N 12(3): 1573-1578, 2017 Save Nature to Survive www.thebioscan.com THE HOSTS, ANATOMY AND UTILIZATION OF THE COMMERCIALLY EXPLOITED PARASITIC HERB fungosa var. INDICA IN THE WESTERN GHATS REGION OF KERALA, INDIA

ASHISH ALEX* AND K. VIDYASAGARAN Department of Forest Management and Utilization, College of Forestry, Kerala Agricultural University, Thrissur-680 656, Kerala e-mail: [email protected]

KEYWORDS ABSTRACT Balanophora fungosa Balanophora fungosa var. indica is a dioecious holoparasitic herb found attached to the roots of higher . parasite The transect method was used to locate the parasite. In Silent valley it was seen in the tropical west coast wet hosts evergreen forest and west coast semi-evergreen forests. The major host in evergreen forest is Polyalthia fragrans and in semi-evergreen forest it is Agrostistachys longifolia. The spheroidal shaped grains area ranged from anatomy 700-1200 micrometers, perimeter ranged 94-161 micrometers and mean length through major and minor axis Received on : were 32.32 micrometers and 30.44 micrometers respectively. The fresh tubers weighed on an average 0.5 kg and 31.03.2017 when 5 kg of fresh Balanophora fungosa was dried, 1kg of dried matter was obtained. The tuber consists of the parenchyma tissues, which are filled with dark brown coloured sticky resinous substance. The host root consists Accepted on : of the vacuolate parenchyma cells and phenolic compounds filled cells. The tribes of Attappady usually applied 20.05.2017 the paste of Balanophora fungosa on the swellings on their body and marketed it through the tribal cooperative society to the pharmaceutical companies. Unfortunately it is used as an adulterant, so further investigations are *Corresponding required to know more about its pharmacognostical characters. author

INTRODUCTION material may be tapped, and for this they get clues from rural or tribal men (Hiremath et al., 2007). For skin diseases, Paliars The herbaceous vegetation is a vital component of the forests, apply the paste of whole over the infected part typically comprising the largest proportion of forest diversity (Ignacimuthu et al., 1996). Such documented information on (Jhariya and Oraon, 2012). Balanophora fungosa is a parasitic forest flora may serve as the potential sources which could be herb found attached to the roots of higher plants. It is a utilized for the benefit of the whole human society (Singh and commercial non-timber forest product, collected and marketed Kumar, 2012). to the pharmaceutical companies in the state. The genus Eventhough various studies have reported the occurrence of consists of several species of fleshy herb this species in Kerala forests. None of them have mentioned parasites which are considered to be rare (Yadagiri et al .,1984). about the hosts associations, use and anatomy of this species. The genus Balanophora (15 species) inhabits the tropical and Since moisture is a must for the growth of the plant; it is expected sub-tropical belts of the world (Hansen, 1972). The distribution to be seen in evergreen forest. This article attempts to document of the plant species depends on many factors such as host plants, pollinators, dispersers and moisture (Rao et al., 2015). the habitat, host association, collection and utilization of This species is found in India, East Malaysia, Taiwan, the Pacific Balanophora fungosa var. indica in Kerala part of Western Islands and northeastern Australia (Hsiao et al., 2010). Ghats. Balanophora species are among the most unusual of all higher plants and are known to parasitize at least 74 species in 35 MATERIALS AND METHODS families (Mabberley, 1987). The color of plant ranges from pale yellow and brown through various hues of pink and red The study was conducted in the buffer zone of Silent Valley to deep purple, but no green due to lack of (Kim National Park. The transect method was used to locate the and Won, 2013).The male flowers are comprised of fused parasite. The methodology stated by Hansen (1972) was (synandria) that usually produce white pollen followed to identify the host of the parasite. The habitat was (Barkman et al., 2003). Balanophora is listed under the App II identified based on the vegetation present. The pollen grains of the CITES (Convention on International Trade in Endangered were observed under microscope and measurements were Species of wild flora and fauna) and is prohibited for export taken using image analysis system.To known about the from India (Kipgen and Singh 2010). anatomy of rhizome, hand sections were taken . The sections Ethno botanists bring out suggestions as to which raw plant of the haustorial tissues were taken using rotary microtome.

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These sections were analyzed under compound microscope. existence of this species. In Silent Valley National Park The tribal settlements were visited to know about the collection Balanophora fungosa var. indica the dioecious holoparasite method and utilization of Balanophora fungosa var. indica. herb is seen in tropical west coast wet evergreen forest and The marketing information about this species was collected west coast semi-evergreen forests. In evergreen forest the major from the Kurumba tribal cooperative society at Chindakki, host of the parasite is Polyalthia fragrans which belongs to the Attappady. family Annonaceae. In semi-evergreen forest the major host is Agrostistachys longifolia which belongs to the family Euphorbiaceae. Among the seven hosts (Table 1), majority of RESULTS AND DISCUSSION the host belongs to the family Euphorbiaceae. In Kerala no Habitat and associated trees studies have reported the host species of the parasite. The In Kerala Balanophora fungosa is distributed in the evergreen host trees reported elsewhere includes Macaranga tanarius forests of Pathanamthitta, Malapuram, Kannur, Palakkad, and Diospyros philippensis (Mabberley, 1987). This species Trivandrum and Idukki districts (Sasidharan, 2004). The is distinctly dioeciously parasitizing the roots of Coffea Arabica, distribution of the plant species depends on many factors Elettaria cardamomum and Acacia decurrens (Shivamurthy such as host plants, pollinators, dispersers and moisture (Rao et al., 1981). This parasite was not found to be so specific to et al., 2015). The moisture is very much essential for the any particular host. The elongation capacity of the host root is

Plate 1. Male inflorescence Plate 2: Female inflorescence

Table 1: The host trees of Balanophora fungosa in Silent Valley National Park Sl.No. Type of forest Host Local name Family Habit 1 West Coast Wet Polyalthia fragrans Nedunar Annonaceae Tree Evergreen Forests Mallotus albus Vattakumbil Euphorbiaceae Tree (1A/C4) Buchanania axillaris Kozhivalan “ Tree 2 West Coast Semi- Agrostistachys longifolia Kulamavu Anacardiaceae Tree Evergreeen forests Macaranga peltata Vatta Euphorbiaceae Tree (2A/C2) Callicarpa tomentosa Cheruthekku Verbenaceae Tree Xanthophyllum arnottianum Mottal Xanthophyllaceae Tree

Table 2: Morphometric parameters of pollen grains of Balanophora fungosa Sl.No. Area* Perimeter* Major Axis* Minor Axis* 1 779.26 94.31 31.03 26.89 2 895.92 109.65 30 28.27 3 907.28 109.65 33.44 28.62 4 1183.31 121.03 34.75 33.1 5 835.97 104.87 29.96 28.65 6 790.54 96.27 35.51 32.75 7 830.25 98.27 32.75 32.06 8 946.95 108.68 32.41 32.06 9 704.12 92.41 29.65 28.96 10 805.38 160.04 33.79 33.1 Mean 867.89 109.51 32.32 30.44 * All measurements in Micrometers

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Plate 3: A White Pollen deposit on Male inflorescence Plate 4.Microscopic view of pollen grains(40x)

Plate 6: Connection between host root and haustoria, dichotomous Plate 5:TS section of parenchyma cells in tuber with brown branching pattern of composite conducting strands coloured resinous substance (40X) lost through the infection of the parasite. small grains (<10 µm), 2) Small pollen grains (10-25 µm), 3) Reproduction in Balanophora Medium size (25-50 µm), 4) Large pollens (50-100 µm), 5) Very large pollen grains (100-200 µm), 6) Gigantic The inflorescences are yellow to pinkish orange in colour, pollen grains (>200µm). The pollens of Balanophora fungosa visible only during the flowering season (during, December to comes under the class medium size pollens. The area of pollen February). The inflorescences (plate 1 and 2) emerge from grains comes in the range 700-1200 micrometers and warty tubers that are attached to their host plants. Pale yellow perimeter in the range 94 -161 micrometers. colored modified leaves are seen below the male and female inflorescence. The female flowers are one among the most Anatomical features reduced in the angiosperms with more than one million hair- Tuber like flowers estimated from a single inflorescence (Hansen, The Tubers are sub-spherical in shape and consist of 1972). The male flowers are comprised of fused stamens parenchyma cells (plate 5). There are many parenchyma cells (synandria) that usually produce white pollen (Barkman et al, that are filled with dark brown or black coloured sticky resinous 2003). The Pollen grains are too small and seen as white substance (Ekambaram and panje, 1935) called balanophorin powdery deposit on the male inflorescence (Plate 3 and 4). (Goeppert, 1841) which attains dark colour when stained. No The male inflorescence is completely covered with the pollen vascular tissue of the parasite was developed in the tuber as sacs. The sacs at the bottom got matured first and opened up long as it continues to be in the vegetative phase (Shivamurthy very quickly. The pollen grains are spheroidal in shape with et al., 1981). At the point of infection on the adventitious root length through major axis ranging from 29.5-35 micrometers of the host, the tuber of the parasite clusters around it. and through minor axis ranging from 26-34 micrometers (Table 2). The average length through major axis is 32.33 micrometers Adventitious Root of host plant and average length through minor axis is 27.24 micrometers. Following infection, the rootlet ceases to grow in length. The Erdtman (1952) has classified the pollen grains into various terminal meristem of the rootlet was rendered functionless classes based on their measurements. The classes are 1) Very (Shivamurthy et al., 1981). Adventitious roots (plate 7 and 8)

1575 ASHISH ALEX AND K. VIDYASAGARAN

Plate 7: Parasite attached to the adventitious root of the host Plate 8: Dead portion of host root due to parasite infection

Plate 9: The unstained section showing tissues inside the host root Plate 10 : The unstained section showing haustorial connection (40x) between host and the parasite (40x) were formed from the host root on which the tuber gets relationship, exist between them. attached. Due to the infestation of the parasite, modifications Extraction and Utilization have occurred in host roots. Some of the cells in the host became meristematic and turned into vacuolate parenchyma The entire plant of Balanophora fungosa is collected during and cells filled with phenolic compounds (plate 9). The January to February. It is composed of an underground tuber adventitious root of the host plant consists of both host tissues which attaches to host roots and inflorescences which are and parasite tissues. only visible during the flowering season (Hsiao et al., 2010). This item was collected only by the Kurumba community. Haustorial tissues Within the Kurumba community those people residing in the The inward growing haustorium penetrating the host and the interior settlements like Anavayi and Thuddukki are mainly outward growing tissues of host root amidst haustorial tubes, involved in the collection. The entire plant is collected using form composite conducting strands (Shivamurthy et al., 1981). sickle. After removing the soil around the plant, using sickle, These strands are referred to as intra-matrical roots (Ekambaram the portion of the tree root to which the Balanophora fungosa and Panje, 1935) or podium strings (Fagerlind, 1948) which tuber got attached was cut. After collection it is washed properly does the absorptive function. The composite conducting to remove the soil particles on the tuber. Then cut into small strands in Balanophora fungosa has dichotomous branching pieces and dried for 10-12 days in open sun (plate 11). When pattern (plate 6). The division of the apical meristem into two 5 kg of fresh Balanophora fungosa was dried, 1kg of dried independently functioning axes is defined as dichotomous matter was obtained. The fresh tubers weighed on an average branching (Gola, 2014). The haustorial organ in Balanophora is a tuberous structure, ramifying to form a complex branch 0.5 kg and were having average height and length of 37 and system (Kuijt and Dong 1989). The host tissues are filled with 20 cm respectively. The collectors travel 10-15 km and collect phenolic compound, which appeared in red colour and the 20-25 kg of Balanophora fungosa per day. tuber tissues are having the balanophorin deposit, which gives The tribes of Attappady usually applied the paste of them brown colour. The plate 10 shows the clear connection Balanophora fungosa on the swellings on their body. For skin of the host and tuber tissues, which indicate a strong haustorial diseases, Paliars apply the paste of whole plant over the

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Table 3: The price spread and collector’s share analysis of the medicinal plant Balanophora fungosa sold through the society Sl.No. Years 2008-09 2009-10 2010-11 2011-12 2012-13 Factors 1 Quantity procured (kg) 38 381 6728 10042 106 2 Procurement price (Rs) 10 17.5 47.5 47.5 50 3 Quantity sold by society (kg) 38 381 1814 Nil 3190 4 Sales price (Rs)A 22 30 80 - 70 5 Price spread 12 12.5 32.5 - 20 6 Collector’s share 45 58 59 - 71

knowledge of Balanophora. I. The morphology and parasitism of the tuber. Proc. Ind. Acad. Sci. B, 1,PP. 452-470. Fagerlind, F. 1948. Bau und Entwicklung der vegetativen organe von Balanophora. Kongl. Svenska vetenskapsakad. Hand!. 25: 1-72. Ganesan, S., Suresh, N. and Kesaven, L. 2004. Ethnomedicinal survey of lower Palni Hills of Tamil Nadu. Indian J. Traditional Knowledge. 3(3): 299-304. Goeppert, H. R. 1841. Bau der Balanophoren. Nova Ada Phys. Med. Acad. Caes. Leop. Carol. Nat. Cur. 18: 229-72. Gola, E.M. 2014. Dichotomous branching: the plant form and integrity upon the apical meristem bifurcation. Front Plant Sci. 5: 263. Hansen, B. 1972. The genus Balanophora: A taxonomic monograph. Dansk Botanisk Arkiv. 28: 1-188. Hsiao, S., Huang, W, and Lin, M. 2010. Genetic diversity of Balanophora fungosa and its conservation in Taiwan. Botanical studies, 51: 217-222. Plate 11: Dried Balanophora fungosa Hiremath, S. G., Lakshmanan, C. G, Byatanal, M. B, Sabannavar, S. J., Bheemareddy, V. S. and Charantimath, A. S. 2007. A check list of infected part (Ignacimuthu et al., 1996). The tribes of Kerala indigenous medicinal plants of Dharwad and their traditional uses. used this plant for piles (whole plant is dried, powdered and The Bioscan. 2(2): 131-134. taken internally with honey) and also used along with other herbs in many preparations for curing internal hemorrhages Ignacimuthu, S., Ayyanar, M. and Sivaraman, K.S. 1996. (Pushpangadan and Atal, 1984). The plant paste is used for Ethnobotanical investigations among tribes in Madurai district of eczema (Ganesan et al., 2004; Samuel and Andrews, 2010). Tamilnadu (India). Journal of Ethnobiology and Ethnomedicine, 2: Though it is not an official drug in any of the indigenous 25. systems of medicine in India, it is used in tribal medicines in Jhariya, M. K. and Oraon, P. R. 2012. Analysis of herbaceous diversity South India. However, it is found in crude drug markets as in fire affected areas of Bhoramdeo wildlife sanctuary, Chhattisgarh. substitute/adulterant for the Ayurvedic drug Gajapippali The Bioscan. 7(2): 325-330. (Scindapsus officinalis) (Kannan and Babu, 2014). Kannan, R. and Babu, U. V. 2011. Pharmacognostical studies on Marketing Balanophora fungosa –a negative listed plant. Ancient Science of Life. Local name of Balanophora fungosa is nilabombu and in trade 31: 22-25. it is known as Athithippali. The quantity of Balanophora Kim, J. H. and Won, H. 2013. Identification of host plant species of collected over the years has shown an increase except in Balanophora fungosa var. indica from Phnom Bokor National Park of 2012-13 (Table 3). Because in 2011-12 the society could not Cambodia using DNA barcoding technique. Korean J. Pl. Taxon. 43(4): 252-262. sell the product, so they have reduced the quantity collected in the following year. The procurement price also had shown Kipgen, L. and Singh, K. J. 2010. Balanophora: a rare and endangered plant found in North-East India. Ecotone. 2(1):6-9. an increase over the years. The sales price has also showed an increasing trend except in 2011-12 when no sales occurred. Kujit, J. and Dong, W. X. 1989. Surface features of the leaves of The price spread has shown a varying trend over the years. Balanophoraceae- a family without stomata? Plant systematic and Evolution. 170: 29-35. The highest collector’s share was obtained in the year 2012- Mabberley, D. J. 1987. The plant book. Cambridge University Press, 13 (71%). Cambridge. P.73. Pushpangadan, P. and Atal, C. K. 1984. Ethno-medico-botanical REFERENCES investigations in Kerala. Some primitive tribals of Western Ghats and their herbal medicine. J. Ethnopharmacology. 11: 59-77. Barkman, T. J., Emoi, B. E. and Repin, R. 2003. The genus Balanophora Rao, J. P., Satish, K. V., Sankar, B. S., Reddy, C. S., and Kumar, O. A. (Balanophoraceae) in Sabah, Malaysia. Blumea. 48: 465-474. 2015. On the occurrence of parasitic plant Balanophora fungosa J.R. Erdtman, G. 1952. Pollen Morphology and Plant — Forster and G. Forster (Balanophoraceae) in AndraPradesh, India. J Angiosperms:AnIntroduction to Palynology vol.I. Threatened Taxa. 7(2): 6943-6946. Chronica. Botanica Co., Stock holm. P.539. Samuel, J. K. and Andrews, B. 2010. Traditional medicinal plant Ekambaram, M. A and Panje, R. R. 1935. Contributions to our wealth of Pachalur and Periyur hamlets Dindigul district, Tamilnadu.

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