GJRMI - Volume 3, Issue 6 - June 2014 by Dr. Hari Venkatesh.K.Rajaraman

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ISSN 2277 â&#x20AC;&#x201C; 4289 www.gjrmi.com Editor-in-chief Dr Hari Venkatesh K Rajaraman

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INDEX – GJRMI - Volume 3, Issue 6, June 2014 MEDICINAL PLANTS RESEARCH Micro-biology ANTIBACTERIAL ACTIVITY OF ESSENTIAL OILS OF ROSMARINUS OFFICINALIS FROM EASTERN ALGERIA Takia Lograda, Messaoud Ramdani, Pierre Chalard, Gilles Figueredo

232–242

Ethno-Botany ETHNOBOTANICAL PLANTS USED BY THE MALAYALI TRIBES IN YERCAUD HILLS OF EASTERN GHATS, SALEM DISTRICT, TAMIL NADU, INDIA Rekha R, Senthil Kumar S

243–251

Review Article MICROPROPAGATION: AN ESSENTIAL TOOL TO FLOURISH ENDANGERED MEDICINAL PLANTS Sharma Rohit

252–262

Review Article A REVIEW ON MICROBIAL ENDOPHYTES FROM PLANTS: A TREASURE SEARCH FOR BIOLOGICALLY ACTIVE METABOLITES Ruby Jalgaonwala, Raghunath Mahajan

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF ANKOLA – ALANGIUM SALVIIFOLIUM (L.F.) WANG., OF THE FAMILY CORNACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA

263–277

Research Article ANTIBACTERIAL ACTIVITY OF ESSENTIAL OILS OF ROSMARINUS OFFICINALIS FROM EASTERN ALGERIA Takia Lograda1*, Messaoud Ramdani2, Pierre Chalard3, Gilles Figueredo4 1 ,2

Laboratory of Natural Resource Valorisation, SNV Faculty, Ferhat Abbas University, 19000 Setif, Algeria Clermont Université, ENSCCF, Institut de Chimie de Clermont-Ferrand, BP 10448, F-63000 CLERMONTFERRAND, France 3 CNRS, UMR 6296, ICCF, F-63171 Aubière, France 4 LEXVA Analytique, 460 rue du Montant, 63110 Beaumont, France. 3

*Corresponding author: Email- tlograda63@yahoo.fr; Phone:(213)36835894;Fax: (213) 36937943. Received: 05/05/2014; Revised: 25/05/2014; Accepted: 02/06/2014

ABSTRACT The hydro-distillation of the essential oil of Rosmarinus officinalis gave a viscous liquid with a whitish colour. The average yield of essential oil of the samples is 0.23%. This investigation allows us to support that R. officinalis includes six chemotypes in eastern Algerian. The difference of these chemotypes variants is the concentration of Eucalyptol, campene, α-pinene and camphor. The antibacterial activity of the essential oils chemotypes was evaluated against nine bacteria (Enterobacter cloacae ATCC 13047, MRSA (Methicillin-resistant Staphylococcus aureus), Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas syringae, Salmonella sp, Serratia liquefaciens ATCC 27592, Serratia marcescens ATCC 14756 and Shigella sp). The results showed that the essential oils have inhibited growth of bacterial strains. The essential oils chemotypes of Kherrata and the Bibans regions, generally exhibit antibacterial activity against the microorganisms’ tested. The camphor chemotype with three variants exhibits a moderate antibacterial activity. KEYWORDS: Rosmarinus officinalis, Lamiaceae, Chemotype, antibacterial activity, Algeria

Cite this article: Takia Lograda, Messaoud Ramdani, Pierre Chalard, Gilles Figueredo (2014), ANTIBACTERIAL ACTIVITY OF ESSENTIAL OILS OF ROSMARINUS OFFICINALIS FROM EASTERN ALGERIA, Global J Res. Med. Plants & Indigen. Med., Volume 3(6): 232–242

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 232–242

INTRODUCTION The essential oil of Rosmarinus officinalis contains mainly monoterpenes (Angioni et al., 2004; Diaz-Maroto et al., 2007). Principal volatile compounds are camphor and eucalyptol, followed by borneol, verbenone, αpinene and camphene (Pino et al., 1998; Zaouali et al., 2005; Diaz-Maroto et al., 2007; Calin-Sanchez et al., 2011; Apostolides et al., 2013; Lograda et al., 2013). The volatile compounds from rosemary samples could be grouped in chemical families; therefore, the predominant group was monoterpenoids (Bozin et al., 2007; Szumny et al., 2010; CalinSanchez et al., 2011). The chemical composition and seasonal variations in rosemary oil were reported from southern Spain (Salido et al., 2003; Jordan et al., 2011, 2013). All the samples studied by Salido et al. (2003) belonging to chemotypes (α-pinene - 1, 8-cineole - camphor). Jordan et al. (2013) identified five chemotypes based on α-pinene - 1, 8-cineole - camphor. The chemical polymorphism of the essential oil of wild rosemary Spanish populations has been reported and different chemotypes were defined according to the geographical area (Varela et al., 2009). In a previous work five chemotypes distributed in eastern Algeria were identified (Lograda et al., 2013). Rosmarinus officinalis oil is widely used in cosmetic, food and pharmaceutical industries as a fragrance component of soaps, creams, lotions, and perfumes. Although it is popular, potential harmful side-effects of this oil have been described. The genotoxicity and mutagenicity are confirmed by Maistro et al. (2010). The rosemary leaves are used in fried chicken (Viuda-Martos et al., 2010). The rosemary essential oils have a number of beneficial properties as natural preservatives in cosmetics, toiletries, drugs and food products (Bakkali et al., 2008; Reichling et al., 2009) Sienkiewicz et al., 2012).

The anti-inflammatory property of rosemary extracts was reported by Masuda et al. (2001); Bozin et al. (2007); Altinier et al. (2007); Poeckel et al. (2008); Viuda-Martos et al. (2010). The interest was also generated due to the anticarcinogenic activity (Cheung and Tai, 2001). The essential oil of R. officinalis showed antimytotic and antifungal activity (Yang et al., 2011; Mugnaini et al., 2012). All extracts of R. officinalis were effective in inhibiting bacterial growth (Abutbul et al., 2004; Bozin et al., 2007). Antimicrobial and antioxidant activities of rosemary are demonstrated (Faixova and Faix, 2008; Kadri et al., 2011). According to Lopez et al. (2005), the oils from R. officinalis have an antibacterial potential against the Gram-positive and Gramnegative bacteria. The authors presented a detailed analysis of the tested oils and their ability to inhibit the growth of bacteria. The rosemary oil has an antibacterial effect on a number of microorganisms responsible for respiratory infections (Fabio et al., 2007). Rosemary oil was found to demonstrate antibacterial activity against Escherichia coli strains with different patterns of resistance (Probuseenivasan et al., 2006; Sienkiewicz et al., 2013). The essential oil from R. tournefortii exhibited strong antibacterial activity against Escherichia coli and Pseudomonas aeruginosa and Staphylococcus aureus (Bendeddouche et al., 2011). All chemotypes of R. officinalis studied, possessed antibacterial activities (Wang et al., 2012). Though the flowering aerial part of this plant commonly is used because of its antiseptic properties, heretofore there is no report that investigated the antimicrobial activity of this plant in eastern Algeria. This study aimed to evaluate the antimicrobial activities of the chemotype essential oils of wild plants of R. officinalis, largely used in Algeria, obtained from samples grown in Eastern Algeria, as well as to validate its traditionally uses.

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 232–242

Figure 1: Populations of Rosmarinus officinalis studied

MATERIALS & METHODS

Antibacterial Activity

Plant material

The antimicrobial activities of the essential oils were evaluated against three Gram positive (Enterobacter cloacae ATCC 13047, MRSA (Methicillin-resistant Staphylococcus aureus), Staphylococcus aureus ATCC 25923) and six Gram negative bacteria (Escherichia coli ATCC 25922, Pseudomonas syringae, Salmonella sp, Serratia liquefaciens ATCC 27592, Serratia marcescens ATCC 14756, Shigella sp). The bacterial inoculums was prepared from overnight broth culture in physiological saline (0.8 % of NaCl) in order to obtain an optical density ranging from 0.08–01 at 625 nm. Muller-Hinton agar (MH agar) and MH agar supplemented with 5 % sheep blood for fastidious bacteria were poured in Petri dishes, solidified and surface dried before inoculation. Sterile discs (6 mm Φ) were placed on inoculated agars, by test bacteria, filled with 10 μl of mother solution and diluted essential oil (1:1, 1:2, 1:4, and 1:8 v:v of DMSO). DMSO was used as negative control. Bacterial growth inhibition was determined as the

Rosmarinus officinalis is collected from five localities in eastern Algeria, Kherrata (Bedjaia), Boutaleb (Setif), Bibans (Bourdj Bou-Arriridj), Agmeroual and N’gaous (Batna), and Boussâada (M’Sila) (Figure 1). The plant identification was performed by Dr. Lograda Takia. The voucher specimens are preserved in the Herbarium at the Department of Biology and Ecology Vegetal, Setif University, Algeria. Aerial parts were collected during the flowering stage in October 2013. Extraction of the essential oil The air-dried aerial parts of the six populations were subjected to hydro-distillation for 3 h with distilled water using a Clevengertype apparatus. The oil obtained was collected and dried over anhydrous sodium sulphate and stored in screw capped glass vials in a refrigerator at 4–5°C, prior to the analysis.

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 232–242

diameter of the inhibition zones around the discs. All tests were performed in triplicate. Then, Petri dishes were incubated at 37°C during 18 to 24h aerobically. After incubation, inhibition zone diameters were measured and documented. RESULTS The hydro-distillation of the essential oil of R. officinalis gave a viscous liquid with a whitish colour. The average yield of essential oil of the samples is 0.23%, the highest rate was observed in the essential oil of Kherrata population (0.35%), while the population of Agmeroual was characterised by the lowest yield (0.10%). R. officinalis of eastern Algeria includes several chemotypes (Table 1). The first chemotype to eucalyptol has two variants; the first variant (E-Ca-αP-C) characterizes Kherrata population; the second chemotype (EαP-Ca-C) characterizes the Bibans population. The difference of these two chemotype variants is the concentration of Eucalyptol and camphor. The second chemotype to camphor had three variants, the variant (Ca-C-αP-E) characterized Boussâada population. Agmeroual population contained the variant (Ca-αP-C-E), while the third variant (Ca-αP-E-C) of this chemotype was found in the region of N'gaous. The third chemotype to α-pinene (αP-Ca-E-C), in R. officinalis of eastern Algeria, characterized the population of Boutaleb. The antibacterial activity of the essential oil chemotypes was evaluated against nine bacteria (Enterobacter cloacae ATCC 13047, MRSA (Methicillin-resistant Staphylococcus aureus),

Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas syringae, Salmonella sp, Serratia liquefaciens ATCC 27592, Serratia marcescens ATCC 14756 and Shigella sp). The disc diameters of inhibition zone of essential oils for the microorganisms tested are grouped in Table 2. The results showed that the essential oils have inhibited the growth of bacterial strains. The diameters of the inhibition zone are between (0 and 40 mm); these diameters depended on the sensitivity of the bacteria tested. The essential oil chemotypes of Kherrata and the Bibans regions, generally exhibited antibacterial activity against the microorganisms’ tested, with an inhibition diameter between 7 and 15 mm. The chemotype of Bibans populations had a high activity, with a dilution ½, on Staphylococcus aureus with a diameter of 40 mm (Figure 2). The camphor chemotype with three variants exhibits a moderate antibacterial activity with an inhibition diameter (7–20 mm). The essential oil is active on Enterobacter cloacea with different dilutions. The dilution 1/2 of Agmeroual chemotype inhibited Escherichia coli strongly with a diameter of 20 mm, and no effect was seen on Pseudomonas syringae (Figure 3). The chemotype of Boutaleb (α-pinene variant) had an inhibitory effect with an inhibition diameter (7–15 mm). This chemotype is active on Enterobacter cloacae and no activity on Pseudomonas syringae in the ¼ dilution (Figure 4).

Table 1: Volatile profile of Rosemary essential oils selected by chemotype (Lograda et al., 2013) Populations Yield (v/v) Chemotypes Eucalyptol α-pinene Camphor Camphene

Kherrata Bibans 0.35 0.3 Eucalyptol E-Ca-αP- E-αP-Ca-C C 35.5 42.2 11.4 13.8 14.5 9.1 8.0 5.3

Agmeroual 0.2 Ca-αP-C-E 5.4 16.9 38.8 13.8

N'gaous Boussaâda 0.1 0.25 Camphor Ca-αP-E-C Ca-C-αPE 12.1 6.6 13.6 15.1 16.9 42.7 3.7 17.7

Boutaleb 0.15 α-pinene αP-Ca-CE 8.8 25.2 24.1 22.7

Chemotype classification in relation with the chemical compounds found in rosemary samples. E = eucalyptol; αP = α-pinene; Ca = camphor; C = camphene. Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 232–242

Table 2: Inhibition diameter (mm) of Rosmarinus officinalis essential oil chemotype Populations

Kherrata

Bibans

Boussâada

Agmeroual

N’gaous

Boutaleb

Chemotypes

E-Ca-αp-C

E-αp-Ca-C

Ca-C-αp-E

Ca-αp-C-E

Ca-αp-E-C

αp-Ca-E-C

Bacteria

14 0 14 10 16 10 12 15 25

1 2 3 4 5 6 7 8 9

Dilution Dilution ½ ¼ 1/8 ½ ¼ 1/8 ½ ¼ 1/8 ½ ¼ 1/8 15 13 11 20 10 11 10 9 11 20 9 8 12 12 9 15 11 10 10 8 9 11 10 8 10 9 9 11 15 10 9 7 12 12 8 8 12 13 12 12 9 10 19 7 10 0 0 0 9 8 10 13 7 12 10 10 10 7 7 7 11 8 14 15 9 13 15 10 15 13 10 11 8 8 10 10 9 8 9 12 10 15 11 10 11 9 12 40 15 10 10 15 15 8 7 0 10 9 12 12 13 13 20 15 12 20 12 15

½ 8 8 10 10 10 10 8 15 10

Dilution ¼ 1/8 ½ ¼ 1/8 8 12 10 12 10 7 14 13 10 10 12 14 15 8 11 9 9 7 0 11 12 11 10 8 10 8 9 14 13 12 7 7 12 10 13 12 10 10 8 10 13 11 11 11 10

1 : Escherichia coli ATCC 25922; 2 : Enterobacter cloacae ATCC 13047; 3 : MRSA = Methicillin-resistant Staphylococcus aureus; 4 : Pseudomonas syringae; 5 : Salmonella sp; 6 : Serratia liquefaciens ATCC 27592 ; 7 : Serratia marcescens ATCC 14756; 8 : Staphylococcus aureus ATCC 25923; 9 : Shigella sp. G. = Gentamicine; E = eucalyptol; αP = α-pinene; Ca = camphor; C = camphene.

Figure 2: Antibacterial activity of Kherrata and Bibans chemotypes 45 Escherichia coli Enterobacter cloacae MRSA Pseudomonas syringae Salmonella sp Serratia liquefaciens Serratia marcescens Staphylococcus aureus Shigella sp

40 35

Inhibition diameter (mm)

30 25 20 15 10 5

E-Ca-ap-C Kherrata

E-ap-Ca- C Bibans

-5 G

1/8

Dilution

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1/8

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Figure 3: Antibacterial activity of Boussaada, Agmeroual and N’gaous Chemotypes 26 Escherichia coli Enterobacter cloacae MRSA Pseudomonas syringae Salmonella sp Serratia liquefaciens Serratia marcescens Staphylococcus aureus Shigella sp

24 22 20

Inhibition diameter (mm)

18 16 14 12 10 8 6 4 2 0

Ca-C-ap-E; Boussâada

Ca-ap-C-E; Agmeroual

-2 Gentamicine

¼ ½

½ 1/8

Ca-ap-E-C; N’gaous

1/8 ¼

¼ ½

1/8

Dilution

Figure 4: Antibacterial activity of Boutaleb Chemotype 26 Escherichia coli Enterobacter cloacae MRSA Pseudomonas syringae Salmonella sp Serratia liquefaciens Serratia marcescens Staphylococcus aureus Shigella sp

24 22 20

Inhibition diameter (mm)

18 16 14 12 10 8 6 4 2 ap-Ca-E-C; Boutaleb

0 -2 G

¼ Dilution

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1/8

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DISCUSSION Our returns of the essential oil are low compared to those of the literature. This yield is between 0.6 and 0.8% (Atti-Santos et al., 2005; Bekkara et al., 2007). Ayadi et al. (2011) found a yield (0.71–2%) which indicates that the yield of rosemary varies according to geographical location. The chemical components of the essential oil of R. officinalis showed that α-pinene, camphene, eucalyptol and camphor are the major products (Jordan et al., 2013; Mugnaini et al., 2012; Sienkiewicz et al., 2013; Matsuzaki et al., 2013; Derwich et al., 2011; Fadli et al., 2011; Jalali-Heravi et al., 2011; Issabeagloo et al., 2012; Hussain et al., 2011; Mudasir et al., 2012; Verma et al.. 2011; Yang et al., 2011; Sui et al., 2012; Aderiana et al., 2013; Lograda et al., 2013). Based on the majority of Rosemary components the authors identified three chemotypes (Issabeagloo et al., 2012; Mudasir et al., 2012; Sui et al., 2012; Mugnaini et al., 2012; Jordan et al., 2013; Sienkiewicz et al., 2013; Matsuzaki et al., 2013; Aderiana et al., 2013), the same chemotypes are found in our samples with the presence of five variants. The eastern Algeria samples showed an average antibacterial activity on all tested bacteria, the same observations were mentioned in the literature (Lopez et al., 2005; Probuseenivasan et al., 2006; Fabio et al., 2007; Jiang et al., 2011; Sienkiewicz et al., 2013). This may be partly due to the fact that the essential oils contained more oxygenated compounds and these classes of compounds have been proved

to possess strong antibacterial activities (Deba et al., 2008). Generally the essential oils of R. officinalis have shown broad spectra of activity against the tested microorganisms, the same findings were made by Gachkar et al., (2007) on some species of the genus. The antimicrobial property of the essential oil of R. officinalis is attributed to the presence of α-pinene, 1,8cineole, camphor, verbinone and borneol (Santoyo et al., 2005), the quantities of these compounds were very high in our oils. CONCLUSION Six variants of Rosemary chemotypes, localized in eastern Algeria, were used in the determination of the antimicrobial activity, against the gram positive and negative bacteria by the disc diffusion method. The essential oil obtained by hydrodistillation was more active against microorganisms than the antibiotic gentamicine. The essential oils showed inhibition zones against all micro-organisms tested. The Results indicate that essential oils of Rosmarinus officinalis present significant antimicrobial activity. This antimicrobial activity can be linked to its higher content of oxygenated compounds. This study provides additional data of antibacterial activity of the essential oils of R. officinalis growing in Algeria. ACKNOWLEDGEMENT The work was supported by Algerian MESRS and Chemical Laboratory of carbohydrates Heterocyclic of Clermont Ferrant. France.

REFERENCES Abutbul S, Golan-Goldhirsh A, Barazani O, Zilberg D (2004). Use of Rosmarinus officinalis as a treatment against Streptococcus iniae in tilapia (Oreochromis sp.), Aquaculture, 238: 97–105.

Adriana M, Ojeda-Sana C M, van Baren M A, Elechosa M A, Juarez S M (2013). New insights into antibacterial and antioxidant activities of rosemary essential oils and their main components, Food Control, 31: 189– 195.

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Altinier G, Sosa S, Aquino RP, Mencherini T, Loggia RD, Tubaro A (2007). Characterization of topical antiinflammatory compounds in Rosmarinus officinalis L., J. Agric. Food Chem., 55: 1718–1723.

Bekkara F Atik, Bousmaha L, Taleb Bendiab S A, Boti J B, Casanova J (2007). Composition chimique de l’huile essentielle de Rosmarinus officinalis L. poussant à l’état spontané et cultivé de la région de Tlemcen, Biologie & Santé, 7(1): 6–11.

Angioni A, Barra A, Cereti E, Barile D, Coisson JD, Arlorio M, Dessi S, Coroneo V, Cabras P (2004). Chemical composition, plant genetic differences, antimicrobial and antifungal activity investigation of the essential of Rosmarinus officinalis L., Journal of Agricultural and Food Chemistry, 52: 3530–3535.

Bendeddouche M S, Benhassaini H, Hazem Z, Romane A (2011). Essential oil analysis and antibacterial activity of Rosmarinus tournefortii from Algeria, Nat. Prod. Commun., 6: 1511–1514.

Apostolides Nelly Arnold, Marc El Beyrouthy, Wissal Dhifi, Samir Najm, Fabrice Cazier, Wafaa Najem, Madonna Labaki, Antoine AbouKaïs (2013). Chemical Composition of Aerial Parts of Rosmarinus officinalis L. Essential Oil Growing Wild in Lebanon, Journal of Essential Oil Bearing Plants, 16(2): 274–282. Atti-Santos Ana Cristina, Marcelo Rossato, Gabriel Fernandes Pauletti, Luciana Duarte Rota, Juarez Ciro Rech, Marcia Regina Pansera, Fabiana Agostini, Luciana Atti Serafini and Patrick Moyna (2005). Physico-chemical Evaluation of Rosmarinus officinalis L. Essential Oils, Brazilian Archives of Biology and Technology, 48(6): 1035– 1039. Ayadi S, Jerribi C, Abderrabba M (2011). Extraction et étude des huiles essentielles de Rosmarinus Officinalis cueillie dans trois régions différentes de la Tunisie, Journal de la Société Algérienne de Chimie, 21(1): 25–33.

Bozin B, Mimica-Dukic N, Samojlik I, Jovin E (2007). Antimicrobial and antioxidant properties of rosemary and sage (Rosmarinus officinalis L. and Salvia officinalis L., Lamiaceae) essential oils, Journal of Agricultural and Food Chemistry, 55: 7879–7885. Calin-Sanchez Angel, Antoni Szumny, Adam Figiel, Klaudiusz Jałoszynski, Maciej Adamski, Angel A, CarbonellBarrachin (2011). Effects of vacuum level and microwave power on rosemary volatile composition during vacuum–microwave drying, Journal of Food Engineering, 103: 219–227. Cheung S, Tai J (2001). Anti-proliferative and antioxidant properties of rosemary Rosmarinus officinalis, Oncol. Rep., 17: 1525–1531. Deba F, Xuan TD, Yasudaa M, Tawata S (2008). Chemical composition and antioxidant, antibacterial and antifungal activities of the essential oils from Bidens pilosa Linn. var. Radiata, Food Control 19, 346–352 Derwich E, Benziane Z, Chabir R (2011). Aromatic and Medicinal Plants of Morocco: Chemical Composition of essential oils of Rosmarinus officinalis and Juniperus Phoenicea, International Journal of Applied Biology and Pharmaceutical Technology, 2(1): 145– 153

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Diaz-Maroto MC, Perez-Coello MS, SanchezPalomo E, Gonzalez-Vinas MA (2007). Impact of drying and storage time on sensory characteristics of rosemary (Rosmarinus officinalis L.), Journal of Sensory Studies, 22: 34–48. Fabio A, Cermelli C, Fabio G, Nicoletti P, Quaglio P (2007). Screening of the antibacterial effects of a variety of essential oils on microorganisms responsible for respiratory infections, Phytother. Res., 21: 374–377. Fadli Mariam, Jacqueline Chevalier, Asmaa Saad, Nour-Eddine Mezrioui, Lahcen Hassani, Jean-Marie Pages (2011). Essential oils from Moroccan plants as potential chemosensitisers restoring antibiotic activity in resistant Gramnegative bacteria International Journal of Antimicrobial Agents, 38: 325–330. Faixov ZR, Faix S (2008). Biological effects of Rosmary (Rosmarinus officinalis) essensial oil (A Review), Folia Veterinaria, 5(3-4): 135–139. Gachkar L, Yadegari D, Rezaei MB, Taghizadeh M, Astaneh SA, Rasooli I (2007). Chemical and biological characteristics of Cuminum cyminum and Rosmarinus officinalis essential oils, Food Chemistry, 102: 898–904. Hussain Abdullah I, Farooq Anwar, Poonam S Nigam, Satyajit D Sarker, John E Moore, Juluri R Rao, Anisha Mazumdar (2011). Antibacterial activity of some Lamiaceae essential oils using resazurin as an indicator of cell growth, LWT. Food Science and Technology, 44: 1199–1206. Issabeagloo Eilyad, Parviz Kermanizadeh, Mohammad Taghizadieh, Reza Forughi (2012). Antimicrobial effects of rosemary (Rosmarinus officinalis L.) essential oils against Staphylococcus spp., African Journal of Microbiology Research, 6(23): 5039–5042.

Jalali-Heravi Mehdi, Rudabeh Sadat Moazeni, Hassan Sereshti (2011). Analysis of Iranian rosemary essential oil: Application of gas chromatography– mass spectrometry combined with chemometrics, Journal of Chromatography A, 1218: 2569–2576. Jiang Y, Wu N, Fu Y-J, Wang W, Luo M, Zhao CJ, Zu YG, Liu XL (2011). Chemical composition and antimicrobial activity of the essential oil of Rosemary, Environ. Toxicol. Pharmacol., 32: 63– 68. Jordan MJ, Lax V, Martinez C, Aouissat M, Sotomayor JA 2011. Chemical intraspecific variability and chemotypes determination of Rosmarinus officinalis L. in the region of Murcia, Acta Horticulturae, 925: 109–114. Jordan MJ, Vanesa Lax, Maria C Rota, Susana Loran,. José A. Sotomayor (2013). Effect of bioclimatic area on the essential oil composition and antibacterial activity of Rosmarinus officinalis L.. Food Control., 30: 463– 468. Kadri Adel, Zied Zarai, Ines BenChobba, Ahmed Békir, Néji Gharsallah, Mohamed Damak, Radhouane Gdoura (2011). Chemical constituents and antioxidant properties of Rosmarinus officinalis L. essential oil cultivated from the South-Western of Tunisia, Journal of Medicinal Plants Research, 5(29): 6502–6508. Lopez P, Sanchez C, Battle R, Nerin C (2005). Soilid- and vapour-phase antimicrobial activities of six essential oils: Susceptibility of selected foodborne bacterial and fungal strains, Agric. Food Chem., 53: 6939–6946.

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Maistro EL, Mota SF, Lima EB, Bernardes BM, Goulart FC (2010). Genotoxicity and mutagenicity of Rosmarinus officinalis (Labiatae) essential oil in mammalian cells in vivo, Genetics and Molecular Research, 9(4): 2113–2122. Martins MR, Tinoco MT, Almeida AS, CruzMorais J (2012). Chemical Composition, Antioxidant and Antimicrobial properties of three Essential oils from Portuguese Flora, Journal of Pharmacognosy, 3(1): 39–44. Masuda T, Inaba Y, Takeda Y (2001). Antioxidant mechanism of carnosic acid: structural identification of two oxidation products, J. Agric. Food Chem., 49: 5560–5565. Matsuzaki Yusuke, Toshiyuki Tsujisawa, Tatsuji Nishihara, Mari Nakamura, Yasuaki Kakinoki (2013). Antifungal activity of chemotype essential oils from rosemary against Candida albicans, Open Journal of Stomatology, 3: 176–182. Mudasir A, Tantry Syed Shabir, Reehana Khan, Afsha Habib, Seema Akbar (2012). Determination of Essential Oil Composition of Rosmarinus officinalis Growing as Exotic species in Kashmir Valley, Chemistry of Natural Compounds, 47(6): 1012–1014. Mugnaini L, Nardoni S, Pinto L, Pistelli L, Leonardi M, Pisseri F, Mancianti F (2012). In vitro and in vivo antifungal activity of some essential oils against feline isolates of Microsporum canis, Journal de Mycologie Médicale, 22: 179–184. Pino JA, Estrarron M, Fuentes V (1998). Essential Oil of Rosemary (Rosmarinus Officinalis L.) from Cuba, J. Essent. Oil Res., 10: 111.

Poeckel D, Greiner C, Verhoff M, Rau O, Tausch L, Heornig C, Steinhilber D, Schubert-Zsilavecz M, Werz O (2008). Carnosic acid and carnosol potently inhibit human 5-lipoxygenase and suppress pro-inflammatory responses of stimulated human polymorphonuclear leukocytes, Biochem. Pharmacol., 76: 91–97. Prabuseenivasan S, Jayakumar M, Ignacimuthu S (2006). In vitro antibacterial activity of some plant essential oils, BMC Complement. Altern. Med., 6: 39. Reichling J, Schnitzler P, Suschke U, Saller R (2009). Essential oils of aromatic plants with antibacterial, antifungal, antiviral and cytotoxic properties-An overview, Forsch Komplementmed, 16: 79–90. Salido S, Altarejos J, Nogueras M, Sanchez A, Luque P (2003). Chemical composition and seasonal variations of rosemary oil from Southern Spain, Journal of Essentials Oil Research, 15: 10–14. Santoyo S, Cavero S, Jaime L, Ibanez E, Senorans FJ, Reglero G (2005). Chemical composition and antimicrobial activity of Rosmarinus officinalis L. essential oil obtained via supercritical fluid extraction, Journal of Food Protection, 68(4): 790–795. Sienkiewicz M, Kowalczyk E, Wasiela M (2012). Recent patents regarding essential oils and the significance of their constituents in human health and treatment, Recent Pat. Anti Infect. Drug Discov., 7: 133–140. Sienkiewicz Monika, Monika Lysakowska, Marta Pastuszka, Wojciech Bienias, Edward Kowalczyk (2013). The Potential of Use Basil and Rosemary Essential Oils as Effective Antibacterial Agents, Molecules, 18: 9334–9351.

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Sui Xiaoyu, Tingting Liu, Chunhui Ma, Lei Yang, Yuangang Zu, Lin Zhang, Hua Wang (2012). Microwave irradiation to pretreat rosemary (Rosmarinus officinalis L.) for maintaining antioxidant content during storage and to extract essential oil simultaneously, Food Chemistry, 131: 1399–1405. Szumny Antoni, Adam Figiel, Antonio Gutierrez-Ortiz, Angel A, Carbonell Barrachina (2010). Composition of rosemary essential oil (Rosmarinus officinalis) as affected by drying method. Journal of Food Engineering, 97: 253–260. Takia Lograda, Messaoud Ramdani, Pierre Chalard, Gilles Figueredo (2013). Characteristics of Essential oils of Rosmarinus officinalis from eastern Algeria. Global J Res. Med. Plants & Indigen. Med., 2(12): 794–807. Varela F. Navarrete P. Cristobal R. Fanlo M. Melero R. Sotomayor J A (2009). Variability in the chemical composition of wild Rosmarinus officinalis L., Acta Horticulturae, 826: 167–174. Verma Ram S, Laiqur Rahman, Sunita Mishra, Rajesh K V, Amit Chauhan, Anand Singh (2011). Changes in essential oil content and composition of leaf and leaf powder of Rosmarinus officinalis cv. CIM-Hariyali during storage, Maejo Int. J. Sci. Technol, 5(02): 181–190.

Source of Support: Algerian MESRS and Chemical Laboratory of carbohydrates Heterocyclic of Clermont Ferrant. France

Viuda-Martos M, El Gendy A E-N. G S, Sendra E, Fernandez-Lopez J, Abd El Razik K A, Omer E, A., Jose A. PerezAlvarez (2010). Chemical composition and antioxidant and anti-Listeria activities of essential oils obtained from some Egyptian plants, Journal Agriculture Food Chemistry, 58(16): 9063–9070. Wang Wei, Nan Li, Meng Luo, Yuangang Zu, Thomas Efferth (2012). Antibacterial Activity and Anticancer Activity of Rosmarinus officinalis L. Essential Oil Compared to That of Its Main Components, Molecules, 17: 2704– 2713. Yang Jiang, Nan Wu, Yu-Jie Fu, Wei Wang, Meng Luo, Chun-Jian Zhao, YuanGang Zu, Xiao-Lei Liu (2011). Chemical composition and antimicrobial activity of the essential oil of Rosemary, Environmental Toxicology and Pharmacology, 32: 63– 68. Zaouali Y, Messaoud C, BenSalah A, Boussaid M (2005). Oil composition variability among populations in relationship with their ecological areas in Tunisian Rosmarinus officinalis L., Flavour Fragrance J., 20: 512–520.

Conflict of Interest: None Declared

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Research Article ETHNOBOTANICAL PLANTS USED BY THE MALAYALI TRIBES IN YERCAUD HILLS OF EASTERN GHATS, SALEM DISTRICT, TAMIL NADU, INDIA Rekha R1*, Senthil Kumar S2 1, 2

PG and Research Department of Botany, Vivekanandha College of Arts and Science for Women (Autonomous), Elayampalayam, Tiruchengode, Namakkal(DT), Tamil Nadu, India *Corresponding Author: E- Mail: rekha87raja@gmail.com

Received: 26/04/2014; Revised: 24/05/2014; Accepted: 30/05/2014

ABSTRACT An ethno-botanical survey was carried out among the Malayali tribals in Yercaud Hills, Southern Eastern Ghats, Salem district, Tamil Nadu, India during November 2012–March 2014. This study mainly focused on the plants used by the Malayali tribes for various purposes (Construction materials, house hold implements, brushing, fuel wood, agriculture tools, religious, decorative to ward off evil spirits) through standardized questionnaires, interviews and discussions with very old and knowledgeable tribals. A total of 84 plant species belonging to 70 genera and 42 families were recorded in the present study. These extremely important plants should be taken into account and steps have to be taken to increase their production for future benefit. KEY WORDS: Yercaud hills, Malayali tribes, Ethno-botany.

Cite this article: Rekha R, Senthil Kumar S (2014), ETHNOBOTANICAL PLANTS USED BY THE MALAYALI TRIBES IN YERCAUD HILLS OF EASTERN GHATS, SALEM DISTRICT, TAMIL NADU, INDIA, Global J Res. Med. Plants & Indigen. Med., Volume 3(6): 243–251

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 243–251

INTRODUCTION Plants are used against a number of diseases by many indigenous communities in traditional medicine; plants are also used in building materials, fodder, weapons and other commodities of economical importance. The tribal people are economically backward ethnic groups and constitute separate socio-cultural groups. Many local and indigenous communities in Asian countries meet their basic needs from the forest products they manufacture and sell based on their traditional knowledge (John Kennedy, 2006). Yercaud hills have been gifted with enormous number of plant species. Tribal people living in Yercaud hills depend on these plants for their survival. These plants play a vital role in their life as these people depend on the forest wealth to meet their needs. Their economic condition is determined by these plants. Yercaud hill range is situated to the north-east part of Eastern Ghats, Salem district, Tamil Nadu, India with rich vegetation and it covers an area of 150 square miles (390 Sq. km). It lies between 11°45’56” N latitude and 78°17’55” E longitude. The temperature ranges from 13°C to 29°C on the peaks and 25°C to 40°C at the foot hills. The average annual rainfall is around 1500–1750 mm. The indigenous people inhabiting Yercaud hill are called Malayali, the oldest group of the branch of the ethnic group in South India. Malayali simply means a hill person an appellation distinguishing them from the people of plains. In physical appearance they scarcely differ from the people of plains. They speak Tamil dialect of their own. They are supposed to be descendants of Kanchipuram vellalar. They appear to have migrated from Kanchipuram (a town near Chennai, Tamil Nadu, India) between seventh and eleventh centuries. The tribals are mostly working as casual laborers in coffee estates. They are cultivating food grains, fruits and vegetable (Alagesaboopathi, et al., 1996). Malayali tribals use more number of plants for various purposes like, for making construction materials, house hold implements, brushing plants, fuel wood, agriculture tools, religious, decorative to ward off evil spirits etc. This

study enumerates such useful plants which are used by Malayali tribals in Yercaud Hills, Eastern Ghats, Tamil Nadu, India. Data Collection Frequent field surveys were carried out in Yercaud hills in different seasons during November 2012–March 2014. Information on the plants was collected through personal interview with village headman, farmers and other knowledgeable tribals. The interviews were conducted in the local language (Tamil), the information includes local names, plant parts used, and method of utilization was gathered from them with regard to each plant. The collected information was recorded on field note books and plants were identified using the Flora of the Presidency of Madras (Gamble, 1935) and Flora of Tamil NaduCarnatic (Matthew, 1983). Plant specimens were deposited for future references in the Botany department, Vivekanandha College of Arts and Sciences for women (Autonomous), Elayampalayam, Tiruchengode, Namakkal (DT), Tamil Nadu, India. RESULTS AND DISCUSSIONS The present study focused mainly on the role of non timber forest products in the livelihood of Malayali tribes of Yercaud hills, Salem district, Tamil Nadu, India. During the study period, 84 plant species belonging to 70 genera and 42 families were identified to be utilized by Malayali tribes (Table 1). The reported plants were arranged to their Botanical Name, Vernacular name as recorded during field work, habit and uses. Plants utilized by Malayali tribes can be classified under various categories like fuel wood plants, house construction and house hold implements, agriculture tools, religious importance plants, plants used to ward off evil afflictions, hair care and toothbrush. The plants listed mainly belong to Rubiaceae (6 species) followed by Fabaceae and Moraceae (5 species), Mimosaceae, Rutaceae and Poaceae (4 species), Combretaceae, Anacardiaceae, Euphorbiaceae, Apocynaceae (3 species), Asparagaceae, Alangiaceae, Asclepidaceae,

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Meliaceae, Caesalpiniaceae, Verbenaceae, Arecaceae, Ebenaceae, Lamiaceae, Tiliaceae, Myrtaceae and Dipterocarpaceae (2 species), Araceae, Acanthaceae, Annonaceae, Papaveraceae, Bambusaceae, Borangiaceae, Cyperaceae, Droseraceae, Proteaceae, Convolvulaceae, Gyrocarpeaceae, Ulmaceae, Malvaceae, Lythraceae, Nyctaginaceae, Moringaceae, Salicaceae, Araliaceae, Asteraceae and Rhamnaceae (1 species). From the study, it was observed that 51 taxa are trees (60%), 23 taxa are herbs (20%) and 10 taxa are shrub (12%). Fire wood plants: One of the most important non timber forest products for daily life is fuel wood, the only means of energy source of Malayali tribes. The fuel wood species are collected from the forest near to the hamlets. The species preferred for fire wood to their comfortable availability and inflammability. According to Malayali tribes which are not considered as a source of good timber are treated as fire wood. The fire wood utilized by Malayali tribes belong to 15 species, 15 genera and 10 families, they are Albizzia lebbeck Benth, Bauhinia tomentosa L, Buchanania angustifolia Roxb, Chomelia astiactica O.Kze, Diospyros montana, Roxb, Gyrocarpus americanus Jacq, Holopetela integrifolia (Roxb) Planch, Mallotus philippensis, M. Arg. Moringa oleifera Lamk, Plectronia didyma, Kurz, Salix tetrasperma Roxb, Shorea robusta Roth, Syzygium cumini (L) Skeels, Terminalia bellerica Roxb, and Zanthoxylum budrunga Wall. House construction implements:

and

house

hold

Malayali tribes have good Knowledge to use natural resources specifically plants for their day-to-day life. Malayali tribes use Gyrocarpus americanus Jacq leaves to make meal plate, Phoenix sylverstris Roxb leaves to make broom stick, Cyperus rotundus, L, Ophiuros exalatus O.Ktz, Oryza sativa L, Sorghum vulgare L are used for roofing and thatching, Mature stem of Cassia fistula Linn,

Chloroxylon swietenia DC, Dalberiga latifolia Roxb, Grewia tiliaefolia Vahl and Terminalia chebula Retz were used to make pounder, Mature stem of Artocarpus hirsutus Lam is used to make churn-staff, woody stem of Mangifera indica L is used to make wood grinder, leaves of Cassia fistula Linn, is used for ripening of fruits, Gmelina arborea Roxb mature stem is used as a stick for musical instrument, the fibers from the leaf fibers of Agave americana, L and Agave angustifolia, Haw are used for making coir, the culm of Bambusa arundinacea Willd is used for making various types of baskets, leaves of Corypha umbracalifer L is used for making baskets and decoration in ceremonies. Wood forms an important construction material for houses, cattle sheds and temporary settlements in the area. Windows, doors and cots were chiefly made up of Albizia odoratissima, Benth, Cleistanthus Collins Benth, Cordia wallichii, G. Don, Dalberiga lanceolaria, L, Diospyros edenum J.Koeing, Gardenia resinifera, Roth, Grevillea robusta A.Cunn. ex R. Br, Melia composite, Willd, Shorea roxburghii Roxb, Tectona grandis Linn, Zanthoxylum budrunga Wall and Zizyphus mauritiana Lamk. Agricultural implements: For making agricultural implements four species were used such as Chloroxylon swietenia DC and Bumbusa arundinacea Willd were used for making handle of axe and woody stem of Anogeissus latifolia, Wall and Tectona grandis Linn is made in the shape of a harrow by Malayali tribes. Plants used for Religious purposes: Ceremonial plant use is of principal importance in daily Malayali tribe life and many species have a specific ceremonial significance, generally associated with blessings, age-rites and witchcraft. Some plants are used as offerings to god among these tribes. Asclepias curassavica L, Cassia montana Heyne, Clerodendrum serratum L, and Plumeria rubra, L flowers are used in religious

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practice to worship God. Holarrhena pubescens (Buch-Ham.) Wall.ex.G.Don, Wrightia tinctoria R.Br mature stem are used in all traditional religious festivals and religious ceremonies.

Plants used as Toothbrush: Malayali tribals generally use the young twigs of Alangium salvifolium Linn, Azadirachta indica A.Juss, Ficus glomerata Roxb, Ficus religosa Linn, Jatropha curcas L and Psidium guajava Linn, areial root of Ficus benghalensis Linn, Mirabilis jalaba, Linn rhizome powder and Terminalia chebula, Retz fruit powder are used as tooth powder.

Plants used to ward off Evil afflictions: Malayali tribes believe that some of the plants bring good fortune and keep off evil spirits. The exudates of Pterocarpus marsupium, Roxb and Semecarpus anacardium, L are used for marking their childrenâ&#x20AC;&#x2122;s forehead to protect them from evils afflictions, brush body with leaves and root of Toddalia asiatica Lamk to protect from snake bite, brush body with leaves of Schefflera racemosa Harms. protect from evils afflictions, roots of Calotropis gigantea (L) Ait.f is stitched together and then worn around the hip to protect from evils afflictions, rhizome of the Acorus calamus, Linn is made into pieces and stitched together and worn around the neck of a new born baby to get rid from evils afflictions. Malayali tribes use some plants to stimulate sexual desire ie. Abrus pulchellus Wall, Alangium hexapetalum Lam, Canthium parviflorum, Lam, Drosera indica L, Evolvulus alsinoides Linn, Ficus bengalensis Linn, Grewia tenax (forssk.) fiori, Mimosa pudica Linn, Mirabilis jalaba Linn, Ocimum canum, Sims and Randia dumetorum, Lam

Plants used in Hair Nourishment: Heredity, older age, lack of nutrition, infections (such as worms, lice, scabies, dandruff and eczema) and use of synthetic products (soaps, shampoos and hair oils) may cause hair loss, dandruff, discoloration of hairs. Women in Malayali tribes use the natural plant resources for hair disorders. For example Albizzia amara leaves powder was used as a shampoo, seeds of Argemone mexicana, root of Cynodon dactylon were used for blackening the hairs, flowers of Annona squamosa & fruit of Citrus medica were used to remove dandruff and kill lice, whole plant parts of Andrographis echioides, flowers of Hibiscus rosa-sinensis, leaves of Lawsonia inermis and Wedelia calendulacea prevented hair loss.

Table: 1 List of plants used by Malayali Tribal people for NTFP in Yercaud hills. S. No 1 2

3 4 5 6

Botanical Name

Vernacular Name (Tamil)

Family

Parts used

Purposes

Whole plant parts Rhizome

Abrus pulchellus, Wall. Acorus calamus, Linn.

Vellaikuntumani

Fabaceae

Vasambu

Araceae

Agave americana, L. Agave angustifolia, Haw. Alangium hexapetalum, Lam. Alangium salvifolium, Linn.

Aanaikattrazhai Aanaikattrazhai

Asparagaceae Asparagaceae

Leaf fibers Leaf fibers

Brush body with whole plant parts to stimulates sexual desire. Rhizome made into pieces and stitched together and then worn around the neck of new born baby to get rid from evils afflictions. Leaf fibers used for making Coir. Leaf fibers used for making Coir.

Earualangi

Alangiaceae

Alangal

Alangiaceae

Whole plant parts Young twigs

Brush body with whole plant parts to stimulates sexual desire. Young twigs are used as tooth brush.

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 243â&#x20AC;&#x201C;251 Albizia odoratissima, Benth. Albizzia amara, Boivin. Albizzia lebbeck, Benth. Andrographis echioides, Nees.

Poosilai maram

Mimosaceae

Wood

Arappu

Mimosaceae

Leaves

Vakaimaram

Mimosaceae

Gopuramthangai

Acanthaceae

Woody stem Whole plant part

Annona squqmosa, L.

Seetha pazham

Annonaceae

Flowers

Anogeissus latifolia, Wall. Argemone mexicana, L.

Namai

Combretaceae

Wood

Bhrahmadandu

Papaveraceae

Whole plant parts

Artocarpus hirsutus, Lam. Asclepias curassavica, L. Azadirachta indica, A.Juss. Bambusa arundinaceae, Willd.

Kattuppala

Moraceae

Mookuthepoovu

Asclepidaceae

Mature stem Flowers

Vembu

Meliaceae

Mungil

Bambusaceae

Bauhinia tomentosa, L.

Pathini maram Seeramaram

Fabaceae Anacardiaceae

Erukkan

Asclepidaceae

Canthium parviflorum, Lam. Cassia fistula, Linn.

Karamullu

Rubiaceae

Kondrai

Caesalpiniaceae

Cassia monatana, Heyne. Chloroxylon swietenia, DC.

Kallipoo

Caesalpiniaceae

Flowers

Purusa maram

Rutaceae

Mature stem

Chomelia astiatica, O.Kze. Citrus medica, L.

Therani

Rubiaceae

Elamachi

Rutaceae

Woody stem Fruit

Cleistanthus collinus, Benth. Clerodendrum serratum, L.

Woodan

Euphorbiaceae

Wood

Kappukattu sedi

Verbenaceae

Flowers

7 8 9 10

14 15 16 17 18 19 20

21 22

23 24

25 26

27 28

Buchanania angustifolia, Roxb. Calotropis gigantea (L.) Ait.f.

Young twigs Culms Woody stem

Woody stem Root

Whole plant parts Mature stem

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Wood is used to make doors cot and windows. Leaves powder is used as shampoo. Woody stem is used as a fuel. Whole plant parts made into powder. This preparation is applied on scalp to wash hairs it prevent the hair loss. The fresh flowers are made into paste. This preparation is applied on scalp to wash hairs it remove the dandruff. The wood is used to make in the shape of harrow. Brush body with whole plant parts to stimulates sexual desire. Seeds are soaked in oil; the oil is applied on scalp. It used to blacken the hairs. Mature stem is used to make churn-staff. Flowers are used in religious practices to worship to God. Young twigs are used as tooth brush. Split culms are woven into mats, baskets and fans. Woody stem is used as a fuel. Woody stem is used as a fuel. Roots are made into pieces and stitched together and then worn around the hip to protect from evils afflictions. Brush body with whole plant parts to stimulates sexual desire. Mature stem is used to make pounder. Leaves are used for ripening of fruits. Flowers are used in religious practices to worship to God. Mature stem is used to make pounder. The wood is used to make handle of axe. Woody stem is used as a fuel. The fruits are soaked in coconut oil for half an hour. The oil is applied on scalp to wash hairs it remove the dandruff and kill lice. Wood is used to make doors and windows. Flowers are used in religious practices to worship to God.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 243â&#x20AC;&#x201C;251 Cordia wallichii, G. Don. Corpha umbracalifer, Linn. Cynodon dactylon (Linn.) Pers.

Paantheakku

Boranginaceae

Wood

Kuthapannai

Palmaceae

Leaves

Arugampullu

Poaceaea

Root

Cyperus rotundus, L.

Koraipullu

Cyperaceae

Dalberiga lanceolaria, L. Dalberiga latifolia, Roxb. Diospyros edenum, J.Koeing. Diospyros montana, Roxb. Drosera indica, L.

Pulavai maram

Fabaceae

Leaves & branches Wood

Eetimaram

Fabaceae

Karugaali

Ebenaceae

Vellisuli

Ebenaceae

Panithangi

Droseraceae

Evolvulus alsinoides, Linn. Ficus benghalensis, Linn. Ficus benghalensis, Linn. Ficus glomerata, Roxb. Ficus religiosa, Linn. Gardenia resinifera, Roth. Gmelina arborea, Roxb Grevillea robusta, A.Cunn. ex R. Br Grewia tenax (forssk.) fiori. Grewia tiliaefolia, Vahl. Gyrocarpus americanus, Jacq.

Vishnukirandhi Alamaram

Convolvulace ae Moraceae

Woody stem Whole plant parts Whole plant parts Aerial root

Alamaram

Moraceae

Root

Attimaram

Moraceae

Young twigs

Arasa maram Kambaimaram

Moraceae Rubiaceae

Young twigs

Young twigs are used as tooth brush.

Wood

Kumizha maram

Lamiaceae

Savukkumaram

Proteaceae

Mature stem Wood

Atchumullu

Tiliaceae

Thadachimaram

Tiliaceae

Thanakkumaram

Gyrocarpeaceae

Wood is used to make doors and windows. The mature stem is used as stick for musical instruments. Wood is used to make doors and windows. Brush body with whole plant parts to stimulates sexual desire. Mature stem is used to make pounder. Woody stem is used as a fuel. Leaves make meal plate.

Hibiscus rosasinensis, Linn.

Semparuthi

Malvaceae

Holarrhena pubescens, Buch.Ham.)Wall.ex.G.Don. Holopetela integrifolia (Roxb) Planch. Jatropha curcas, L. Lawsonia inermis, L.

Kudasapalai

Apocynaceae

Mature stem

Avali

Ulmaceae

Woody stem

Paikkottai Marudondri

Euphorbiaceae Lythraceae

Young twigs

Young twigs are used as tooth brush.

Leaves

Leaves are crushed and boiled in oil for one hour. The oil is applied on hairs regularly it prevent the hair loss.

29 30 31

34 35 36 37 38 39 40 41 42 43 44 45 46 47 48

52 53

Mature stem Wood

Whole plant parts Mature stem Woody stem and leaves Flowers

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Wood is used to make doors cot and windows. Split leaves are woven into baskets. Roots are soaked in oil. The oil is applied on scalp. It used to blacken the hairs. Leaves and branches are used for roofing and thatching. Wood is used to make doors and windows. Mature stem is used to make pounder. Wood is used to make doors and windows. Woody stem is used as a fuel. Brush body with whole plant parts to stimulates sexual desire. Brush body with whole plant parts to stimulates sexual desire. Aerial root is used as tooth brush. Brush body with whole plant parts to stimulates sexual desire. Young twigs are used as tooth brush.

Flowers are crushed and boiled in oil for one hour. The oil is applied on hairs regularly it prevent the hair loss. Mature stem is used in all traditional religious festivals and religious ceremonies. Woody stem is used as a fuel.

Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 243–251 Mallotus philippensis, M. Arg. Mangifora indica, L.

Thirichichilai maram Mamaram

Euphorbiaceae Anacaradiaceae

Woody stem Wood

Melia composita, Willd. Mimosa pudica, Linn.

Malaivembu

Meliaceae

Wood

Thottal surungi

Mimosaceae

Mirabilis jalaba, Linn.

Anthimantharai

Nyctaginaceae

Whole plant parts Whole plant parts

Moringa oleifera, Lamk. Ocimum canum, Sims. Ophiuros exalatus, O.ktz. Oryza sativa, L.

Murungai

Moringaceae

Ganjamkori

Lamiaceae

Kenangupullu

Poaceae

Nellu

Poaceae

Phoenix sylvestris, Roxb. Plectronia didyma, Kurz. Plumeria rubra, L.

Icham

Palmaceae

Nikkanai maram

Rubiaceae

Paalarali

Apocynaceae

Psidium guajava, Linn.

Koiya

Myrtaceae

Young twigs

Pterocarpus marsupium, Roxb.

Vangai

Fabaceae

Exudates

Randia dumetorum, Lam. Randia malabaricum, Lam Salix tetrasperma, Roxb. Schefflera racemosa Harms. Semecarpus anacardium, L. f.

Aaichumulu

Rubiaceae

Kaarakkai

Rubiaceae

Whole plant parts Wood

Vanji

Salicaceae

Peiviratti

Araliaceae

Woody stem Leaves

Saramaram

Anacardiaceae

Exudates

73 74

Shorea robusta, Roth. Shorea roxburghii, Roxb.

Salamaram Kunkilium

Dipterocarpaceae

Woody stem

Dipterocarpaceae

Wood

Sorghum vulgare, L.

Solam

Poaceae

Syzygium cumini, (L.) Skeels. Tectona grandis, Linn.

Naaval

Myrtaceae

Thekku

Verbanaceae

Leaves & branches Woody stem Wood

54 55 56 57

60 61 62 63 64 65

69 70 71 72

Woody stem Whole plant parts Leaves & branches Leaves & branches Leaves Woody stem Flowers

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Woody stem is used as a fuel. Wood is used to make wood grinder. Wood is used to make doors cot and windows. Brush body with whole plant parts to stimulates sexual desire. Brush body with whole plant parts to stimulates sexual desire. Rhizome powder is used as tooth powder. Woody stem is used as a fuel. Brush body with whole plant parts to stimulates sexual desire. Leaves and branches are used for roofing and thatching. Leaves and branches are used for roofing and thatching. The leaves are used to make broom stick. Woody stem is used as a fuel. Flowers are used in religious practices to worship to God. Young twigs are used as tooth brush. Woody stem is used as a fuel. Exudates used for marking their children’s forehead to protect from evils. Brush body with whole plant parts to stimulates sexual desire. Wood is used to make windows. Woody stem is used as a fuel. Brush body with leaves to protect from evils afflictions. Exudates used for marking their children’s forehead to protect from evils. Woody stem is used as a fuel. Wood is used to make doors cot and windows. Woody stem is used as a fuel. Leaves and branches are used for roofing and thatching. Woody stem is used as a fuel. Wood is used to make doors cot and windows. Woody stem is used as a fuel. The wood is used to make in the shape of harrow.

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Terminalia bellerica, Roxb.

Thanrikkai

Combretaceae

Woody stem

Woody stem is used as a fuel.

Terminalia chebula, Retz.

Kadukkai

Combretaceae

Wood and fruit

Wood is used to make doors cot and windows. Fruit powder is used as tooth powder. Woody stem is used as a fuel.

Toddalia asiatica, Lamk. Wedelia calendulacea, Less.

Mulaikaradan mullu Manjakarisalangan i

Rutaceae

Leaves and root Leaves

Wrightia tinctoria, R.Br.

Palai

Apocynaceae

Mature stem

Zanthoxylum budrunga, Wall

Karunjoori

Rutaceae

Wood

Zizyphus mauritiana, Lamk.

Yellandai

Rhamnaceae

Wood

Brush body with leaves and root to protect from snake bite. Leaves are crushed and boiled in oil for one hour. The oil is applied on hairs regularly it prevent the hair loss. Mature stem is used in all traditional religious festivals and religious ceremonies. Wood is used to make doors cot and windows. Woody stem is used as a fuel. Wood is used to make doors cot and windows.

Asteraceae

CONCLUSION The present investigation revealed that significant role of plants used by Malayali tribes in Yercaud hills. It is clear that these products are extremely important and significant component of the household livelihood of Malayali tribes. The information collected from tribals is useful for carrying out further research in the field of ethnobotany, taxonomy. These indigenous plants must be taken into consideration and treated with equal importance as that of other plant species, many research works must be carried out on these plants to increase their productivity which will help in increasing our country’s economy. The diversity of species used by Malayali tribes is

incredible and this sound knowledge has been documented through this study. ACKNOWLEDGEMENT We the authors extend our special thanks to Mr. N. Thangaraju, I.F.S., District Forest Officer, Salem Division, Salem, Tamil Nadu, India for giving permission to carry out this Research work in Yercaud hills area. Its our pleasant duty to express our gratitude to the local people in the Yercaud hills for sharing their knowledge on plants. We gratefully acknowledge Mr. R. Prabakaran, Department of Botany, Vivekanandha College of Arts and Sciences for Women (Autonomous), Elayampalayam for his help in identification of the Plant species.

REFERENCES Anthony P Cavender and Manuel Alban. (2009). The use of Magical plants by curanderos in the Ecuador highlands. Journal of Ethnobiology and Ethnomedicine. 5 (3): 1–9.

Ayyanar, M and Ignacimuthu, S. (2010). Plants used for non-medicinal purposes by the tribal people in Kalakad Mundanthurai Tiger Reserve, Southern India. Indian Journal of Traditional Knowledge. 9 (3): 515–518.

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Gamble, JS and Fischer, CEC. (1935). Flora of Presidency of Madras, London (Issued in II part: 1–7 By Gamble, 8-11 by Fischer), Vol. 1–3, Calcutta. John Kennedy, S M. (2006). Commercial Nontimber forest products collected by tribals in the Palni hills. Indian Journal of Traditional Knowledge. 5 (2): 212– 216. Mathew, KW. (1983). Flora of Tamil Nadu Carnatic, the Rapinat Herbarium, Tiruchirapalli, India. 3 vol. Prabakaran, R, Senthil Kumar, T and Ravo, M.V. (2013). Role of Non Timber Forest Products in the Livelihood of Malayali tribe of Chitteri hills of Southern Eastern Ghats, Tamil Nadu. Journal of Applied Pharmaceutical Sciences. 3 (05): 056–060.

Source of Support: NIL

Sanjay Kr Uniyal, Anjali Awasthi and Gopal S Rawat. (2002). Traditional and ethnobotanical uses of plants in Bhagirathi Valley (Western Himalaya). Indian Journal of Traditional Knowledge. 1 (1): 7–19. Subbaiah Muruganandam, Singaram Rathinakumar and Arunachalam Selvaraju. (2012). Plants used for nonmedicinal purposes by Malayali tribals in Jawadhu Hills of Tamil Nadu, India. Global Journal of Research on Medicinal plants & Indigenous Medicine. 1 (12): 663–669. Vijay V. Wagh, Ashok K. Jain and Chitralekha Kadel. (2010). Role of forest products in the livelihood of tribal community of Jhabua district (M.P.). Biological Forum – An International Journal, 2 (1): 45–48.

Conflict of Interest: None Declared

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Review Article MICROPROPAGATION: AN ESSENTIAL TOOL TO FLOURISH ENDANGERED MEDICINAL PLANTS Sharma Rohit1* 1

Plant Biotechnology Laboratory, R&D Division, Tropilite Foods Pvt. Ltd., Davars Campus, Tansen Road, Gwalior-474002 (M.P.), India *Corresponding Author: E-mail: accessrohit25@gmail.com; Mob: +919755594040

Received: 10/05/2014; Revised: 22/05/2014; Accepted: 28/05/2014

ABSTRACT Micropropagation or tissue culture of plants holds tremendous potential for the production of high-quality plant-based medicines. This technique is of vital use in germplasm preservation of various useful endangered medicinal plants species with a multiplication benefit of production of large number of plants starting from single explant. Even temperature dependent species can be maintained throughout the year provided with specific media and environment. Tissue cultured plants are generally free from fungal and bacterial diseases. Virus eradication and maintenance of plants in a virus-free-state are also readily achieved in tissue culture. Propagating media holds a key aspect for micropropagation. The paper reviews the recent advances, achievements and potential of tissue culture for in vitro regeneration with a special elucidation of media salt and hormonal requirements of different medicinal plants. KEY WORDS: Micropropagation, tissue culture, saponin, breeding, propagation, in vitro. ABBREVIATIONS: BA = 6-Benzylaminopurine NAA = a-Naphthalene acetic acid IAA = Indole-3 acetic acid 2iP = 6-(g -Dimethylallylamino) purine 2, 4-D = 2,4-Dichlorophenoxyacetic acid KN = Kinetin TZD = Thiazolidinedione

Cite this article: Sharma Rohit (2014), MICROPROPAGATION: AN ESSENTIAL TOOL TO FLOURISH ENDANGERED MEDICINAL PLANTS, Global J Res. Med. Plants & Indigen. Med., Volume 3(6): 252â&#x20AC;&#x201C;262

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INTRODUCTION The creation of nature is thoughtful and meaningful. It is important to maintain the balance between plant and animal kingdom. But because of various reasons, the plant resources were destroyed in last few decades, and many species were rendered threatened or extinct. Medicinal plants play a key role in world health care systems. Herbal medicine is one of the most remarkable uses of plant based biodiversity. These plants constitute an important natural wealth of a country. They play a significant role in providing primary health care services to people and serve as therapeutic agents as well as important raw materials for the manufacture of traditional and modern medicine (Debnath et al., 2010). Reserves of herbs and stocks of medicinal plants in developing countries are diminishing and in danger of extinction as a result of growing trade demands for cheaper healthcare products and new plant-based therapeutic markets in preference to more expensive targetspecific drugs and biopharmaceuticals. Indian continent is the repository of large number of medicinal plants, and most of these are available as wild plants in the forests of hills and planes. The biotechnological tools are important to select, multiply and conserve the critical genotypes of medicinal plants. Disturbances of the natural habitats of these plants, as a result of anthropogenic activities and invasion of the exotic species has resulted in the drastic decline in the population of these important plant species and many of these species are now listed among the rare, critically rare and endangered category (Thakur et al., 2009). Conventional plant-breeding methods can improve both agronomic and medicinal traits. In vitro propagation or tissue culture of plants holds tremendous potential for the production of high-quality plant-based medicines. This can be achieved through different methods including micropropagation. The evolving commercial importance of secondary metabolites has in recent years resulted in a great interest in secondary metabolism, particularly in the possibility of altering the

production of bioactive plant metabolites by means of tissue culture technology. Plant cell culture technologies were introduced at the end of the 1960's as a possible tool for both studying and producing plant secondary metabolites. Different strategies, using an in vitro system, have been extensively studied to improve the production of plant chemicals (Debnath et al., 2006). Micropropagation has many advantages over conventional methods of vegetative propagation, which suffer from several limitations. There has been significant progress in the use of tissue culture and genetic transformation to alter pathways for the biosynthesis of target metabolites. Obstacles to bringing medicinal plants into successful commercial cultivation include the difficulty of predicting, which extracts will remain marketable and the likely market preference for what is seen as natural source of extracts. Unlike the conventional methods of plant propagation, micropropagation of even temperature species may be carried out throughout the year and the produced tissue culture plants are generally free from fungal and bacterial diseases. Virus eradication and maintenance of plants in a virus-free-state are also readily achieved in tissue culture. The high multiplication rate also permits the production of pathogen free material. SELECTED MEDICINAL PLANTS AND THEIR MICROPROPAGATION PROTOCOL 1. Chlorophytum borivilianum Chlorophytum borivilianum Santapau & Fernandes (Liliaceae) also known as â&#x20AC;&#x17E;Safed Musliâ&#x20AC;&#x; is a traditional rare Indian medicinal herb which has many therapeutic applications in Ayurvedic, Unani, Homeopathic and Allopathic systems of medicine. Its roots (tubers) are widely used for various therapeutic applications. It is used to cure physical illness and weakness, as an aphrodisiac agent and revitalizer, as a general sex tonic, remedy for diabetes, arthritis and increasing body immunity, curative for natal and postnatal problems, increase lactation in feeding mothers, as antimicrobial, anti-inflammatory, antitumor

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agent, also used in diarrhea, dysentery, gonorrhea, leucorrhea etc (Thakur et al., 2009). Novel propagation techniques like tissue culture can play an important role in the rapid multiplication of elite clones and germplasm conservation of C. borivilianum. Plant micropropagation is an efficient method of breeding and propagating disease free, genetically uniform and massive plants in vitro. Rapid micropropagation procedure for this endemic medicinal plant from various explants in solid media (Dave et al., 2004) and in liquid media (Rhizvi et al., 2007) has been reported. Leaf base segments, stem disc with shoot meristem, sliced root tuber, young shoot buds, immature floral buds and inflorescence axis were used as explants. Debnath et al. described that regeneration of C. borivilianum from both young shoot buds as well as inflorescence axis bearing an axillary bud gave good results. Regeneration was described successfully both from the inflorescence axis and young shoot buds on MS basal medium supplemented with (1–5 mg/l) BAP. Extensive work has been done on the micropropagation of C. borivilianum including somatic embryogenesis, callus culture, encapsulation of young shoot buds and plantlets regeneration, in situ Hybridization (FISH) technique (Debnath et al., 2007). Stem disc explant of C. borivilianum transferred to MS medium supplemented with 5 mg/L BAP gave the maximum shoot proliferation and shoot bud initiation (14.83 shoots) as compared to different concentration of BAP. BAP at 5 mg/L after sub-culturing produced highest significant value of shoot numbers per explant (14.83) with no callusing in the cultures (Purohit et al., 1994b). Thus, there was no chance of genetic variability. Kn (kinetin) showed no increment in shoot number. The observations revealed that different concentration of cytokinins influenced the shoot length of the in vitro growth of C. borivilianum. It was further observed that Kn at 5 mg/L gave the optimum shoot length (4.11 cm) and BAP at 5 mg/L gave the maximum (4.51 cm) shoot length. Contrary to the reports of Purohit et al. (1994a), the interaction of BAP and Kn showed no significant results in shoot proliferation and

shoot elongation. On sub-culturing, shoot proliferation and shoot elongation was not affected by different strengths of MS media and maximum shoots (4.33) were observed in the full strength MS media but ½ MS medium showed significant shoot length (7.51 cm) elongation (Thakur et al., 2013). Auxins showed a significant effect on supplementation in MS medium on the root initiation, proliferation and growth. For MS medium supplemented with IBA at 2 mg/L, numbers of roots/explant were less (3.58) compared to IAA at 2 mg/L (3.67). The results obtained with the studies on combination of IBA and IAA supplementation in MS medium was not significant. Observations on the effect of different strengths of medium concentration (Full MS, 1/2 MS, ¾ MS) on root number/explant and root length/explant revealed that maximum number of roots were obtained in ½ MS medium (6.42), optimum response in full MS (3.67) and minimum in ¾ MS (3.08). The maximum lengths of roots were also obtained in ½ MS medium (7.24 cm). Handling of liquid medium is easier in comparison to solid medium (Rhizvi et al., 2007) and so the response of rooting was observed using liquid media. The plant yields a flavonone glycoside, which is a powerful uterine stimulant, steroidal saponins which have muscle building properties and their structure is similar to male anabolic hormones testesterone. Roots of Chlorophytum contain 42% carbohydrate, 80–89% protein, 3–4% fiber and 2–17% saponin. Research studies on Chlorophytum conducted in India and elsewhere indicate that saponins are responsible for medicinal properties. C. borivilianum produces the maximum root tuber along with the highest saponin content (Attele et al., 1999). Traditionally, roots of these species are reputed to possess various pharmacological utilities having saponins as one of the important phyto-chemical constituents (Sharma et al., 2012). Furthermore saponin from Chlorophytum is a hidden gift from nature which is now proving its efficacy and potential as a miracle herb for biopharmaceutical and neutraceutical attention for human welfare (Sharma et al., 2014).

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Fig. 1: Micropropagation of Chlorophytum borivilianum

(A) Initiation in MS media, (B, C) Shoot multiplication in MS media supplemented with 5 BAP, (D, E, F) Multiple shoot and root indcution in MS media supplemented with 5 BAP + 1 K + 2 IBA.

2. Momordica balsamina Momordica balsamina also known as „Balsam apple‟ (African pumpkin), is an important medicinal and nutritional plant of Cucurbitaceae. It is an annual to perennial tendril-bearing herb native to tropical regions of Africa. In India, it occurs naturally in forest, in the rainy season. The leaves, fruits, seeds, and bark are reported to have various medicinal and nutritional importance and called „Hidden gift of Nature‟ (Thakur et al., 2009). The fruit extract of M. balsamina shows anti-HIV property (Bot et al., 2007). „Momordins‟ are capable of inhibiting the growth of HIV and other viruses. The leaves and fruit extracts of this plant shows antiplasmodial activity and is being used against malaria in African traditional medicine. The extract of various parts of this plant shows shigellocidal, antidiarrhoeal, antiseptic, antibacterial, antiviral, antinflammatory, hypoglycemic and antimicrobial properties (Hassan and

Umar, 2006; Akinyemi et al., 2005; Jigam et al., 2004). The whole plant is used as a bitter stomachic and an infusion is used as a wash in the management of fevers and yaws. A macerate of the whole plant is also used as a galactogogue and to massage the chest to relieve intercostals pains. The plant is sometimes used as an ingredient in aphrodisiac preparations. The wallops in Senegal have used the fruits as purgative agents and vermifuge. The fruits and leaves are used for treatment of wounds in Nigeria and in Syria as hemostatic antiseptic. The whole plant is used as sponge in treating skin disease such as scabies and as tranquilizer in the treatment of mental illness (Akinniyi et al., 1986). The aqueous leaf extract of M. balsamina has also been used in reducing and relieving period pain in young girls. The natives also use the seed of the plant in arrow poison. The whole plant extract has insecticidal properties.

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The pulverized plant is applied externally against malignant ulcers. Young peeled fruits are cooked and eaten; they are often steeped in salt water after peeling and before cooking to remove its bitter taste (Thakur et al., 2009). The fruits are common ingredients in Indo Pakistan pickles and are often used in curries and meat dishes. Tender shoots are usually consumed with Okra soup by the Kanuris of Borno State where the plant is locally known as dagdawu. Phytochemical screening of M. balsamina Linn has revealed the presence of tannins, saponins and lectins (Akinniyi et al., 1986). High frequencies of multiple shoot regeneration were achieved from auxillary buds of nodal explants by Thakur et al. 2011. The bud explants were cultured on MS media supplemented with different concentration of BAP. 1 mg/L BAP stimulated proliferation of the bud meristems to form bud clusters and the co-efficient reached 6–8. The maximum survival percentage of explant was also observed in the same medium compared to different concentrations of BAP. It is clear that MS medium supplemented with BAP at 1 mg/L was most effective in multiplication of shoots but BAP at different concentrations showed high callusing. Activated charcoal at 0.2% was found to be most effective to inhibit the callus growth (Thakur et al., 2011). However, BAP at 1 mg/L with 0.2% activated charcoal achieved most significant value of shoot. The shoot elongation was prominent in MS medium supplemented with 1 mg/L BAP and 1 mg/L Kn, 0.2% activated charcoal and 0.01% glutamine. Regeneration of Momordica dioica on MS medium supplemented with 1 mg/L BAP and 0.1 mg/L NAA from node shoot tip, leaf and cotyledon explants was reported earlier. Contrary to the studies on M. balsamina, multiple shoot regeneration of Cucumis melo using shoot tips as explant was found at 2.5 mg/L NAA and 1 mg/L BAP (Thakur et al., 2009). 3. Bacopa monnieri Bacopa monnieri, also referred to as Bacopa monniera, Herpestis monnieri, water

hyssop, and “Brahmi”, has been used in the Ayurvedic system of medicine for centuries. Bacopa monniera, a member of the Scrophulariaceae family, is a small, creeping herb with numerous branches, small oblong leaves, and light purple flowers. In India and the tropics it grows naturally in wet soil, shallow water, and marshes. The herb can be found at elevations from sea level to altitudes of 4,400 feet, and is easily cultivated if adequate water is available. Flowers and fruit appear in summer and the entire plant is used medicinally. Brahmi may be useful for people who want to improve mental function and concentration particularly under pressure or in stressful conditions. Research on anxiety, epilepsy, bronchitis and asthma, irritable bowel syndrome, and gastric ulcers also supports the Ayurvedic uses of Bacopa. Bacopa’s antioxidant properties may offer protection from free radical damage in cardiovascular disease and certain types of cancer. An effective protocol for mass propagation of Bacopa monnieri (L.) Pennell, an important medicinal plant (Shalini et al., 1998) was developed using shoot tips and nodal segments as explants (Debnath et al., 2006). Shoot regeneration and somatic embryogenesis from different explants of Brahmi. The explants were cultured on Murashige and Skoog's medium supplemented with various auxins, cytokinins either alone or with coconut milk and auxins plus cytokinins. Multiple shoots were obtained on MS medium supplemented with auxins or/and cytokinins with or without coconut milk. Maximum number of plants were obtained on medium containing KN/2-ip (0.1 mg/l) and KN (1 mg/1) in shoot tip and nodal cultures, respectively. The regenerated shoots developed roots on the same medium. In our lab, similar observation was noticed. The initiation of shoot proliferation from nodal explants was observed on MS medium supplemented with BAP (1 mgl-1) and KN (4 mgl-1). When these were subcultured on MS medium supplemented with BAP (5 mgl-1) multiple shoot. Bud proliferation was further enhanced. They were found to originate, associated with or without roots. Shoot elongation was observed on the same medium. These shoots when transferred to 1\2

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MS medium supplemented with IBA (2 mgl-1) resulted in rooting. Regenerated plantlets were transferred to soil after a very brief period of

hardening. The regenerated plants are dark green in colour and more robust in their growth than the normal plants.

Fig. 2: Micropropagation of Bacopa monniera and Momordica balsamina

(A) Initiation stage of Bacopa monniera in MS media, (B, C) Shoot and root multiplication of Bacopa monniera in MS media supplemented with 1 BAP + 1K + 0.5 IAA, (D) Hardening of Bacopa monniera in pots., (E, F) Shoot multiplication of Momordica balsamina in MS media supplemented with 1 BAP + 0.5K

4. Tinospora cordifolia Tinospora cordifolia is a deciduous climbing shrub described as „the one who protects the body against diseases‟. It is one of the most versatile rejuvenating shrub also known as ’Giloya’ in Indian vernacular having many therapeutic applications. The pharmaceutical significance of this plant is mainly because of the leaves, barks and roots contain various bioactive compounds such as alkaloids, glycosides, lactones, steroids, polysaccharides and aliphatic compounds having various medicinal importance viz.

immuno-modulatory or immuno-stimulatory, antitumor, cognition, anti-inflammatory, antineoplastic, anti-hyperglycemia, antihyperlipidemia, antioxidant, anti-tubercular, gastrointestinal and hepatoprotection, antiosteoporotic, anti-angiogenic, anti-malarial and anti-allergic. The bitter principles present in the leaves, stems, roots and barks viz. tinosporine, tinosporide, tinosporaside, cordifolide, cordifol, berberine, cordifolioside A, B. C, amritosides A, B, C, and columbin which act as therapeutic agents and play vital role in many therapeutic applications (Pandey et al., 2012). The nodal segments inoculated in MS basal medium

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supplemented with 0.5 mg/l IBA + 0.2 mg/l NAA produced multiple shoots with an average height of 9.98 ± 0.88 after 2 weeks of incubation. Regenerated shoots were rooted on half strength MS basal medium containing 1.0 mg/L BA = 0.2 mg/L IAA. Rooted plantlets were transferred to pots containing soil for acclimatization, for a period of three weeks and were successfully established in soil. After acclimatization and transplantation 100% acclimatized plantlets were found healthy in ex-vivo conditions (Bhat et al., 2013). 5. Calotropis procera Calotropis procera a giant milk weed, is known for its pharmacological importance for centuries. The coarse shrub is a very promising source of anticancerous, ascaricidal, schizonticidal, anti-microbial, anthelmintic, insecticidal, anti-inflammatory, anti-diarrhoeal, larvicidal with many other beneficial properties. Plant is described as a golden gift for human kind containing calotropin, calotropagenin, calotoxin, calactin, uscharin, amyrin, amyrin esters, uscharidin, coroglaucigenin, frugoside, corotoxigenin, calotropagenin and voruscharine used in many therapeutic applications. Different compounds like norditerpenic esters, organic carbonates, the cysteine protease procerain, alkaloids, flavonoids, sterols and numerous cardenolides made this plant of scientific attraction for centuries. Plant is not only a great source of natural hydrocarbons but also contains several metabolites used as folk medicine for the treatment of leprosy, elephantiasis, fever, menorrhagia, malaria and snake bite (Sharma et al., 2012). In addition, latex-derived extracts induce selective cytotoxicity and anti-tumor activity (Mathur et al., 2009). The latex of C. procera has been shown to protect against gastric ulcers in rats, and latex extracts protected against inflammation and oxidative stress in arthritic rats (Bharti et al., 2010). Healthy seeds of C. procera were treated with 70% (v/v) ethanol for 2 min before surface sterilization by dipping into 33% (v/v) sodium hypochlorite solution (v/v) for 15 min, followed by five rinses in sterile distilled water. Thereafter, five seeds were germinated in each

glass flask containing 30 ml of 1/2 MS medium salts, supplemented with 100 mg/L myoinositol, and 3% (w/v) sucrose (pH 5.8) that was solidified with 0.7% (w/v) agar. Cultures were maintained at a photoperiod of 12 h and illuminated with cool white fluorescent lamps at a light intensity of 10–15 μmol/m2/s. The culture room temperature was kept at 28 ± 2°C. After 30 days under these culture conditions, plantlets reached ca. 5 cm in height and were used as an explant source. In vitro-grown plantlets (30 days after germination) were used as the source of hypocotyl and cotyledon explants. Hypocotyls were aseptically removed and cut longitudinally and then cut into slices approximately 10 mm in length. Callus induction medium consisted of complete MS basal formulation as prepared for seed germination added of 3 μM 1-naphthylacetic acid (NAA) and 4.6 μM kinetin (KIN) (Teixeira et al., 2011). 6. Gymnema sylvestre Gymnema sylvestre also known as „gurmar’ or „sugar destroyer‟ is a woody, climbing traditional medicinal herb which has many therapeutic applications in Ayurvedic system of medicine. It is used for lowering serum cholesterol, triglycerides and blood glucose level (hypoglycemic or antihyperglycemic), hypolipidaemic, weight loss, stomach ailments, constipation, water retention and liver diseases, either high or low blood pressure, tachycardia or arrhythmias, and used as aperitive, purgative, in eye troubles, anti-inflammatory, smooth muscle relaxant, prevention of dental caries, cataract and as anticancer-cytotoxic agent (Thakur et al., 2012). Its flowers, leaves, and fruits contains alkaloids, flavones, saponins, sapogenins, anthraquinones, hentriacontane, pentatriacontane, α and βchlorophylls, phytin, resins, d-quercitol, tartaric acid, formic acid, butyric acid, lupeol, β-amyrin related glycosides and stigmasterol having main principle bioactive compounds viz. gymnemic acids, gymnemasides, gymnemagenin, gurmarin, gymnemosides, gymnemanol, gymnemasins, gypenoside, and conduritol which act as therapeutic agent and play vital role in many therapeutic applications. Gymnemic acids are thought to be responsible

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for its antidiabetic activity and it is the major component of an extract shown to stimulate insulin release from the pancreas. Nodal segments were washed thoroughly under running tap water for 30 minutes, followed by treatment with Bavistin solution 1% for 15 minutes. It was further washed under tap water thoroughly for pre-sterilization and then rinsed with distilled water thrice and finally surface sterilization was carried out for 3 minutes with 0.01% mercuric chloride (HgCl2) and then explants were washed 3 times with sterile distilled water. After surface sterilization, the explants were inoculated on MS medium 4 containing 3% (w/v) sucrose, various concentrations of cytokinin (BAP) (0.5, 1.0, 1.5, 2.0, 3.0 and 4.0 mg/l) and auxin

(NAA) (0.5, 1.0, 1.5 and 2.0 mg/l). The pH of the medium was adjusted to 5.7 before gelling with 0.8% (w/v) agar and autoclaved at 121°C for 15 minutes. Among the various concentrations (0.5, 1.0, 1.5, 2.0, 3.0 and 4.0 mg/l) of BA used, the best response was observed at 1.0 mg/l. A maximum number of 7 shoots (Plate 1b) was observed at 1.0 mg/l BA with the average length of 3.4 cm The multiple shoots were harvested and transferred to rooting medium with different concentrations of NAA (0.5–2.0 mg/l). A maximum number of roots were observed at 1.0 mg/l NAA with the average length of 2cm. BAP is said to be the inducer of multiple shooting (Manonmani and Francisca, 2012).

Table 1: Micropropagation media of different medicinal plants with their potent media, active metabolite and specific action Plant

Active Metabolite Camptothecin

antitumor

vincristine, vinblastine

anticancerous

Coleus forskohlii

Forskolin

cardiovascular

Heliotropium indicum

Heliotrine

antitumor, hypotensive

Momordica charantia Saussurea lappa Simarouba glauca

Charantin

antidiabetic

Saussurine

bronchiorelaxant

Glaucarubin

antiamebic

Trichosanthin

abortifacient

Camptotheca acuminata Catharanthus roseus

Trichosanthes kirilowii

Action

Micropropagation media WPM + 4 μM BAP MS + 1 mg/l BAP +0.2 mg/l αnaphthaleneacetic acid MS + 0.57 μM IAA + 0.46 μM kinetin MS + 1mg/l Kinetin + 0.5 mg/l BAP + 0.05 mg/l IAA MS + 2 mg/l BAP + 0.5 mg/l Kinetin MS+ 4.4 μM BAP + 0.45 μM TDZ MS + 11.1 μM benzyladenine + 13.42 μM α-naphthaleneacetic acid. MS + 2 mg/L BA + 0.5 mg/L NAA

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Reference Linebereger et al., 1998 Kumar et al., 2013 Bhattacharya and Bhattacharya, 2001 Kumar and rao, 2007 Agarwal and Kamal, 2004 Johnson et al., 1997 Rout and Das, 1994

Yang et al., 2006

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7. Catharanthus roseus Catharanthus roseus originated from Madagascar, but has now been spread throughout the tropics and subtropics by human activity. This plant is cultivated as an ornamental plant throughout tropical and subtropical areas and many parts of the world. Catharanthus is a plant belonging to the family, well known for being rich in alkaloids. All parts of plant, especially, the leaves contain antineoplastic bisindole alkaloids, among those are vinblastine and vincristine, which are widely employed as chemotherapeutic agents against cancer. The absolute levels of vinblastine and vincristine are considered far too low to explain the activity of crude extracts of Catharanthus. Various studies show the presence of other antineoplastic alkaloids in the plant (Piovan and Filippini, 2000). Callus induction from nodal explants was observed on Murashige and Skoog by Debnath et al. 2006 MS medium supplemented with NAA (0.2 mgl1 ) and KN (2 mgl-1). Multiple shoot proliferation and shoot elongation was observed on MS medium supplemented with NAA (0.5 mgl-1) and KN (2 mgl-1). These shoots when transferred to MS medium supplemented with IBA (2 mgl-1) resulted in

rooting. For alkaloid extraction, in vitro plant multiplication is an ideal approach to produce leaf material in large quantity. In vitro heavy metal stress on callus culture and regeneration of pathogen free healthy plants through callus culture have been reported (Debnath et al., 2006). CONCLUSION Micropropagation of medicinal plants with enriched bioactive principles and cell culture methodologies for selective metabolite production is found to be highly useful for commercial production of medicinally important compounds. The efficient techniques plant tissue culture are playing a crucial role in serving the life science and opening new horizons in preserving endangered species compiling our green world. Tissue culture protocols have been developed for several plants but there are many other species, which are over exploited in pharmaceutical industries and need conservation. These new technologies will serve to extend and enhance the continued usefulness of higher plants as renewable sources of chemicals, especially medicinal compounds.

REFERENCES Agarwal, M,, Kamal, R., (2004) In vitro clonal propagation of momordica charantia L. Indian Journal of Biotechnology, 3, 426–430. Akinniyi, J.A., Sultanbawa, M.U.A., Manawaku, D, (1986) In: The State of Medicinal Plant Research in Nigeria. University Press: lbadan Nigeria, pp. 154–165 Akinyemi, K.O., Mendie, U.E., Smith, S.T., Oyefolu, A.O., Coker, A.O. (2005) Screening of some medicinal plants used in south-west Nigerian traditional medicine for anti-salmonella typhi activity. J Herb Pharmacother. 5(1): 45–60.

Attele, A.S., Wu, J.A., Yuan, C.S. (1999). Ginseng pharmacology: Multiple constituents and multiple actions. Biochemical. Pharmacology, 58, 1685– 1693. Bhat, T.M., Singh, M., Malik, T. (2013) Micropropagation of Tinospora cordifolia (willd) Miers ex Hook. F. Thoms – A threatened medicinal plant. IAJPR, 3(5), 3636–3641 Bhattacharyya, R., Bhattacharyya, S. (2001) In vitro multiplication of Coleus forskohlii Briq.: An approach towards shortening the protocol. In Vitro Cellular & Developmental Biology – Plant, 37(5), 572–575

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Bot, Y.S., Mgbojikwe, L.O., Nwosu, C., Abimiku, A., Dadik, J., Damshak, D. (2007) Screening of the fruit pulp extract of Momordica balsamina for anti HIV property. African journal of Biotechnology, 6, 047–052. Dave, A., Joshi, N., Purohit, S.D. (2004). In vitro propagation of Chlorophytum borivilianum using encapsulated shoot buds. European Journal of Horticulture Science, 69(1), 37–42. Debnath, M., Malik, C.P., Bisen, P.S. (2006) Micropropagation: A tool for the production of high quality plant-based medicines. Current Pharma Biotechnology, 7, 33–49 Debnath, M., Malik, C.P., Bisen, P.S. (2007) Clonal propagation of Chlorophytum borivilianum- An endangered medicinal plant. Phytomorphology, 57, 117–121. Debnath, M., Pandey, M., Sharma, R., Thakur, G.S., Lal, P. (2010) Biotechnological intervention of Agave sisalana: A unique fiber yielding plant with medicinal property. Journal of Medicinal Plants Research, 4(3), 177– 187.

Kumar, A., Prakash, K., Sinha, R.K., Kumar, N. (2013) In vitro plant propagation of Catharanthus roseus and assessment of genetic fidelity of micropropagated plants by RAPD marker assay. Applied Biochemistry Biotechnology, 169(3), 894–900. Kumar, M.S., Rao, M.V. (2007) In vitro micropropagation of Heliotropium indicum Linn. An ayurvedic Herb. Indian journal of Biotechnology, 6, 245–249 Limburger, D., Reed, D., Rumpho, M. (1998) Micropropagation of Camptotheca acuminata. Hort Science, 33, 604. Manonmani, R., Francisca, P. (2012) In vitro multiplication of Gymnema sylvestre R. BR. Through nodal explants. International Journal of Pharma and Bio Sciences, 3, 2. Pandey, M., Chikara, S.K., Vyas, M.K., Sharma, R., Thakur, G.S., Bisen, P.S. (2012) Tinospora cordifolia: A climbing shrub in health care management. International Journal of Pharma and Bio Sciences, 3(4), 612– 628.

Hassan, L.G., Umar, K.J. (2006) Nutritional value of balsam apple leaves. Pakistan Journal of Nutition, 5, 522–529.

Piovan, A., Filippini, R. (2000) Somatic embryogenesis and indole alkaloid production in Catharanthus roseus. Plant Biosystems, 134(2), 179–184.

Jigam, A.A., Akanya, H.O., Adeyemi, D.J. (2004) Antimicrobial and antiplasmodial effects of Momordica balsamina. Niger J Nat Prod Med. 8, 11–12.

Purohit, S.D., Dave, A., Kukda, G. (1994a) Micropropagation of Safed Musli (Chlorophytum borivilianum), a rare Indian medicinal herb. Plant Cell Tissue Organ Culture, 39, 93–96.

Johnson, T.S., Narayan, S.B., Narayana, D.B.A (1997) Rapid in vitro propagation of Saussurea lappa, an endangered medicinal plant, through multiple shoot cultures. In Vitro Cellular & Developmental Biology – Plant, 33(2), 128–130

Purohit, S.D., Dave, A., Tiagi, Y.D. (1994b) Chlorophytum borivlianum Sant. and Fern. An (Liliaceae) interesting species from the Aravallis in Rajasthan. Rheedea, 4, 113–115.

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Rhizvi, M.Z., Kukreja, A.K., Knanuja, S.P.S. (2007) In vitro culture of Chlorophytum borivilianum Sant .et. Fernand .in liquid culture medium as a cost effective measure. Current Science, 92(1), 87–90. Rout,

G.R., Das, P. (1994) Somatic embryogenesis in Simarouba glauca. Plant Cell, Tissue and Organ Culture, 37(1), 79–81.

Shalini, Mathur and Sushilkumar. (1998) Journal of Medicinal and Aromatic Plant Science, 20, 1056–59. Sharma, R., Saxena, N., Thakur, G.S., Sanodiya, B.S., Jaiswal, P (2014) Conventional method for saponin extraction from Chlorophytum borivilianum Sant. et Fernand, Global Journal of Research in Medicinal Plants & Indigenous Medicine, 3(2), 33–39. Sharma, R., Thakur G., Sanodiya, B.S., Pandey, M., Bisen, P.S. (2012) Saponin: A wonder drug from Chlorophytum species. Global Journal of Research in Medicinal Plants & Indigenous Medicine, 1(10), 503–515. Sharma, R., Thakur, G.S., Sanodiya, B.S., Savita, A., Pandey, M., Sharma, A., Bisen, P.S. (2012). Therapeutic potential of Calotropis procera: a giant milkweed. IOSR-Journal of Pharmacy and Biological Science, 4(2), 42–57. Teixeira, F.M., Ramos, M.V., Soares, A.A. (2011) In vitro tissue culture of the medicinal shrub Calotropis procera to produce pharmacologically active proteins from plant latex. Process Biochemistry, 46, 1118–1124

Source of Support: NIL

Thakur, G.S., Bag, M., Sanodiya, B.S., Debnath, M., Bhadouriya, P., Prasad, G.B.K.S., Bisen, P.S. (2009) Momordica balsaminaa medicinal and neutraceutical plant for health care management. Current Pharma Biotechnology, 10(7), 667–682. Thakur, G.S., Pandey, M., Sharma, R., Sanodiya, B.S., Prasad, G.B.K.S, Bisen, P.S. (2011) Factors affecting in vitro propagation of Momordica balsamina: a medicinal and nutritional climber. Physiology and molecular biology of Plants. 17(2) 193–197 Thakur, G.S., Sharma, R., Sanodiya, B.S. et al. (2013) In vitro induction of tuber formation for the synthesis of secondary metabolites in C. borivilianum sant ed. Fernand. African journal of Biotechnology, 12(20), 2900–2907. Thakur, G.S., Sharma, R., Sanodiya, B.S., Pandey, M., Baghel, R,, Gupta, A,, Prasad, G.B.K.S., Bisen, P.S. (2011) High frequency in vitro shoot regeneration of Momordica balsamina, an important medicinal and nutritional plan. African Journal of Biotechnology, 10(70), 15808–15812. Thakur, G.S., Sharma, R., Sanodiya, B.S., Pandey, M., Prasad, G.B.K.S, Bisen, P.S. (2012) Gymnema sylvestre: An Alternative Therapeutic Agent for Management of Diabetes. Journal of Applied Pharmaceutical Science, 2 (12), 001–006. Yang, X.L., Jin, G.R., Yang, D.P., Li, S., Zhu, Y.G. (2006) Tissue culture and rapid propagation of Trichosanthes kirilowii. Zhong Yao Cai, 29(11), 1129–1130

Conflict of Interest: None Declared

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Review Article A REVIEW ON MICROBIAL ENDOPHYTES FROM PLANTS: A TREASURE SEARCH FOR BIOLOGICALLY ACTIVE METABOLITES Ruby Jalgaonwala1*, Raghunath Mahajan2 1,2

Department of Biotechnology M.J.College, North Maharashtra University, Jalgaon.M.S.India *Corresponding Author: E-mail: r_jalgaonwala@yahoo.co.in; Mobile: +919725400205

Received: 23/04/2014; Revised: 27/05/2014; Accepted: 30/05/2014

ABSTRACT Microbial endophytes normally reside asymptomatically in the tissues of higher plants and act as source of original organic metabolites. In recent years, a great deal of information on the role of endophytes in host plants has been collected. Many important chemotherapeutics from endophytic metabolites could be used in medicine, agriculture and industry. With the intention to provide studies on endophytic microbes, this review focuses on the role of endophytes with respect to production of anticancer, antimicrobial, antioxidant and other biologically important compounds. The main topics addressed are plant-endophyte relationship, potential in drug discovery, host-endophyte relationship, diversity, distribution and natural products from endophytic microbes. KEY WORDS: Endophytes, anticancer, antimicrobial, antioxidation, secondary metabolites

Cite this article: Ruby Jalgaonwala, Raghunath Mahajan (2014), A REVIEW ON MICROBIAL ENDOPHYTES FROM PLANTS: A TREASURE SEARCH FOR BIOLOGICALLY ACTIVE METABOLITES, Global J Res. Med. Plants & Indigen. Med., Volume 3(6): 263–277

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INTRODUCTION The word endophyte is derived from Greek „endo‟>< „endon‟ meaning within, and „phyte‟>< „phyton‟ meaning plant. The word “endophyte” was introduced by de Bary and was for some time applied to all “organisms occurring within plant tissues” (de Bary, 1866) or “all organisms inhabiting plant organs that at some time in their life, can colonize in internal plant tissues without causing apparent harm to the host” (Petrini, 1991). An endophyte by definition is one which resides in the tissues beneath the epidermal cell layers and causes no apparent harm to the host or fungi (Schulz and Boyle, 2005). Generally several hundred endophyte species can be isolated from a single plant (Tan and Zou, 2001). Mostly Ascomycetes, Deutromycetes, and Basidiomyctes classess of fungi are reported as endophytic fungi. The class and species of fungi depends upon the host plants. Recent studies suggested that endophtytic fungi are not host specific and generally have widespread host range. Earlier studies lead to the conclusion that fungal endophytes are ubiquitous in plant species (Huang et al., 2008; Wu et al., 2006). Numerous reports infer that endophytic actinomycetes play roles in plant protection against pathogens and their metabolic products have influence on plant growth and physiology (Katznelson and Cole, 1965; Strobel and Daisy, 2003). The fungi like mycorrhizae are symbiotic associations in between fungi and roots of majority of plants. The external hyphae of mycorrhizae spread out in to soil surrounding the infected root tips, and as a result mycorrhizal fungi reside only partly inside the plant tissues. In this way, they are different from typical endophytes. Arnold et al., 2007 explored the difficulty of endophytes versus mycorrhizae and exemplify that the mycorrhizae found in roots are mostly different in taxonomic composition to those endophytes found in leaves. On one hand, endophytes can produce analogous or the same biologically active constituents as its host, such as an endophytic fungus producing taxol (Strobel et al., 1993).

Many important chemotherapeutics are either microbial metabolites or their semisynthetic derivatives. Investigating the metabolites of endophytes can boost the chance of finding novel compounds so; an intensifying stream of attention is being directed to the endophytes and biomass can be accumulated by large scale fermentation (Tan and Zou, 2001). This review aims to provide an overview of the endophytic natural products, along with potential applications particularly in the area of agriculture, medicinal industry and biological diversity with respect to microbial endophytes. The plant endophyte relationship The host-endophyte relationship is supposed to be complex and different from host to host and microbe to microbe. The fungus passes from one generation to next through the seeds (Boursnell, 1950; Bultman and Murphy, 2000). The fungus enters the seedling from the seed and spreads through out, enters new tissues as they arise for this plant. Germination and subsequent development of the seedling depends on the presence of the fungus, devoid of it the plant ceases to grow beyond certain stages. During the fall, the plant digests the swollen, hyphae of the fungus that are found in the roots, and obviously benefits nutritionally from the microbe. The relationship is truly mutalistic because the fungus must obtain nourishment from the plant since it does not have contact with the soil. Endophytes that inhabit foraging grasses e.g rye grasses, do not leave their plant host and can only reproduce by invading seed tissue of the plant (Stone et al., 2000). Endophytes could be involved in pathogenecity of the host plants. The endophyte population in abnormally developed plant tissues, such as in galls and cysts are often quite different from healthy secretions (Stone et al., 2000). Environmental conditions such as soil temperature and humidity etc.would also are expected to affect the nature and the population of endophytes (Hata et al., 1998). Plants in unique environments that fight to compete with other living organisms or that

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need as much resistance as possible to survive, are probable candidates to host endophytes, which generate secondary metabolites that will assist the plants (Strobel et al., 2004). Many researchers hold that plants growing in tropical forest rainforest where competition for light and nutrient is intense, are most likely to host the greatest number of bioactive endophytes. Several studies note that endophytes from tropical regions produced significantly more bioactive secondary metabolites than those from temperate parts of the world (Bills et al., 2002). Most of the innovation of surfeit of microbes for applications that span a wide range of efficacy in medicine, agriculture and industrial area is currently useful. However, considerable work has also been carried out on a variety of different plants example Musa acuminata, Glycine max, Sorgum bicolour, Triticum aestivum, Zea mays, Agropyron elongatum, Sorghastrum nutans, Sweet potato, Ocimum sanctum, Ocimum bacilicum, Leucas aspera and Azadirachta indica. (Wipornpan et al., 2001; Khan et al., 2010; Banerjee et al., 2009; Kharwar et al.,2008; Kumaresan et al., 2001) Endophytic microbes are the source of natural products, for optimizing the search for new bioactive secondary metabolites. It is also relevant to consider that, i. The secondary metabolites synthesis may correspond with its respective ecological niche. OR ii. Continual metabolic interaction between endophytes and plant may enhance the synthesis of secondary metabolites. Studies have proved that they have the capacity to produce toxins in response to infection that benefits the host plants (Stone et al., 2000). Unlike the plant host, many endophytes are able to survive under quite extreme and inhospitable conditions. In one study, hyphae within stored mycorrhizal roots survived for six years in dry soil (Stone et al., 2000; Strobel et al., 2004) and endophytes can

be extracted from plant samples long after the plant tissues has died. Endophytes and their potential in drug discovery Endophytic microbes from medicinal plants are good source of functional metabolites (Tejesvi et al., 2007; Bailey et al., 2006). Endophyte infections have been found to alter pattern of gene expression in the host plant (Baily et al., 2006). Endophytes from Angiosperms as well as Gymnosperms have been studied for presence of novel secondary metabolites. The natural products produced by endophytes have vast range of bioactivities, representing a vast reservoir offering an enormous potential for exploitation in medicinal, agricultural and industrial uses (Tan and Zou, 2001). Crude extracts from culture broth of endophytes found to show antibacterial, antifungal, antiviral, antiinflammatory and antitumor activities (Silva et al., 2007). Therefore endophytes open up new areas for the biotechnological exploitations. Host endophyte relationship and effect on metabolites production Microorganisms are likely to harbor metabolic pathways that lead to the production of novel secondary metabolites. Many of the important secondary metabolites have been extracted and characterized from endophytic microbes (Tan and Zou, 2001), which includes alkaloids, steroids, terpenoids, peptides, polyketones, flavenoids, quinols and phenols. In addition, natural products also often serve as lead structures whose activity can be enhanced by exploitation through synthetic chemistry (Strobel and Daisy, 2003). Endophytes are able to increase host fitness and competitive abilitity, by increasing nutritional uptake, resistance to seed predators, seed germination success, tolerance to heavy metals, high salinity and good growth rate through biochemical pathways such as phytohormone indole acetic acid (IAA) from fungal endophytes Acremonium coenophialum, Aureobasidium pullulans, Epicoccum purpurascens and Collectotrichum sp along with IAA, cytokinins

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were also produced by an endophytic Hypoxylon serpens (Tan and Zou, 2001). Plants provide spatial arrangement, shelter, nutrients and distribution to the next generation of microbes (Rudgers, 2000). Plant may provide vital compounds for the completion of the life cycle of the endophytes (Strobel, 2002). Current research suggested that endophytes and plant genotypic combinations together with environmental conditions are an important source of variation in endophyte host interactions (Faeth and Fagan, 2002). Many factors such as season, age, environment and location may contribute and influence the biology of endophyte (Strobel and Daisy, 2003). Endophytes derive nutrients from the plant without killing host, endophytic microbes such as Phomopsis, Phoma, Colletotrichum and Phyllosticta have wide host range and found to colonize numerous taxonomically distinct plants (Murali et al., 2006; Sieber, 2007) developing adaptations to overcome different types of host defence. Endophyte infection affects the concentration of abscisic acid in leaves of drought stressed grasses and this helps in the recovery of endophytes infected plant in water deficient conditions also. Endophytic microbes residing in the host tissue some time turn in to a pathogen in response to some environmental signal (Hendry et al., 2002). Such a change in the nature of the endophytes would also result in a change in its metabolite profile (Suryanarayanan and Murali, 2006). Endophytic microbes associated with traditionally used medicinal plants particularly of the tropics could be a rich source of functional metabolites (Tejesvi et al., 2007). Diversity and distribution of microorganisms recovered as endophytes There are some major points represented with respect to diversity and distribution of endophytes as follows i. Individual endophytes can switch symbiotic lifestyles and the result of symbiosis is influenced by host genotypes ii. Mutalistic benefits conferred by endophytes are also influenced by plant genotypes iii. The host range of endophytes is inadequately defined

and which includes both monocot and dicot species and iv. Endophyteâ&#x20AC;&#x;s host plant describes adaptive symbiosis. Some endophytes have evolved with a high degree of suppleness to enter between genetically distinct plant species which provides endophytes an option to develop habitat range. Endophytic microbes can have intense effects on plant ecology, their fitness and are able to produce number of bioactive agents. The fossil proof shows that fungal symbionts have been associated with plants from the Ordovician period of approximately 400 million years ago, when plants first became established on land (Redecker et al., 2000), migrating from aquatic to terrestrial habitats. There are two major classes of fungal symbionts associated with internal plant tissues such as i. Fungal endophytes residing entirely within host plants and associated with roots, stems, leaves, and flowers. ii.

Mycorrhizal fungi that are residing only in roots but extend out into the rhizosphere.

In count to this, fungal endophytes also are divided into two classes: i.

A comparatively minute number of fastidious species limited to a few monocot host plants (Clay and Schardl, 2002).

ii. A huge number of tractable species with broad host ranges, together with monocots and eudicots (Stone, 2000). Considerable research have been done in class I endophytes as compared to class II endophytes, corresponding largest group of fungal symbionts. This is because the class II endophytes have only been elucidated in recent times and shown to be responsible for the adaptation of some plants to high-stress environments (Suryanarayanan and Murali, 2006). Endophytic fungi may express different symbiotic lifestyles in response to the host genotypes and environmental factors. Lifestyle expression of endophytes is a post colonization phenomenon which involves biochemical and genetic communications between endophytic microbes and host. Basically grass species,

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have been entirely studied in relation to their endophytic biology (Tan and Zou, 2001). Clavicipitaceous endophytes represents Class I and are small number of phylogenetically related Clavicipitaceous species which are fastidious in culture and also limited to some cool and warm seasonal grasses, (Stone et al., 2004). Transmission of these class I endophytes is mostly vertical, with maternal plants passing fungi on to offspring by means of the seed infections (Leuchtman, 2003). Class II endophyte generally comprises diverse species, all of which in general are members of the Dikarya (Ascomycota or Basidiomycota) having ability to give habitat specific stress tolerance to host plants.The main established hypothesis says that Clavicipitaceous endophytes are defensive mutualists of host grasses and play role during their evolution (Tanaka et al., 2005). Class III endophytes are basically distinguished on the basis of their occurrence and horizontal transmission, including vascular, nonvascular plants, some woody and herbaceous angiosperms in tropical forest and antarctic plant communities. These endophytes are especially known for their huge diversity within individual host tissues, plants and also populations. Class IV endophytes contains darkly melanized septa and they are restricted to plant roots. Generally these are Ascomycetous fungi conidial or sterile and forming melanized structures and also found in non mycorrhizal plants from antarctic, arctic, tropical ecosystems and temperate zones. Diversity of endophytic microbes shows to protect plants from the herbivores and is responsible for the production of novel secondary metabolites. Fungal endophytes those vertically transmitted are sexual and transmit via fungal hyphae penetrating the hostsâ&#x20AC;&#x; seeds for e.g Neotyphodium, these fungi are frequently mutualistic and on the contrary, endophytes transmitting horizontally are sexual and transmit via spores which can be spread by wind and insect vectors also. The endophytic microbes possibly adopt the same strategy as that of plant pathogenic fungi in order to enter the host plant (Sieber, 2007).

Research emphasize that endophytes are usually not host specific, single endophyte can have wide host range. Same microbe isolated from different tissue or part of the same host plant differs in their abilities for utilization of different substances, endophytic organisms associated with plants are varied and complex. Subsequent identification of potential genes provides evidence of specific pathway for known alkaloids synthesis by endophytes (Tanaka et al., 2005). Consequently if endophytes can produce the same bioactive compounds as their host plants this would reduce the need to harvest slow growing rare plants and also help to preserve the worlds diminishing biodiversity. Plant growth promoting endophytes Endophytes show numerous direct and indirect mechanisms to promote plant growth and health. Direct plant growth promoting mechanisms from endophytic suppression of the production of stress ethylene by 1aminocyclopropane-1-carboxylate (ACC) deaminase activity (Dellâ&#x20AC;&#x;Amico et al., 2005) and alteration of sugar sensing mechanisms in plants. Non reducing disaccharide such as trehalose is main storage carbohydrate of bacteria; it is also produced in plants but in lesser extent as compaire to sucrose. This sugar thought to play a vital role in plants for controlling their partitioning of carbon into cell wall biomass (Ramon and Rolland, 2007). Alteration in biosynthesis and metabolism of trehalose also increase tolerance to drought, salt, and cold. Therefore several endophytic bacteria from poplar tree were able to metabolize trehalose for example Plasmodiophora brassicase. Plant-associated bacteria benefits plant by preventing the growth of pathogens through, antibiosis (Zhang et al., 2004). Plant-growth-promoting endophytic bacteria were isolated from Brachiaria hybrid CIAT 36062 and introduced into Brachiaria hybrid cv. Mulato, positive for nif H gene sequences and inoculated Mulato plants showed higher chlorophyll and total nitrogen contents in leaves, DNA sequence analysis demonstrated that the nif H gene found were highly similar to Klebsiella pneumoniae and

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some other N2-fixing organisms identical to those of other N2-fixing bacteria. For this reason plant research area are now diverted to use endophytes in development of agriculture crops and forest regeneration. Natural Products from Endophytes The requirement for new antimicrobial agents generally, comes from the increasing resistance of pathogenic microbes towards antibiotics. Many microorganisms are known to acquire resistance to commonly used antimicrobial chemical compounds. So the interest in natural methods of pathogen control through new, eco-friendly agents has increased. The biologically active natural products from endophytes are excellent resources for medicine, agriculture and industry (Guo et al., 2008). About 51% of biologically active substances from fungal endophytes were previously unknown. Amines and amides are very common metabolite products from endophytes and have shown to be toxic to insects but not to mammals. Bioactive metabolites, such as steroids, terpenoids and diterpenes also are generated by endophytes. Endophyte also produces extracellular hydrolyases to establish a resistance mechanism against plant invasion which includes some of the extracellular enzymes like cellulases, proteinase, lipases and esterases. The actions of these enzymes found to support the hypothesis of co-evolution between endophytes and their hosts (Tan and Zou, 2001). Number of secondary metabolites produced by fungal endophytes is larger than Figure 1.Vincristine

that of any other endophytic microorganisms. Endophytic fungi are a promising source of novel compounds (Redecker et al., 2000). Role of Endophytes in the discovery of anticancer agents Endophytes hold main position in drug discovery as it has antibiotic, antiviral and anticancer properties, due to their ability to produce novel chemicals which can be used as drugs. Pacli taxel was first found in plants and later on reported from fungal endophyte. It is the first major group of anticancer agents which is produced by endophytes and now much research has been conducted on endophytes to determine its anticancer activity. Production of taxol was also done from endophytic fungi, Lasiodiplodia theobromae isolated from Morinda citrifolia with its cytotoxicity against human breast cancer cell line. Other important anticancer agents from the fungal endophytes were reported including camptothecin and several analogues (Redman et al., 2001) vincristine (Figure 1.Chemdraw) and podophyllotoxin. Subsequently one hundred anticancer compounds which belong to different chemical classes with activity against 45 different cell lines have been isolated from different fungal species belonging to different groups, out of which 57% were novel oranalogues of known compounds. Endophytic fungi was isolated and identified from Juniperus communis L. Horstmann, as a novel producer of deoxypodophyllotoxin (Kusari et al., 2009).

Figure 2. Sterigmatocystin

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Secondary metabolites from endophytes as antibiotics Plants with ethno botanical history are generally expected to be powerful source of endophytes producing active natural products. As more than 3000 diseases are clinically described today less than one third of these can be treated symptomatically. And even a less than that need needs new therapeutic agent with infectious diseases control (Strobel and Daisy, 2003). Tan and Zou in recent times isolated secondary metabolites of endophytes are synthesizing via variety of metabolic pathways (Tan and Zou, 2001) e.g polyketide, isoprenoid or amino acid derivation and belonging to different structural groups such as phenols, isocoumarins steroids, xanthones, perylene derivatives, depsipetides quinines, furandiones, terpenoids, and cytochalasines. Fungus Podospora Sp endophytic from the plant Laggera alata (Asteraceae) shows presence of xanthones sterigmatocystin (Figure 2.Chemdraw). Also chaetoglobosin A and rhizotonic acid were reported from endophytic Chaetomium globosum in Maytenus hookeri and Rhizoctonia Sp. in cynodon dactylon correspondingly to be active against the gastric ulcer. Altersetin from endophytic Alternaria Sp shows potent activity against pathogenic Gram positive bacteria. Fungal genus Cinnamomum zeylanicum found to be producing extremely bioactive volatile organic compounds (VOCs). Endophytic Muscodor albus produces a mixture of VOCs consists primarily of various alcohols, acids, esters, ketones and lipids. Cryptocandin A, an antifungal lipopeptide was isolated from endophytic Cryptosporiopsis quercina containing a number of unusual hydroxylated amino acids and 3 hydroxy-4hydroxymethyl proline which founds to be active against some fungal pathogens like Candida albicans, Trichophyton Sp., Sclerotinia Sclerotiorum and Botrytis cinerea (Wipornpan et al., 2001). The endophytic Chloridium Sp. from A.indica produces Javanicin (Figure 3. Chemdraw) which is highly active against Pseudomonas Spp. (Tejesvi et al., 2007). A tetramic acid cryptocin

(Figure 4. Chemdraw) was obtained from endophytic microbe, strong activity against Pyricularia oryzae plant pathogenic fungi. Endophytic fungus initiate production of Ambillic acid which is highly functionalized cyclohexenone with strong antifungal activity. A strain of Pestalotiopsis microspora, isolated from the tree Torreya taxifolia, produces a compound pestaloside having antifungal activity. Pestalotiopsis jester is an endophytic fungi produces the extremely functionalized cyclohexenone epoxides jesterone and hydroxy jesterone, exhibiting excellent antifungal activities against a variety pathogenic fungi of plants. Fungal endophyte isolated from Acalypha indica species shows potent antibacterial activity against human pathogenic bacteria such as Bacillus subtilis, Klebsiella pneumoniae and Staphylococcus aureus. The mechanism of antibiosis includes production of antibiotic compounds, bioactive volatile organic compounds (VOCs) and some enzymes (Ownley et al., 2010). Fungal endophyte Phomopsis Sp YM 311483 produces four new ten membered lactones activite against Aspergillus niger, Botrytis cinere and Fusarium spendophytic (Strobel, 2002). Jesterone synthesis was reported with potent antifungal activity from endophytic Pestalotiopsis jesteri. Endophytic fungi isolated from Rhizophora mucronata, Avicenna officialis and Avicenna marina and their ethyl acetate extract showed maximum antibacterial activity against bacterial pathogens and anticancer activity for Hep2 and MCF7 cell line In Vitro. Endophytic Gram positive bacteria like Bacillus Sp have also been isolated from cotton, cucumber root and citrus plant. Coronamycin characterize a complex peptide antibiotic with activities against pythiaceaus fungi, human fungal pathogen Cryptococcus neoformans and also against the malarial parasite Plasmodium falciparum was produced by a Verticillate Streptomyces spendophyte from an epiphytic vine Monstera Sp (Ezra et al., 2004). During the isolation of endophytes, actinomycetes generally appeared much later than endophytic bacteria and fungi and are also able to produce various metabolites.

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Streptomyces Sp. NRRL30566, from a fernleaved grevillea (Grevillea pteridifolia) tree, reported to produce original kakadumycin chemically related to echinomycin. Biologically dynamic species of Streptomyces were isolated from species of Nothofagus and Figure 3. Javanicin

some other plants from the southern reaches Patagonia. Having activity against plant pathogens, like Pythium ultimum, Sclerotinia sclerotiorum, Mycosphaere llafijiensis and Rhizoctonia solani (Wu et al., 2007; Castillo et al., 2007). Figure 4. Cryptocin

Endophytes have also been studied for their antiviral activity, the emergence of multiresistance against existing drugs, and high cost of current therapies as well as the AIDS associated opportunistic infections, such as Cytomegalo virus and Polyoma virus needs essential antiviral agent. Cytonic acid A and B were recognized as human cytomegalo virus protease inhibitors from endophytic fungus Cytonaema Sp. isolated from Quercus Sp. (Guo et al., 2008). A novel quinine related metabolites xanthoviridicatins E and F was also produced by an endophytic Penicillium chrysogenum able to inhibit the cleavage reaction of HIV-1 integrase

microspora from host Terminalia morobensis (Strobel, 2002). About 12 endophytes from Trachelospermum jasminoides were assayed for more potent free radicals scavenging activities using 1, 1, diphenyl,-2-picrylhyrazyl (DPPH) and hydroxyl radicals assay. Endophytes from medicinal plants are main resources for antioxidant metabolites helps to study relationship between total antioxidant capacity (TAC) and total phenolic content (TPC). The antioxidant capacities of the endophytes were significantly correlated with their total phenolic contents, suggesting that phenolics are the key antioxidant constituents of endophytic microbes. Metabolites produced by fungal endophyte can be a good source of novel natural antioxidant compounds (Wu et al., 2007).

Some antioxidant compounds produced by endophytes

Secondary metabolites from endophytes as antimycotic agents

Free radicals are atoms causing damage to body cells and harmful to our immune system leading to many of degenerative diseases. Antioxidant donates electron to free radicals and converts them to harmless molecules, protecting cells from oxidative damage aging and various diseases. Antioxidant are habitually produced many endophytes. Pestacin and Isopestacin were produced by Pestalotiopsis

Fungal infections are now becoming difficult problem as a result of the bigger numbers organ transplants patients with weakened immume systems so required new antimycotic agent to contest these problems (Strobel, 2002). A unique peptide antimycotic, Cryptocandin A was isolated from Cryptosporiopsis quercina, endophyte of medicinal plant Tripterigeum wilfordii.

Secondary metabolites from endophytes as antiviral agent

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Secondary metabolites from endophytes with further intersting Pharmacological activities Compounds with immuno-suppressive activities were obtained from endophytic fungi such as subglutinols A and B which are non cytotoxic diterpene pyrones from Fusarium subglutinans, endophyte from Triptergium wilfordii. Aurasperone A (Figure 5. Chemdraw) from endophytic Aspergillus niger isolated from Cynodon dactylon is xanthine oxidase inhibitor. 3 Hydroxypropionic acid was isolated from endophytic fungi showing nematicidal activity against the plant-parasitic nematode such as Meloidogyne incognita with the Lithal dose 50 values of 12.5â&#x20AC;&#x201C;15Âľg/ml.

This was the first report of 3-hydroxypropionic acid from endophytic fungi with the nematicidal activity (Michael et al., 2004). Endophytes do produce secondary metabolites when placed in culture, however, the temperature ,the composition of the medium and the degree of aeration will affect the amount and kind of compound that are produced by an endophytic fungus (Strobel et al., 2004). The host endophyte interaction provides nutrients and shelter for endophytes, which in substitute improve plant growth and health. Many endophytic bacteria are closely related to environmental and clinical isolates whose genomes have been or are in the process of being sequenced.

Figure 5. Aurasperone

Plant endophyte interactions metabolite production

affect

Plants have been viewed as a major source of new lead compounds for drug discovery, attention has more recently turned to endophytes as these microorganisms have great potential as sources for new bioactive compounds (Jalgaonwala and Mahajan, 2011). This may be the case because endophytes may have developed close biological associations with and inside their host, leading to the production of high number and diversity of classes of biological activities. Thus they represent an under-utilized resource in the search for new compounds. Studies of these organisms indicate that they are prolific producers of compounds that can be exploited as both agrochemical and medicinal agents (Jalgaonwala and Mahajan, 2011; Jalgaonwala

et al., 2011). Jalgaonwala and Mahajan, (2011) made investigation on different tissues of selected fifteen indigenous medicinal plants such as Aloe vera, Curcuma longa, Azadirachta indica, Coriandrum sativam, Eucalyptus globules, Hibiscus rosa sinensis, Ixora coccinea, Murrayo koenginii, Musa paradiasica, Ocimum sanctum, Pongamia glabra, Sphaeranthus indicus, Vinca rosea, Vitex nigundo and Withania somniphera. The research provided by Jalgaonwala and Mahajan (2011) evidence that isolated endophytes such as bacteria, fungi and actinomycetes are capable to survive inside medicinal plants. The endophytic diversity from selected plant species was rich. About 50% of test isolates exhibit potent antimicrobial activity and metabolite were partially characterized with attempts to identify potent

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microbes.The search for new compounds is certainly of equal importance however, has been the discovery that some endophytes produce compounds that have been exclusively isolated from higher plants. Described below in

Table 1 are some examples of bioactive products from endophytic fungi and their potential in the pharmaceutical and agrochemical arenas.

Table 1. List of natural products produce by endophytes Microbial strain

Plant host

Natural product(s)

Biological activity

Acremonium zeae (NRRL 13540) (mitosporic Hypocreales) Aspergillus clavatus strain H037 (Trichocomaceae)

Zea maydis L. (maize) (Poaceae)

Pyrrocidine A Pyrrocidine B

Antibacterial Antifungal

Brefeldin A

Antifungal; Antiviral Anticancer

Aspergillus fumigatus CY018 (Trichocomaceae)

Taxus mairei (Lemée & Lév.) and Torreya grandis Arn. (Taxaceae); Cynodon dactylon (L.) Pers. (Poaceae)

Asperfumoid , Asperfumin Monomethylsulochrin

Aspergillus niger IFB-E003 (Trichocomaceae)

Cynodon dactylon (L.) Pers. (Poaceae)

Rubrofusarin B,Fonsecinone A Aurasperone A,Asperpyrone B

Aspergillus parasiticus RDWD1-2 (Trichocomaceae) Aspergillus sp. (Strain #CY725) (Trichocomaceae)

Sequoia sempervirens (D. Don) Endl.(Taxodiaceae) Cynodon dactylon (L.) Pers. (Poaceae)

Sequoiatone C Sequoiatone D

Antifungal Mycotoxin Antifungal Mycotoxin Cytotoxic; Xanthine oxidase Inhibitor Antifungal; Xanthine oxidase inhibitor toxic to brine shrimp toxic to brine shrimp Antibacterial; Eosinophil inhibitor Antibacterial

Botrytis sp. (Sclerotiniaceae)

Taxus brevifolia Nutt. (Taxaceae)

Cephalosporium sp. IFB-E001 (mitosporic Hypocreales)

Trachelospermum jasminoides Lemoire (Apocynaceae) Baccharis cordifolia L. (Asteraceae);

Ceratopycnidium baccharidicola (Ascomycetes, Incerte sedis) Chaetomium chiversii CS-3662 (Chaetomiaceae) Chaetomium globosum (Chaetomiaceae) Cladosporium herbarum IFBE002 (Mycosphaerellaceae) Diaporthe sp. CR 146 (Valsaceae) Fusarium oxysporum strain 97CG3 (mitosporic Hypocreales) Fusarium sp. IFB-121 (mitosporic Hypocreales) Hormonema sp. ATCC 74360 (Dothioraceae) Melanconium betulinium (Melanconidaceae)

Monomethylsulochrin Helvolic acid Ergosterol 5α,8α-Epidioxyergosterol Ramulosin 6hydroxyramulosin 8dihydroramulosin Graphislactone A (23)

Antibiotic Antibiotic Antibiotic Antioxidant

Rodicins Verrucarins

toxic to livestock toxic to livestock

Ephedra fasciculata A. Nels (Ephedraceae); Ephedra fasciculata A. Nels (Ephedraceae)

Radicicol Orsellinic acid Globosumone A

cytotoxic; Hsp90 inhibitor Cytotoxic Cytotoxic

Cynodon dactylon (L.) Pers. (Poaceae) Forsteronia spicata G. Meyer (Apocynaceae) Catharanthus roseus (L.) G. Don (Apocynaceae) inner Quercus variabilis L. (Fagaceae) Juniperus communis L. (Cupressaceae) Betula pendula Roth; B. pubescens Ehrh. (Betulaceae)

Aspernigrin A Rubrofusarin B Cytosporone A Cytosporone B Vincristine

Cytotoxic; xanthine oxidase inhibitor Antifungal; cytotoxic Antibacterial Anticancer

Cerebroside fusaruside Enfumafungin

Antibacterial; xanthine Oxidase inhibitor Antifungal

3-hydroxypropionic acid

Nematocidal

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 263–277 Pilgerodendron uviferum (D. Don) Florine (Cupressaceae) [Gymnosperm] Buxus sempervirens L. (Buxaceae)

7-hydroxy-2,4-dimethyl3(2H)-benzofuranone Enalin Graphislactone A Botrallin Lactone S 39163/F-I

acetylcholinesterase (AChE) inhibitor acetylcholinestase (AChE) inhibitor Antimicrobial; Antiviral

several rain forest plants

Ambuic acid

Antimycotic

Cinnamomum zeylanicum Schaelter. (Lauraceae) Atropa belladonna L. (Solanaceae)

Volatile antibiotics

Antibiotic

Preussomerin G Preussomerin H Preussomerin I Nodulisporic acid A Nodulisporic acid A1

Antibacterial; Antifungal; FPTase inhibitor Insecticidal Insecticidal

Taxus mairei (Lemée & Lév) and Torreya grandis Arn. (Taxaceae) Diphylleia sinensis H. L. Li (Berberidaceae)

Brefeldin A

Antifungal; Antiviral; Anticancer; weed management Anticancer

Prumnopitys andina (Endl.) Laubenf. (Podocarpaceae)

Peniprequinolone Gliovictin Mellein

Periconia sp. OBW-15 (Halosphaeriaceae)

Taxus cuspidata Siebold & Zucc. (Taxaceae)

Periconicin A Periconicin B

Pestalotiopsis jesteri (Amphisphaeriaceae) Pestalotiopsis microspora (Amphisphaeriaceae)

Fragraea bodenii Thunb. (Gentianaceae) Terminalia morobensis L. (Combretaceae)

Jesterone hydroxyjesterone Pestacin Isopestacin

Pestalotiopsis spp. (Amphisphaeriaceae) Phomopsis phaseoli (Valsaceae) Phomopsis sp. (Valsaceae)

several rain forest plants

Ambuic acid

Nematicidal; root growth accelerator; weakly cytotoxic antibacterial; antiviral Antimycotic; Hypocotyl elongation and root growth Antifungal; Antimycotic Antimycotic; Antioxidant Antifungal Antimycotic

tropical

3-hydroxypropionic acid

Nematicidal

Erythrina crista-galli L. (Fabaceae)

Phomol

Pseudomassaria sp. ATCC 74411 (Hyponectriaceae)

unidentified plant (collected near Kinshasa, Democratic Republic of Congo) Rhyncholacis penicillata Tul. (Podostemaceae); Daphnopsis americana (Miller) J. S. Johnson (Thymelaeaceae)

Demethylasterriquinone B1 (DMAQ-B1) asterriquinone

Antibacterial; Antifungal; anti-inflammatory Insulin receptor activator

Picea glauca (Moench) Voss. (Pinaceae)

Microsphaeropsis olivacea (mitosporic Ascomycota)

Microsphaeropsis sp. strain NRRL 15684 (mitosporic Ascomycota) Monochaetia sp. (Amphisphaeriaceae) Muscodor albus (mitosporic Xylariales) Mycelia sterilia (Ascomycota)

Nodulisporium sp. MF 5954, ATCC 74245 (microsporic Xylariales) Paecilomyces sp. H-036 and W-001 (Trichocomaceae) Penicillium implicatum (isolate SJ21) (Trichocomaceae) Penicillium janczewskii (Trichocomaceae)

Serratia marcescens MSU-97 (Enterobacteriaceae) unidentified fungus CR115 (90% similarity to an uncharacterized oat root Basidiomycete) Unidentified fungus strain SWS 2611L (DAOM 229664)

Bontia daphnoides L. (Scrophulariaceae)

Substance analogous to podophyllotoxin

(–)-oocydin A

Antifungal

guanacastepene A guanacastepene B

Antibacterial Antibacterial

6,7-dihydroxy-2-propyl-2,4octadien-4-olide 5,6,8trihydroxy-4-(1′-hydroxyethyl)

Toxic to spruce budworm, cell line CF1

*Source: Gunatilaka, 2006

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CONCLUSION Endophytes comprise a diverse group of species existing in several ecosystems. In present report, we summarise the study of endophytes, their diversity and bioprospecting. Endophytic microbes are omnipresent within all identified plants in various bionetworks, but the geographical differences in their diversity, composition, host and tissue specificity have not been well documented. A powerful and good sequencing technology will make the global assessment of endophyte diversity. The above discussed novel bioactive compounds are only a few examples of what has been found after the isolation and culturing of individual

endophytic fungi followed by purification and characterization of some of their natural products. The prospects of finding new drugs that may be affective for treating newly developing diseases in humans, plants and animals are great their applications in industry agriculture may also be discovered among the novel products produced by endophytes. ACKNOWLEDGMENT We are thankful to principal Moolji Jaitha College, Jalgaon for providing laboratory as well as library facilities to complete the research work.

REFERENCES Arnold, A.E (2007). Understanding the diversity of foliar endophytic: progress,challenges, and frontier.Fungal Biology Reviews,.21:51–66. Boursnell, B (1950). The symbiotic seed borne fungi in the Cistaceae distribution and function of the fungus in the seedling and the tissues of the mature plant.Annals of Botany.,54:217–234. Bultman, T.L and Murphy, J.C (2000). Do fungal endophytes mediate woundinduced resistance? In Microbial Endophytes. Bacon C.W.and White J.F (eds). Marcel Dekker: New York.16:421–452. Bills, G., Dombrowski A., Pelaez, F., Polishook, J and An, Z (2002). Recent and future discoveries of pharmacologically active metabolites from tropical fungi.Tropical Mycology., CABI Publishing:New York.l2:65–194.

Bailey, B.A., Bae, M.D., Strem, D.P., Roberts, S.E.,Thomas, J., Crozier, G.J., Samuels, I.Y.,Choi and Holmes, K. A (2006).Fungal and plant gene expression during the colonization of cacao seedlings by endophytic isolates of four Trichoderma species. Planta., 224:1449– 1464. Banerjee, D., Manna, S., Mahapatra, S and Pati, B.R (2009).Fungal endophytes in three medicinal plants of Lamiaceae. Acta. Microbiol. Immunol. Hung.56:3:243–50. Clay, K and Schardl, C. (2002).Evolution origins and ecological consequences of endophyte symbiosis with grasses.American Naturalist.160:99–127. Castillo, U.F., Browne, L., Strobel, G., Hess, W.M., Ezra, S., Pacheco, G and Ezra, D (2007) Biologically active endophytic Streptomycetes from Nothofagus Spp. and other plants in Patagonia. Microbial Ecology..53:12–19.

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 263–277

Bary, A (1866). Morphologie and physiologie der Pilze, Flechten, and Myxomycten. In Hofmeister‟s. Handbook of Physiological Botany.2:Leipzig

Hendry, S.J., Boddy ,L and Lonsdole, D. (2002).Abiotic variable effect differential expression of latent infections in beech (Fagus sylvatica). New phytologist.,155:449–460.

Dell‟Amico, E., Cavalca, L and Andreoni, V (2005). Analysis of rhizobacterial communities in perennial Graminaceae from polluted water meadow soil, and screening of metal-resistant, potentially plant growth-promoting bacteria. FEMS Microbiol Ecol., 52: 153–162.

Huang, W.Y., Cai, Y.Z., Hyde, K.D., Corke, H and Sun, M (2008). Biodiversity of endophytic fungi associated with 29 traditional Chinese Medicinal Plants.Fungal Diversity.,33:61–75.

Ezra, D., Uvidello, F., Castillo, Strobel, G.A., Hess, W.M., Porter, H., Jensen, J.B., Margaret, A.M.,Condron, D.B.,Teplow, J.S., Maranta, M., Maranta, M.H., Weber, B and Yaver, D (2004).Cronomycins, peptide antibiotics produced by a verticillate Streptomyces Sp (MSU-2110) endophytic on Monstera Sp. Microbiology.,150:785– 793 Faeth, S.H and Fagan, W.F (2002). Fungal endophytes:Common host plant symbionts but uncommon mutualists. Integrative and Comparative Biology., 42(2):360–368. Gunatilaka, A.A.L. (2006). Natural products from plant-associated microorganisms : distribution, structural diversity, bioactivity, and implications of their occurrence. J. Nat. Prod., 69(30): 509– 526. Guo, B., Wang, Y., Sun, X and Tang, K (2008). Bioactive natural products from endophytes: A review. Applied Biochemistry and Microbiology., 44 (2):136–14. Hata, K., Futai, K and Tsuda, M (1998). Seasonal and needle age-dependent changes of the endophytic mycobiota in Pinus thunbergii and Pinus densiflora needles.Can. J. Bot.,76: 245–250.

Jalgaonwala, R.E and Mahajan, R.T. (2011).Isolation and characterization of endophytic bacteria from roots of Pongamia glabra vent. International Journal of Pharma and Bio Sciences., 2( 1): 280–287. Jalgaonwala, R.E and Mahajan, R.T (2011).Evaluation of hydrolytic enzyme activities of endophytes from some indigenous medicinal plants. Journal of Agricultural Technology., 7(6):1733– 1741. Jalgaonwala, R.E and Mahajan, RT (2011). Proteolytic activity of endophytic bacteria from leaves of Eucalyptus globules.Bionano Frontier., 4(2): 287– 289. Jalgaonwala, R.E.,Mohite, B.V and Mahajan, R.T (2011).A Review: Natural products from plant associated endophytic fungi. Journal of Microbiology and Biotechnology Research.1(2):21–32. Jalgaonwala, R.E and Mahajan, R.T (2011).A Review: Bacterial endophytes and their bioprospecting. Journal of Pharmacy Research.,4(3):795–799. Katznelson, H and Cole, S.E (1965).Production of giberellin like substances by bacteria and actinomycetes. Can. J. Microbiol.,11:733–741.

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 263–277

Kumaresan, V and Suryanarayanan, T.S (2001).Occurrence and distribution of endophytic fungi in a mangrove community. Mycological Research., (105):388–1391. Kharwar, R.N., Verma, V.C., Kumar, A., Gond, S.K., Harper, K.J., Hess, W.M., Lobkovosky, E., Ma, C., Ren, Y and Strobel, G.A (2008).Javanicin, an antibacterial naphthaquinone from an endophytic fungus of neem, Chloridium Sp. Current Microbiol.,58:233–238. Kusari, S., Lamshoft, M and Spiteller, M (2009).Aspergillus fumigatus Fresenius, an endophytic fungus from Juniperus communis L. Horstmann as a novel source of the anticancer pro-drug deoxypodophyllotoxin. Journal of Applied Microbiology.,107:1019–1030. Khan, R., Shahzad, S., Chaudhary, I.M., Khan, S,A and Ahmad, A (2010).Communities of endophytic fungi in medicinal plant Withania somnifera. Pak.J.Bot. 42(2):1281–1287. Leuchtman, A (2003). In,.White JF, Bacon CW, Hywel-Jones NL, Spatafora JW:eds. New York, USA. MarcelDekker.341–175. Michael, S., Barbel, K., Roland, W.S., Weber Lov, O.S and Heidrum A (2004) -3 Hydroxypropionic acid was a nematicidal principle in endophytic fungi.Phytochemistry.65: 2239–2245. Murali, T.S.,Suryanarayanan, T.S and Geeta, R. (2006). Endophytic Phomopsis species:host range and implications for diversity estimates.Canadian J. of Microbiology.52:673–680. Ownley, B.H., Gwinn, K.D and Vega, F.E (2010). Endophytic fungal entomopathogens with activity against plant pathogens: ecology and evolution. Review Biocontrol. 55(1):113–128.

Petrini, O (1991). Fungal endophytes of tree leves. In :Microbial ecology of leaves (edn) by Andrews J.H. and Hiran S.S.Springer Verlag.NewYork.USA. Redecker, D., Kodner, R and Graham, L.E (2000). Glomalean fungi from the Ordovician. Science., 289:1920–192. Redman, R.S., Dunigan, D.D and Rodriguez, R.J (2001). Fungal symbiosis: from mutualism to parasitism, who controls the out come, host or invader? New Phytologist.,151:705–716. Rudgers, J.A., Kaslow, J.M and Clay, K (2000). Endophytic fungi alter relationships between diversity and ecosystem properties. Ecology Letters.,7(1):42–51. Ramon, M and Rolland, F. (2007). Plant development: Introducing trehalose metabolism. Trends Plant Sci.12:185– 188. Strobel, G.A., Stierle, A., Stierle, D and Hess, W.M (1993). Taxomyces a proposed new taxon for a Bulbilli ferrous hyphomycete associated with Pacific yew. Mycotaxon.,47:71–78. Stone, J.K., Bacon, C.W and White, J.F (2000). An overview of endophytic microbes: endophytism defined. In Microbial Endophytes, Bacon C.W. and White J.F (eds), Marcel:New York. 1:3–29. Saikkonen, K., Ion, D and Gyllenberg, M (2002). Proceedings of the Royal Society B. Biological Sciences.269:1397–1403. Strobel, G.A (2002a). Microbiol gift from rain forest. Can.J.Plant Pathol. 24(1):14–20. Strobel, G and Daisy, B (2003). Bioprospecting for microbial endophytes and their natural products. Mol.biol.Rev,.67: 491– 502.

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Global J Res. Med. Plants & Indigen. Med. | Volume 3, Issue 6 | June 2014 | 263–277

Strobel, G., Daisy, B., Castillo, U and Harper, J (2004). Natural products from endophytic microorganisms. J Nat Prod.,67: 257–268. Stone, J.K., White, J.F and Polishook, J.D (2004). Endophytic fungi.In: measuring and monitoring biodiversity of fungi. Inventory and monitoring methods. Mueller G., Bills G., Foster M. (eds.). Elsevier Academic Press, Boston. MA. 241–270. Schulz, B and Boyle, C. (2005). The endophyte continuum. Mycological Research., 109:661–686. Suryanarayanan, T.S and Murali, T.S (2006). Incidence of Leptosphaerulina crassiasca in symptomless leaves of peanut in southern India.Journal of Basic Microbiology.,46:1003–1006. Sieber, T. (2007). Endophytic fungi in forest trees: are they mutualists? Fungal Biology Reviews., 21:75–89. Silva, G.H., Teles, L.H., Zanardi, L.M., Young, M.C., Eberlin, M.N., Hadad, R., Pfenning, L.H.,Costa-Neto, C.M., Gamboa, I., Bolzani, V and Arau, A.R. (2007). Cadinane sesquiterpenoids of Phomopsis cassiae an endophytic fungus associated with Cassia spectabilis (Leguminosae). Phytochemistry.,67.1964:1969. Tan, R.X and Zou, W.X (2001). Endophytes: a rich source of functional metabolites. Natural Product Reports.,18(4):448– 459.

Source of Support: NIL

Tanaka, A., Tapper, B.A., Popay, A., Parker, E.J and Scott, B (2005). A symbiosis expressed non ribosomal peptide synthetase from a mutualistic fungal endophyte of perennial ryegrass confers protection to the symbiotum from insect herbivory. Mol. Microbiol.,57:1036– 1050. Tejesvi, M.V., Kini, K.R., Prakash, H.S., Subbiah, V and Shetty, H.S (2007). Genetic diversity and antifungal activity of species of Pestalotiopsis isolated as endophytes from medicinal plants. Fungal Diversity.,24:37–54. Wipornpan, P., Saisamorn, L., Pipob, L and Hyde, K.D. (2001). Endophytic fungi of wild banana (Musa acuminata) at Doi Suthep Pui National Park, Thailand. Mycological Research.,105(121):508– 1513. Wu, W.J, Nong, J.Y and Shi, B.J (2006). New pesticides from natural productsprinciple. Method (In Chinses) Beijing Chem.Ind Press.301–302. Wu,Y.H., Yi, Z.C., Jie, X., Harlod, C and Meisun (2007). A potential Antioxidant Resources: Endophytic fungi from medicinal plants. Economic Botany.7(61):1:14–30. Zhang, S., Reddy, M. S and Kloepper, J.W. (2004). Tobacco growth enhancement and blue mold disease protection by rhizobacteria: Relationship between plant growth promotion and systemic disease protection by PGPR strain 90– 166. Plant and Soil., 262: 277–288.

Conflict of Interest: None Declared

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Call for Papers â&#x20AC;&#x201C; Vol. 3, Issue 8, August 2014 Submit your manuscripts (Research articles, Review articles, Short Communications, Letters to the Editor, Book Reviews) to Global Journal of Research on Medicinal plants & Indigenous medicine â&#x20AC;&#x201C; GJRMI Submit it online through www.gjrmi.com or mail it to submitarticle@gjrmi.com on or before July 10th 2014.

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