CURRENT STATUS AND CHALLENGES FOR CONSERVATION AND SUSTAINABLE USE OF BIODIVERSITY

234 Current status and challenges for conservation and sustainable use of biodiversity dominant over the shrubs and herbs. But the conservation. Journal of Bombay Natural History Society, 72(2): 313-320 no. of shrubs are greater than that of trees and Gadgil, M. and Vartak, V.D.1976. The sacred herbs. Chassalia curvifolia is the most groves of Western Ghats in India. Economic Botany, 30:152-160. common plant in the sacred grove. Hopea Induchoodan, N.C. 1998. Ecological Studies parvifolia is on the endangered species , that of the sacred Groves of Kerala. Ph.D .Thesis. Central University, Pondicherry, 110-198. was also present in the sacred grove. Joshi, N.V. and Gadgil , M. 1991. On the role Chassalia curvifolia, Pothos scandens, of refugia in promoting prudent use of biological resources. Theor. Popul. Bio., Curcuma longa, Tabernamontana 40:211-229 alternifolia, Arbus precatouris, Alstonia Mohanan Nair, M. and Nair, N .C. 1981. Kunstleriaprain- an new genus record of scholaris etc. are the most common species India and a new species in the genus. Proceedings of Indian Academy of Science, on the sacerd grove. The study was based on 90: 207-209. the specie area estimation and quadrant Rajendraprasad, M. 1995. The Floristic, structural and Functional Analysis of Sacred analysis. After the analysis it results that, groves of Kerala. Ph.D. Thesis, University of Kerala, Thiruvanthapuram, India, 42-35. Chassalia curvifoliawas the most frequent, Ramachandran, K.K. and Mohanan, C.N. abundant and richest plant species in the 1991. Studies on Sacred Groves of Kerala. Final report submitted to Ministry of sacred grove and followed by Arbus environment and Forests, Government of India, 337-352 precatorius and Pothos scandens. Most of the Schaaf, T.1998. Sacred groves in Ghana: tree species having frequency about 20% in Experiences from an integrated study project. In: Ramakrishnan, P.S., Saxena, K.G and the sacred grove. But there different varieties Chadrashekara, U.M(Editors) Conserving the Sacred for Biodiversity Management. of the tree species are found on grove. UNESCO and Oxford- IBH Publishing, New Delhi, 145-150 Biophytum senitivum was the most abundant Unnikrishnan, 1995. Sacred Groves of North plant. Kerala-An Ecofolklore Study (in Malayalam ). Jeevarekha, Thrissur, Kerala, 55-89 References Vartak, V.D. and Gadgil, M. 1981. Studies Amirthalingam, M. 2016, Sacred grove of on Sacred groves along the Western Ghats India – An overview. Int. Journal curr. Res. from Maharashtra and Goa: Role of beliefs and folklores. In: Jain, S.K (Ed.) Glimpses of Biosci. Plant Biol., 3(4):64-74 Ethnobotany. Oxford University Press, Bombay, 272-278 Bash, S. C 1998. Conservation and management of sacred groves in Kerala. In: P.S. Ramakrishnan., K.G.Saxena and U.M. Chandrashekara (Eds). Conserving the Sacred forBiodiversity Management. Oxford and IBH Publishing Co. Pvt . Ltd., New Delhi, 32337-348 Chandrasekara, U.M. and Sankar, S.1998.Ecology and Management of sacred groves in Kerala, India Forest Ecology and Management, 112:165-177 Gadgil, M. and Chandran, M.D.S. 1992. Sacred groves of India In: Sen, G. (Editor) Indigenous Vision. Sage Publications (India) and International Centre, New Delhi,183-187 Gadgil, M. and Vartak, V.D. 1975. Sacred grove of India -A plea of the continuous Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.229-234 © Principal, Sree Narayana College, Kollam, Kerala, India

ISBN 978-93-5396-871-7 CHAPTER 37 A PRELIMINARY ASSESSMENT ON THE DIVERSITY OF GENUS FICUS L. (MORACEAE) IN KERALA Sreehari S Nair* and Amitha Bachan K H Research Department of Botany, MES Asmabi College, P. Vemballur – 680 671, Kerala *Correspondence E-mail: [email protected] ABSTRACT The current estimate accounts for the presence of about 750 species of FicusL. in the world. 115 species have been so far reported within the political boundary of India. The plants are grouped into 6 subgenera and 12 sections. Majority of the species belongs to the sub genus Urostigma. Among them, only 10 species are reported as endemic. North eastern parts of India are considered as the hot spot of figs with a total diversity of 43 species. The current study provides a preliminary assessment on the diversity of figs in Kerala. The assessment revels the identity of 33 species within the Kerala state. Plants belonging to 5 sub genera are only reported from Kerala. Members belonging to the subgenus Ficus are not reported from Kerala. Five species are endemic to Western Ghats one each being in the endangered and Near Threatened category. Four among the species are exotic. In Kerala subgenus Urostigma is dominant with a total of 19 species, in which 11 and 8 species in the subsection Conosycea and Urostigma respectively almost similar to the Indian distribution. Key words: Diversity, Distribution, Endemic, Ficus Introduction works of Sasidharan and Jomy Augustine (1999) and Priyadharsanan (1999). No complete revisionary work The term ‘Figs’ commonly refers to the genus FicusL., was done on the genus in Kerala. Hence being a belonging to the family Moraceae. It is one among the keystone species, supporting a wide range of flora and most advanced and abundant plants among the fauna, a check list of the Ficus species in Kerala is angiosperms. The genus is characterised by the inevitability. The current study provides a checklist of presence of simple alternate leaves, latex, the species of figs from Kerala. hypanthodium inflorescence with minute flowers embedded within the cup shaped structure. They are Materials and Methods considered as a keystone species in many ecosystems (Vanitharani et al., 2009; Kumar et al., 2011), with a Extensive field trips, herbarium visits and literature diversified habitat and life forms. The highly nutritional survey were done for the study. Fields works were done attractive figs, fruiting the year around are a source of within the Kerala state for the collection of specimens. nutrition to a wide variety of birds and animals. 27 live samples were collected during the study period. The collected samples were identified using literatures The genus has worldwide distribution (Corner, 1965; and regional floras. Confirmation of the specimens was Lambert and Marshall, 1991; Lomáscolo et al., 2010) done by herbarium consultation in CALI, TBGT, KFRI and with majority of the species in the subtropical and and MH. Additional samples reported from Kerala were tropical regions. According to the current status, around recorded by literature reference and observed during the 750 species of figs were reported from the world herbarium visits for confirmation of the species. (Chaudhary et al., 2012). The variable habitat pattern Relevant sources including International Plant Name and life forms could be the reason for the diversification Index (IPNI) and Plant list were used for the updated of the genus. The recent systematic account for the nomenclature and for checking the protologue of the genus states the presence of 115 species of figs within species. the political boundary of India. The taxa being problematic with respect to the taxonomic characters Results and identification difficulties, remain unnoticed. The comprehensive studies at global level, done on the According to the current estimate, 33 species of figs genus accounts for the work of King (1887-1888), have been reported from Kerala. Among them five Corner (1958, 1965, 1981) Berg and Corner (2005) etc species are endemic to Western Ghats. As per the (Chaudhary et al., 2012). In India the revisionary study current system of classification, the genus Ficusis on the genus was initiated by Botanical Survey of India divided into six subgenera and 12 sub sections. (BSI) in 2008. Only minor regional works were done on Members belonging to six subgenera including Ficus, the genus in Kerala and the studies accounts for the Pharmacosycea, Sycidium, Sycomorus, Synoecia, Urostigma is present in India. No species belonging to Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.235-242 © Principal, Sree Narayana College, Kollam, Kerala, India

236 Current status and challenges for conservation and sustainable use of biodiversity the subgenus Ficus is reported from Kerala so far. Ficus peepul Griff. Not. Pl. Asiat.4: 393.1854. Urostigma is the most dominant subgenus with 19 species. Urostigma religiosum(L.) Gasp. Ficus genus, dissertation botanica 82, pl. 7.1844. Systematic Treatment Distribution: All throughout India, China, Sri Lanka, Subgenus: Urostigma, Section: Urostigma, Subsection: Myanmar. Distributed above sea level to 1025 m. Urostigma 5. Ficus rigidaJacq. var. bracteata(Corner) Bennet, 1. Ficus amplissima Smith in Rees, Cycl. xiv. n. 1. Inidan J. For. 5(4): 326. 1982; Vajr., Fl. Palghat Dist. 1810; Gamble, Fl. Pres. Madras. 1362. 1928; Manilal, 449. 1990; Mohanan and Sivadasan., Fl. Agasthyamala Fl. Silent Valley. 259. 1988; Sasidh and Sivar., Fl. Pl. 636.2002; Ratheesh Narayanan, Fl. Stud. Wayanad Thrissur For. 427.1996; Anilkumar et al., Fl. Dist. 774. 2009; Sasidharan and Sivarajan., Bio. Docu. Pathanamthitta. 462. 2005; Sunil and Sivadasan, Fl. Kerala part 6: 441. 2004; Nayar et al., Fl. Pl. Kerala Alappuzha Dist. 660. 2009; Sasidharan and Sivarajan, 436. 2006. 2004 2. Ficus indica Wild. Sp. Pl. 4: 1146. 1806; Linn, Sp. F. glaberrima Blume var. bracteataCorner, Gard. Bull. Singapore 17: 388. 1960. Pl. 2: 1060. 1753. F. lawesiiKing, J. As. Soc. Bengal 55 (2): 403. 1887. Ficus tjiela Miq. Ann. Mus. Bot. Lugduno – Batavi. 3: 286. 1867. F. travancorica King, Ann. Roy. Bot. Gard. (Calcutta) 1: 28, t. 26, 82o. 1887 and in Hook. f., Fl. Brit. India 5: Ficus tsiela Roxb.Hort. Bengal.66; Hook.of., Fl. Brit. 503. 1888. India.5: 515. 1888. Distribution: Kerala and Tamil Nadu. Distribution: Distributed in elevation up to 1000 m from sea level. Native to India. 6. Ficus superba(Miq.) Miq. Ann. Mus. Bot. Lugd.-Bat. 3: 264, 287. 1867. 2. Ficus arnottiana (Miq.) Miq.., Ann. Mus. Bot. Lugd- Bat. 3: 287.1897; Gamble, Fl. Pres. Madras. 1363. Urostigma superbum Miq., Pl. Jungh. 46. 1851; Fl. Ind. 1928; Sasidharan and Sivarajan Fl. Pl. Thrissur Bat., 1, 2: 334. 1859. For.428.1996; Anilkumar et al., Fl. Pathanamthitta.462. 2005; Mohanan and Sivadasan, Fl. Agasthyamala. 629. Ficus tenuipes S. Moore, J. Bot. 63, Suppl. 107. 1925. 2002; Sasidharan and Sivarajan., Bio. Docu. Kerala part 6: Flow. Plant. 438. 2004. Distribution: India (Kerala), Indochina, Malesia, Thailand. Ficus courtallensis (Miq.)Baill, Hist. Pl. (Baillon).6: 7. Ficus tsjahela Burm. f. Fl. Indica. 227. 1768; Hook. 176.1875. f., Fl. Brit. India 5: 514. 1888; Gamble, Fl. Pres. Ind. 1362.1928; Anilkumar et al., Fl. Pathanamthitta. 466. Ficus populeasterDesf, Tabl.Ecole Bot, 3: 412.1829. 2005; Sunil and Sivarajan, Fl. Alappuzha Dist. 666. 2009; Sasidharan and Sivarajan, 2004; Mohanan and Distribution: Usually fond above 700 m from sea level, Sivadasan, Fl. Agasthyamala. 637.2002. in rocky places. 3. Ficus caulocarpa (Miq.) Miq., Ann. Mus. Bot. Ficus venosa Aiton. Hort. Kew. 3: 451. 1789. Lugd.-Bat. 3: 268, 287. 1867; Khanna et al., Suppl. Fl. Madhya Pradesh 162.2001; Fl. Kasaragod Div. 354. Distribution: Peninsular India, Sri Lanka. 1985; Sasidharan and Sivarajan, 2004. 8. Ficus virensAit. var.virens Hort. Kew (ed. 1) 3: 451. Urostigma caulocarpumMiq., Hook. London J. Bot. 6: 1789, var. virens; Sasidharan and Sivarajan (2004). 568. 1847. Ficus infectoria Roxb.Fl. Ind. 3: 551. 1832, non Ficus infectoria Roxb.var. caulocarpa(Miq.) King, Willd.1806; Hook. f., Fl. Brit. India 5: 515. 1888; Ann. Roy. Bot. Gard. (Calcutta) 1: 63, t. 79. 1888. Gamble, Fl. Pres. Madras 1362(953). 1928. Distribution: India (Karnataka, Kerala, Madhya Distribution: Peninsular India. Indo Malaysia. Pradesh). Subgenus: Urostigma, Section: Urostigma, Subsection: 4. Ficus religiosaL. Sp. Pl. 1059. 1753; Hook, Fl. Brit. Conosycea Ind. 5: 512. 1888; Gamble, Fl. Pres. Ind. 1363.1928; Anilkumar et al., Fl. Pathanamthitta. 465. 2005; Sunil 9. Ficus beddomei King. Ann. Roy. Bot. Gard. and Sivadasan, Fl. Alappuzha Dist. 665. 2009; (Calcutta) 1. 1888 26, t. 24; Hook. f., Fl. Brit. India.5: Sasidharan and Sivarajan, 2004. 502.1888; Sasidharan and Sivarajan.Fl. Pl. Thrissur For.428.1996; Mohanan and Sivadasan, Fl. Ficus caudata Stokes. Bot. Mat. Med. 4: 358. 1812; Agasthyamala.630. 2002; Sasidharan and Sivarajan., King,Ann. Roy. Bot. Gard. (Calcutta) 1: 87, 154, 162. Bio. Docu. Kerala part 6: Flow. Plant. 438. 2004. 1888. Ficus rama-varmae Bourd. J. Bombay Nat. Hist. Soc. 13: 155. 1994 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.235-242 © Principal, Sree Narayana College, Kollam, Kerala, India

A preliminary assessment on the diversity of genus Ficus l. (moraceae) in Kerala 237 Distribution: Endemic to the Western Ghats. Seen in Sivarajan., Fl. Pl. Thrissur For. 428. 1996; Mohanan altitude ranging from 200 – 1200 m. and Sivadasan, Fl. Agasthyamala. 631.2002; Sunil and Sivadasan, Fl. Alappuzha Dist. 661. 2009; Sasidharan 10. Ficus benghalensisL.sp. Pl. 1059. 1753; Gamble, Fl. and Sivarajan., Bio. Docu. Kerala part 6: 439. 2004. Pres. Madras. 1361. 1928; Manilal and Sivar., Fl. Calicut. 277. 1982; Mohanan and Sivadasan, Fl. Ficus mysorensis B. Heyne ex Roth.Nov. Pl. Sp. Agasthyamala. 630. 2002; Anilkumar et al., Fl. 390.1821; Gamble, Fl. Pres. Madras. 1361. 1928; Hook. Pathanamthitta.462. 2005; Sunil and Sivadasan, Fl. f., Fl. Brit. India.5: 500. 1888. Alappuzha Dist. 661. 2009; Sasidharan and Sivarajan., Bio. Docu. Kerala part 6: Flow. Plant. 438. 2004. Urostigma mysorense Miq. London. J. Bot. 6. 574. 1847. Ficus umbrosa Salisb, Prodr. Strip. Chap. Allerton 16.1796. Distribution: India, China, Sri Lanka, Myanmar. Found up to 1025 m altitude. Native to India. Urostigma benghalense(L.) Gasp, Nov. Gen. Fic. 7. 1844. 15. Ficus elastic Roxb. exHornem. Hort. Bot. Hafn. Suppl. 7.1818; Hook.of., Fl. Brit. India.5: 508.1888; Distribution: Find wild in the sub- Himalayan forest Gamble, Fl. Pres. Madras. 1369. 1928; Sasidharan and and on the lower slopes of Deccan Hills. Distributed Sivarajan., Bio. Docu. Kerala part 6: 439. 2004. from sea level to 700m altitude. Urostigma elasticum (Roxb. ExHornem.) Miq.London. 11. Ficus benjaminaL. Mant. Pl. 129. 1767; King,Ann. J. Bot. 6: 578. 1847; King, Ann. Roy. Bot. Gard. Roy. Bot. Gard. (Calcutta) 1:50. 1888; Hook.f., Fl. Brit. (Calcutta) 1:45. 1888. India.5: 508.1888; Gamble, Fl. Pres. Madras. 1367. 1928; Sasidharan and Sivarajan, 2004. Distribution: Common ornamental tree. Widely cultivated. Cultivated in India from 1874. Ficus comosaRoxb.Pl. Coromandel. 2(1): 14, t. 125. 1799. 16. Ficus krishnae C.DC. Bot. Mag. 132. t. 8092. 1906; Sudhakar et al., Figs of Eastern Ghats, India. 118. 2017. Ficus nuda (Miq.)Miq. Ann. Mus. Bot. Lugduno – Batavi. 3(9): 288. 1867. Ficus bengahlensis var. Krishnae DC., Bull. Herb.Boissier 760. 1902. Urostigma benjaminum (L.)Miq.London J. Bot. 6: 583. 1847. Distribution: Cultivated variety. Native to India. Urostigma nudumMiq.London J. Bot. 6(72): 584 (-585). 17. Ficus microcarpaL.f. Suppl. Pl. 442. 1782; Manilal 1847. and Sivar., Fl. Calicut 276. 1982; Sasidharan and Sivarajan., Fl. Pl. Thrissur For. 429. 1996; Mohanan Distribution: Usually cultivated. Distributed from 25 m and Sivadasan., Fl. Agasthyamala 634. 2002; Anil – 900m above sea level. Kumar et al., Fl. Pathanamthitta. 465. 2005; Sunil and Sivadasan, Fl. Alappuzha Dist. 664. 2009; Sasidharan 12. Ficus costata Aiton, Hort. Kew 3: 452. 1789; and Sivarajan., Bio. Docu. Kerala part 6: 440. 2004. Sasidh., Fl. Periyar Tiger Reserve 392. 1998; Sasidh., Fl. Parambikulam WLS 311. 2002; Sasidharan and Ficusretusa King, Ann. Roy. Bot. Gard. (Calcutta) 1: Augustine, Rheedea 9 (1): 79. 1999. 50. tt.61 and 84 P.1887, non L. 1767; Hook. f., Fl. Brit. India 5: 511. 1888; Gamble, Fl. Pres. Madras. 1362. F. caudiculata Trim., J. Bot. 23: 242. 1885 and Handb. 1928. Fl. Ceylon 4: 88. 1898. Urostigma microcarpum Miq. London. J. Bot. 6: 583. F. mooniana King, Ann. Roy. Bot. Gard. (Calcutta) 1: 1847. 48, t. 58A. 1887. Distribution: Throughout in the Western Ghats, China, Distribution: India (Andaman & Nicobar Islands, Sri Lanka. Distributed up to 1050 m from sea level. Kerala), Sri Lanka. 18. Ficus mollis Vahl.,Symb. Bot. 1: 82. 1790; Dass. 13. Ficus dalhousiae (Miq.) Miq.Ann. Mus. Bot. and Forsb., Rev. Handb. Fl. Ceylon 3: 249, f. 12. 1981.. Lugduno - Batavi.3: 285. 1867; Hook.f., Fl. Brit. India.5: 499.1888; Gamble, Fl. Pres. Madras. 1364. F. tomentosa Roxb.exWilld., Sp. Pl. 4 (2): 1136. 1806; 1928; Sasidharan and Sivarajan., Fl. Pl. Thrissur For. Hook. f., Fl. Brit. India 5: 501. 1888. 428. 1996; Sasidharan and Sivarajan., Bio. Docu. Kerala part 6: 439. 2004. Distribution: Central and Southern provinces India. Distribution: Endemic to the Western Ghats. 19. Ficus talbotii King. Ann. Roy. Bot. Gard. (Calcutta) Distributed up to 3000 m altitude. 1: 51. 1888; Hook, Fl. Brit. Ind. 5: 512. 1888; Gamble, Fl. Pres. Madras. 1363. 1928; Anilkumar et al., Fl. 14. Ficus drupacea Thunb. var. Pubescens(Roth) Pathanamthitta. 465. 2005. Corner, Gard. Bull. Singapore 17: 381. 1960; Manilal and Sivar., Fl. Calicut. 276. 1982; Sasidharan and Distribution: Peninsular India, Sri Lanka, Myanmar. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.235-242 © Principal, Sree Narayana College, Kollam, Kerala, India

238 Current status and challenges for conservation and sustainable use of biodiversity Subgenus:Pharmacosycea Distribution: India, China, Myanmar, Sri Lanka. Found up to 1200 m from sea level. 20. Ficus anamalayana Sudhakar and G. V. S. Murthy. Rheedea.25(1): 1- 8. 2 0 1 5; Sudhakar et al., Figs of Subgenus:Sycomorus,Section: Sycomorus, Subsection: Eastern Ghats, India. 28. 2017. Neomorphe Distribution: Endemic to the Southern Western Ghats. 25. Ficus auricularta Lour, Fl. Cochinch. 666.1790; King, Ann. Roy. Bot. Gard. (Calcutta) 1: 179. 1888; Subgenus:Pharmacosycea, Section: Oreosycea, Sasidh.&Sivar., Bio. Docu. Kerala part 6: Flow. Plant. Subsection: Grandulosae 438. 2004. 21. Ficus nervosa B.Heyne ex Roth.Nov. Pl. Sp. Covellia macrophylla Miq. London. J. Bot. 7:465, 1848. 388.1821; Hook. Fl. Brit. India. 5: 512.1888; Gamble, Fl. Pres. Madras. 1364. 1928; Sasidharan and Ficus regia Miq.Ann. Mus. Bot. Lugd-Bat. 3: 230.1867. Sivarajan., Fl. Pl. Thrissur For. 429.1996; Mohanan and Sivadasan, Fl. Agasthyamala.634.2002; Sasidh.&Sivar., F. roxburghiiWall.exSteud., Nom. Bot. ed. 2, 1: 637. Bio. Docu. Kerala part 6: 440. 2004. 1840. Urostigma modestum Miq. London J. Bot. 6: 586. 1847. Distribution: Commonly cultivated. Seen wild almost throughout the Eastern Ghats. Distribution: Peninsular India, Sri Lanka, China, Myanmar, to elevations up to 1200m. 26. Ficus guttata (Wight) Wightex King.Ann. Roy. Bot. Gard. (Calcutta) 1:167. 1888; Hook, f. Fl. Brit. Ind 5: Subgenus:Pharmacosycea, Section: Oreosycea, 534, 1888; Mohanan and Sivad., Fl. Agasthyamala 633. Subsection: Pedunculatae 2002 Sasidharan and Sivarajan., Bio. Docu. Kerala part 6: 439. 2004. 22. Ficus callosa Wild.Mem. Acad. Roy. Sci. Hist. (Berlin) 1798: 102 1798; Hook.f., Fl. Brit. India.5: Distribution: Endemic to Southern Western Ghats from 516.1888; Gamble, Fl. Pres. Madras. 1364. 1928; 900 – 2200 m altitude. Endangered. Anilkumar et al., Fl. Pathanamthitta. 463. 2005; Mohanan and Sivadasan, Fl. Agasthyamala. 631.2002; Subgenus:Sycomorus,Section: Sycomorus, Subsection: Sasidharan and Sivarajan., Bio. Docu. Kerala part 6: Sycomorus 438. 2004. 27. Ficus racemosaL. Sp. Pl. 1060.1753; King, Ann. Ficus cinerascens Thwaites.Enum.Pl. Zeyl. [Thwaites] Roy. Bot. Gard. (Calcutta) 1: 183. 1888; Sunil and 266.1861. Sivadasan, Fl. Alappuzha Dist. 664.2009; Mohanan&Sivadasan, Fl. Agasthyamala. 635.2002; Ficus scleroptera Miq.Pl. Jungh. 63. 1851. Sasidh. &Sivar., Bio. Docu. Kerala part 6: 441. 2004. Distribution: In the Western Ghats, from elevations of Covellia glomearta(Roxb.)Miq. London J. Bot. 7: 600 to 1200 m. 465.1847. Subgenus:Sycomorus Ficus glomerata Roxb.Pl. Coromandel.2(1).13, t. 123.1799; Hook.f., Fl. Brit. India.5: 535. 1888. 23. Ficus binnendijkii (Miq.) Miq. Ann. Mus. Bot. Lugduno – Batavi. 3: 288.1867; King,Ann. Roy. Bot. Distribution: Indo Malaysia, Sri Lanka, Myanmar in Gard. (Calcutta) 1:41. 1888; Sudhakar et al., Figs of elevations up to 1200m. Eastern Ghats, India.106. 2017. Subgenus: Sycidium,Section: Sycidium Distribution: usually cultivated. Find wild in the forests of Sumatra, Java, Borneo etc. 28. Ficus exasperate Vahl. Enum. Pl. 2: 197. 1805; Manilal and Sivar., Fl. Calicut 275. 1982; Manilal, Fl. Subgenus:Sycomorus,Section: Sycocarpus, Subsection: Silent Valley 259.1988; Sasidharan and Sivarajan., Fl. Sycocarpus Pl. Thrissur For. 429. 1996; Mohanan and Sivad., Fl. Agasthyamala 632. 2002; Anil Kumar et al., Fl. 24. Ficus hispidaL.f. Suppl. Pl. 442.1782; Hook. Fl. Pathanamthitta 464. 2005; Sunil and Sivadasan, Fl. Brit. India. 5: 522.1888; Fischer in Gamble, Fl. Pres. Alappuzha Dist. 662. 2009. Madras.1367.1928; Sasidh.&Sivar., Fl. Pl. Thrissur For. 429.1996; Anilkumar et al., Fl. Pathanamthitta. 464. Ficus asperrima Roxb., Fl. Ind. 3: 554. 1832; Hooker. 2005; Sunil &Sivadasan, Fl. Alappuzha Dist. 663. 2009; London. J. Bot. 7: 230. 1848; Hook. f., Fl. Brit. India 5: Mohanan&Sivadasan, Fl. Agasthyamala. 629. 2002; 522. 1888; Gamble, Fl. Pres. Madras 1366(955). 1928. Sasidh. &Sivar., Bio. Docu. Kerala part 6: 440. 2004. Ficus hispidissima Wight ex Miq. London. J. Bot. 7: CovelliaassamicaMiq. London. J. Bot. 7: 464. 1848. 229. 1848. Covelliahispida(L.f.) Miq. London. J. Bot. 7: 462. 1848. Distribution: Peninsular India, Sri Lanka, East Africa. Found from sea level to 1500 m. FicusoppositifoliaWild.Sp. Pl., ed. 4 [Wildenow], 4 (2): 1151.1806. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.235-242 © Principal, Sree Narayana College, Kollam, Kerala, India

A preliminary assessment on the diversity of genus Ficus l. (moraceae) in Kerala 239 29. Ficus heterophylla L.f. Suppl. Pl. 442. 1782; Distribution: Cultivated throughout the world. King,Ann. Roy. Bot. Gard. (Calcutta) 1:75. 1888; Hook. Fl. Brit. India. 5:518.1888; Gamble, Fl. Pres. Subgenus: Synoecia, Section: Rhizocladus, Subsection: Madras.1366.1928; Anilkumar et al., Fl. Pogonotrophe Pathanamthitta. 464. 2005; Sunil and Sivadasan, Fl. Alappuzha Dist. 663. 2009. 33. Ficus amplocarpa Govind. and Masil., Proc. Indian Acad. Sci. (Pl. Sci.) 91: 117. 1982; Chithra in A.N. Ficus aquatica K.D. Koenig ex Wild. Sp. Pl., ed. 4 Henry et al., Fl. Tamil Nadu 2: 251. 1987; T.S. Nayar et [Wildenow], 4 (2): 1133.1806. al., Fl. Pl. Kerala: 433. 2006. Ficusbiglandula Blume, Bijdr. Fl. Ned. Ind. 9: F. macrocarpa (Miq.) King, Ann. Roy. Bot. Gard. 475.1825. Calcutta 1: 166, t. 208. 1888 and in Hook.f., Fl. Brit. India 5: 534. 1888, non Blume (1823); C.E.C. Fisch.in Ficus elongataMiq. London. J. Bot. 7: 231.1848. Gamble, Fl. Madras: 1365. 1928; Vajr., Fl. Palghat: 447. 1990. Distribution: Peninsular India, Sri Lanka, Myanmar, China. Distribution: Endemic to Southern Western Ghats. Evaluated as Near Threatened according to IUCN Red Subgenus:Sycidium,Section: Palaeomorphe List. 30. Ficus tinctoriaG. Frost.ssp. gibbosa(Blume) Corner Acknowledgements var.cuspiderifera(Miq.) Chithrain Henry et al., Fl. Tamil Nadu 2: 255. 1987; Sivar., Bio. Docu. Kerala Authors are thankful to Mr.Ebin P. J. and Dr.Lesly part 6: 441. 2004. Augustine, Assistant Professors, Department of Botany, Sacred Herat College, Thevara for the support. We are FicuscuspidiferaMiq. Hook, London. J. Bot. 7: 434. grateful to the curators of CALI, KFRI, TBGT and MH 1848. for the help in completion of the work. Ficus gibbosa Blume.King. Ann. Roy. Bot. Gard. Reference (Calcutta) 1: 4. 1888. Amitha Bachan, K. H. and Jisy, E. D. 2016. A Ficus gibbosa Blumevar. cuspiderifera(Miq.)King. Preliminary Assessment of The Diversity of The Genus Ann. Roy. Bot. Gard. (Calcutta) 1: 6, t.2A. 1887; 'Ficus' In the Vazhachal Forests, Western Ghats, India. Gamble, Fl. Pres. Madras. 1366.1928. Meridian, 5 (1): 45- 54. Distribution: Throughout India. Sri Lanka, China, Berg, C. C. and Corner, E. J. H. 2005. Moraceae - Myanmar. Ficus. 17: 1-730. Flora Malesiana Series I (Seed Plants), 17: 1–730. 31. Ficus tinctoria G. Frost.ssp. parasitica (Koen. Ex Wild.)Corner, Gard.Bulll. Singapore 17: 476. 1960; Bowles, J. M. 2004. Guide to Plant Collection and Manilal and Sivar., Fl. Calicut 276. 1982; Sasidharan Identification. Plant Evolutionary Ecology. and Sivar., Fl. Pl. Thrissur For. 430. 1996; Mohanan and Sivadasan., Fl. Agasthyamala 636. 2002; Anil Chaudhary, L. B., Sudhakar, J. V., Kumar, A., Bajpai, Kumaret al., Fl. Pathanamthitta 466.2005; Sunil and O. and Tiwari, R. 2012. Synopsis of The Genus Ficus L. Sivadasan, Fl. Alappuzha Dist. 666. 2009;Sivar., Bio. (Moraceae) In India. Taiwania., 57(2): 193–216. Docu. Kerala part 6: 441. 2004. Corner, E. J. H. 1965. Check list of Ficus in Asia and Ficus parasitica Koenig ex Willd.,Mem. Acad. Roy. Australasia With Keys To Identification. Gardens Sci. Hist. (Berlin) 2: 102. 1798. Bulletin, Singapore.21: 1–186. Ficus gibbosa Blume var. parasitica (Koenig ex Willd.) Corner, E.J.H. 1965. Ficus. Philosophical Transactions King, Ann. Roy. Bot. Gard. (Calcutta) 1: t.2,16.1887. of the Royal Society of London. 255(800): 567–570. Distribution: Throughout India. Sri Lanka, China, Gamble, J. S.; Dunn S. T. and Fischer, C. E. C. 1967. Myanmar. Flora of the Presidency of Madras. Adlard and Son Ltd., London. Subgenus: Synoecia, Section: Rhizocladus, Subsection: Plagiostigma Http:// Apps.Kew.Org/Herbcat/Navigator.Do 32. Ficus pumilaL. Sp. Pl. 1060.1753; King, Ann. Roy. Http://Ipni.Org. Bot. Gard. (Calcutta) 1: 124. 1888; Lour, Fl. Cochinch.2: 667. 1790; Sp. Pl., ed. 4 (Wildenow), 4 (2): Http://Www. Kfriherbarium.In/ 1140.1806; Linn, Sp. Pl. 2: 1060. 1753. Http://Www.Theplantlist.Org Ficus stipulate Thunb. Dissert. Ficus. 8. 1786. Http://Www.Keralaplants.In. Ficus repens Rottler, Ges. Naturf. Freunde Berlin Neue Schriften 4: 208. 1803. Kumar, A., Bajpai, O., Mishra, A. K., Sahu, N., Behera, S. K. and Chaudhary, L. B. 2011. Assessment of Diversity In The Genus Ficus L. (Moraceae) of Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.235-242 © Principal, Sree Narayana College, Kollam, Kerala, India

240 Current status and challenges for conservation and sustainable use of biodiversity Katerniaghat Wildlife Sanctuary, Uttar Pradesh, India. American Journal of Plant Sciences., 2: 78-92. Lambert, F. R. and Marshall, A. G. 1991. Keystone Characteristics of Bird-Dispersed Ficus in A Malaysian Lowland Rain Forest. The Journal of Ecology., 79: 793- 809. Lansky, E. P. and Paavilainen, H. M. 2011. Figs The Genus Ficus. Taylor and Francis Group, Llc. Lomáscolo, S. B.; Levey, D. J.; Kimball, R. T.; Bolker, B. M. and Alborn, H. T. 2010. Dispersers Shape Fruit Diversity in Ficus (Moraceae). Proceedings of The National Academy of Sciences., 107: 14668-14672. Loutfy, M. H. A.; Karakish, E. A.; K., Khalifa, S. F. and Mira, E. R. A. 2005. Numerical Taxonomic Evaluation of Leaf Architecture of Some Species of Genus Ficus L. International Journal of Agriculture and Biology., 7: 352-357. Priyadarsanan, D.R. 2000. Fig Insects of Kerala, Rec. Zool. Survev. India, Occ. Paper No. 182: I-Iv, 1-175 (Published: Director, ZSI, Calcutta). Sasidharan, N. and Sivarajan, V. V. 2004. Biodiversity Documentation of Kerala. Part 6.Kfri Hand Book. Sudhakar, J. V. and Murthy, G. V. S. 2015. Ficus Anamalayana ( Moraceae ): A New Species From South India. Rheedea.,25(1): 1–8. Sudhakar, J.V., N. C. M. Reddy. and Murthy,G. V. S. 2017. Figs of Eastern Ghats, India. Nbri and Pragati Printers, Hyderabad. Vanitharani, J.; Bharathi, B. K.; Margaret, I. V.; Malleshappa, H.; Ojha, R. K. and Naik, K. G. A. 2009. Ficus Diversity In Southern Western Ghats: A Boon For Biodiversity Conservation. Journal of Theoretical Experimental Biology., 6: 69-79. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.235-242 © Principal, Sree Narayana College, Kollam, Kerala, India

Short-term temporal variation in coastal phytoplankton of Saurashtra coast: 243 ISBN 978-93-5396-871-7 CHAPTER 38 SHORT-TERM TEMPORAL VARIATION IN COASTAL PHYTOPLANKTON OF SAURASHTRA COAST: INFLUENCE OF DISSOLVED NUTRIENTS Ambili Nath* and Suresh Balakrishnan** *Department of Zoology, Pazhassi Raja N.S.S College, Mattanur, Kannur, Kerala **Department of Zoology, Faculty of Science The Maharaja Sayajirao University of Baroda, Vadodara – 390 002L; *Correspondence E-mail: [email protected] ABSTRACT Surface samples of phytoplankton were collected by filtering 100L of water through a bag net of pore size 70µm. The samples representing winter, premonsoon, monsoon and postmonsoon seasons were collected, from three study stations Diu, Veraval and Alang situated along the coasts of Saurashtra. Short term changes in the community structure (in terms of species composition and abundance) were investigated with respect to seasonal variations in the dissolved nutrient concentration. In order to comprehend the influence of dissolved nutrients on phytoplankton communities that bring in temporal and spatial variation and to further understand the significant seasonal difference in plankton diversity, one way ANOVA without replication and one way ANOSIM were performed. The oxidized nitrogen and phosphate phosphorous seems to have a substantial influence on the overall diversity. Silicates appear to be a key nutrient influencing the relative abundance of diatoms at Diu, Veraval and Alang. ANOSIM revealed a significant similarity between the three stations, with pair wise comparison of the nutrients and plankton diversity. The R static (relative measure of the degree of separation of samples) revealed that total oxidized nitrogen and phosphate were more involved in bringing temporal distribution among the species. Furthermore, the assessment based on the multivariate non-metric multidimensional (NMDS) ordinations, indicated maximum similarity accounting high temporal variability during winter and pre monsoon in Diu and Veraval respectively. However, Alang showed proximity in phytoplankton community during monsoon and winter thus retaining a temporal pattern of assemblages. Key words: Temporal variations, Phytoplankton, Dissolved nutrients, Statistical analysis Introduction community. However, this fluctuates with the variant environmental conditions, thus Temporal variability is an essential function exhibiting some common pattern of of significance in the aquatic ecosystem. distribution. Generally, the dynamics of the Temporal variation is one of the important temporal variations mainly depend upon processes characterized by the phytoplankton Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.243-250 © Principal, Sree Narayana College, Kollam, Kerala, India

244 Current status and challenges for conservation and sustainable use of biodiversity frequent reorganization of relative abundance studied for several coastal and oceanic water and plankton species composition, as a result bodies’ world over (Eskinazi et al., 1997; of interaction between physical, chemical and Webber, 1998; Koening et al., 2003; Linton biological variations. These environmental and Warnce, 2003), in India, Gujarat state variables fluctuate with time, as with all which has got the longest coastal area, this planktonic organisms inhabiting different aspect of plankton biology has been largely niche in marine ecosystem leading to decline neglected. Hence, the present study was in the population count (Kimmer and Orsi, aimed at evaluating the effectiveness of 1996; Kimmerer, 2002). Further, these measuring the association of dissolved variations are considered as an essential nutrients and phytoplankton communities prerequisite for assessing alterations caused during different seasons, between stations viz. by perturbations in the biome (Underwood, Diu, Veraval and Alang thereby studying the 1992). Amongst the various environmental different level of temporal variations among variables in marine environment, dissolved the phytoplankton communities. nutrients( NO3  N , NO2  N , PO34  P Materials and Methods and SiO 4  Si ) are considered to contribute The samplings for physicochemical and to a significant extent in brining spatio- biological parameters were done from the temporal variations in the plankton surface zone of the lower littoral or the low community. This documented nutrient level, tide region of the intertidal area of Diu in addition to their role in dictating (Position: N 20° 42' 260 E 70° 58' 683 ), competition, can be a major factor in defining Veraval, (Position: N 21° 52' 945 E 70° 21' the community structure. Therefore, the 231 ) and Alang-Sosiyo for a period of one evaluation and systemization of year from 2016-2017, covering all the major phytoplankton community structural analyses seasons’ viz. pre monsoon, monsoon, post and its associated spatio-temporal variations monsoon and winter. All the have been supported extensively by statistical physicochemical analysis of the water analytical procedures. These analytical samples were estimated as per the treatise, procedures included One Way ANOVA, One ‘Standard Methods for the examination of Way ANOSIM and Multivariate non-metric water and wastewater’, prepared and multidimensional ordinations (NMDS). published jointly by the American Public Consequently, these analytical procedures for Health Association (APHA), American the study of phytoplankton community Water Works Association (AWWA) and structure take into account both the Water Environment Federation (WEF). The significance of the information of seasonal plankton data were quantitatively analyzed variations in the nutrient concentration and using standard analytical and statistical its associated spatio-temporal variations in methods with computer software packages the phytoplankton community. viz. programme PRIMER version 5.2.4, Origin ver.7 (Clark and Warwick, 1994) Though, spatial and temporal variation in plankton community structure has been Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.243-250 © Principal, Sree Narayana College, Kollam, Kerala, India

Short-term temporal variation in coastal phytoplankton of Saurashtra coast: 245 Map of Gujarat – Depicting the Saurashtra Region & the Sampling Sites therein Saurashtra Region Sampling Sites Alang Veraval Diu Arabian Sea N Results increase was evident during winter Seasonal concentration of dissolved NO2  N mg/L (0.232 ± 0.02) and nutrients NO3  N mg/L (2.135 ± 0.05). The The hydrographic conditions revealed a phosphate phosphorous concentration in Diu seasonal fluctuation in the concentration of reached a maximum 1.2mg/L in post dissolved nutrient in the coastal waters of monsoon. Veraval and Alang showed an Diu, Veraval and Alang. The total oxidized increasing trend for the same in monsoon nitrogen seems to be influenced by the with values 1.08mg/L and 0.99mg/L seasons, significant variations in values were respectively. The concentration of silicates observed during the three successive season’s exhibited maximum values after showers in viz.winter, pre monsoon, monsoon and post Diu (8.79 ± 0.14) and Alang (9.89 ± 0.09). monsoon. The total oxidized nitrogen, nitrate to nitrite proportion showed a hike in Temporal variations in phytoplankton concentration in monsoon season, at Veraval abundance and diversity NO2  N mg/L (1.49 ± 0.00) and Thirty species of phytoplankton belonging to NO3  N mg/L (0.149 ± 0.00). Whereas in 3 divisions, 3 classes, 6 orders and 17 families were recorded during the study Alang total oxidized nitrogen concentration period. Chrysophyta were the most dominant Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.243-250 © Principal, Sree Narayana College, Kollam, Kerala, India

246 Current status and challenges for conservation and sustainable use of biodiversity group (23 species), followed by, Cyanophyta during winter. The maximum abundance of (3 species) and Phyrophyta (4 species). The phytoplankton species was observed during Coscinodiscus was the most abundant species the monsoon. Centric diatoms, Coscinodiscus and was found persistent throughout the janesianus and Coscinodiscus sp. both were seasons. equally dominant. Thalassiosira and Melosira also showed dominance with 19.16 The phytoplankton density increased and 11.79%. However, Thalassionema gradually from 8440 cells to 59040 cells/L showed the maximum percentage of with an average of 20705 cells/L and 41423 abundance during post monsoon. cells/L in Alang and Diu respectively. The phytoplankton abundance showed temporal Phytoplankton community similarity trends at all three stations during different indices seasons. The overall abundance in all the three stations was primarily due to The application of the similarity indices to Crysophyta. The phytoplankton community data matrix at all the three study station was in Diu area is dominated by Thalassiosira performed. This in fact was meant to sp., Coscinodiscus sp. and Coscinodiscus comprehend, the influence of dissolved janesianus during winter. Navicula and nutrients on phytoplankton community Pleurosigma showed equal distribution, structure and also to understand whether this while centric diatom, Biddulfia sp. showed relationship is getting altered through time abundance in summer months. However, the and space. This representation involves three monsoon seemed to be very assuring with indices: One Way ANOVA without maximum species abundance and diversity. replication with nutrients and phytoplankton The observed high density of phytoplankton as selected factors. in monsoon could be a result of increased nutrient content in the seawater. Pleurosigma Secondly, ANOSIM, to test the significant numerically dominated the samples on all seasonal difference in plankton diversity occasions in Veraval. Pre monsoon though between the three stations, in relation to that indicated a low abundance of the of the nutrients. The third is non-metric phytoplankton population their diversity multidimensional scaling (NMDS) however, reached its maximum value of 2.4 ordinations using non-transformation to in this season. High values of diversity could assess the temporal variations among the be a result of low nutrient availability and plankton communities. In the present study, grazing pressure (Fredy and Ferdinand, analysis of variance explained, that the 2006). Among the seasonal variation seasonal variations between the stations Thalassionema (22.3%), Thalassiosira (space) was dependent on its hydrological (20.44%), Fragilaria sp. (11.15%), parameters, showing significant interaction Fragilaria hyaline (9.44%) and between the nutrients and phytoplankton Coscinodiscus gigas with 16.72% of total species. The Tukeys test revealed significant abundance prevailed during post monsoon .In effect on the phytoplankton diversity with all, 19 diatoms genera were identified at respect to variations in the nutrient contents Alang. Amongst all Treubaria exhibited the like total oxidized nitrogen, nitrate to nitrite maximum abundance followed by proportion and phosphate phosphorous in Diu Coscinodiscus janesianus and Triceratium and Veraval. Moreover, in Alang silicate exhibited significant effect on the community Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.243-250 © Principal, Sree Narayana College, Kollam, Kerala, India

Short-term temporal variation in coastal phytoplankton of Saurashtra coast: 247 diversity and it seemed likely that the other 0.25 at Diu to large value of 1 at Alang, and nutrients were insignificant in the temporal the global R value indicated a distinction distribution of phytoplankton communities at between the three stations i.e. Veraval Alang. Such relationships well documented (0.222) and Alang (0.333), while Diu with R in the literature and is related to the extreme values of 0. restrictive environmental conditions associated with the eutrophication process ANOSIM randomization test to confirm Moreover; in Alang silicate exhibited statistically significant differences between significant effect on the community diversity the hydrochemical parameters and and it seemed likely that the other nutrients phytoplankton community in each station at were insignificant in the temporal distribution Saurashtra Table 1, 2, 3 and 4. of phytoplankton communities at Alang. Table 1. DIU Further, analysis of similarities (ANOSIM) SAMPLE FACTORS was used as a conservative test to determine any significant difference between the Total oxidized nitrogen C nutrient concentration and community structure (Table 1). However, the results of PO4-p mg/L and SiO3-Si µg/L L the R statistics (relative measure of the degree of separation of samples) were Phytoplankton Diversity H different (Clarke and Warwick, 1994). In pair wise comparison phosphate and total *C, L & H are factors for identification. oxidized nitrogen seemed to be more involving in bringing temporal distribution Global Test among the species. The values of the R statistics in the global test for difference Number of permutations: 15 (All possible between the groups increased strongly from permutations) Table 2. DIU. Sample statistic (Global R): 0. Significance level of sample statistic: 53.3% Groups R Statistic Significance Possible Actual Number level % observed C,L permutations permutations C,H 3 L,H -0.25 100 3 3 2 2 0.25 66.7 3 3 0 66.7 3 3 Table 3. VERAVALSample statistic (Global R): 0.222 Significance level of sample statistic: 53.3% Groups R Significance Possible Actual Number observed C,L Statistic level % permutations permutations C,H 3 L,H -0.25 100 3 3 1 2 0.75 33.3 3 3 0 66.7 3 3

248 Current status and challenges for conservation and sustainable use of biodiversity Table 4. ALANG Sample statistic (Global R): 0.333 Significance level of sample statistic: 33.3% Groups R Statistic Significance Possible Actual Number level % permutations permutations observed C,L C,H 0 100 3 3 3 L,H 1 1 33.3 3 3 2 0 66.7 3 3 The pair wise R values give absolute measure of how separated the groups are R > 0.75: groups being well separated R > 0.5: groups overlapping but clearly different R > 0.25: groups barely separable at all. Discussion that such rises in the phytoplankton might Hydrodynamic characteristics of the marine occur without reducing the phosphate content environment are the most relevant factor, of the water and that on occasions both which regulates the structure of phosphate and plankton might rise together. phytoplankton communities (Legendre and Nutrients such as nitrates and phosphates are Rassoulzadegan, 1995). The importance of comparatively high creating a discordant note these factors in controlling phytoplankton in the normal planktonic rhythm and what distribution is recognized. In the present combination of environmental parameters study phytoplankton showed temporal triggers this abnormal bloom is a matter of variation in community structure in further research. Besides, the statistical accordance with the changes in the hydro- analyses also reveal notable difference chemical (dissolved nutrients) concentration between the species diversity and nutrient in the seawater. Seasonal difference in the concentration along the three stations of the plankton diversity occurred in all the three Saurashtra coast. But the aggregation of all stations of the Saurashtra coast. Many factors produced apparent changes in the opined that available food production; community structure, thus, brining temporal grazing, competition and predation are the amendments within the community. This has essential factors that regulate plankton been to some extent depicted by ANOSIM. compositions (Wooldridge and Melville Temporal and spatial pattern are often Smith, 1979, Koening et.al., 2003). Evidence interdependent. The phytoplankton species from the analysis of variance (ANOVA) compositions relates not only to local revealed that total oxidized nitrogen and hydrodynamic features but also to biological phosphate phosphorus were significantly processes, inducing primary productivity and influencing the phytoplankton diversity in the migratory behaviors of organisms. Hence, two stations viz. Diu and Veraval. The it could be concluded that plankton phytoplankton also exhibited an increase in communities exhibit temporal variation with abundance in accordance with that of total respect to the alterations in the oxidized nitrogen and phosphate phosphorus. physiochemical characteristic of marine Similarly, elevated concentrations of nitrates waters. The temporal distributions of the and phosphates favor the development of plankton species were largely influenced by diatoms. Kimmerer and Orsi, (1999) noted the changes in the dissolved nutrients level in Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.243-250 © Principal, Sree Narayana College, Kollam, Kerala, India

Short-term temporal variation in coastal phytoplankton of Saurashtra coast: 249 Coastal waters. However, nutrients solely Francisco Bay Estuary since 1987. In: J. I. cannot be considered as a complete Hollibaugh JI (Ed) San Francisco Bay: the contributing factor for the determination of ecosystem. 403--414. The American temporal variations in a community. Besides, Association for the Advancement of Science. from the statistical analysis it is clear that no San Francisco. factor acts in isolation and any variations in the biological and physical variables, such as Kimmerer, W.J. 2002. Effects of freshwater food quality, turbidity and salinity may flow on abundance of estuarine organisms: influence the plankton community as a Physical effects or trophic linkages? Marine whole, contributing to the spatial and Ecology Progress Series, 243: 39-55. temporal variations among the community. Knatz, G. 1978. Succession of Copepod Acknowledgment species in a Middle Atlantic estuary. Estuaries, 1: 61-68. The authors are highly appreciative of, the Department of Zoology, Faculty of Science, Koening, L.M, Eskinazi, L.E., Neumann- The M.S. University of Baroda, for support. Leitão, S. and Maced J.S. 2003. Impacts of We are thankful to Gujarat Ecological the construction of the Port of Suape on Society, Baroda for their assistance in data phytoplankton in the Ipojuca River estuary collection and to National Institute of (Pernambuco- Brazil). Brazilian Archives of Oceanography, Dona Poula, and Goa for Biology and Technology, 46(1): 73-81 their timely guidance. Legendre, L. and Rassoulzadegan, F. 1995. References Plankton and nutrient dynamics in marine waters. Ophelia, 41: 153-172. APHA, AWWA, WEF. 1998. Standard method for the examination of water and Linton, D.M. and Warner, G.F. 2003. wastewater (American Public Health Biological indicators in the Caribean coastal Association, Washington DC, 1998) zone and their role in integrated coastal management. Ocean and Coastal Clarke, K. R. and Warwick, R. M. 1994. Management, 46: 261-276 Similarity–based testing for community pattern: Two ways lay out with no Orsi, J..J. and Oh Tsuska, S. 1999. replication. Marine Biology, 118: 167–176. Introduction of Asian copepods. Acratiella sinensis, Tortanus dex trilobatus ( Copepod: Desikachery, T.V. 1959. ICAR monographs Calanoida) and Limnoithona tetraspina on algae ( Cyanophyta 1959), 2: 686. (Copepoda: Cyclopoida) to the San Francisco Estuary, California, USA. Plankton Biology Eskinazi, L. E., Silva-Cunha, M.G.G., and Ecology, 46: 128-131. Koening, M.L, Macedo, S. J. and Costa, K.M.P. 1997. Variacao especial e temporal Santhanam, R. and Srinivasan, A. 1994. A do fitoplancton no plataforma continental de manual of marine zooplankton (Oxford and perambuca- Brazil. Trabalhos do Instituto de IBH Publishing Co Pvt Ltd, New Delhi, Ocenografa da Universidade Federal de India, Pernambuco., 25:1-16. (Spanish) Underwood, A.J. 1992. Competition in Kimmerer, W. and Orsi, J..J. 1996. Causes of marine plant- animal interactions. In: Plant- long term decline in zooplankton in the San Animal Interactions in the Marine Benthos.

250 Current status and challenges for conservation and sustainable use of biodiversity Edited by John DM, Hawkins SJ and Price JH. Oxford University Press, 443-475 Wabber, D.F. and Webber, M.K.. 1998. The water quality of Kingston Harbour: evaluating the use of the Planktonoic community and traditional water quality indices, Chemistry and Ecology, 14: 357-374. Wooldridge, T. and Melville Smith, R. 1979. Copepod succession in two African estuaries. Journal of Plankton Research, 1: 329-341 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.243-250 © Principal, Sree Narayana College, Kollam, Kerala, India

251 Current status and challenges for conservation and sustainable use of biodiversity ISBN 978-93-5396-871-7 CHAPTER 39 ANALYSIS OF MORPHOLOGICAL VARIABILITY IN TWO DIFFERENT VARIETIES OF CARICA PAPAYA (L.) Remya R and Nisha A P * Post graduate and Research Department of Botany Sree Narayana College, Kollam, Kerala. *Correspondence E- mail: [email protected] ABSTRACT Carica papaya (L.) is widely grown throughout the tropics. It consists of different varieties. The objective of the present study is to analyze the morphological variability among two varieties (C. papaya var. yellow and C. papaya var. red lady) of papaya cultivated in Kerala. Four accessions from each variety were collected from different localities of Thiruvanathapuram and Kollam districts. Observations on twenty four qualitative and ten quantitative characters were taken from all accessions. Quantitative characters were subjected to one way ANOVA. Qualitative and quantitative characters showed significant variations among the two varieties except fruit length, fruit width and petiole length. In principal component analysis, the first principal component accounted for 68.88% variation followed by 11.73% and 9.92%. The highest loaded variables in PC1 are colour of petiole, colour of fruit, colour of sepal, colour of petal, colour of fruit in young condition and odor of leaf. The highly loaded characters in PC2 and PC3 are fruit length and fruit width respectievely. Major trait that accounted for more variability in PC1, PC2 and PC3 is the color of petiole. UPGAMA cluster analysis showed two principal clusters. The accessions of two varieties form separate clusters due to dissimilarity in most of the characters. It shows that the two varieties of Carica papaya have distinct morphological characters. These characters will help in the proper identification of these two varieties for large scale cultivation and in crop improvement programmes. Key words: Carica papaya, Caricaceae, ANOVA, Principal Component Analysis, UPGMA Introduction confined to the top of the trunk. The lower trunk is conspicuously scarred where leaves Carica papaya (L.), belongs to the family and fruits are borne. All parts of the plant Caricaceae, is widely grown throughout the contain latex in articulated laticifers. Carica tropics. It is a native of Central and South papaya consists of different varieties. America. It is appreciated not only for its Morphological characters are powerful tool delicious and nutritive fruits, but also it for the identification and characterization of contains the enzyme papain, which is plant species and varieties. There are several extensively used in medicines, as meat reports about the characterization of tenderizer, for softening textiles, silk and germplasm using morphological markers. leather and in beer production. The papaya Suvalaxmi et.al (2019), assessed the genetic plant is a small, sparsely branched tree, diversity on the basis of morphological and usually with a single stem growing from 5 to molecular characterization among 12 popular 10m tall, with spirally arranged leaves Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.251-256 © Principal, Sree Narayana College, Kollam, Kerala, India

252 Current status and challenges for conservation and sustainable use of biodiversity papaya varieties of India. The objective of accessions. Vegetative, floral and fruit the present study is to analyze the characters were studied in all the 8 accessions morphological variability among two collected. In all accessions, ten observations varieties (C. papaya var. yellow and C. were taken for each qualitative and papaya var. red lady) of papaya cultivated in quantitative character. The quantitative and Kerala. qualitative characters selected for the study were recorded in Table 2 and Table 3 Materials and Methods respectively. The measurements were taken using standard rulers. The plants selected for the present study are Carica papaya var. yellow and Carica For morphometric analysis, the quantitative papaya var. red lady. It belongs to the genus data were subjected to one way ANOVA Carica in the family Caricaceae. Four using SPSS Version 16. Multivariate accessions from each species were collected analyses were performed using the procedure from different localities of of principal component analysis (PCA). Data Thiruvananthapuram and Kollam districts. was subjected to cluster analysis based on Specific codes were allotted to each UPGMA method to find out the similarity accession. The details of collection of and dissimilarity among the 8 accessions. different accessions were represented in Table 1. Observations on 24 qualitative and 10 quantitative characters were scored in all Table 1. Accessions collected from different localities Sl. No Accession code Place of collection Genetic group 1 CPOV Varkala Carica papaya var.yellow.. 2 CPOC Chirayinkizhu Carica papaya var.yellow.. 3 CPOK Kallambalam Carica papaya var.yellow.. 4 CPOKo Kollam Carica papaya var.yellow.. 5 CPRV Varkala Carica papaya var. red lady 6 CPRC Chirayinkizhu Carica papaya var. red lady 7 CPRK Kottarakkara Carica papaya var. red lady 8 CPRKo Kollam Carica papaya var. red lady Results and Discussion lady. The observations on different quantitative characters are represented in Morphological analysis Table 2. The qualitative and quantitative vegetative, Vegetative morphology floral and fruit morphological characters were studied in different accessions of C. In the present study the qualitative vegetative papaya var. yellow and C. papaya var. Red characters of C. papaya var. yellow and C. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.251-256 © Principal, Sree Narayana College, Kollam, Kerala, India

Analysis of morphological variability in two different varieties of Carica papaya (L.) 253 papaya var. red lady showed some variations accession CPOV (0.33cm) and shortest sepal and more similarity. C. papaya var. yellow length was found in accession CPOC and C. papaya var. red lady are trees, leaf (0.195cm). In variety red lady, the accession type is simple, phyllotaxy is alternate and CPRV showed the longest sepal length venation is reticulate. The leaf shape is (0.31cm) and shortest sepal length was found similar in all accessions. Leaf shape, leaf in accessions CPRC and CPRK (0.21cm). In base, leaf apex and leaf surface are star the case of variety yellow, the accession shaped, acute, parted and smooth CPOV possesses highest petal length and respectively. In C.papaya var. yellow. the shortest petal length in accession CPRKo. In petiole color is green and in C. papaya var. variety red lady, the accession CPRK possess Red lady is brown. Leaves possess medium highest petal length and CPRV showed aroma in variety red lady and mild aroma in shortest petal length. variety yellow. Fruit morphology Quantitative characters showed variations. From the quantitative characters highest leaf The fruit is a berry. The unripe fruit color in length in C. papaya var. yellow was found in yellow variety is dark green and in variety accession CPOV(130cm) and shortest leaf red lady, it is green. The ripened fruit color length was found in CPOC (112cm). In Red in C. papaya var. yellow is orange and in C. lady highest leaf length was found in CPRV papaya var. red lady, it is dark red in some (98.75cm), shortest leaf length was found in accessions and light red in certain accessions CPRC (95.75cm). In yellow variety highest . The shape of the fruit is pyriform in variety petiole length was found in CPOV (86.25cm) yellow and elongated in variety red lady. and shortest petiole length was found in Shape of the seed and color are same in all CPOK (77.25cm). In the case of variety red accessions. The fruits of yellow variety lady highest petiole length was found in possess mild aroma and variety red lady CPRV (67.5cm) and shortest petiole length possess medium aroma. was observed in CPRK and CPRC(65cm). So the leaf length and petiole length were The quantitative characters also showed highest in yellow variety when compared to variations in two species. In C. papaya var. the variety red lady. yellow, longest fruit length was found in accession CPOKo (19.25cm) and shortest Floral morphology fruit length was found in accessions CPOK (17.75cm). In the case of C. papaya var. red Floral morphology shows minute variations lady, the highest fruit length was found in among the two varieties. Inflorescence is CPRC (18.35cm) and shortest fruit length solitary type, aestivation of sepal is valvate was found in CPRK and CPRKo (16.27cm). and petals are twisted. The flower colour in In variety red lady, highest fruit diameter was C. papaya var. yellow is light yellow and C. observed in CPRV (9.27cm) and lowest papaya var. red lady is cream . In C. papaya diameter was found in CPRKo (7.65cm). In var. yellow sepal colour is light green and in yellow variety, highest fruit diameter was C. papaya var. red lady, it is light yellow. found in accession CPOC and lowest Petal colour in variety red lady is cream and diameter in CPOV. In yellow variety, the light yellow in variety yellow. highest fresh fruit weight was found in accessions CPOV and lowest in accession Quantitative floral morphological characters CPOKo. In the case of variety Red lady also showed significant variations. In yellow highest and lowest fruit weight are found in variety, longest sepals were found in accessions CPRK and CPRKo respectively. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.251-256 © Principal, Sree Narayana College, Kollam, Kerala, India

254 Current status and challenges for conservation and sustainable use of biodiversity In C. papaya var. yellow highest numbers of characters in diversity analysis, cultivar seeds are present in CPOV and lowest in identification etc. Ruquale et al. (2016) CPOK. In the case of Red lady highest reported the morphological analysis in Olive numbers of seeds were observed in CPRC cultivar. Nishimwe (2019) reported the and lowest in CPRK.There are several reports morphological analysis of four varieties of about the significance of morphological papaya cultivar. Table 2. Quantitative morphological characters observed in C.papaya var. yellow and C. papaya var. red lady Name of LL PL SL PTL NOS FL FRW FD accession (cm) (cm) (cm) (cm) 532.5 ± (cm) (g) (cm) CPOV 130 ± 86.25 0.33 ± 5.15 ± 8.8 17.77 ± 1015.7 7.8 ± CPOC 2.20 ± 1.07 0.126 0.311 493.2 ± 1.48 ± 1.29 1.24 CPOK 112.2 78 ± 0.195 ± 5± 4.2 19.07 ± 925 ± 9.4 ± CPOKo ±1.08 1.16 1.73 0.65 452.7 ± 2.45 2.50 2.33 CPRV 121.75 77.25 0.320± 5± 4.2 17.75 ± 770 ± 7.75 ± CPRC ± 3.46 ± 2.18 0.102 0.26 469.25 0.96 16.71 0.92 CPRK 116.5 ± 78.25 0.210 ± 4.95 ± ± 4.18 19.25 ± 660 ± 8± CPRKo 3.06 ± 3.96 1.41 0.574 230 ± 2.06 5.96 0.587 98.75 ± 67.5 ± 0.31 ± 4.52 ± 8.13 17.67 ± 823.75 9.27 ± 2.98 2.65 0.5 0.330 259 ± 0.39 ± 1.52 1.18 95.75 ± 65 ± 0.21 ± 4.8 ± 5.4 18.35 ± 626.25 8.6 ± 3.14 2.45 0.5 0.35 163.5 ± 0.574 ± 8.16 0.483 98 ± 65 ± 0.21 ± 4.95 ± 5.68 16.27 ± 872.5 ± 8.77 ± 2.83 2.45 1.70 0.21 168.25 0.56 4.03 0.699 96.75 ± 66 ± 0.23 ± 4.7 ± ± 3.98 16.27 ± 505 ± 7.65 ± 2.83 3.0 2.16 0.163 0.56 2.78 0.532 LL- Leaf length, PL- petiole length, SL- sepal length, PTL- Petal length, NOS-Number of seeds, FL- Fruit length, FRW-Fruit weight, FD-Fruit diameter Morphometric analysis component had traits with highest loadings Analysis of variance are leaf length, petiole length, sepal length, petal length, number of seed, color of petiole, Analysis of variance carried out in different color of flower, sepal color, petal color, color quantitative characters showed significant of fruit in young, fruit pulp color, presence of variation ( p<0.05) in the most of the odor in leaves.(Table 6). characters except fruit length, fruit width and petiole length. The second principal component accounted for 11.73% of variation with highest loadings Principal component analysis are leaf length, number of seed, length of petal, fruit length, width and weight, color of In principal component analysis, the first petiole, and shape of fruit. The third principal principal component accounted maximum component accounted for 9.921% of variation (68.88%). The first principal variations with number of seed, fruit width and weight, color of petiole, and shape of Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.251-256 © Principal, Sree Narayana College, Kollam, Kerala, India

Analysis of morphological variability in two different varieties of Carica papaya (L.) 255 fruit. Major trait that accounted for more among the two varieties. The most widely variability in PC1, PC2 and PC3 is the color used analysis methods in characterization of of petiole. The highest loaded variables in plant genetic resources are principal PC1 are colour of petiole, colour of fruit, component analysis (PCA) and cluster colour of sepal, colour of petal, colour of analysis. PCA enables visualization of fruit in young condition and odor of leaf, in differences among individuals, identification PC2, it is fruit length and in PC3 it is fruit of possible groups and relationships among width. individuals and variables. Principal component analysis is a way to highlight Cluster analysis similarity and differences (Mattos et al., 2010). Cluster analysis has been employed to UPGAMA cluster analysis showed two assess similarities among genotypes in plant principal clusters (Fig 1). All accessions of C. breeding programmes (Rakonjac et al., papaya var. red lady are grouped in the first 2014). In the present study, PCA helps to principal cluster. It consists of two sub identify the characters which show more clusters. The first sub cluster consists of variations in the two species studied and the CPRKO and CPRC and second sub cluster cluster analysis helps to assess the similarity consists of CPRK and CPRV. All accessions and dissimilarity among different accessions of C. papaya var. yellow form the second of the same variety and between two principal cluster. It consists two sub clusters. varieties. The character that showed more The first sub cluster consists of CPOKO and variability in PC1, PC2 and PC3 is the color CPOK and the second sub cluster consists of of petiole. The highest loaded variables in CPOC and CPOV. The accessions of two PC1 are colour of petiole, colour of fruit, varieties form separate clusters due to colour of sepal, colour of petal, colour of dissimilarity in most of the characters. It fruit in young condition and odour of leaf, in shows that the two varieties of Carica PC2, it is fruit length and in PC3 it is fruit papaya have distinct morphological width. So we can use this characters as characters helps in the identification of these morphological markers in C. papaya for two varieties. varietal identification. This will be helpful for farmers and plant breeders. In the present study, principal component analysis was used to analyze the diversity Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.251-256 © Principal, Sree Narayana College, Kollam, Kerala, India

256 Current status and challenges for conservation and sustainable use of biodiversity Conclusion Ruqaie-AL and AL-khalifah, A.E. 2016. Morphological cladistics analysis of eight The qualitative vegetative, floral and fruit popular Olive cultivars grown in Saudi characters showing variations in leaf petiole Arabia using numerical taxonomic system for color, color of flower, color of sepal, color of personal computer to detect phyletic petal, unripe fruit color, fruit pulp color, relationship and their proximate fruit shape of fruit, odor of leaves and fruits. The composition. Saudi Journal of Biological quantitative vegetative, floral and fruit Science., 5(1): 115-121. characters showing significant variations are length of leaf petiole, weight of fruit and Suvalaxmi, P., Rout, G.R. and Dilipkumar, moisture content. In principal component D. 2019. Molecular and morphological analysis, the first principal component assessment of papaya (Carica accounted for 68.88% of variation, second papaya).Research Journal of Biotechnology., principal component accounted for 11.73% 14 (1): 63-70. of variation and third principal component accounted for 9.921% of variation. Major trait that accounted for more variability in PC1, PC2 and PC3 is the color of petiole. The highest loaded variables in PC1 are colour of petiole, colour of fruit, colour of sepal, colour of petal, colour of fruit in young condition and odor of leaf, in PC2, it is fruit length and in PC3 it is fruit width. In cluster analysis, the accessions of the two varieties are grouped in two separate principal clusters. References Mattos, A.L., Amorim, P.E., Amorim, O.B.V., Cohan, O.K., Lodo, S.A.C. and Siha. 2010. Agronomical and molecular characterization of Banana Germplasm. Pesq. agropec. Bras., 45(2): 146-154. Nishimwe, G., Janet Chepng’etichKosgei, Everlyn Musenya Okoth, George Ochieng Asudi, and Fredah Karambu Rimberia. 2019. Evaluation of the morphological and quality characteristics of new papaya hybrid lines in Kenya. African Journal of Biotechnology., 18: 58-67. Rankonjac, V., Aksic, F.M., Nikolic, D., Milatovic, D. and Colic, S. 2010. Morphological characterization of Oblacinka sour cherry by multivariate analysis, Scientia. Horticulture., 125: 679-684. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.251-256 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 257 ISBN 978-93-5396-871-7 CHAPTER 40 BIOSYTEMATICS OF SIDA ACUTA, Burm.f Chithra Vijayan*, Amina S and Sreeja S** *Post Graduate and Research Department of Botany, Sree Narayana College, Kollam **Department of Botany, University of Kerala, Kariavattom, Thiruvananthapuram,Kerala Correspondence Email: [email protected] ABSTRACT The present study was carried out to characterize the subspecies of Sida acuta viz., Sida acuta ssp acuta and Sida acuta ssp carpinifolia. In order to characterize the two subspecies, five accessions each, of the subspecies were subjected to morphological, foliar epidermal, cytological, palynological, biochemical and molecular analyses. The data were analyzed carefully to find out significant and consistent character- istic features to identify the two subspecies. Morphological features were found to be providing useful contributions in the species level systematics. Among the morphological features leaf shape, leaf base, leaf colour, size and colour of flower were found good for the delimitation of two subspecies. SDS- PAGE revealed the two subspecies as two distinct groups. RAPD analysis also proved genetic diversity in the two subspecies. The whole data from morphological, palynological and molecular analyses have been subjected to various statistical and morphometric analysis. The morphometric analyses characterize the subspecies of Sida acuta as two distinct groups. Based on the present study it is concluded that ex- treme morphological variations in the two subspecies of Sida acuta is attributed by genetic or environ- mental factors or by their interaction. Key words: Morphological, Genetic diversity, Palynological, Sida acuta Introduction tips of the branches. One of the important aspects of systematics is that it may utilize Systematics is the science of organismal di- data from all fields of biology: morphology, versity. It entails the discovery, description anatomy, embryology, ecology, cytology, and interpretation of biological diversity, as palynology, genetics and cell or molecular well as the synthesis of information on diver- biology. sity in the form of predictive classification systems. According to paleontologist George The study of systematics provides the scien- Gaylord Simpson (1961) “Systematics is the tific basis for defining or delimiting species scientific study of the kinds and diversity of and infraspecific taxa (subspecies or varie- organisms and of any and all relationships ties) and for establishing that these are dis- among them”. The fundamental aim of sys- tinct from others, closely related and similar tematics is to discover all the branches of the taxa. Earlier taxonomic classification of plant evolutionary tree of life, to document chang- species was mainly based on morphological es that have occurred during evolution and to characteristics though there may existgeneti- the greatest extent describe all species-the cal, cytological, palynological and molecular relations also. But present day systematist Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

258 Current status and challenges for conservation and sustainable use of biodiversity takes into consideration all the characteristics genetic polymorphism using RAPD (Random that is from morphology to molecular in tax- Amplified Polymorphic DNA). onomic for diversity studies. The present Materials and Methods study was undertaken to analyze the diversity among the two subspecies of Sida acuta The materials for the present study were two Burm.f. The main objectives of the study were 1.To characterize the two subspecies of subspecies of Sida acuta viz., Sida acuta ssp Sida acuta by using morphological, cytologi- acuta and Sida acuta ssp carpinifolia.Five cal and foliar epidermal characters. 2. To accessions each, of the subspecies were col- determine the extent and magnitude of pollen lected from different locations of Thiruvan- diversity in the two subspecies of Sida acuta. anthapuram and Kollam districts of Kerala 3. To assess the genetic diversity in the two and Kanyakumari district of Tamil Nadu subspecies of S. acuta using biochemical (Table 1). Potted plants were maintained in green house for easy availability of experi- marker (polypeptide). 4. To ascertain the mental material. Table 1. List of taxa under study and their place of collection. Accession No Name of taxon Place of collection Nedumangad 1 S.acuta ssp. acuta 2 S.acuta ssp. acuta Nagercoil 3 S.acuta ssp. acuta Kariavattom 4 S.acuta ssp. acuta Kollam 5 S.acuta ssp. acuta Kayamkulam 6 S.acuta ssp. carpinifolia Nedumangad 7 S.acuta ssp. carpinifolia Nagercoil 8 S.acuta ssp. carpinifolia Kariavattom 9 S.acuta ssp. carpinifolia Kollam 10 S.acuta ssp. carpinifolia Kayamkulam The various methods adopted in the present 2. Foliar epidermal studies study are explained below. For foliar epidermal studies, mature leaves 1. Morphological studies. were collected and washed well in running water. Thin epidermal peelings from the Fresh plant materials were used for morpho- adaxial and abaxial surfaces of the leaves logical analysis. A datasheet was designed were taken at around 11 am. The peelings for collecting information in 10 qualitative were stained with 1% aqueous solution of and 10 quantitative characters. Morphologi- saffranin as suggested by Ahamed (1964). cal features were observed and recorded ac- The stained peelings were washed well in cording to the system suggested by Hickey distilled water for uniform staining and (1971). Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 259 mounted in glycerine on a clean glass slide 4. Palynological studies and observed under 10×20 magnification of a binocular research microscope. Frequency of For the study of pollen morphology the ace- stomata was calculated as the number of sto- tolysis method described by Nair (1970) was mata per unit area by taking the mean from employed. Average pollen grain size and 10 fields of an Olympus binocular research other measurements were taken from random microscope. The length and breadth of sto- samples of 100 pollen grains in each taxon. matal complex and guard cells were meas- The size classification proposed by Erdtman ured using an ocular and stage micrometer. (1960) was adopted for constructing the fre- quency table. Stomatal index was calculated using the for- mula, Morphometric and Statistical analysis SI = S / S+E x 100 Qualitative characters were subjected to vari- ous morphometric analyses such as UPGMA where, clusteringand Principal Component Analysis. Morphometric analyses were performed by SI -Stomatal index Multi Variate Statistical Package (MSVP) version 3.1 (Kovach computing services, S - Number of stomata per unit area Wales, UK). ANOVA was carried out by the SPSS 7.5 (SPSS Inc., Chicago, IL, USA). E - Number of epidermal cells in the same unit area 5. Biochemical Marker Analysis The terminology by Metcalfe and Chalk Proteins of all the 10 accessions were ana- (1950) was used to describe foliar features. lyzed by SDS-PAGE. The presence or absence of trichomes was recorded under different categories as de- i) Protein extraction: Mature leaves were scribed by Fahn (1989). Photomicrographs selected for extracting the proteins. The were taken with the help of an Image Ana- leaves were collected from the plants main- lyzer (Olympus BX, 51. Japan). tained in the green house inorder to eliminate the influence of different edaphic and envi- 3. Cytological studies ronmental factors on the protein quality and composition. Meiotic chromosome studies were made from immature flower buds. The age and size of Protein extraction was done in 0.2 M TrisHCl the buds were decided by trial and error (pH 7.4) containing 1% PVP, 0.9% sodium method. The buds were collected between chloride and 0.5% β - mercaptoethanol. 100 8.30 and 9.30 am, fixed in Sem- mg of fresh leaf sample was homogenized men’sfluid(3absolute alcohol:1acetic acid: with 1ml extraction buffer in a pre-chilled 1chloroform) without any pretreatments and mortar and pestle. The homogenate was cen- kept in refrigerator for about 24 hours. About trifuged at 12,000 rpm for 10 minutes in a 2 hours before the smear preparation were refrigerated table top centrifuge (Eppendorf, made; the flower buds were transferred to Germany). The supernatant was collected and fresh fixative with 2 or 3 drops of ferric ace- stored at -200C until used as sample for elec- tate. For meiotic studies the aceto carmine trophoresis. (2%) smear technique of Belling (1926) was followed. ii) Protein preparation and electrophore- sis: The protein in the sample was quantified by Bradford’s method (Bradford, 1976). The Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

260 Current status and challenges for conservation and sustainable use of biodiversity sample preparation and SDS-PAGE was car- RAPD profiling was done on the genomic ried out according to Laemmli (1970). DNA of 10 accessions belonging to the two subspecies of Sida acuta. iii) Staining and gel documentation: After run, gel was stained with 0.1% Coomassie i) Isolation of genomic DNA: The genomic Brilliant Blue R-250 overnight and destained DNA from fresh, young leaves of the 10 ac- by water:methanol:acetic acid(6:3:1) solu- cessions was extracted by the modified tion, scanned and photographed with the gel CTAB method (Dellaporta et al., 1983). documentation system (Alpha Innotech Cor- poration, USA). Data Analysis: Only clear and evenly dis- tributed bands were selected and scored as iv) Data analysis : Relative molecular present ‘1’ and their absence were marked as weight of polypeptide bands were calculated ‘0’ in a matrix that was manually construct- by comparing the bands of the protein mo- ed. The reproducibility of the patterns ob- lecular weight marker using Alpha EaseFC served was assessed by conducting the whole version 4 (Alpha Innotech Corporation, RAPD experiments twice for ten accessions. USA). The bands were considered present Cluster analysis and Principal Coordinate when seen on the gel and also as peaks in the Analysis were carried out using the Multi densitrometric graphs. SDS-page analysis Variate Statistical Package (MVSP) version was performed in duplicate and only those 3.1 (Kovach computing services, Wales, bands clearly obtained twice were scored. UK). The bands present were coded as ‘1’ and when it is absent designated as ‘0’. Cluster Results analysis and Principal Coordinate Analysis were performed on the resultant data using 1. Morphological studies the Multi Variate Statistical Package (MVSP ver 3.1) (Kovach computing services, Wales, The subspecies of Sida acuta viz., Sida acuta UK). ssp acuta and Sida acuta ssp carpinifolia shows marked difference in their qualitative 6.RAPD Marker Analysis and quantitative characters (Table 2 ) Table 2: List of characters selected for morphological analysis. Sl. No Qualitative characters Character states 1 Habit ‘0’ herb/ ‘1’ under shrub/ ’2’ shrub/ ’3’ tree 2 3 Leaf arrangement ‘0’ opposite/ ‘1’ alternate 4 Leaf shape ‘0’ lanceolate/ ‘1’ ovate 5 Leaf apex ‘0’ acute/ ‘1’ obtuse/ ‘2’ acuminate 6 Leaf base ‘0’ acute/ ‘1’ obtuse/ ‘2’ rounded 7 Leaf colour ‘0’ green/ ‘1’ reddish green 8 9 Size of flower ‘0’ small/ ‘1’ large 10 Colour of flower ‘0’ pale yellow/ ‘1’ dark yellow Shape of ovary ‘0’ ovoid/ ‘1’ globose Leaf margin ‘0’ entire/ ‘1’ serrate/ ‘2’ revolute Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 261 Quantitative characters Flowers are solitary or paired in the leaf axil 1 Leaf length opening at about 8.30 am and closing at 11 2 Leaf width am. Flowers are small and pale yellow in 3 Length of petiole colour, ovary ovoid, schizocarpic fruit. No 4 Internode length significant differences in qualitative charac- 5 Pedicel length ters were noticed among the five accessions 6 Length of corolla of subspecies acuta (1, 2, 3, 4 and 5). 7 width of corolla 8 Length of calyx Sida acuta ssp carpinifolia 9 width of calyx 10 Length of staminal column It is an undershrub reaching a height of 1m. Its leaves are alternate, ovate, with acute apex Sida acuta ssp acuta and rounded base, margin serrate, reddish green in colour. Flowers are large and dark It is an undershrub growing to a height of 1 yellow in colour, opening at around 10.30 am meter. Leaves alternate, lanceolate, with and closing at around 2 pm. Ovary globose, acute apex and obtuse base, margin serrate. fruit schizocarp. All the five accessions of S.acuta ssp carpinifolia are similar in their qualitative characters (Table 3). Table 3. Variations in quantitative morphological characters Means with similar superscripts are not significantly different (p =0.05) Accessions Leaf length Leaf breadth Length of Internode Pedicel (cm) (cm) petiole (cm) length (cm) length (cm) 1 6.56±0.29bcd 1.96±0.12cd 0.45±0.02ab 2.50±0.23abcd 0.75±0.09cd 2 5.27±0.23cd 2.08±0.26cd 0.45±0.02ab 2.11±0.23cd 0.70±0.06d 3 7.20±0.40a 2.40±0.38bc 0.40±0.04ab 2.70±0.31abc 0.92±0.04bcd 4 5.89±0.34cd 1.62±0.17d 0.38±0.03b 2.32±0.11bcd 0.84±0.10bcd 5 6.90±0.44b 2.28±0.22bcd 0.40±0.03ab 1.98±0.22d 0.80±0.05cd 6 4.89±0.36cd 3.25±0.24ab 0.46±0.24ab 2.16±0.06cd 0.76±0.05cd 7 4.78±0.15d 2.12±0.11cd 0.42±0.03ab 2.84±0.17ab 1.04±0.08ab 8 5.82±0.23bc 3.02±0.19ab 0.46±0.02ab 2.79±0.21abc 1.20±0.707a 9 5.55±0.24bcd 3.45±0.18a 0.46±0.04ab 3.08±0.16a 0.88±0.03bcd 10 5.35±0.13bcd 2.98±0.31ab 0.49±0.03a 3.10±0.26a 0.98±0.06abc Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

262 Current status and challenges for conservation and sustainable use of biodiversity ANOVA differentiating the two subspecies of Sida All the quantitative morphological features acuta Burm. f. and their accessions (Table 4 selected for morphometric analysis except and 5). However quantitative characters length of petiole were found significant for among the accessions of both the subspecies of Sida acuta were found varied (Table 5). Table 4. Variations in quantitative morphological characters Means with similar superscripts are not significantly different (p =0.05) Accessions Corolla length Corolla width Length of calyx Width of calyx Length of stami- (cm) (cm) (cm) (cm) nal column (mm) 1 0.76±0.035c 0.45±0.036e 0.52± 0.028c 0.54±0.036c 3.47±0.19b 2 0.77±0.03c 0.48±0.026de 0.63±0.028bc 0.56±0.017bc 3.1±0.14b 3 0.82±0.04c 0.59±0.04cd 0.68±0.037bc 0.58±0.037bc 3.2±0.20b 4 0.78±0.037c 0.52±0.037cde 0.70±0.037abc 0.57±0.03bc 3.26±0.19b 5 0.85±0.02c 0.60±0.031c 0.68±0.048bc 0.56±0.02bc 3.2±0.20b 6 1.52±0.08ab 0.88±0.037a 0.79±0.033a 0.72±0.037a 4.1±0.08a 7 1.85±0.12ab 0.89±0.03ab 0.84±0.024ab 0.70±0.054bc 4.0±0.00a 8 1.40±0.07b 0.80±0.05b 0.74±0.024ab 0.66±0.04ab 4.0±0.00a 9 1.66±0.092a 0.90±0.031a 0.80±0.031a 0.74±0.04a 4.2±0.09a 10 1.50±0.10ab 0.90±0.031a 0.80±0.031a 0.74±0.024a 4.0±0.00a Table 5: ANOVA of quantitative characters- Morphology Sl.No Characters N F df P 1 Leaf length 10 9.727 9 .000 2 Leaf width 10 6.429 9 .000 3 Length of petiole 10 1.111 9 .375 4 Internode length 10 3.350 9 .003 5 Pedicel length 10 4.420 9 .000 6 Length of corolla 10 31.09 9 .000 7 Width of corolla 10 26.39 9 .000 8 Length of calyx 10 4.22 9 .001 9 Width of calyx 10 5.10 9 .000 10 Length of Staminal column 10 8.068 9 .000 N: Sample size, F: F value, df: degree of freedom, p: Statistical significance at 0.05 level. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 263 PCA Fig.9 UPGMA phenogram showing relation- ship among the accessions of two subspecies Inorder to find out the significant characters of Sida acuta based on morphology (qualita- contributing to the identification of subspe- tive and quantitative characters) cies or accessions, Principal Component Analysis was performed considering 10 LEAF EPIDERMAL STUDIES quantitative and 10 qualitative traits. In the Sida acuta ssp acuta PCA of morphological data, 95.524% of the All the five accessions were uniform in the phenetic variance was accounted by the first leaf epidermal features. Epidermal cells with principal component axis, followed by deeply undulating walls were observed in 2.114% in the second principal axis (Table both adaxial and abaxial surfaces. The leaves 6). All the qualitative traits excluding habit, were amphistomatic and amphitrichomic but leaf arrangement, leaf apex and leaf margin generally the stomata and trichomes were were found principally influential in the more concentrated on the abaxial sur- PCA. The most important quantitative load- face(Plate 3) The stomatal type is anisocytic. ing traits as per first principal axis were leaf Simple, stellate and glandular trichomes were length, leaf breadth, length of corolla, width present in the subspecies acuta. of corolla and length of staminal column. The Sida acuta ssp carpinifolia. highest loading traits in second principal axis Epidermal cells with undulating walls were include leaf length, leaf breadth, internode seen on both adaxial and abaxial surfaces. length, length of corolla. Characters such as The stomatal type is anisocytic and stomata petiole length, pedicel length, length and were more concentrated on the abaxial sur- width of calyx were found insignificant for face than adaxial surface. Simple and stellate designing the principal components. trichomes were present. No significant dif- ferences in the qualitative characters were Cluster analysis noticed among the five accessions (6, 7, 8, 9 and 10). UPGMA phenogram (fig 9) generated from the qualitative and quantitative data together provided two principal clusters. The first one with all the accessions of S.acuta ssp acuta (1, 2, 3, 4 and 5) and the second principal cluster included accessions of subspecies carpinifolia (6, 7, 8, 9 and 10). The first prin- cipal cluster divides into two sub-clusters, the accessions 3 and 5 were grouped in first sub cluster and remaining accessions 1, 4, and 2 were grouped together in the second sub- cluster. Within the second principal cluster accession 7 lies as an outlier. Remaining ac- cessions were placed in two sub clusters, in which accession 9 and 6 were placed in one sub-cluster. Accessions 8 and 10 were pre- sent in second sub-cluster. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

2642B6i4osytematics of Sida AcuCCtauu, rrBrreeunnrmtt ss.ttfaattuuss aanndd cchhaalllleennggeess ffoorr ccoonnsseerrvvaattiioonn aanndd ssuussttaaiinnaabbllee uussee ooff bbiiooddiivv2ee6rrss4iittyy Plate 1 showing distribution of stomata Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 265 Cytological studies: Sida acuta ssp acuta Cytological investigation of the flower buds showed 14 bivalents at first metaphase of meiosis (fig 5.1). Pollen mother cells showed normal meiosis. Fig 5.1 Sida acuta ssp carpinifolia The meiotic study reveals the meiotic chro- Palynological studies mosome number as n = 13. All the five ac- cessions were subjected to meiotic prepara- The pollen grains present in the two subspe- tion, and all of them showed 13 bivalents at cies of Sida acuta are pantoporate, spheroi- dal and spinate. The pollen grains from all first metaphase of meiosis (fig 5.2). the ten accessions showed similarities in qualitative characters such as exine ornamen- tation, shape and aperture type (Plate 6). Di- morphic pollen grains (large and medium size) are present in S. acuta ssp acuta while only one type of pollen (large size) was found in subspecies carpinifolia. Figure 5.2 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

266 Current status and challenges for conservation and sustainable use of biodiversity Plate 6. palynological features SDS-PAGE Cluster analysis All the accessions were subjected to SDS- The UPGMA cluster analysis with SDS- PAGE for revealing the polypeptide compo- PAGE data revealed two principal clusters sition. The figure represents the electropho- (fig 8). The first principal cluster consisted of retic pattern of the soluble proteins of 10 ac- the accessions 6 and 8, while the rest of the cessions by SDS-PAGE. Twenty three bands accessions were grouped in the second prin- were identified in the protein profile ranging cipal cluster. In the second principal cluster, from 260 kDa to 15 kDa. 73.94% polymor- two sub- clusters were distinguished. The phism was observed in the protein profile first sub-cluster comprised of accessions 7, 9 (Fig 7). and 10. Accession 7 lies well apart within this cluster. The second sub-cluster consisted of all the accessions of Sida acuta ssp acuta (1, 2, 3, 4 and 5). In this second sub-cluster accessions 1, 2 and 4 were grouped together while accessions 3 and 5 lies as an outlier. Figure 8. UPGMA phenogram showing rela- tionship among the accessions of two sub- species of Sida acuta based on SDS -PAGE profiling. Fig 7:- SDS PAGE Profile Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 267 cluster consist of accessions 7 and 6. Acces- sions 8, 9 and 10 were grouped together in the second sub-cluster while accession 10 lies as an outlier. Figure 9. UPGMA phenogram showing rela- tionship among the accessions of two sub- species of Sida acuta based on RAPD marker analysis. RAPD Marker Analysis Discussion RAPD analysis was carried out using a set of Morphological studies 20 primers of the S- series. Out of the 20 primers, twelve primers gave reliable ampli- Morphology of an individual is a structural fication profile for obtaining the genetic rela- and functional consequence of the activities tionships between the two subspecies of Sida of the genetic material at all levels from cell acuta. Twelve primers generated a total of 93 to organism (ChayaKumari and Ramesh, amplified products with an average of 7.7 2008). Phenotypic variance among individu- fragments per primer. Out of 93 amplification als is a regular feature in natural populations. products 49 bands were polymorphic. The The plants chosen for the study, S.acuta ssp individual primers produced 4 to 12 amplifi- acuta and S.acuta ssp carpinifolia are under cation products. Primer S143 produced max- shrubs of the family Malvaceae. In the pre- imum bands (12) while the least number was sent investigation, the foliar characters and produced by primers S166 and S119 (4 floral characters were found to have some bands). Polymorphism across the two sub- taxonomic significance. In all the ten acces- species was found to be 52.68%. The per- sions, the phyllotaxy, leaf margin, leaf apex centage of polymorphism for each primer were found uniform i.e., alternate, serrate and ranged from 25.00 to 75%. The primers pro- acute respectively. The leaf characters such duced various level of polymorphism among as leaf base, leaf shape and leaf colour distin- the two subspecies of Sida acuta. guish the two subspecies of Sida acuta. In the infinite range of diversity of form, structure Cluster analysis and size of leaves the flowering plants sur- The UPGMA phenogram (fig 9) showed two principal clusters A and B. Accessions (1, 2, 3, 4 and 5) of S. acuta ssp acuta were grouped in the principal cluster A. Acces- sions of subspecies carpinifolia were grouped in the principal cluster B. cluster A consisted of two sub-clusters. Accession 1, 2 and 3 are grouped together in the first sub-cluster. Re- maining accessions of subspecies acuta (4 and 5) were grouped in the second sub- cluster. In the principal cluster B, two sub- clusters were distinguished. The first sub- Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

268 Current status and challenges for conservation and sustainable use of biodiversity pass all other groups of seed plants. Accord- problems in the species Sida acuta. The pre- ing to Eames (1961) and Stuessy (2002) due sent study only provided some information to obvious differences in size and shape, the on the inter and intraspecific diversity exist- leaf blade has been examined extensively in ing between the accessions of the two sub- taxonomic studies. Levin (1986) pointed out species of Sida acuta. that foliar characters do have systematic val- ue; the value of individual characters varies Foliar epidermal studies considerably. According to Levin (1986), most of the leaf characters, both qualitative The foliar epidermis is one of the most sig- and quantitative, have systematic relevance at nificant taxonomic character from the biosys- lower taxonomic levels. tematic point of view and the taxonomic studies of a number of families are made on ANOVA pointed out significant variations the basis of leaf epidermis (Bhatia, 1984; existed in the quantitative morphological Jones, 1986). Although the foliar epidermal characters of different accessions investigat- anatomy of a number of Malvaceae species ed. The cluster and principal component has been described (Inamdar and Chohan, analysis of morphological characters revealed 1969; Ramayya and Rao, 1976; Adedeji and the existence of variability among the inves- Dloh, 2004; Rudgers et al., 2004; Celka et tigated accessions. Highest loading traits are al., 2006), the emphasis was on general ana- leaf shape, leaf base, leaf colour, size and tomical features, ontogeny of stomata and colour of flower, these traits distinguished the gross morphology of trichomes. accessions of subspecies acuta and subspe- cies carpinifolia. An infra-specific variability In the present study epidermal cells with un- based on morphological characters was high- dulating walls were observed in all the acces- lighted in the present study and all the acces- sions of the two subspecies of Sida acuta. sions clustered in two principal clusters. First Leaves amphistomatic and amphitrichomic, principal cluster with accessions of S.acuta generally stomata and trichomes were more ssp acuta and second principal cluster con- concentrated on the abaxial surface. In both sisted of accessions of S. acuta ssp carpini- subspecies, the stomatal type is anisocytic. folia. Thus the morphometric output in the The quantitative characters like length and form of UPGMA phenogram clearly spelt out width of guard cell, size of stomatal complex, the two distinct groups of plants. Accession 7 stomatal frequency are found to be varied lies as an outlier. This may be due to the var- within and between the accessions (Table 8). iation in the quantitative characters. The The size of stomatal complex and guard cells coordination among the accessions in the observed in S. acuta ssp acuta were compara- scatter plot also supports this. tively larger than those in subspecies carpini- folia.Carlquist (1961) emphasized the contri- Morphological characterization is a prelimi- bution of variation in stomatal size in delimit- nary and easily manageable tool, although ing species within a genus. According to additional information about the genotype of Carpenter and Smith (1975) the variations in the plant may help to resolve taxonomical stomatal frequencies have taxonomic signifi- cance at generic level. In this study the dif- Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 269 ferences in stomatal frequency among the carpinifolia were characterized by only one accessions have been noted. The differences type of pollen (large size). The pollen grains in most of the stomatal features were of little show variations in quantitative characters taxonomic importance to delimit different within and between the accessions. In the two taxa under study. subspecies pollen shape was found to be spherical, the exine ornamentation is spinate The trichome features now have considerable and the aperture type is pantoporate. The data importance in taxonomic studies (Leelavathi of pollen size showed wide range of variation and Ramayya, 1983). The size and morphol- among the two subspecies; it ranged from ogy of trichomes are variable, and provide 53.7 µm - 85.4 µm in the subspecies acuta some taxonomic distinction to the respective and 75 µm -102.09 µm in the studied acces- taxa. The foliar trichomes of Sida however sions of subspecies carpinifolia. possess a remarkable diversity and provide a great deal of systematic evidence. In the pre- Pollen grains are usually classified on the sent study simple, stellate and glandular tri- basis of their shape, size, symmetry, apertural chomes were present in subspecies acuta types and exineornamentation. Variations in while, subspecies carpinifolia has simple and pollen size, aperture and spine features as stellate trichomes only. These characteristics well as exine ornamentation has taxonomic are significant in the identification of the two significance, of which the aperture character subspecies. Gamble (1935); Hutchinson is considered to be of primary importance, (1959) and Metcalfe and Chalk (1950) re- whereas exine sculpturing secondary and the ported that stellate trichomes are characteris- others as tertiary (Nair, 1965; Lakshmi, tic of entire family Malvaceae. 2003). Pore diameter and the appearance of pores on the inner surface of the acetolysed Palynological studies pollen are also of taxonomic significance (El Naggar, 2003). Pore diameter in the studied Pollen morphology has great importance in taxa was in the range of 3.1µm -5.8 µm for taxonomic studies. Pollen morphology can be subspecies acuta and 4.5 µm -6.8 µm for used for the classification of flowering plants subspecies carpinifolia. It serves as a distin- (Erdtman, 1943, 1966; Walker and Doyle, guishing feature at infraspecific level in the 1975). A systematic study of spore morphol- species Sida acuta. Subspecies acuta could ogy was initiated by Nayar (1962). Pollen be delimited from the subspecies carpinifolia grains of the family Malvaceae has been in- on the basis of pore diameter. vestigated by Master (1875), Erdt man (1952), Fryxell and Hashmi (1971), External marking of the pollen grains is de- Perveenet al (2007), El Naggar (2003) and scribed as the best, most constant and distinct Lakshmi (2003). character by which grains may be delimited at different taxonomic levels in case of In the present investigation, the pollen grains stenopalynous families (Pope, 1925; Nair and are seen in monads. Dimorphic pollen grains Sharma, 1965). One of the most prominent (large and medium size) were present in sub- and interesting feature of malvaceous pollen species acuta. The accessions of subspecies Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

270 Current status and challenges for conservation and sustainable use of biodiversity is the echinations or prolongations of the SDS – PAGE profiling exine into definite spines (El Naggar, 2003). The spines show reliable variation in size, The protein banding pattern obtained by gel shape and surface distribution and are used to electrophoresis has been utilized for solving distinguish the two subspecies. problems in the identification of critical taxa, their relationship and taxonomic status. Gar- Cytological studies diner and Forde (1987) stated that electro- phoresis can also be used to characterize the In the present study, the cytological investi- protein profiles of species and cultivars to gation showed difference in chromosome compare plants of different geographical number. 28 chromosomes were present in origin, and also to provide taxonomically subspecies acuta while subspecies carpini- useful descriptors that are substantially free folia has 26 chromosomes. Pushparajan from environmental influence. Protein elec- (1986) reported that subspecies acuta has trophoresis is most useful at the level of pop- n=14 chromosomes. Adhikary (1963) report- ulations, within a species or among closely ed that x = 7 can be taken as the basic chro- related species (Crawford and Julian, 1976). mosome number and that the species have Protein electrophoresis has been utilized as a evolved through polyploidy or occasionally powerful tool in the phylogenetic systematics aneuploidy. In the present study this is found of various plant groups (Ahmad and Slinkard, true only for the subspecies acuta. Hazra and 1992; El Naggar, 2001; Bhargava et al., Sharma (1971) considered that the occur- 2005). Electrophoresis profiles of proteins rence of numbers other than x = 7 in some and isozymes have been successfully used to Sidaspecies is due to accidental aneuploidy. clarify the taxonomy of families such as Po- In the light of available literature it may be aceae (Johnson et al., 1967), Cucurbitaceae assumed that subspecies carpinifolia is a tet- (Pasha and Sen, 1991) and Fabaceae (Misset raploid based on x = 7 by the reduction of and Fontenelle, 1992). two chromosomes. The importance of chro- mosome number as a taxonomic character as In the present investigation, the total protein reported by Davis and Heywood (1963) is polymorphism across the two subspecies of that it is probably one of the most constant Sida acuta is high. It is obvious from the pol- single features employed. They also reported ypeptide pattern of the soluble protein that that all individuals within a species usually the subspecies acuta shows marked differ- have the same chromosome number, alt- ence from subspecies carpinifolia. Also there hough there are exceptions due to somatic exist some similarities in the protein compo- instability. The differences in chromosome nents. number indicate the different genetic stock across the subspecies. It is thus in positive The morphometric output (UPGMA pheno- correlation with the extreme morphological gram) indicate the utility of protein profile in diversity. Based on the present data, the mor- elucidating relationship among the two sub- phological variation in the subspecies may be species of Sida acuta. UPGMA phenogram attributed by the genetic diversity. revealed two principal clusters. Accessions 6 and 8 were grouped in the first principal clus- Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 271 ter. The accessions (7, 9, and 10) of subspe- are grouped together in the first sub-cluster. cies carpinifoliaand all the accessions of sub- This clustering may be due to the same geo- species acuta are seen in the second principal graphical location of the two accessions. Ac- cluster. This indicates that the subspecies has cession 3 from Kariavattom which is placed some similarities in their protein components. well apart within the first sub-cluster but lies The accessions are found scattered in the closer to accession 1. This can also be at- PCoA scatter plot. Clustering in the pheno- tributed to the similarity in geographic condi- gram was able to characterize the two sub- tions. Accessions 4 and 5 from Kollam and species of Sida acuta as two distinct groups. Nedumangadu are grouped together in the It also provides some information on the ex- second sub-cluster. The second principal tent of diversity existing between the acces- cluster is separated into two sub-clusters of sions of two subspecies of Sida acuta. which accessions 6 and 7 from Neyattinkara and Nagercoil are grouped together in first RAPD marker analysis sub-cluster. This grouping is due to geo- graphical similarity. Accessions 8, 9 and 10 RAPD marker analysis is the simplest and are grouped together in the second sub- fastest of DNA based technique in genetic cluster. From the RAPD analysis we can dis- similarity studies (Gwanama et al., 2000). tinguish the two subspecies of Sida acuta. RAPD has a higher resolving power as a fin- The UPGMA cluster analysis provided dis- ger printing technique as it scans the entire tinct species cluster to subspecies acuta and genome for polymorphism (Williams et al., subspecies carpinifolia. The results obtained 1990). This makes RAPD profiling ideal for in the present study confirm the usefulness of studying genetic variation at lower taxonomic RAPD for the assessment of genetic diversity levels, thus we employed RAPD as a tool for among the two subspecies of Sida acuta. The assessing genetic variation in the two subspe- result obtained from RAPD analysis substan- cies of Sida acuta. tiates the cytological data. These results indi- cate the different genetic stock across the two In the present study 10 accessions of the two subspecies of Sida acuta. It is thus in positive subspecies of Sida acuta were examined for correlation with the extreme morphological their RAPD - PCR patterns. RAPD analysis diversity between the two subspecies of Sida was carried out using a set of 20 primers. 12 acuta Burm.f. Based on the present study it primers gave reliable amplification profile. A can assume that, the morphological diversity total of 93 amplicons were produced. The or variation may be attributed by cytological percentage of polymorphism across the sub- and molecular level variations. species was found to be 52.68%. The high polymorphism indicates remarkably high Conclusion genetic variability. The dendrogram analysis indicates that all the accessions of the two The present study was carried out to charac- subspecies of Sida acuta form two principal terize the subspecies of Sida acuta viz., Sida clusters. The first principal cluster consisted acuta ssp acuta and Sida acuta ssp carpini- of accessions of subspecies acuta. Acces- folia. In order to characterize the two subspe- sions 1 and 2 from Parassala and Nagercoil Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

272 Current status and challenges for conservation and sustainable use of biodiversity cies, five accessions each, of the subspecies cies. The whole data from morphological, were subjected to morphological, foliar epi- palynological and molecular analyses have dermal, cytological, palynological, biochemi- been subjected to various statistical and mor- cal and molecular analyses. The data were phometric analysis. The morphometric anal- analyzed carefully to find out significant and yses characterize the subspecies of Sida consistent characteristic features to identify acutaas two distinct groups. Based on the the two subspecies. present study it is concluded that extreme morphological variations in the two subspe- Morphological features were found to be cies of Sida acuta is attributed by genetic or providing useful contributions in the species environmental factors or by their interaction. level systematics. Among the morphological features leaf shape, leaf base, leaf colour, size References and colour of flower were found good for the delimitation of two subspecies. ANOVA out- Ahmed, F. and Slinkard, A.E. 1992. Genetic put pointed out that all the quantitative char- relationships in the genus Cicer L. as re- acters except length of petiole were signifi- vealed by polyacrylamide gel electrophoresis cant for differentiating the two subspecies. of seed storage proteins. Theor. Appl. Genet., When compared to morphological features, 84: 688–692. foliar epidermal features have lesser taxo- nomic significance in the delimitation of two Bhargava, A., Rana, T.S. and Shukla, S. subspecies. Moreover, the variations in the 2005. Seed protein electrophoresis of some quantitative traits were found effective to a cultivated and wild species of Chenopodium . particular extent for the delimitation of the BiolPlant .,49: 505–511. two subspecies. The pollen grains show vari- ations in quantitative characters within and Chayakumari and Ramesh, S. R. 2008. Vari- between the accessions. ANOVA output and ability of morphological traits in wild type PCA analysis also pointed out the signifi- and mutant strains of Drosophila nasuta- cance of all the quantitative traits in delimit- nasuta and Drosophila nasutaalbomicans.J. ing the two subspecies of Sida acuta. Cytol. Genet. (India) 9: 13-19. Cytological analysis was confined to the Crawford, D.J. and Hugh D. W. 1976. “Al- chromosome numbers. Cytological analysis lozyme Variation in ChenopodiumFremontii.” revealed different genetic stock across the Systematic Botany, vol. 2 pp. 180–190. two subspecies or genetic diversity in the two subspecies. To find out the extent of genetic Davis. P. H. and Heywood. V. H. 1963. diversity, molecular analyses was done by \"Principles of Angiosperm Taxonomy\". Oli- RAPD marker profiling of genomic DNA ver and Boyd, Edinburgh. 556 pp. and SDS-PAGE profiling of leaf proteins. SDS-PAGE revealed the two subspecies as El Naggar, S.M. 2001. Implications of seed two distinct groups. RAPD analysis also proteins in Brassicaceae systematics.-Biol. proved genetic diversity in the two subspe- Plant. 44: 547–553. Gardiner, S.E. and Forde, M.B. 1987. SDS polyacrylamide gel electrophoresis of grass Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

Biosytematics of Sida Acuta, Burm.f 273 seed proteins: a method for cultivar identifi- Wight and its taxonomic signifi- cation of pasture grasses. Seed SciTechnol cance.Curr.Sci.55:542. 15: 663–674. El Naggar, S.M.I. 2003. Pollen morphology Gwanama, C., Labuschagne, M.T. and Botha, of Egypyian Malvaceae: an assessment of A.M. 2000. Analysis of genetic variation in taxonomic value. Turk. J. Bot. 28: 227-240. Cucurbita moschata by random amplified polymorphic DNA (RAPD) markers. Euphyt- Nair, P. K. K. and Sharma, M. 1965. Pollen ica. 113: 19–24. morphology of Liliaceae. – J. Palynology 1: 38-61. Johnson, B.L. 1967.Tetraploidwheats: seed protein electrophoresis pattern of the emmer Pope, M.A. 1925. Pollen morphology as an and timopheevi groups. Science., 158: 131– index to plant relationship. I. Morphology of 132 . pollen. Botanical Gazette., 80(1): 63-73. Pasha,M. K. and Sen, S. P. 1991. Seed pro- Lakshmi, K.G. 2003. Palynological studies tein patterns of Cucurbitaceae and their taxo- on certain Malvales. Ph.D. thesis, Mahatma nomic implications. Biochemical Systematics Gandhi University (India). and Ecology,19:569-576. Perveen, A. and Qaiser, M. 2007. Pollen Flo- Misset, M.T. and Fontenelle, C. 1992. Pro- ra of Pakistan-Malvaceae- Grewioide- tein relationships between natural popula- aeLII.Pak. J. of Bot., 39 (1), 1-7. tions ofUlexeuropaeus andU.gallii (Faboide- ae, Genisteae) and their hybrids. Pl SystEvol Fryxell, P.A. and Hashmi, S.H. 1971. The 179, 19–25 . segregation of Raydero from Hibiscus.Bot. Gaz., 132: 57-62. Radharanjan, H. and Archana, S. 1971. Chromosome Studies in Different Species Erdtman, G. 1952. Pollen morphology and and Varieties of Sida with Special Reference plant taxonomy. Geolog.Fören.Stoc.Förhan., to Accessory Chromosomes. Cytologia: 74(4): 526– 527. ISSN 1348-7019.p. 285-297. Master, M.T. 1875. Malvaceae. In: J.D. Hooker Williams, J.G.K., Kubelik, A.R., Livak, K.J., (ed.), Flora of British India. Rafalski, J.A. and Tingey, S.V. 1990. DNA polymorphisms amplified by arbitrary pri- Walker, J. W. and Doyle, J. A. 1975. The mers are useful as genetic markers. Nucleic bases of angiosperm phylogeny: Palynology. Acids Res., 18: 6531–6535. – Ann. Missouri Bot. Gard., 62: 664 –723. Adhikary, A. K.1963. Cytotaxonomical stud- Metcalfe, C. R. and Chalk, L. 1950. Anatomy ies in some species of Sida. Trans. Bose Res. of the Dicotyledons: Leaves, Stem, and Inst. 26: 59-63. Wood in Relation to Taxonomy with Notes on Economic Uses. v.2. Claredon Press, Ox- Pushparajan, G., Kuriachan, P. I. and Ni- ford. nan,C.A. 1986. Cytology of Cullenia excels Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

274 Current status and challenges for conservation and sustainable use of biodiversity Hutchinson, J.1959. Families of the flower- Bhatia, R.C., 1984. Foliar Epidermal Studies of ing plants, 2nd Ed., Clarendon press, oxford. Heliotropiumsupinum L. Folia Geo- bot.Phytotaxon.,19: 381–385. Gamble, J.S. 1935. Flora of the presidency of the Madras. Part I. Jones, J.H., 1986. Evolution of the Fagaceae: the implications of foliar features. Annl. Missouri Bot. Leelavathi, P. and Ramayya, N. 1983. Struc- Gard., 73: 228–275 . ture, distribution and classification of plant trichomes in relation to taxonomy Perveen, A., 1993. A preliminary study of the pol- III.Papilionoideae. Proc. Indian Acad. Sci. len flora of Karachi.Ph.D. Thesis, Department of (Plant Sci.) 92, 421–441 . Botany, University of Karach, Karachi. Carpenter, S.B. and Smith, N.D. 1975. Sto- Perveen, A. and Qaiser, M. 2007. Pollen Flora of matal distribution and size in southern Appa- Pakistan-Malvaceae-Grewioideae-L II.Pakistan J. lachian hardwoods.Canadian Journal of Bota- Bot., 39: 1–7 . ny.,53: 1153–1156. Dellaporta, S., Wood, J. and Hicks, J.B. Carlquist, S. 1961. Comparative Plant Anatomy. 1983. A plant DNA minipreparation: version New York, USA: Holt, Rinehart and Win- II. Plan.Mol Biol. Rept., 1:19–21. ston. Eame,J. 1961. Morphology of the angio- Celka, Z.P., Skudlarz andBiereznoj, U. 2006. sperms.biodiversity. Org. Morphological variation of hairs in Mal- vaalcea L. (Malvaceae).Biodiversity: Re- Hickey, L. J. 1997. Evolutionary significance search and Conservation., 3: 258-261. of leaf architectural features in the woody dicots.Amer. J. Bot.,58: 469. Rudgers, J.A., Strauss,S.Y.and Wendel, J.F. 2004. Am. J. Bot.., 91: 871-88. Adedeji, O., 2004. Leaf epidermal studies of the species of Emilia Cass.(Senecioneae, Asteraceae) in Nigeria.BotanicaLithuanica., 10: 121-133. Ramayya, N. and Rao, R.S. 1976. Morphology phylesis and biology of the peltate scale, stellate and tufted hairs in some malvaceae. J. Indian Bot. Soc., 55: 75–79 . Inamdar, J.A. and Chohan, A.J. 1969. Epidermal structure and ontogeny of stomata in vegetative and floral organs of Hibiscus rosa-sinensis L.Australian J. Bot., 17: 89–95 . Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.257-274 © Principal, Sree Narayana College, Kollam, Kerala, India

A comprehend in to the wing scales of butterfly craea terpsicore 275 978-93-5396-871-7 CHAPTER 41 A COMPREHEND IN TO THE WING SCALES OF BUTTERFLY ‘ACRAEA TERPSICORE’ Amina Thaj, Munisha Murali S* and Sheeba S PG and Research Department of Zoology Sree Narayana College, Kollam, India *Correspondence E-mail: [email protected] ABSTRACT Acraea terpsicore commonly known as ‘Tawny coster’ is a small leathery winged butterfly belonging to the family Nymphalidae and subfamily Heliconiinae. The present study is focused on the morphology of different wing scales of Acraea terpsicore. The different coloured wing scales are scrapped out of the wing to a glass slide and fixed using xylene to observe under light microscope for scrutiny. A total of 99 different scales were studied from the dorsal and ventral side of the wing. However, 30 varieties of single scales were observed from orange coloured region, 24 scales from hindwing termen and 21 scales were noted from ventral region respectively. The isolated scales show diversity in shape, colour, length and width. The scales are isolated from the differently coloured wing areas namely orange, black and hindwing termen having black band with white spots from the upperside of the wing and similar areas from the underside. Examining at the orange coloured region on the dorsal side of the wing exhibited abwing and adwing with a stalk at its base. The scale colour ranges from dark or bright orange to pale orange. The scales have a dimensions ranging from 70µ to 114µ in length and 47µ to 82µ in width. Observation on black scales indicates majority of the scales are similar to orange scales with short black rounded terminus and no dentation. Scales possess abwing, adwing and a stalk. The intensity of the dark colour varies in each. The scales have a dimensions ranging from 78µ to 102.44 µ in length and 51.22µ to 74.86µ in width. The hindwing termen of the butterfly have black bands with small spots embedded in white colour. This area is composed of white scales, brown scales and black scales with different dimensions. White scales have narrow adwing , broader and round abwing with no dentation except for two white scales where there abwing are dentated and structured. Brown scales are short when compared to the former having structured abwing and other brown scales have no dentation. The white scales together with the black and brown scales at the hindwing termen help in the formation of bands.The underside of the Tawny coster wing is dull coloured unlike the upperside. Several pale orange and black colored scales are obtained. Some scales are glassy type and are transparent. Brownish scales differ from each other since some are short and some are tall. Scale count are less on underneath when compared to the upperside. The transparency of the scales depends upon the intensity of the light they absorb. Overall, these slight variations in the scale size, length, structure and color are the basic concept behind the bright and pale coloration of ‘Tawny costers’. Key words: Morphology, Wing, Scales, Butterfly, Tawny costers Introduction characteristic of butterflies. The gorgeous color patterns in the wings are created by the Butterflies are famous for their magnificent microstructure of the scales. This physical beauty as they bear attractive wings of structure of coloration is the outcome of various colors of various shades. The scale- coherent scattering of light by the photonic covered wings provide coloration to crystal nature of the scales (Ding et al., butterflies which are an important Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

276 Current status and challenges for conservation and sustainable use of biodiversity 2009). There is no baseline information on comprising of 30 different scales from the the wing scales of butterflies so the present orange colored area, 24 scales from black, 24 study is focused on the scales of Acraea scales from hind wing termen and 21 scales terpsicore. from the underside respectively (Table 2, Fig.5). Examining at the orange coloured Materials and Methods region of the upper side exemplify several orange coloured scales. The orange scales The butterflies of Acraea terpsicore was were short with rounded terminus (Plate A1, collected from the premises of Sree Narayana Plate A2:1-30). The light micrograph showed College, Kollam (8˚52̍55N̎ 76˚364̍ ̎ E) by that the orange scales of the Acraea using handheld insect net. Orange, black and terpsicore consist of a more or less thin lower white colours were observed from the wings. lamina (adwing) which faces the wing The scales are dislodged from the wing substrate and a broad, round upper lamina surface as per the standard method of (abwing). Also, it contains a stalk at the base Grodnitsky and Kozlov (1991). Scales were of the lower lamina. Strikingly, the thickness shredded from each region of wing separately of the adwing or the lower lamina and the that shows varied colors into a glass slide. A abwing or the upper lamina varied among drop of xylene was used for fixation of these scales. The adwing and abwing shows scalesand the samples were studied under the variations in their length as well as thickness. light microscope. The measurements of the The abwing of some scales are more broader scales were calculated using micrometry. (Plate A1: 5,8,11,13,15,16,17) while some Microscopic photography was used for taking are less broader (Plate A1: 14,24 ). Often, the photomicrographs of the prepared scale some scales are longer (Plate A1: 14, 23) sample for result analysis. while some are shorter (Plate A1: 1, 6, 8). Results Here the orange colour is distributed throughout the abwing and adwing. The scale The upper side of the wing is tawny with color ranges from dark or bright orange to deep orange color while the underside is pale pale orange. The brightly colored scales tawny with pale orange color. Costers have (Plate A1:11) are more striking when elongate forewing and rounded hindwings. compared to some paler colored scales (Plate The forewings have several black spots. A1: 6). Repetition of these scales makes the Costal margin and termen are black. The butterfly wing brighter and attractive. The hindwing also possess black spots. The scales have a dimensions ranging from 70µ termen is edged with a black marginal band to 114µ in length and 47µ to 82µ in width. which has a series of small spots in white Some scales have similar length with color embedded. Underneath, the wings have different width (Plate A1:10, 11) and vice black markings as per the upper side. A series versa (Plate A1: 9,10). Moving to the second of large prominent white spots are embedded region, the black colored area, micrographs in the broad black terminal margin on the shows several black colored scales varying in hindwing. Wingspan ranges in between 1-3 shape and size and possesses slight variation cm for forewing and hindwing (Plate A). in color. On observing the black scales it indicates that majority of them are similar to The wings are scaled smoothly and they tend the orange scales with short black rounded to get rubbed off very easily. Tawny costers terminus and no dentations. Scales have an have a amazingly vibrant dorsal and a abwing, adwing and a stalk. The abwing or gloomy ventral side. A total of 99 different the upper lamina is more broader compared scales were identified from the wing Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

A comprehend in to the wing scales of butterfly Acraea terpsicore 277 to adwing or the lower lamina (Plate A2: 32, and other brown scales have no dentations 33, 35, 36, 38, 52, 54). Some scales are flat (Plate A3: 63,71). They are short when and short (Plate A2: 45,49) while some are compared to the former. The scale no:61 narrow and long (Plate A2: 44,52). The (Plate A3) has 4 buds or tooth on their intensity of the black color vary in each abwing and their adwing seems to be thinner scales. Some are more darker with black than the abwing. Similarly, scale no: 56 (Plate A2:32, 34) while some are brownish in (Plate A3) is long having 4 small blade like color (Plate A2: 42, 43, 48). The scale count buds or tooth. The white scales together with is less when compared to orange scales even the black and brown scales at the hindwing through, they are present in repetitive mode. termen help in the formation of bands. The scales have a dimensions ranging from Underneath, wing scales also possess scales 78µ to 102.44µ in length and 51.22µ to similar to the upper side of the wings. The 74.86µ in width. Length of some scales undersides of the Tawny coster wing are dull seems to be same but their width varies (Plate coloured unlike the upper side. Several pale A2: 33, 39). Some black scales also shows orange and black colored scales are obtained similar dimensions as that of the orange (Plate A4: 91, 92, 96). Scale no: scales. Black scale have similar length like 80,81,87,90,93 (Plate A4) are glassy type and the orange scale (Plate A2: 42, Plate A1: 5) are transparent. Brownish scales differ from and some have similar width (Plate A2: 35, each other since some are short and some are Plate A1: 21). Here the black colour is tall. Abwing flat and broad, becomes narrow uniformly distributed throughout the adwing towards the adwing. Scale no: 91, 92 and 96 and abwing. Hind wing termen of (Plate A4) shows a cluster of scales which is Tawnycoster have black bands which has a similar to the upper side wing scales. Scale series of small spots in white colour counts are less on underneath when embedded (Plate A). This region contains compared to the upper side. In some scales, numerous white scales, brown scales, and long straight lines can also be seen but often black scales that are having different not clear (Plate A4: 86). Glassy type dimensions. Each scale has an adwing, transparent scales cannot be clearly seen on a abwing and a stalk. White scales have narrow micrograph due to their high transparency adwing, broader and round abwing with no (Plate A4: 90, 93). The transparency of the dentations(PlateA3: 55,57,62,68,70,74,75,77) scales depends upon the intensity of the light except for two white scales where there they absorb. abwing are dentated and structured (Plate A3: 56,57). Even though, these scales appear Fig.1 describes the upper side orange colored similar, they vary in their dimensions. An area contributes the majority of the scales in isolate single brown scale are obtained (Plate Tawny coster. Also, the upper side scales are A3: 65) which too have structured abwing more in number when compared to the underside scales Table 1. The number of scales of ‘Tawny coster’ in the respective wing area. Wing Area Color No of Scales Upperside Orange 30 Upperside Black 24 Upperside hindwing termen Black band with white spots 24 Underside - 21 Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

278 Current status and challenges for conservation and sustainable use of biodiversity Fig.1 Number of scales in wing area No: of wing scales in 'Tawny coster' 35 Upperside Orange 30 Upperside black Upperside hindwing termen 30 Underside No: of scales 25 24 24 21 20 15 10 5 0 Underside Upperside Orange Upperside black Upperside hindwing termen Wing area PLATE A Acraea terpsicorem (Tawny coster) Orange Black Low Power View (10x) Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

A comprehend in to the wing scales of butterfly Acraea terpsicore 279 PLATE A1 High Power View (40x) Upperside Wing Scales 1. Orange 1 2 3 4 5 6 Length:78.8µ Length:86.68µ Length:110.32µ Length:90.6µ Length:90.62µ Length:106.38µ Width:47.28µ Width:59.1µ Width:66.98µ Width:63.0µ Width:66.98µ Width:82.74µ 7 8 9 10 11 12 Length:94.56µ Length:78.8µ Length:102.44µ Length:98.5µ Length: 98.5µ Length:110.32µ Width:70.92µ Width:59.1µ Width:70.92µ Width:70.92µ Width: 74.86 µ Width:66.98µ 13 14 15 16 17 18 Length:90.62 Length:98.5µ Length:110.32µ Length:82.74µ Length:86.68µ Length:102.44µ Width:74.86 Width:66.98µ Width:74.86µ Width:63.04µ Width:63.04µ Width:78.8 19 20 21 22 23 24 Length:90.62µ Length:102.44µ Length:110.32µ Length:102.44µ Length:114.26µ Length:106.38µ Width:66.98µ Width:70.92µ Width:70.92µ Width:63.04µ Width:47.28µ Width:59.1µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

280 Current status and challenges for conservation and sustainable use of biodiversity PLATE A2 25 26 27 28 29 30 Length:82.74µ Length:94.56µ Length:98.5µ Length:90.62µ Length:94.56µ Length:90.62µ Width:74.86µ Width:70.92µ Width:70.92µ Width:66.98µ Width:63.04µ Width:63.04µ 2- Black 31 32 33 34 35 36 Length:86.68µ Length:98.5µ Length:78.8µ Length:86.68µ Length:118.2µ Length:94.56µ Width:55.16µ Width:66.98µ Width:59.1µ Width:55.16µ Width:70.92 µ Width:63.04µ 37 38 39 40 41 42 Length:86.68µ Length:78.8µ Length:78.8µ Length:90.62µ Length:86.68µ Length:90.62µ Width:59.1µ Width:55.16µ Width:51.22µ Width:63.04µ Width:63.04µ Width:66.98µ 43 44 45 46 47 48 Length:86.68µ Length:90.62µ Length:98.5µ Length:102.44µ Length:86.68µ Length:90.62µ Width:66.98µ Width:59.1µ Width:74.86µ Width:63.04µ Width:59.1µ Width:59.1µ 49 50 51 52 53 54 Length:82.74µ Length:98.5µ Length:90.62µ Length:98.5µ Length:90.62µ Length:86.68µ Width:70.92µ Width:66.98µ Width:70.92µ Width:47.28µ Width:59.1µ Width:55.16µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

A comprehend in to the wing scales of butterfly Acraea terpsicore 281 PLATE A3 3. Hindwing Termen ( Black band with white spots) 55 56 57 58 59 60 Length:86.68µ Length:106.38µ Length:82.74µ Length:98.5µ Length:94.56µ Length:98.5µ Width:63.04µ Width:43.34µ Width:47.28µ Width:70.92µ Width:74.86µ Width:66.98µ 61 62 63 64 65 66 Length:106.38µ Length:102.44µ Length:82.74µ Length:86.68µ Length:102.44µ Length:102.44µ Width:55.16µ Width:70.92µ Width:59.1µ Width:63.04µ Width:70.92µ Width:51.22µ 67 68 69 70 71 72 Length:94.56µ Length:90.62µ Length:94.56µ Length:90.62µ Length:98.5µ Length:102.44µ Width:66.98µ Width:51.22µ Width:59.1µ Width:47.28µ Width:70.92µ Width:63.04µ 73 74 75 76 77 78 Length:82.74µ Length:86.68µ Length:94.56µ Length:82.74µ Length:106.38µ Length:94.56µ Width:47.28µ Width:51.22µ Width:59.1µ Width:47.28µ Width:51.22µ Width:66.98µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

282 Current status and challenges for conservation and sustainable use of biodiversity PLATE A4 Underside Wing Scales 79 80 81 82 83 84 Length:90.62µ Length:94.56µ Length:98.5µ Length:98.5µ Length:82.74µ Length:86.68µ Width:63.04µ Width:66.98µ Width:63.04µ Width:59.1µ Width:47.28µ Width:55.16µ 85 86 87 88 89 90 Length:90.62µ Length:82.74µ Length:90.62µ Length:86.68µ Length:94.56µ Length:82.74µ Width:51.22µ Width:47.28µ Width:59.1µ Width:47.28µ Width:55.16µ Width:51.22µ 91 92 93 94 95 96 Length:90.62µ Length:94.56µ Length:98.5µ Length:90.62µ Length:94.56µ Length:98.5µ Width:47.28µ Width:66.98µ Width:63.04µ Width:47.28µ Width:66.98µ Width:63.04µ 97 98 99 Length:102.44µ Length:98.5µ Length:102.44µ Width:70.92µ Width:59.1µ Width:66.98µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

A comprehend in to the wing scales of butterfly craea terpsicore 283 Discussion be the source of various technical applications such as photonic devices. In the case of Nymphalids, the butterflies Overall, these slight variations in the scale possess bright and dark brown colored scales size, length, structure and color are the basic as in case of ‘Euploea core’ and concept behind the bright and pale coloration ‘Melanitisleda’. In this category, the scales of ‘Tawny costers’. are being observed as having more or less pointed dentations. Some possesses orange Acknowledgement colored scales with rounded terminus which is fully devoid of dentations as in ‘Acraea The authors are grateful to the Principal, Sree terpsicore’. Often, black scales are also seen Narayana College, Kollam for providing as having dentations while some lack facilities to completing the work. dentations on the upper lamina as in ‘Acraea terpsicore’. The work of Stavenga (2014) Reference reported that the black and brown colors of the scales are due to the presence of pigment Ding, Y., Xu, S. and Z.L. Wang. 2009. ‘melanin. Minor amount of melanin leads to Structural colors from Morphopeleides the production of brown color. The scale butterfly wing scales. Journal of Applied colours and transparency depends upon the Physics, 106: 074702-074706. intensity of light. The typical colours of butterfly wing scales may be solely produced Grodnitzky, D. L. and M. V. Kozlov. 1991. from either micro- and nanoarchitectures, or Evolution and function of wings and their pigments, but most frequently from a scale covering in butterflies and moths combination of both (Kinoshita et al., 2002 (Insecta: Papilionida-Lepidoptera). Biol. and Wilt et al., 2015). Micrometric analyses Zent. Bl., 110:199–206. recorded slight variations in the dimensions of scales. Kinoshita, S., Yoshioka, S. and Kawagoe, K. 2002. Mechanisms of structural colour in the Conclusion Morpho butterfly: cooperation of regularity and irregularity in an iridescent scale. Proc. However, the brightly coloured scales have Biol. Sci., 269: 1417–1421. low transparency due to this fact that they are strikingly attracted to the eye. The wing Wilts, B. D., Matsushita, A., Arikawa, K. and coloration provides several functions such as Stavenga, D. G. 2015. Spectrally tuned mating, camouflage and warning purposes. structural and pigmentary coloration of Butterfly wing scale colour study is found to birdwing butterfly wing scales. Journal of The Royal Society Interface, 12:717. Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

284 Current status and challenges for conservation and sustainable use of biodiversity Stavenga, D. G., Leertouwer, H. L. and Wilts, B. D. 2014. Coloration principles of nymphaline butterflies - thin films, melanin, ommochromesand wing scale stacking. J. Exp. Biol., 217: 2171–2180. Current Status and Challenges for Conservation and Sustainable use of Biodiversity| 2020 | pp.275-284 © Principal, Sree Narayana College, Kollam, Kerala, India

285 A preview on the wing scales of danaus chrysippus using light microscopy ISBN 978-93-5396-871-7 CHAPTER 42 A PREVIEW ON THE WING SCALES OF DANAUS CHRYSIPPUS USING LIGHT MICROSCOPY Amina Thaj, MunishaMurali S* and Sheeba S PG and Research Department of Zoology Sree Narayana College, Kollam, India *Correspondence E-mail: [email protected] ABSTRACT The brush footed butterfly known as Danaus chrysippus or Plain Tiger/African Queen belongs to the family Nymphalidae and subfamily Danainae. The present investigation aimed to study the different patterns in the scales on the wing of Plain Tiger. The study is done by detaching the different coloured scales from the wing to a glass slide and made permanent using xylene to perceive using light microscope. The dimensions of the scales were observed using micrometry. A total of 159 distinctive types of scales were identified. Among them dorsal forewing area comprises of 27 scales from brown area followed by 22 scales from black and 12 scales from white. However, upperside hindwing area comprises of 18 scales from brown, 24 scales from black and 27 scales from white. The ventral area of the wings consists of 29 wing scales. The brown colored area of the upperside forewing includes several dark to pale brown coloured scales. Majority of the scales possesses abwing and adwing with a stalk. These scales also have dentations and some lack. The scale dimensions of this area range from 78.8µ-110.32µ in length and 63.04µ-90.62µ in width. The black coloured region on the dorsal side of the forewing has several black colored scales together with some odd white scales. The dimension of scale ranges from 78.8µ-110.32µ in length and 63.04µ- 78.8µ in width. The white spotted region on the upper side of forewing has dimensions ranging from 78.8µ-102.44µ in length and 59.1µ-78.8µ in thickness. The dorsal side of hindwing contains several brown, black and white colored scales. Brown colour region contains a total of 18 scales that differ from each other. The dimensions of these scales range from 86.68µ-98.5µ in length and 66.98µ- 74.86µ in thickness. The black colored region on the upperside of the hindwing area contains 24 varieties of scales. The black and white scales are intermixed in this region. The dimension of the scales ranges between 86.68µ-98.5µ in length and 63.04µ-74.86µ in width. The white scales of hindwing area possess more transparency than the dorsal part of the forewing. Several grey shaded scales have also been observed in this area. The dimension of white scales ranges between 82.74µ-106.38µ in length and 59.1µ-74.86µ in width. The ventral portion of the wings composed of several black colored, brown colored and white colored scales. The dimensions of the underside scales ranges between 82.74µ-98.5µ in length and 59.1µ-74.86µ in width. The exploration on the scales in the wings of butterfly provides knowledge on colouration pattern. Key words: Danaus chrysippus, Scales, Colouration pattern, Light microscope Introduction self-assembled photonic crystals. It is an ideal example, which can encourage us in design and In nature, a great number of organisms, for example, construction of new photonic structures, and also can some butterflies, beetles, and peacock, possesses serve directly as biotemplates to mimic those strikingly brilliant colors (Kinoshita et al., 2008). structures (Huang et al., 2006). A Danaus These structural colors, are not made from the chrysippus-mimicking nymphalid, is a typical pigments, but from their periodic nature-made example of the female-limited dimorphic mimics photonic crystals, or biophotonic materials. Butterfly (Nishida, 2017). This butterfly is among “poisonous wing scales pattern are outcome of such biologically Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India