In vitro Propagation, Phytochemical Investigations ...

International Journal of Medicinal Plants. Photon xxx (xxxx) xxx-xxx https://sites.google.com/site/photonfoundationorganization/home/international-journal-of-medicinal-plants Review. ISJN: 6672-4384: Impact Index: 3.12

International Journal of Medicinal Plants

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In vitro Propagation, Phytochemical Investigations and Biological Activities of an Endemic Medicinal Plant, Indian Red Wood (Meliaceae): A Review Kishore K. Chiruvellaa,b*, Arifullah Mohammedb,c, Chakradhar Thamminenid, Vikram Paritalac, Rama gopal Ghantab a

Department of Molecular Biosciences, Stockholm University, Sweden Division of Plant Tissue Culture, Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pradesh, India c Faculty of Agrobased Industry, Universiti Malaysia Kelantan, Jeli Campus, Locked bag-100, 17600, Jeli, Kelantan, Malaysia d Project Scientist, International Crop Research Institute for Semi Arid Tropics, Patancheru, Hyderabad, India b

Article history: Received: 24 April, 2014 Accepted: 01 May, 2014 Available online: xx xxxx, xxxx Keywords: Soymida febrifuga, terpenoids, mahogany, meliaceae, phytochemistry, bark Abbreviations: BA: 6, Benzyladenine, CM: Coconut Milk, 2-iP: 2isopentynyl Adenine, IBA: Indole-3-Butyric-Acid, IAA: Indole-3-Acetic-Acid, KN: kinetin, MS: Murashige and Skoog, STN: shoot tip necrosis Corresponding Author: Chiruvella K.K.* Postdoctoral Fellow Email: [email protected] Mohammed A. Lecturer Email: [email protected] Thammineni C. Scientist Email: [email protected] Paritala V. Masters Student Email: [email protected] Ghanta R. G. Professor [email protected]

Abstract This review focuses on cinchona/oak bark substitute, Soymida febrifuga (Meliaceae), also known as Indian red wood whose medical virtues are used for treating various health ailments in folk and traditional medicine. Here we review the ex situ conservation strategies for the mass multiplication of this endemic plant by micropropagation due to its difficulty in rooting of stem cuttings and high seedling mortality rates. Further we also discuss phytochemical investigations from plant parts led to the isolation of tetranortriterpenoids, flavonoids, limonoids, alkaloids, tannins and glycosides. These chemical constituents were reported to be cytotoxic and strikingly are antimalarial, anti-inflammatory, antioxidant, antidiabetic, antiallergic, antifungal, antiulcer, spasmolytic, insect antifeedant, antihelminthic, antibacterial and anticancer in action. Overall this review highlights and compiles the available information of Soymida febrifuga propagation, medicinal importance of secondary metabolites and their role in the treatment of various diseases. Citation: Chiruvella K.K., Mohammed A., Thammineni C., Paritala V., Ghanta R.G., 2014. In vitro Propagation, Phytochemical Investigations and Biological Activities of an Endemic Medicinal Plant, Indian Red Wood (Meliaceae): A Review. International Journal of Medicinal Plants. Photon xxx, xxx-xxx. All Rights Reserved with Photon. Photon Ignitor: ISJN66724384D6965xxxxxxxx

1. Introduction The mahogany (Meliaceae) family covers more than 50 genera with more than 1,000 species distributed in tropical and subtropical regions. In India, the family is represented by 17 genera and 72 species of which 12 species and 2 varieties i.e., approximately 18% are Ph ton

endemic to peninsular India. Mahoganies represent the most important species for the development of the forest industry in Asia, tropical Africa and Latin America. Economically the Meliaceae is the most

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important family of timber trees Swietenia, Cedrela species etc.,).

(eg.,

Trees of genus Swietenia are major afforested trees in tropical zones of the world, and their loss due to pest injuries is a serious problem. Amongst different meliaceae members, successful regeneration of plantlets in Azadirachta indica has been reported by several workers and the explants used were different and substantial work was done in other species (Table 1). Various phytochemicals were isolated from different parts of meliaceous members. Chemically, this family is characterized by synthesis of modified triterpenes known as limonoids. Here we review the distribution, description, plant propagation, phytochemicals and their implications in treatment of various diseases obtained from endemic medicinal tree, Soymida febrifuga. Soymida febrifuga (Roxb.) A. Juss. or Swietenia febrifuga Roxb. (Meliaceae) commonly called Chandravallabha (Sanskrit), Indian red wood, Bastarol cedar (English), Somi, Somidha, Sumi (Telugu) is an indigenous lofty deciduous medicinal tree (Fig.1) and monotypic genus endemic to India (Wealth of India, 1952). It is commonly found in dry forests of western peninsula and Indomalaysia. 1.1 Distribution Soymida is represented by one species from India and East tropical Africa. It is distributed in the dry forests of peninsular India right upto Kerala and also occurs in North west Central India down through peninsular Srilanka, Gujarat, Uttar Pradesh, Bihar and Central India extending northwards to Merwara, the Mirzapur hills and Chota Nagpur, Ceylon and Chengalpattu, Coimbatore, Dharmapuri, Nilgiri, Ramanathapuram, Salem, Tiruchchirapalli-South West India.(Wealth of India (1952); Yoganarasimhan (1996); Kirtikar and Basu (2003).However the plant is threatened in Gujarat and needs care for its revival. 1.2 Botanical Description A lofty deciduous tall tree, grows up to 22-25 mtrs in height and 2.5-3.0 mtrs in girth. Bark is very tough, exfoliating in large plates or scales. Leaves 22-45 cm, imparipinnate, long, crowded at the ends of branches, branchlets with persistent leaf bases. Flowers are bisexual, greenish white in large axillary or terminal panicles. Fruit is a black, obovoid, woody, septifragally 5-valved capsule (Fig.2). Ph ton

Seeds winged at both ends. Sapwood is whitish and heart wood dark blood red to reddish brown with silver streaks with an oily feel, without characteristic odour or taste (Kirtikar and Basu, 2003). 1.3 Ecological Sensitivity The plant is found on dry stony hills and on laterite soils. Prefers absolute maximum temperature ranging between 40° C and 46° C and minimum between 0° C and 10° C. Direct sowing on ridges on 30 cm height, is more successful than transplanting the nursery raised plants. Propagated by seeds and germination is best in porous, well-drained soil but are liable to damp off in badly drained damp soils. Seedling growth is slow and sensitive to frost and growth ceases during the cold season. Tree is subjected to defoliation by insect (Wealth of India, 1952). The tree shed leaves during winter. 1.4 Economic importance The tree has aromatic odour when cut. Bark is very tough and used as rope.The heartwood is dark brown in colour and is hard and durable. The wood is used in house building as posts, rafters and beams, for well work, plough shares, pestles and pounders and furniture. It is also well suited for carving and turnery, wooden flooring, for making furniture for frames and stiles and for framing lighter coloured panelling. Each forest division where species is plentiful can supply 20 - 100 tonnes of timber annually. Bark is also used in tanning and tanned leathers possess good color, good feel and fullness. A clear gum from the bark forms good adhesive mucilage and a strong red fibre from it is used for making ropes (Wealth of India, 1952). 2. Seed Germination Naturally the plant is propagated by seeds and seed germination is best in porous, welldrained soil but is liable to damp off in badly drained damp soils. Seeds are dispersed by wind. The felty cover over the seed acts as protection and also for absorption of moisture. Seed loses viability quickly. It can be raised better by directly sowing of the seed, or by planting out seedlings from a nursery. It is very slow growing, as the plant is drought resistant, it can grown in drought prone areas. Seedling growth is slow and sensitive to frost and growth ceases during the cold season. Attempts were made to understand the seed germination capabilities of S. febrifuga under in vivo and in vitro conditions (Chiruvella et al.,

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2014a). Various factors such as sterilants, type of media, hormones, sucrose concentration and seed viability influenced in vitro seed germination of Soymida. Rate of seed germination in vitro but not in vivo led to the raising of healthy seedlings. MS full strength medium containing sucrose was found to be effective for seed germination with well developed root system (Chiruvella et al., 2014a). These findings have implications for the conservation of recalcitrant seed germplasm. 3. In vitro Propagation Traditional multiplication of Soymida is limited by difficulty in rooting of stem cuttings, high seedling mortality rates, low seed viability period. Plant tissue culture of tree species has been progressing successfully from the past few years to overcome the above problems to a great extent and assessment of the regeneration potential of tissues for the forest tree improvement. Various protocols have been established by our group for mass propagation of S. febrifuga using mature nodes and aspetic seedling explants as outlined (Fig.3). Micropropagation has been established through both direct and indirect organogenesis using aseptic seedling explants (Chiruvella et al., 2011; 2014). Fig. 2 represents the schematic sketch of the in vitro propagation of S. febrifuga. Mature nodal explants showed maximum shoot multiplication on MS medium supplemented –1 –1 with 2.0 mgl BA + 0.2 mgl NAA, whereas explants of aseptic seedlings showed maximum shoot multiplication on MS medium –1 –1 supplemented with 3.0 mgl BA + 0.2 mgl NAA. Axillary buds of aseptic seedlings showed better organogenic response than mature node. The usage of aseptic seedlings for regeneration bypassed the step of explant preparation and simplified micropropagation. Highest numbers of shoots were obtained from cotyledonary nodal explants compared to other explants. Explants such as cotyledonary node, shoot tip and node also showed one stage method of indirect organogenesis whereas cotyledon, leaves and root only showed two stage method of indirect organogenesis (Chiruvella et al., 2014c). BA proved to be efficient for shoot multiplication, though kinetin showed better shoot elongation. Maximum frequency of shoot regeneration was noticed in explants inoculated vertically. Addition of CM enhanced the leaf expansion and also shoot length. Among the various Ph ton

media (MS, WPM and B5) employed, MS medium proved to be efficient for high frequency shoot regeneration and multiple shoot production. Phenotypic abnormalities were associated during In vitro propagation of Soymida febrifuga. High concentrations of cytokinins in cultures induced vitrification, fasciated shoots and development of albinos (Chiruvella et al., 2014b). However shoot tip necrosis was the major obstacle observed in proliferating cultures as well with shoots kept for rooting. We could eliminate this malady by calcium supply to MS medium, suggests shoot necrosis is a physiological disorder caused by calcium deficiency (Chiruvella et al., 2011). Overall micropropagation and callus culture protocols offer the possibility to use the organ/cell culture techniques for vegetative propagation and extraction of compounds throughout the year without any seasonal constraints. 4. Phytochemical Investigations Chemically, Meliaceae is characterized by the occurrence of the gedunin nucleus and modified triterpenes known as limonoids. Previous investigations of the various parts of the Soymida febrifuga led to the isolation of methyl angolensate, deoxyandirobin and two tetranortriterpenoids with a modified furan ring from the bark (Ambaye et al., 1971; Adesida and Taylor, 1972; Purushothaman and Chandrasekharan, 1974; Purushothaman et al., 1977) obtusifoliol and the flavonoids syringetin and dihydrosyringetin from the root heartwood (Pardhasaradhi and Sidhu, 1972) quercetin 3-0-rhamnoside and quercetin 3-0rutinoside from the leaves (Nair and Subramanian, 1975) and the tetranortriterpenoids febrifugin (Rao et al., 1978) and febrinins A and B (Rao et al., 1979) together with the flavonoids naringenin, quercetin, myricetin and dihydromyricetin from the heartwood (Rao et al., 1979), three new tetranortriterpenoids, epoxyfebrinin- B, 14,15dihydroepoxyfebrinin B and febrinolide together with deoxyandirobin, 17β-hydroxy-6αacetoxyzadiradione (Connolly et al., 1979) methyl angolensate and sitosterol from the fruits (Mallavarapu et al.,1984). The bark contained approximately 0.1% of methyl angolensate, identical with an authentic specimen (Adesida, 1971). These results are in agreement with botanical conclusion that Soymida is closely related to the African genus Khaya (Adesida and Taylor, 1972). The chemical structures are depicted in (Fig.4).

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Figure 1: (A) Natural habitat of S. Febrifuga (B, C) Inflorescence and fruits of S. febrifuga

Figure 2: Morphology of S. febrifuga (A) A twig with flowering, inset: flowers in close up (B) Immature fruit (C)Mature fruit about to dehisce (D,E) Fruits showing dehiscence (F) Winged seeds (G) Woody 5-celled septifragal capsule after complete seed dispersal (Chiruvella et al., 2014)

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Figure 3: A schematic diagram of In vitro propagation of Soymida febrifuga using mature node and aseptic seedling explants by direct and indirect organogenesis (Chiruvella et al., 2011; 2013; 2014).

Figure 4: Chemical structures of phytochemical active constituents of Soymida febrifuga

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5. Biological activities Ayurvedic properties like Rasa-Kasaya, Katu; Guna-Laghu, Roksa; Virya-Sita; Vipaka-Katu which are Stambhana, Vranaropana, Grahl, Paustika in action are well described for S. febrifuga (Joshi, 2000). This tree well known substitute for cinchona/oak bark. The medical virtues of S. febrifugafor treating intermittent fevers like cinchona bark substitute is well known from 1791 onwards (Pratik Chakrabart 2010).

Seed oil consists of linolenic, linoleic, oleic, palmitic and stearic acids, lupeol and sitosterol (Yoganarasimhan, 1996). The leaves were found to contain Quercetin-3-O-L-rhamnoside and 3-O-rutinoside (Rastogi and Mehrotra, 1993). Using HPTLC technique, Attarde et al.2008 have quantified and estimated betasitosterol, lupeol, quercetin and quercetin glycosides from leaflets and carried out pharmacognostic studies of Soymida febrifuga leaflets (Attarde et al.2010). Our successive hexane, ethyl acetate and methanol root callus extracts of S. febrifuga were tested for their phytochemical constituents (Fig.5). Preliminary phytochemical screening revealed the maximum diversity of chemical constituents in ethyl acetate extract. The ethyl acetate extract was found active against most of the pathogenic microorganisms tested as they showed potential phytochemical constituents. Hence this extract was subjected to silica gel column chromatography. From ethyl acetate callus extract two compounds, methyl angolensate and luteolin-7-O– glucoside were isolated and separated (Fig.5) (Chiruvella et al., 2007). Phytochemical investigation led to the isolation of 27 compounds including four new compounds [(3R)-6,4'-dihydroxy-8methoxyhomoisoflavan, (2R)-7,4'-dihydroxy-5methoxy-8-methylflavan,7-hydroxy-6-methoxy3-(4'-hydroxybenzyl) coumarin and 6-hydroxy7-methoxy-3-(4'-hydroxybenzyl) coumarin (Awale et al.,2009). Two novel phragmalin type limonoids have been isolated from the bark of Soymida febrifuga (Yadav, 2012). Phytochemical screening of root bark showed the presence of alkaloid, tannins, glycosides etc chemical constituents (Palei, 2013).

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5.1 Bark Bark contains a resinous bitter principle and is acrid, refrigerant, antihelminthic, aphrodisiac, laxative; good for sore throat; removes vata; cures tridosha fevers, cough, asthma (Kirtikar and Basu, 2003). Bark crushed with water and administered for cough and dysentery (Murthy et al. 2007;Prayaga Murty Pragada 2012). Bark is used in Ayurveda to remove blood impurities; good for ulcers, leprosy, dysentery and anti-inflammatory in action (Diwan and Singh, 1993).Decoction of stem bark (½ cup) is given orally in snake bite (Chiranjibi Pattanaika et al., 2008). Fresh or dried bark is boiled in water to make paste, it is tied on swelling for 3 days. The bark is credited with antiperiodic properties, astringent to the bowels and useful in fevers (Unani), bitter tonic, febrifuge, particularly prescribed in malaria. The decoction of the bark is well adapted for gargles, vaginal infections, enemata, rheumatic swellings and stomach pain. The bark is said to be used for blood coagulation, wounds, dental diseases, uterine bleeding and haemorrhage (Yoganarasimhan, 1996).The powdered bark is applied as a poultice and is used for leucorrhoea and leucoderma (Maurya and Dongarwar, 2012). Remarkably S. febrifiuga is well known to treat fever/malaria among 80 plants analyzed for antiplasmodial activity (Simonsen et al., 2001; Kantamreddi and Wright, 2012). Decoction of inner bark possess in vivo antitumour activity against transplantable rat carcinosarcoma and rat yoshida sarcoma (ascites) (Ambaye, 1971). Extract of Soymida febrifuga were cytotoxic and was found to kill human pancreatic cancer cells (Awale, 2009; Balachandran and Govindarajan, 2005). Antioxidant, 5-lipoxygenase inhibitory and anticancer activities of the bark extracts were recently reported (Karunasree, 2012). Further recent reports also demonstrate that bark extracts were found to be hypoglycemic and

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Figure 5: A schematic of phytochemical and antimicrobial studies of S. febrifuga root callus (Chiruvella et al., 2007)

Table 1: In vitro propagation of Meliaceae members Plant Explant Results Mature embryo Organogenesis Stem bark Stem, leaf, Axillary buds Leaf Stem bark Cotyledons /leaf/ Stem Cotyledon

Azadirchta indica

Leaf Cotyledon Nucellar tissue and cotyledon Axillary buds Axillary buds Anther Immature embryo Axillary buds Cotyledons or Hypocotyl Leaflets, Rachis, Cotyledon, Inflorescence axis Dried cells Embryo, leaf and ovary Leaf Axillary buds Axillary buds of mature trees Immature embryos

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Organogenesis Organogenesis Organogenesis Organogenesis Callus Morphogenesis

References Rangaswamy and Promila, 1972 Sanyal et al., 1981 Jaiswal and Narayan, 1983

Morphogenesis Embryogenesis Embryogenesis

Narayan and Jaiswal, 1985 Sanyal and Datta, 1986 Rao et al., 1988 Muralidharan and Mascarenhas, 1989 Ramesh and Padhya, 1990 Shrikhande et al., 1993 Joarder et al., 1993

Plantlets Plantlets Organogenesis Plantlets Plantlets Somatic embryogenesis Indirect organogenesis

Joshi and Thengane,1993 Drew, 1993 Gautam et al., 1993 Thiagarajan and Murali, 1993 Joshi and Thengane, 1996 Su et al., 1997 George and Kulkarni, 1997

Azadirachtin Azadirachtin Morphogenesis Morphogenesis/Embryoge nesis Plantlets Azadirachtin and Nimbin

Jarvis et al., 1997 Singh and Chaturvedi, 2013 Eeswara et al., 1998 Murthy and Saxena, 1998 Venkateshwarulu et al., 1998 Srividya et al., 1998

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Callus cultures Seeds

Azadirachta excelsa

Callus cultures leaf, hypocotyl, epicotyl, cotyledons, root Nodes, apical meristem Callus cultures Flower bud

Azadirachtin production Callus induction

Veeresham et al., 1998 Venkateswarlu Mukhopadhyaya, 1999 Babu and Nair, 2004 Srikanth et al., 2006

Micropropagation Azadirachtin production Oraganogenesis

Biswas and Gupta, 2007 Sujanya et al., 2008 Srivastava et al., 2009

Plant cells Axillary buds Leaf

biotransformation Micropropagation indirect organogenesis, embryogenesis Direct organogenesis

Saifullah et al., 2013 Liew and Teo, 1998 van der Esch and Stephen et al., 2001 Chin Chiew Foan and Rofina Yasmin Othman , 2006 Nunes et al., 2002

Leaf Nodal segments of aseptic seedlings Axillary bud, seeds

Cedrela fissilis Cotyledonary and epicotyl nodes,Zygotic seeds Zygotic embryo Zygotic embryo Twigs, fruits Cedrela odorata

production Azadirachtin content Azadirachtin comparision

Seeds, Buds Nodes, seeds

Direct organogenesis

and

Germplasm conservation, Plant retrieval, Seed Cryopreservation Micropropagation

Nunes et al., 2003

Somatic embryogenesis Embryogenesis Micropropagation, Embryogenesis Seed germination, bud sprouting Morphogenesis

Vila et al., 2009 Stewart and Cameron, 2010 Yuri Peña-Ramírez et al., 2010 Valverde-Cerdas et al., 2008

Nunes et al., 2007

Immature embryo

Embryogenesis

Cedrela Montana

Trees and juvenile plants Seeds

Cedrela sinensis Khaya grandifoliola

Stem segments

Organogenesis Clonal Propagation and Germplasm Conservation Embryogenesis

Rolando García-Gonzáles et al., 2011 Yuri Peña-Ramírez et al., 2011 Sofia Basto et al., 2012 Gabriela Díaz-Quichimbo et al., 2013 Choi et al., 1986

Seed embryo

Micropropagation

Okereand Adegeye, 2011

Axillary buds of mature trees Internode of mature trees Hypocotyl segments

Plantlets organogenesis In vitroPrecocious Flowering Plantlets Direct and Indirect organogenesis In vitro storage or conservation Indirect organogenesis Somatic embryogenesis Direct organogenesis Clonal Propagation Embryogenesis Organogenesis, Micropropagation In vitro propagation High frequency of shoot regeneration

Dhingra et al., 1991 Ahmad et al., 1990 Alice Sato and Maria Apparecida Esquibel, 1995 Thakur et al., 1998 Shahzad and Siddiqui, 2001

Melia azedarach

Axillary buds Mature nodal segments Apical meristem tips Leaves Seeds Mature node Immature seeds Immature zygotic embryos Axillary buds

Melia dubia Melia volkensii Munoria pinnata

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Mature seedlings Zygotic embryos

Apical buds, nodal segments, petioles, floral parts, leaf discs

In vitro mass propagation

Scocchi and Mroginski, 2004 Vila et al., 2004 Sharry et al., 2006 Hussain and Anis, 2009 Antara Sen et al., 2010 Vila et al., 2010 Mroginski and Rey, 2013 Bhimi Ram et al., 2012 Mulanda et al., 2012

Senerath et al., 2007

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Munronia pumila

Swietenia macrophylla

Leaf, petiole, seed, floral parts Axillary buds Nodal explants Mature node Seed germination Seedling explants Seedling explants

Soymida febrifuga

Immature cotyledons Juvenile shoots Mature node, seedling explants, cotyledonary node, shoot tip, cotyledon and leaf

Indirect organogenesis

Hirimburegama et al., 1994

Organogenesis Organogenesis Somatic Embryogenesis epicotyl

Lee and Rao, 1988 Tacoronte et al., 2004 Maruyama and Ishii, 1999 Juliana Margarido Fonseca Couto, 2004 Maruyama 2006

Shoot tip culture Somatic embryognesis Nodal culture Somatic embryogenesis Direct organogensis Direct and indirect organogenesis, abnormalities

antihyperglycemic suggesting its mode of action as hepatoprotective and antidiabetic nature (Karunasree, 2013). Strikingly, decoction of stem bark is used to increase sexual vitality in women and mixture along with sugar cane juice is used to regularize menstruation (Jain A,Katewa et al., 2004). Bark decoction (50ml, daily 2 times) significantly reduced the CPK levels in two patients suffering from DMD (Breton, 1821). 5.2 Root Callus extracts of S. febrifuga root was found to exhibit growth inhibition. Pronounced activity is observed against Aspergillus niger and Alternaria rather than Aspergillus fumigatus by ethyl acetate extract. Methyl angolensate isolated from the former extract showed effect on Bacillus while luteolin-7-Oglucoside showed considerable growth inhibition of Salmonella typhimurium. MA displayed the maximum zone of inhibition against Aspergillus niger while the luteolin-7O-glucoside exhibited maximum zone of inhibition on Alternaria alternata. MA is tetranortriterpneoid isolated from root callus extracts of S.febrifuga and from other members of meliaceae (Fig.5) (Chiruvella et al., 2007). MA is known to be antiinflammatory (Thioune et al., 1994), antiulcer (Njar et al., 1995), antimalarial (Bickii et al., 2000; Kantamreddi et al., 2009), spasmolytic (Orisadipe et al., 2001), antiallergic (Penido et al., 2006), insect antifeedant (Samir et al., 2001), sedative in nature (Amos et al., 2002), antifungal (Abdelgaleil et al., 2005) properties. Strikingly it is shown that MA is cytotoxic and induces apoptosis in various cancer cells (Chiruvella et al., 2008; 2010; 2011b). Root is used for cough, cold (Bahraich) (Vedavathy et al., 1997).

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Eliane de Souza Schottz et al., 2007 Raúl Collado et al., 2010 Mona, 2012 Chiruvella et al., 2011; 2013;2014

5.3 Flowers Flowers along with fruit juice are used to cure earache (Vedavathy et al., 1997). Flowers along with leaves of Ocimum tenuiflorum, fruits of Embelia ribes,a pinch of asafoetida and about 5-10 drops of castor oil is given for relief from headache due to constipation (Vedavathy et al., 1997). 5.4 Leaves Leaf extracts were found to be potent antioxidant, antimicrobial, antihelminthicand anti-ulcerogenic potential (Reddy et al., 2008; Prasad and Kumar, 2012; (Gangurde et al., 2008)). Extracts of leaf, bark, root and root bark induced oxidative damage in HepG2 cells (Reddy et al., 2008). Ethanobotanical studies have shown S. febrifuga in the management of leucorrhoea, menorrahagia and dysmenorrheal muscular dystrophies (DMD). Recently it has been reported against periodontal disease causing microorganisms (Moon et al., 2013). Leaf juice is used to control over bleeding in menstruation (Vedavathy et al., 1997). Conclusion We believe this combination of insights would provide information of S. febrifuga containing various secondary metabolites which might be effective in controlling infectious diseases. Besides it provides as good source of timber and afforestation programmes. It is crucial to develop biotechnological methods to improve the cultivation of S. febrifuga to minimize the pressure on wild plants. This review might be of great interest for researchers for further studies in principle bioactive compounds which have not been investigated for the invention of their potential pharmacological benefits.

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Acknowledgements We thank Gayathri Dampuri for critical reading and help. The authors are highly grateful to the UMK for their logistical support under Grant No. R/SGJP/ A07.00/00710A/001/2012/000081). Conflicts of Interest The authors declare no conflicts of interest References Anita Jain S.S., Katewa P.K., Pallavi Sharma., 2005. Medicinal plant diversity of Sitamata wildlife sanctuary, Rajasthan, Indian Journal of Ethnopharmacology, 102(2), 143–157. Attarde D.L., Chaudhari B.J., Kale S.S., Bhamber R.S., Pal S.C., 2010. Pharmacognostic Studies on leaflets of Soymida febrifuga Adr. Juss. Family: Meliaceae. Journal of Pharmacy Research, 3(10), 2435-2440. Ambaye R.Y., Indap M.A., Panse T.B., 1971. Identification of methyl angolensate in the bark of Soymida febrifuga (Roxb.) A. Juss. Current Science, 7: 158. Adesida G.A and Taylor D.A.H., 1972. Extractives from Soymida febrifuga. Phytochemistry, 11, 15201524.

Anonymous., 1952. The Wealth of India. Publication and Information Directorate CSIR, New Delhi 9, 471-472. Alice S., and Maria A.E., 1995. In Vitro precocious flowering of Melia Azedarach hypocotyl segments. Revista Brasileira de Fisiologia Vegetal, 7(1), 107110. Antara S., Amla B., Rao D.V., 2010. Pivotal role of plant growth regulators in clonal propogation of Melia azedarach L. International Journal of Pharmaceutical Sciences Review and Research, 5(2), 43-49. Ahmad Z., Zaidi N., Shah F.H., 1990. Micropropagation of Melia azedarach from mature tissue. Pakistan Journal of Botany, 22, 172-178. Babu V.S., and Nair G.M., 2004. Bioproduction of Azadirachtin from Callus Cultures of Neem (Azadirachta indica A. Juss.), IUPAC: International Conference on Biodiversity and Natural Products Chemistry and Medical Applications, New Delhi, held on 26-31, Jan, p. 52. Bhimi R., Trilok S.R., Gillela R.S.R., 2012. In vitro propagation of Melia dubia Cav. from seedling explants. Biotechnology, Bioinformatics and Bioengineering, 2(1), 610-616. Balachandran P., Govindarajan R., 2005. Cancer ayurvedic perspective. Pharmacological Research, 51: 19–30.

Adesida G.A., Adesogan E.K., Okorie D.A., Taylor D.A.H., 1971. The limonoid chemistry of the genus Khaya (Meliaceae). Phytochemistry, 10, 1845-1853.

Bickii J., Njikam N., Foyere J.A., Basco L.K., Ringwald P., 2000. In vitro antimalarial activity of limonoids from Khaya grandifolia. Journal of Ethnopharmacology, 69, 27-33.

Attarde D., Aurangabadkar V., Belsare D., Pal S., 2008. Quantitative estimation of beta-sitosterol, lupeol, quercetin and quercetin glycosides from leaflets of Soymida febrifuga using HPTLC technique. Pakistan Journal of Pharmceutical Sciences, 21(3), 316-319.

Biswas B.K., Gupta S.C., 2007. Micropropagation of EliteNeem Tree (Azadirachta indica A. Juss.).S.M. Jain and H. Häggman (eds.), Protocols for Micropropagation of Woody Trees and Fruits, pp, 345–359.

Awale S., Miyamoto T., Linn T.Z., Li F., Win N.N., Tezuka Y., Esumi H., Kadota S., 2009. Cytotoxic constituents of Soymida febrifuga from Myanmar. Journal of Natural Products, 72(9),1631-1636. Ashok Yadav P., Suresh G., Rajendra Prasad K., Suri Apparao M., Suresh Babu K., 2012. New phragmalin-type limonoids from Soymida febrifuga. Tetrahedron Letters, 53(7), 773–777. Abdelgaleil S.A., Hashinaga F., Nakatani M., 2005. Antifungal activity of limonoids from Khaya ivorensis. Pest Management Science, 61, 186-90. Amos A., Orisadipe A., Binda L., Emeje M., Okogun J., Akah P., 2002. Behavioural effects in rodents of methyl angolensate: a tetranortripernoid isolated from Entandrophragma angolense. Basic Clinical Pharmacological Toxicology, 91, 71-76.

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Breton P., 1821. On the Efficacy of the Bark of the Swietenia Febrifuga, as a Substitute for that of the Cinchona. Medico-Chirurgical Transactions, 11(2), 310–329. Chiranjibi P., Sudhakar Reddy C., Murthy M.S.R., 2008. An ethnobotanical survey of medicinal plants used by the Didayi tribe of Malkangiri district of Orissa, India. Fitoterapia 79, 67–71. Chiruvella KK., Mohammed A., Gayathri D., Ghanta R.G., Raghavan S.C., 2007. Phytochemical and antimicrobial studies of methyl angolensate and luteolin-7-0-glucoside isolated from callus cultures of Soymida febrifuga. International Journal of Biomedical Science, 3,269-278. Chiruvella K.K., Kari V., Choudhary B., Nambiar M., Ghanta R.G., Raghavan S.C., 2008. Methyl angolensate, a natural tetranortriterpenoid induces

557

intrinsic apoptotic pathway in leukemic cells, FEBS Letters, 582, 4066-4076. Chiruvella K.K., Kuppusamy P., Choudhary B., Joy O., Raghavan S.C., 2010. Methyl angolensate from callus of indian redwood induces cytotoxicity in human breast cancer cells, International Journal of Biomedical Science, 6,182-194. Chiruvella K.K., Raghavan S.C., 2011. A natural compound, methyl angolensate, induces mitochondrial pathway of apoptosis in Daudi cells. Investigational New Drugs, 4,583-92. Chiruvella K.K., Mohammed A., Ghanta R.G., 2013. Utilization of aseptic seedling explants for in vitro propagation of Indian red wood. Notulae Scientia Biologicae, 5, 1-6. Chiruvella K.K., Mohammed A., Ghanta R.G., 2014a. Factors influencing the seed germination of Soymida febrifuga (Roxb.) A. Juss. (Meliaceae). Trakia Journal of Biosciences, 13(2), 121-131. Chiruvella K.K., Mohammed A., Ghanta R.G., 2014b. Phenotypic aberrations during micropropagation of Soymida febrifuga (Roxb.) Adr. Juss. Notulae Scientia Biologicae, 6(1), 99-104. Chiruvella K.K., Mohammed A., Gayathri D., Ghanta R.G., 2011a. In vitro shoot regeneration and control of shoot tip necrosis in tissue cultures of Soymida febrifuga (Roxb.) A. Juss. Plant Tissue Culture & Biotechnology, 21(1), 11-25. Chiruvella K.K., Mohammed A., Gayathri D.,Ghanta R.G., 2014c. In vitro callus induction and plant regeneration in Indian red wood, Soymida febrifuga A. Juss (Roxb.) Journal of Medicinal Plants Research, 2014 Chin C.F., Rofina Y.O., 2006. In vitro direct shoot organogenesis and regeneration of plantlets from leaf explants of eentang (Azadirachta excelsa). Biotechnology, 5, 337-340. Dhingra M., Sujatha M., Ranganatha A.R.G., 1991. Rapid multiplication of Melia azedarach Linn. through tissue culture. Journal of Oil Seeds Research, 8, 215-219. Drew R.A., 1993. Clonal propagation of neem by tissue culture. Paper presented at the World Neem Conf., February 24-28, Bangalore. pp. 999-1005. Paper Presented at the World Neem Conf., Feb. 2428, Banglore. Diwan P.V., Singh A.K., 1993. Anti-inflammatory activity of Soymida febrifuga (Mansa rohini) in rats and mice from bark powders. Phytotherapy Research 7(3), 255-256. Eeswara J.P., Stuchbury T., Allan E.J., Mordue A.J., 1998. A standard procedure for the micropropagation of neem tree (Azadirachta indica A. Juss). Plant Cell Reports, 17, 215-219.

Ph ton

Eliud S.M., Mark O.A., Nelson O.A., Elijah A., Jenesio I.K., 2012 High frequency regeneration of the drought tolerant tree Melia volkensii gurke using low-cost agrochemical thidiazuron. Biotechnology Research International, doi:10.1155/2012/818472. Eliane S.S., Antonio N.K.F., Anderson L.T., Henrique K., Luciana L.. Ribas F., Marguerite Q., 2007. In vitro multiplication of Swietenia macrophylla King (Meliaceae) from juvenile shoots. Ciência Florestal, Santa Maria, 17(2), 109-117. Gautam V.K., Nanda K., Gupta S.O., 1993. Callus induction and plant regeneration from anthers of Azadirachta indica. Plant Cell Tissue Organ Culture, 34, 13-18. George I.A., Kulkaarni A.R., 1997. Micropropagation of Neem- Azadirachta indica A Juss. Annals of Forestry, 50, 50-54. Gabriela D.Q., Ruth P.A., Julia M.P., Darlin G.Z., Consuelo R.I., Guillermo E.D.P., 2013. In vitro clonal propagation and germplasm conservation in the tropical timber tree spanish white cedar (C. Montana Moritz Ex Turcz.) (Meliaceae). eJournal of Biological Sciences, 7 (1), 59-69. Gangurde S.A., Pal S.C., Yeole D.U., Wagh A., Potawale S.E., Deshmukh R.S., 2008. In vitro evaluation of antioxidant and antihelminthic activity of different extracts of Soymida febrifuga. Pharmacologyonline, 2, 726-732. Hirimburegama K., Seneviratne A.S., Gamage N., Ganegama J., 1994. In vitro propagation of Munronia pumila (Binkohomba). Journal of the National Science Council of Sri Lanka, 22, 253-260. Hussain M.K., Anis M., 2009. Rapid in vitro multiplication of Melia azadarach L. (a multipurpose) woody tree. Acta Physiologiae Plantarum, 31, 765-772. Joshi S.G., 2000. Medicinal Plants, Oxford and IBH publishing Co. Pvt. Ltd., New Delhi p. 263. Jain A., Katewa S.S., Chaudhary B.L., Galav P., 2004. Folk herbal medicines used in birth control and sexual diseases by tribals of southern Rajasthan, India. Journal of Ethnopharmacology, 90(1), 171–77. Joarder N., Islam R., Joarder O.I., 1993. Micropropagation of Azadirachta indica through axillary bud culture. Paper Presented at the World Neem Conf., Feb. 24-28, Bangalore pp961-966. Joshi M.S., Thengane S.R., 1993. Potential application of in vitro methods for propagation of neem, Paper presented at the World Neem Conf., Feb. 24-28, Bangalore. Joshi M.S., Thengane S.R., 1996. In vitro propagation of Azadirachta indica A. Juss. (neem) by shoot proliferation. Indian Journal of Experimental Biology, 34, 480-482.

558

Comment [P1]: Volume/ serial no./ Page no. Should be edit

Jaiswal V.S., Narayan P., 1983. Regeneration of plantlets from the somatic tissue of some Indian trees. In: Proc Natl Symp Adv Front Plant Sci, Jodhpur. Journal of Experimental Biology, 22, 138139. Jarvis A.P., Morgan E.D., Vanderesch S.A., Vitali F., Ley S.V., Pape A.,1997. Identification of azadirachtin in tissue-cultured cells of Neem (Azadirachta indica). Natural Product Letters, 10, 95-98. Juliana M.F.C., Wagner C.O., Antônio L.P., Ésio F., 2004. In vitro sterilization and germination of mahogany seeds (Swietenia macrophylla King). Revista Árvore, 28(5), 633-645. Kantamreddi V.S., Wright C.W., 2012. Screening Indian plant species for antiplasmodial propertiesethnopharmacological compared with random selection. Phytotherapy Research, 26(12), 1793-99. Kirtikar K.R., Basu B.D., 2003. Indian Medicinal Plants. Oriental Enterpri,Dehradun, 1,2,559-560, 778-780. Kantamreddi V.S., Parida S., Kommula S.M., Wright C.W., 2009. Phytotherapy used in Orissa state, India for treating malaria. Phytotherapy Research, 23, 1638–1641. Karunasree V., Veeresham C., Rao K.,, Asres K., 2012. Evaluation of antidiabetic actity of column fractions obtained from bark extract of Soymida febrifuga. Pharmacognosy Journal, 4 (31), 37-43. Kantamreddi V.S., Lakshmi Y.N., Kasapu V.V., 2010. Preliminary phytochemical analysis of some important Indian plant species. International Journal of Pharma and Biological Sciences 1(4), 35-58. Lee S.K., Rao A.N., 1988. Plantlet production of Swietema macrophylla King through tissue culture. Gardens’ Bulletin, 41, 11-18. Liew T.K., Teo C.K.H., 1998. Multiple shoot production in vitro of the tropical timber tree, Sentang (Azadirachta excels Linn.). Horticultural Science, 33(6), 1073-1075. Luis A., Mroginski., Hebe Y.R., 2013. Micropropagation of paradise tree (Melia azedarach) by in vitro culture of axillary buds. Maurizio Lambardi et al. (eds.), Protocols for Micropropagation of Selected EconomicallyImportant Horticultural Plants. Methods in Molecular Biology, vol. 994, DOI 10.1007/978-1-62703-0748_17, New York.

Murthy B.N.S., Saxena P.K., 1998. Somatic embryogenesis and plant regeneration of neem (Azadirachta indica A. Juss.). Plant Cell Reports, 17, 469-475. Muralidharan E.N., Mascarenhas A.F., 1989. In Vitro Morphogenesis in Azadirachta indca A. Juss. And Eucalyptus citriodora Hook, In: Tissue culture and biotechnology of medicinal and aromatic plants. (eds.) Kukreja AK, Mathur AK, Ahuja PS and Thakud RS, Paramoam Pub, New Delhi. pp 49-55. Maruyama E., 2006. Tissue culture of Swietenia macrophylla King (Big-Leaf Mahogany)”, In: Plantation technology in tropical forest science, K. Suzuki, K. Ishii, S. Sakurai, and S. Sasaki (eds.), Springer-Verlag, Tokio, Japan, pp. 131-136. Murthy E.N., Reddy C.H.S., Reddy K.N., Raju V.S., 2007. Plants used in ethnoveterinary practices by koyas of pakhal wildlife sanctuary. Ethnobotanical Leaflets, 11, 1-5. Ninad M., Prasad S.S.V., Guduguntla S., Rajender A., Archana M., Srilakshmi S., 2013.The multidrug resistance of peridontitis causing microorganisms with phytochemicals derived from Soymida febrifuga. Journal of Research and Advancement in Dentistry, (2)3, 13-21. Njar V.C., Adesanwo J.K., Raji Y., 1995. Methyl angolensate: The antiulcer agent of the stem bark of Entandrophragma angolense. Planta Medica, 61: 91-92. Narayan P., Jaiswal V.S., 1985. Plantlet regeneration from leaflet callus of Azadirachta indica Juss. Journal of Tree Science, 4, 65-68. Nunes E.C., Castilho C.V., Moreno F.N., Viana A.M., 2002. In vitro culture of Cedrela fissilis Vellozo. (Meliaceae). Plant Cell Tissue Organ Culture, 70: 259-268. Nunes E.C., Laudano W.L.S., Moreno F.N., Castilho C.V., Mioto P., Sampaio F.L., Bortoluzi J.H., Benson E.E., Pizolatti M.G., Carasek E., Viana A.M., 2007. Micropropagation of Cedrela Fissilis Vell. (Meliaceae). S.M. Jain and H. Häggman (eds.), Protocols for Micropropagation of Woody Trees and Fruits, 221–235. Nunes E.D., Erica E.B., Ana C.O., Patricia S.A., Juliana R.M., Ana M.V., 2003. In vitroconservation of Cedrela fissilis Vellozo (Meliaceae), a native tree of the Brazilian Atlantic Forest. Biodiversity and Conservation, 12, 837–848.

Maruyama E., Ishii K., 1999. Somatic embryogenesis in big-leaf mahogany (Swietenia macrophylla king). Forestry Sciences, 59, 45-62.

Okere A.U., Adegeye A., 2011. In vitro propagation of an endangered medicinal timber species Khaya grandifoliola C. Dc. African Journal of Biotechnology, 10(17), 3335-3339.

Mona A.A., 2012. In vitro propagation of Swietenia macrophylla king. Research Journal of Agriculture and Biological Sciences, 8(2), 282-287.

Orisadipe A., Amos S., Adesomoju A., Binda L., Emeje M., Okogun J., Wambebe C., Gamaniel K., 2001. Spasmolytic activity of methyl angolensate: a

Ph ton

559

triterpenoid isolated from Entandrophragma angolense. Biologia Pharma Bulletin 24, 364-367. Prayaga M.P., Duvvada S.R., Malleboyina V., 2012. Study of some ethnomedicinal plants for treatment of dysentery of North Coastal Andhra Pradesh, India. International Journal of Biosciences, 2 (1), 18-24. Penido C., Costa K.A., Costa M.F., Pereira J.F., Siani A.C., Henriques M.G., 2006. Inhibition of allergen-induced eosinophil recruitment by natural tetranortriterpenoids is mediated by the suppression of IL-5, CCL11/eotaxin and NFkappaB activation. International Immunopharmacology, 6, 109-21. Palei A.K., Harisha C.R., Shukla V.J., 2012. Detailed pharmacognostical and phytochemical investigation on Soymida febrifuga Roxb. (Root bark). International Journal of Pharmacology & Biology Archives, 3(5), 1180-83.

Rangaswamy N., Promila P., 1972. Morphogeneis of adult embryo of Azadirchta indica A. Juss. Z. Pflazenphysiol, 67, 377-379. Rao M.V., Rao Y.V., Rao Y., Manga V., 1988. Induction and growth of callus inAzadirachta indica A. Juss. Crop Improvement, 15(2), 203–205. Ramesh K., Padhya M.A., 1996. In vitro propagation of neem, Azadirachta indica (A. Juss.), from leaf discs. Indian Journal of Experimental Biology, 28, 932-935. Raúl C., Raúl B., Daniel A., Felipe J.T., Martha P.O.G., 2010. Indirect somatic embryogenesis of Swietenia macrophylla King in semisolid culture medium. Biotecnología Vegetal, 10 (3), 177-84. Rolando G.G, Miladys D., Yailín G., Aníbal G., Miguel G., Peter D.S., Caligari, Basilio C., Karla Q., 2011. In vitro propagation of cedar (Cedrela Odorata L.) from juvenile shoots. Chilean Journal of Agricultural Research, 71(3), 376-382.

Padal S.B., Vathi K., Sathya, Sandhyasri B., 2013. Ethnomedicinal survey for important plants of gudem Kotha Veedi Mandalam, Visakhapatnam district, Andhra Pradesh, India. ZENITH. International Journal of Multidisciplinary Research, 3 (6), 175-181.

Simonsen H.T., Nordskjold J.B., Smitt U.W., Nyman U., Palpu P., Joshi P., Varughese G.G., 2001. In vitroscreening of Indian medicinal plants for antiplasmodial activity. Journal of Ethnopharmacology, 74, 195-204.

Pardhasaradhi M., Sidhu G.S., 1972. Obtusifoliol, syringetin and dihydrosyringetin from Soymida febrifuga. Phytochemistry, 11: 1520-1522.

Samir A.M., Abdelgaleil O.H., Iwagawa T., Sato A., 2001. Antifeedants from Khaya senegalensis. Tetrahedron, 57, 119-126.

Purushothaman K.K., Chandrasekharan S., 1974. Occurrence of methyl angolensate and deoxyandirobin in Soymida febrifuga A.Juss. Indian Journal of Chemistry, 12, 207-208.

Singh M., Chaturvedi R., 2013. Sustainable production of azadirachtin from differentiated in vitro cell lines of neem (Azadirachta indica). Annals of Botany PLANTS, 5, plt034; doi:10.1093/aobpla/plt034.

Purushothaman K.K., Chandrasekharan S., Conolly J.D., Rycroft D.S., 1977. Tetranortriterpenoids and related substances. New tetranortriterpenoids with a modified ring from bark of Soymida febrifuga A.Juss (Meliaceae) Journal of the Chemical Society, Perkin Transactions, 1, 1873-1882. Palei A.K., Nishteswar K., Shukla V.J., 2013. Phytochemical screening of Soymida febrifuga Roxb. (Meliaceae). International Journal of Pharmacological & Life Sciences, 4(2), 2371-2374. Rastogi R.P., Mehrotra B.N., 1993. Compendium of Indian Medicinal Plants, vol. 2, CDRI, Lucknow; Publications and Information Directorate, New Delhi. Reddy B.S., Reddy B.P., Raghavulu S.V., Ramakrishna S., Venkateswarlu Y., Diwan P.V., 2008. Evaluation of antioxidant and antimicrobial properties of Soymida febrifuga leaf extracts. Phytotherapy Research, 22(7), 943-47. Reddy B.S., Reddy R.K., Reddy B.P., Ramakrishna S., Diwan P.V., 2008. Potential in vitro antioxidant and protective effects of Soymida febrifuga on ethanol induced oxidative damage in HepG2 cells. Food & Chemical Toxicology. 46(11), 3429-3442.

Ph ton

Sharry S., Cabrera P.J., Herrera E.L., Rangel C.R., Lede S., Abedini W., 2006. An alternative pathway for plant in vitro regeneration of chinaberry -tree Melia azedarach L. derived from the induction of somatic embryogenesis. Electronic Journal of Biotechnology, North America, 915-16. Scocchi A.M., Mroginski L.A., 2004. In vitro conservation of apical meristem tip of Melia azedarach L. (Meliaceae) under slow growth conditions. International Journal of Experimental Botany, 137-143. Srividya N., Poorna S.D. B., Satyanarayana P., 1998. Azadirchtin and nimbin content in invitro cultures shoots and roots of Azadirchta indica. Indian Journal of Plant Physiology, 3, 129-134. Silvia V., Gonzalez A., Hebe R., Mroginski L., 2010. Effect of morphological heterogeneity of somatic embryos of Melia azedarach on conversion into plants. Biocell. 34(1), 7-13. Senerath W.T.P.S.K.,Chandrasena K.G.P.H., Fernando K.M.E.P., 2007. In vitro mass propagation and greenhouse establishment of Munroniapinnata (Wall) Theob. (Binkohomba). Journal of National Science Foundation Sri Lanka, 35(3), 181-189.

560

Shahzad A., Siddiqui S.A., 2001. Micropropagation of Melia Azedarach L. Phytomorphorphology, 51 (2), 151-154. Sanyal M, Das A, Bennerjee M and Datta PC (1981).In vitro hormone induced chemical and histological differentiation in stem callus of neem, Azadirachta indica A. Juss. Journal of Experimental Biology, 19, 1067-1068. Sanyal M., Datta P.C., 1986. Nutritional environment and differentiation from cultured tissues of Azadirachta. Acta Horticulturae, 188, 99103. Shrikhande M., Thengane S.R., Mascarenhas A.F., 1993. Somatic embryogenesis and plant regeneration in Azadirachta indica A. Juss. In Vitro Cell Developmental Biology, 29, 38-42. Su W.W., Hwang W.I., Kim S.Y., Sagawa Y., 1997. Induction of somatic embryogenesis in Azadirachta indica. Plant Cell Tissue Organ Culture, 50, 91-95. Stewart IC., 2010. Plant regeneration in Spanish cedar, Cedrela odorata L. Using zygotic embryo explants from mature seed and improvement of embryogenic nodule intiation by heat shock. In Vitro Cell Developmental Biology Plant, 46,126-133. Srikanth K.N.D., Raghupati R., Parvathi C., 2006. In vitro response of select regions of Azadirachta indica A. Juss (Meliaceae) as elucidated by biochemical and molecular variations. Current Science, 91(6), 770-76. Sujanya S., Poornasri D.B., Isha S., 2008. In vitro production of azadirachtin from cell suspension cultures of Azadirachta indica. Journal of Bioscience, 33(1),113–120. Srivastava P., Singh M., Mathur P., Chaturvedi R., 2009. In vitro organogenesis and plant regeneration from unpollinated ovary cultures of Azadirachta indica. Biological Plantarum, 53 (2), 360-364. Saifullah K., Azizuddin K., Sobia A.H., Muhammad K., Almas J., Muhmammad A., Muhammad A.M., Zaheer U.H., Ahsana D., Muhammad I.C., 2013. In vitro immunomodulatory and anti-cancerous activities of biotransformed products of Dianabol through Azadirachta indica and its molecular docking studies. Chemistry Central Journal, 7:163, 1-11. Sofia B., Carlos S., Elizabeth H.J., 2012. Effects of donor plant age and explants on in vitro culture of Cedrela montana Moritz ex Turcz. Universitas Scientiarum, 17(3), 263-271. Tacoronte M., Vielma M., Mora A., Valecillos C., 2004. In vitro propagation of mahogany tree (Swietenia macrophylla King) from axillary buds. Acta Cient Venez, 55(1),7-12.

Thakur R, Rao P.S., Bapat V.A., 1998. In vitro plant regeneration in Melia azedarach L. Plant Cell Reports, 18, 127-131. Thiagarajan N., Murali M., 1993. Paper Presented at the World Neem Conf., Feb. 24-28, Bangalore 975-79. Thioune O., Pousset J.L., Lo I., 1994. Antiinflammatory activity of the bark of Khaya senegalensis (A Juss). Preliminary research of structure/activity relationship. Dakar Medical, 44, 12-15. Valverde C.L., Rojas V.A., Hine G.A., 2008. In vitro propagation of Albizia guachapele, Cedrela odorata, Platymiscium pinnatum and Guaiacum sanctum. Plant Tissue Culture & Biotechnology, 18,151-156. Vila S.K., Rey H.Y., Mroginski L.A., 2004. Influence of genotype and explant source on indirect organogenesis by in vitro culture of leaves of Melia azedarach L. Biocell. 8(1), 35-41. Veeresham C., Raj Kumar M., Sowjanya D., Kokate C.K., Apte S.S., 1998. Production of Azadirachtin from Callus Cultures of Azadirachta indica. Fitoterapy, 69, 423-424. Venkateswarlu B., Katyal J.C., Choudhuri J., Mukhopadhyay K., 1998. Micropropagation of plus neem (Azadirachta indica A. Juss) and evaluation of field transferred plants. Indian Forestry, 124: 537543. Yoganarasimhan S.N., 1996. Medicinal plants of India, Vol.1 Interline Publishing Pvt. Ltd., Bangalore, pp1. Vander E.S., Maccioni O.V., Fabio G.G., Pasqua G, Monacelli, B., (2001). Organogenesis and embryogenesis from callus cultures of Azadirachta excels. Plant Biosystems, 135 (1), 13-18. Vila S., Gonzalez A., Rey H., Mroginski L., 2009. Somatic embryogenesis and plant regeneration in Cedrela fissilis. Biologia Plantarum, 53 (2), 383-386. Vedavathy S., Sudhakar A., Mrdula V., 1997. Tribal medicinal plants of chitoor. Ancient Science Life, 16, (4), 307-331. Yadav P.A., Suresh G., Rao M.S., Shankaraiah G., Usha Rani P., Babu K.S., 2014. Limonoids from the leaves of Soymida febrifuga and their insect antifeedant activities. Bioorganic Medicinal Chemistry Letters, 24(3), 888-92. Yuri P. R., Juan J.G., José A., González R., Manuel L.R., 2012. Tissue Culture Methods for the Clonal Propagation and Genetic Improvement of Spanish Red Cedar (Cedrela odorata). Plant Cell Culture Protocols. Methods in Molecular Biology, 877, 129141. Yuri J.P.R, Israel G.S., Ángel H.E, Alfredo D.H., Felipe A.BP., José A., González R., Manuel L.R.,

Ph ton

561

(2011). Induction of somatic embryogenesis and plant regeneration in the tropical timber tree Spanish red cedar [Cedrela odorata L. (Meliaceae)]. Plant Cell Tissue and Organ Culture, 105 (2), 203209.

Ph ton

562