.93 (5.68) 34.00 (9.40) 17.49 Sunderdhunga 6.57 (1.18) 9.93 (2.57) 12.03 (3.77) 11.23 (2.76) 7.20 (2.29) 17.23 (4.92) 15.12 Kaphani 9.83 (2.32) 16.73 (3.50) 7.87 (1.57) 19.80 (3.70) 11.03 (2.04) 25.50 (5.90) 10.59 Site mean 8.52 14.06 9.69 16.51 12.05 25.58

Figure 2. Habitat preference of Nardostachys jatamansi in selected sites of Kumaun Himalaya. MRC, Moist rocky crevices; DMLR, Dripping moss-laden rocks; MAS, Moist-alpine-slope; DRB, Dry rocky boulders; DAS, Dry alpine slope and DRC, Dry rocky crevices.

S1, < 3400 m; S2, 34013600 and S3, > 3600; E, east facing slope and W, west facing slope. Value in parenthesis is relative density. LSD (< 0.05) for site mean and catchment mean are 3.57 and 5.98 respectively. CURRENT SCIENCE, VOL. 79, NO. 10, 25 NOVEMBER 2000

P < 0.001) and organic carbon (r = 0.892, P < 0.001). The density of N. jatamansi showed significantly positive correlation with some of the substrate components like organic carbon (r = 0.613, P < 0.01), moisture content (r = 0.611, P < 0.01) and nitrogen (r = 0.795, P < 0.001). In general, while comparing the means, the site on the west slope at the highest elevation (> 3600 m asl) showed high moisture content, organic carbon, and nitrogen, compared to low elevation (< 3600 m asl) sites on the east slope (Table 2). Across sites, a considerable variation in mean biomass (above and below) of N. jatamansi (20.72 81.25 g/m2) was revealing (Table 3). The sites on west facing slopes had higher biomass (40.3181.25 g/m2) and the mean biomass of two high elevation sites (> 3400 m asl) on this slope was significantly higher (P < 0.05) than that at the other sites (Table 3). Similar is the case of below ground biomass (9.8430.64 g/m2).
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Table 3. Species performance in different sites of Nardostachys jatamansi (values represent means of all catchments) Biomass (g/m2) Plant height (cm) 10.68 12.24 12.61 14.57 11.28 11.60 3.75 No. of leaves 12.17 12.05 13.26 14.17 11.36 11.99 5.33 Above ground 10.88 17.08 13.49 31.46 15.28 50.40 13.19 Below ground 9.84 23.43 13.18 30.08 18.79 30.64 9.16

commercial exploitation. In this regard it will be pertinent to consider the specific preferred environmental/ edaphic requirement of the species before developing improved agrotechnologies for cultivation of the species.
1. Dhar, U., Rawal, R. S. and Upreti, J., Biol. Conserv., 2000, 95, 5765. 2. Samant, S. S., Dhar, U. and Palni, L. M. S., Medicinal Plants of Indian Himalaya Diversity, Distribution and Potential Values, Gyanodaya Prakashan, Nainital, 1998. 3. Nayar, M. P. and Sastry, A. R. K., Red Data Book of Indian Plants, B.S.I. Calcutta, 1988, vol. II. 4. CAMP, Workshop Proceedings, WWF India, Z00/CBSG, UP Forest Department, 1997. 5. Dhar, U., Rawal, R. S. and Samant, S. S., in Biodiversity Conservation in Managed and Protected Areas (eds Kotwal, P. C. and Banerjee, S.), IIFM, Bhopal, 1998, pp. 128142. 6. Prakash, V., Indian Valerianaceae, Scientific Publishers, Jodhpur, 1999. 7. Hara, H. and Williams, L. H. J., An Enumeration of the Flowering Plants of Nepal, Trustees of British Museum (Natural History), London, 1979. 8. Anonymous, The Wealth of India: Raw Material, CSIR, New Delhi, 1966, vol. VII. 9. Anonymous, Medicinal Plants Source Book of India, Natraj Publishers, Dehradun, 1996. 10. Dweck, A. C., in Valerian The Genus Valeriana (ed. Houghton, P. J.), Harwood Academic Publishers, UK, 1997, pp. 120. 11. Rani, P. U. and Naidu, M. U. R., Phytomedicine, 1998, 5, 253 257. 12. Bagchi, A., Oshima, Y. and Hikino, H., Planta Medica, 1991, 57, 9697. 13. Mathur, J. and Mathur, J., Ann. Bot., 1992, 70, 419422. 14. Mathur, J. and Mathur, J., Plant Cell Org. Cult., 1993, 33, 163 169. 15. Vijay, K. and Vashist, D. P., J. Hill Res., 1998, 11, 202206. 16. Rucker, G., Paknikar, S. K., Mayer, R., Breitmaier, E., Will, G. and Wiehl, L., Phytochemistry, 1993, 33, 141143. 17. Girgune, J. B., Jain, N. K. and Garg, B. D., Curr. Sci., 1978, 47, 454455. 18. Mishra, D., Chaturvedi, R. V., Tripathi, S. C. and Mishra, D., Tropical Agriculture, 1995, 72, 4852. 19. Horster, H., Rucker, G. and Tautges, J., Phytochemistry, 1977, 16, 10701071. 20. Vohra, S. B. and Dandiya, P. C., Fitoterpia, 1992, 63, 195 207. 21. Bagchi, A., Oshima, Y. and Hikino, H., Tetrahedron, 1990, 46, 15231530. 22. Airi, S., Rawal, R. S., Dhar, U. and Purohit, A. N., Plant Genet. Res. Newslett., 1997, 110, 2934. 23. Jackson, M. L., Soil Chemical Analysis, Prentice Hall, New Delhi, 1967. 24. Allen, S. E., Chemical Analysis of Ecological Materials, Blackwell Scientific Publication, Oxford, 1989. 25. Wilkinson, L., SYSTAT: The System for Statistics, Systat. Inc., Evanston IL, 1986. 26. Snedecor, G. W. and Cochran, W. G., Statistical Methods, Lowa State University Press, Ames, 1980, 7th edn. 27. Druitt, D. G., Enright, N. J. and Ogden, J., J. Biogeography, 1990, 17, 205220. 28. Singh, J. S. and Singh, S. P., Forests of Himalaya: Structure, Functioning and Impact of Man, Gyanodaya Prakashan, Nainital, 1992. 29. Rawat, G. S. and Rodgers, W. A., Proceedings of the National Rangeland Symposium, 1987, pp. 119137. CURRENT SCIENCE, VOL. 79, NO. 10, 25 NOVEMBER 2000

Site S1 S1 S2 S2 S3 S3 E W E W E W

Total 20.72 40.51 26.67 61.54 34.07 81.04 16.34

LSD (P < 0.05)

Site details as in Table 1.

Increase in plant density (r = 0.60, P < 0.01), plant height (r = 0.389, P < 0.1) and above ground biomass (r = 0.706, P < 0.001) positively correlate with below ground biomass. This suggests that overall robustness of the individuals is an indicator of increased below ground biomass. Also, significant increase in below ground biomass was observed with increasing soil nitrogen (r = 0.510, P < 0.05) and soil moisture content (r = 0.575, P < 0.01). The advantage of high water and nutrient supply for rhizome and root production has also been reported elsewhere for related valerian species31. Interestingly, unlike above ground biomass and density which show significant increase with altitude (r = 0.433, P < 0.05), below ground biomass does not show such relationship. This does not match with the observations on Podophyllum hexandrum, another rhizomatous medicinal plant of the region, which shows significant increase in below ground biomass at lower elevations22. Variations in response to environmental stresses are species specific and therefore must be considered while developing strategies for sustainable harvest and conservation. However, ecological implications can be meaningful if the correlations between secondary compounds (active ingredients), morphological and environmental parameters are also taken into consideration. Such type of information is presently lacking. On the basis of the present investigations, it is concluded that (i) N. jatamansi is restricted to some specialized habitats and is subject to destructive harvesting (removal of root/rhizome) from the wild. In this context, the quantified data on its occurrence and availability and relationship of utilizable part (below ground biomass) with various biological and environmental parameters can be gainfully utilized for identification of potential habitats and expected yield per unit area. (ii) Ex situ conservation of the species assumes greater significance especially in a scenario when harvesting potential from the wild falls short of the demand for
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30. Ram, J. and Singh, S. P., in High Altitudes of the Himalaya (eds Pangtey, Y. P. S. and Rawal, R. S.), Gyanodaya Prakashan, Nainital, 1994, pp. 3355. 31. Bernath, J., in Valerian the Genus Valeriana (ed. Houghton, P. J.), Harwood Academic Publishers, UK, 1997, pp. 77100. ACKNOWLEDGEMENTS. We are grateful to the Director, G. B. Pant Institute of Himalayan Environment and Development, Kosi Katarmal, Almora for constant help and support. The study was carried out with financial assistance from the Department of Biotechnology (BT/R&D/19/02/92), New Delhi. Received 4 August 2000; accepted 12 September 2000

Decaying wood as a natural habitat of melanin-forming (Mel+) variant of Cryptococcus laurentii

A. Y. Mussa*, H. S. Randhawa*, and Z. U. Khan**
*Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, P. O. Box No. 2101, Delhi 110 007, India **Department of Microbiology, Faculty of Medicine, University of Kuwait, Kuwait 13110

The association of a melanin-forming (Mel+) variant of Cryptococcus laurentii with decaying sal wood, Shorea robusta, is reported here. The variant was cultured from 8 of the 14 samples of the wood detritus collected over a period of two and a half years. The average population density of the fungus was found to be 1 10 4 colony forming units (CFU)/g of the wood substrate. The variant was not detected in any of the 702 samples of miscellaneous plant materials, such as bark of Eucalyptus camaldulensis, E. tereticornis, Eugenia jumbolina, Syzygium cumini, Ficus religiosa, etc., slime flux of various trees and Sphagnum moss. Likewise, it was not detected in any of the 181 samples of bat guano and 137 of soil. The repeated isolation of the Mel+ variant in considerable numbers from the S. robusta wood detritus indicates that decaying wood may serve as a natural habitat for this fungus. This observation is of considerable epidemiologic importance since phenoloxidase (melanin-forming) activity is a virulence factor in Cryptococcus neoformans, a well known pulmonary and meningeal yeast pathogen of humans and animals. Also, it was noteworthy that the Mel+ variant incited macroscopic lesions on the liver and spleen of intraperitoneally infected cortisone-treated (immunosuppressed) white mice, indicating that the fungus is a potential opportunistic pathogen. CRYPTOCOCCUS laurentii (Kufferath) C.E. Skinner (1950), an encapsulated yeast-like fungus, has been oc casionally implicated in the etiology of cryptococcosis,

a pulmonary and meningeal fungal infection of humans and animals15. It has a melanin-forming variant which is of clinical interest because melanin formation or phenoloxidase activity has been reported as a virulence factor for Cryptococcus neoformans6,7, the etiologic agent of cryptococcosis. The natural habitat of the variant, as also of the parent species, C. laurentii, is as yet unknown. In the course of an environmental survey of clinically important yeast-like fungi, a number of isolates of the C. laurentii variant were recovered. In this communication we report that decaying sal wood, Shorea robusta, may serve as its natural habitat. The material investigated was collected from the hollowed part of an old decaying beam of sal wood, S. robusta, kept outside the boundary wall of the Engineering Department, University of Delhi, North Campus. The beam measured 0.25 0.3 4.7 m with a hollow at each end and one in the middle (Figure 1). The timber beam had been removed in 1993 from a demolished British Army barrack constructed way back in 1911. The wood samples were collected in polythene bags using disinfected spoons and forceps and stored at room temperature in the laboratory until processed; they were apparently free from avian or any other animal excreta. About 5 g of each sample was suspended in 50 ml of sterile physiological saline, containing chloramphenicol (0.05 mg ml1). The suspension was shaken for 5 min on a vortex mixer and allowed to settle up to 60 min. The supernatant was collected by a sterile Pasteur pipette and 0.5 ml aliquots were streaked in triplicate on plates of Staibs niger seed (Guizotia abyssinica) medium of the following composition: Niger seed, 70 g; glucose, 1 g; chloramphenicol, 40 mg; gentamycin, 25 mg; diphenyl solution (1 g 10 ml1 of 95%

For correspondence. (e-mail: vpci@delnet.ren.nic.in)

Figure 1. Part of an old decaying timber beam of sal wood, Shorea robusta, showing an apical hollow at one end. The wood detritus collected from inside this and other hollows in the beam yielded repeated isolation of the Mel+ C. laurentii variant. 1471

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