• Priyom Bose
  • Uma Gowrie Ethiraj College for Women (Autonomous) Chennai 600008.


Objective: Casuarina junghuhniana Miq is a well known multipurpose tree belonging to family Casuarinaceae. This tree has the ability to sustain in different edaphic and extreme environmental conditions. The root extract of this tree is a rich source of potential secondary metabolites. Therefore, the main objective of this study is to isolate fungal root endophyte from this tree and to determine its bioactive compound which can be utilized in agriculture and pharmaceutical industries.

Methods: Casuarina junghuhniana Miq root samples were collected from the State Forestry Research Institute, Neyveli, Tamil Nadu. Isolation of fungal endophyte was carried out using Potato Dextrose Agar medium. The qualitative and quantitative phytochemicals screening and enzyme assays were carried out using standard procedures. Antimicrobial assay was tested against different pathogens using well diffusion method. Further, the phytochemicals in the root extract was evaluated using FTIR, HPLC and GC MS studies.

Result: Among the isolated endophytes, the dominant isolate (CJN5) was identified as Aspergillus sp which showed positive result in the production of industrially important enzymes, namely, cellulase, lipase and amylase The qualitative screening revealed the presence of various phytoconstituents Total phenolic content of CFE (208.20±2 mg/g) was found higher than CME(165.5±3mg/g). The total flavonoid content of CFE (25.95±2 mg/g) was also found higher than CME (3.59±2 mg/g) using respective standards. Antimicrobial activity showed maximum zone of inhibition against Pseudomonas aeruginosa (bacterial pathogen) (24±0.1cm) and Fusarium oxysporum (fungal phytopathogen) (34 ± 0.2cm).FTIR analysis showed the presence of varied functional groups. HPLC study revealed the presence of Gallic acid acid and Salicylic acid. Several peaks were obtained in GCMS analysis which indicates the presence of different secondary metabolites.

Conclusion: Aspergillus sp, isolated from the root of Casuarina junghuhniana Miq is a promising source of bioactive compounds which can be utilised in agriculture and pharmaceutical industries.

Key words: endophyte, Casuarina junghuhniana, phytochemical, antimicrobial, HPLC, GC MS

Author Biography

Uma Gowrie, Ethiraj College for Women (Autonomous) Chennai 600008.
associate Professor, Department of plant Biology and Plant biotechnology, Ethiraj College for Women, Chennai 600008.


1. Srivastava A, Anandrao RK. Antimicrobial potential of fungal endophytes isolated from leaves of Prosopis juliflora (SW.) DC, an important weed. Int J Pharm Pharm Sci 2015;7(12):128-36.
2. Xing YM, Chen J, Cui JL, Chen XM, Guo SX. Antimicrobial activity and biodiversity of endophytic fungi in Dendrobium devonianum and Dendrobium thyrsiflorum from Vietnam. Curr Microbiol 2011;62:1218-24.
3. Teiten MH, Mack F, Debbab A, Aly AH, Dicato M, Proksch P, et al. Anticancer effect of altersolanol A, a metabolite produced by the endophytic fungus Stemphylium globuliferum, mediated by its pro-apoptotic and anti-invasive potential via the inhibition of NF-k B activity. Bioorg Med Chem 2013;21(13):3850-8.
4. Purwantini I, Wahyono, Mustofa, Asmah R. Isolation of endophytic fungi from L. and identification of their antimicrobial compounds using bioaautography method. Int J Pharm Pharm Sci 2015;7(12):95-9.
5. Jose AC, Christy PH. Assessment of antimicrobial potential of the endophytic bacteria isolated from Rhizophora mucronata. Int J Curr Microbiol Appl Sci 2013;2(10):188-94.
6. Radu S, Kqueen CY. Preliminary screening of endophytic fungi from medicinal plants in Malaysia for antimicrobial and antitumor activity. Malays J Med Sci 2002;9(2):23-33.
7. Zhao J, Shan T, Mou Y, Zhou L. Plant-derived bioactive compounds produced by endophytic fungi. Mini Rev Med Chem 2011;11(2):159-68.
8. Saikkonen K, Wäli P, Helander M, Faeth SH. Evolution of endophyte-plant symbioses. Trends Plant Sci 2004;9(6):275-80.
9. Strobel G, Daisy B, Castillo U, Harper J. Natural products from endophytic microorganisms. J Nat Prod 2004;67(2):257-68.
10. Phongpaichit S, Nikom J, Rungjindamai N, Sakayaroj J, Hutadilok-Towatana N, Rukachaisirikul V, et al. Biological activities of extracts from endophytic fungi isolated from Garcinia plants. FEMS Immunol Med Microbiol 2007;51(3):517-25.
11. Dreyfuss MM, Chapela IH. Potential of fungi in the discovery of novel low-moleculae weight pharmaceuticals. In: Gullo VP, editor. The Discovery of Natural Products with Therapeutic Potential. London, UK: Butterworth-Heinemann; 1994. p. 49-80.
12. Schluz B, Boyle C, Draeger S, Rommert A, Krohn K. Endophytic fungi: A source of novel biologically active secondary metabolites. Mycol Res 2002;106(9):996-1004.
13. Thomas SN. Endophytic fungi in forest tree: Are they mutualists? Fungal Biol Rev 2007;21:75-89.
14. Al-Snafi AE. The Pharmacological importance and chemical constituents of Arctium Lappa. A review. IJPRS 2014;3(1-1):663-70.
15. Murugesan S, Sumathi R, Senthilkumar N, Rajeshkannan C, Karthikeyan A. Biotransformation of some secondary metabolite of Frankia-Casuarina relationship. Jayaraj RS, Warrier RR, Nicodemus A, Kumar, NK, editors. Advances in Casuarina Research in India-Proceedings of the 2nd National Seminar on Casuarinas; 2012. p. 271-6.
16. Jayaraj RS. Casuarina junghuhniana (Casuarinaceae) in India. Aust J Bot 2010;58(2):149-56.
17. Kannan CS Warrier EV, Anoop B, Singh G. Screening of clones of Casuarina equisetifolia for pulping traits using wood fibre characteristics. Int J Curr Res Rev 2015;7(12):12.
18. Fisher PJ, Petrini O, Sutton BC. A comparative study of fungal endophyte in leaves, xylem and bark of Eucalyptus in Australia and England. Sydowia 1993;45:338-54.
19. Suryanarayan TS, Senthilarasu G, Muruganandam V. Endophytic fungi from Cuscuta rejlexa and its host plant. Fungal Divers 2000;4:117-23.
20. Domsch KH, Gamas W, Anderson TH. Compendium of Soil Fungi. New York: Academic Press; 1980. p. 168-9, 540, 559-60.
21. Raaman N. Phytochmeical Techniques. Ch. 5. New Delhi: New India Publishing Agency; 2006. p. 19-25.
22. Srinivasan K, Jagadish LK, Shenbhagaraman R, Muthumary J. Antioxidant activity of endophytic fungus Phyllosticta sp. Isolated from Guazuma tomentosa. J Phytol 2010;2(6):37-41.
23. Chang C, Yang M, Wen H, Chern J. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 2002;10:178-82.
24. Senthilmurugan VG, Sekar R, Suresh K, Balamurugan S. Phytochemical sceening, enzyme and antibacterial activity analysis of endophytic fungi Botrytis sp isolated from Ficus benghalensis(L). Coden IJPRNK 2013;2(4):264-73.
25. Perez C, Pauli M, Bezevque P. An antibiotic assay by agar well diffusion method. Acta Biol Med Exp 1990;15:113-5.
26. Boyle C, Gotz M, Dammann-Tugend U, Schulz B. Endophyte-host interaction III. Local vs. Systemic colonization. Symbiosis 2001;31:259-281
27. Pandit S, Lawrence K, Singh A, Singh S, Lawrence R. Cellulase production by Aspergillus flavus and saccharification of wheat straw. Int J Sci Eng Res 2013;4(6):1965-71.
28. Hasan F, Shah AA, Hamed A. Industrial application of microbial lipases. Enzyme Microb Technol 2006;39(2):235-50.
29. Sharma R, Chisti Y, Banerjee UC. Production, purification, characterization, and applications of lipases. Biotechnol Adv 2001;19(8):627-62.
30. Jain P, Aggarwal V, Sharma A, Pundir RK. Screening of endophytic fungus Acremonium sp. For amylase production. J Agric Sci Technol 2012;8(4):1353-64.
31. Burhan A, Nisa U, Gokhan C, Ashabil A, Osmair G. Enzymatic properties of a novel thermostable thermophilic alkaline and chelator resistant amylase from analkaphilic Bacillus sp Isolate ANT-6. Process Biochem 2003;38:1397-403.
32. Tran HB, McRae MJ, Lynch F, Brett CT, Waldron K, Unwin, Hyman, Palombo EA, editors. Identification and bioactive properties of endophytic fungi isolated from phyllodes of Acacia species. Current Research, Technology & Education Tropics. Applied Microbiology and Microbial Biotechnology. Spain: Formatex; 2010.
33. Muthukrishnan SD, Subramaniyan A. Phytochemical constituents of Gloriosa superb seed, tuber and leaves. RJPBCS 2012;3(3):111-7.
34. Jack IR, Okorosaye-Orubite K. Phytochemical analysis and antimicrobial activity of the extract of leaves of fleabane (Conyza sumatrensis). J Appl Sci Environ Manage 2008;12(4):63-8.
35. Segismundo AB, Florendo PE, Roman PA. In vitro antifungal activity and phytochemicalscreening of Gouania javanica Miq. Leaves. UNP Res J 2008;17:1-10.
36. Stefanovic O, Radojevic I, Comic L, Vasic S. Antibacterial activity of naturally occurring compounds from selected plants. Antimicrobial Agents. Rijeka, Croatia: InTech; 2012.
37. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12(4):564-82.
38. Wu T, He M, Zang X, Zhou Y, Qiu T, Pan S, et al. A structure-activity relationship study of flavonoids as inhibitors of E. coli by membrane interaction effect. Biochim Biophys Acta 2013;1828(11):2751-6.
39. Tsuchiya H, Sato M, Miyazaki T, Fujiwara S, Tanigaki S, Ohyama M, et al. Comparative study on the antibacterial activity of phytochemical flavanones against methicillin-resistant Staphylococcus aureus. J Ethnopharmacol 1996;50(1):27-34.
40. Alcaráz LE, Blanco SE, Puig ON, Tomás F, Ferretti FH. Antibacterial activity of flavonoids against methicillin-resistant Staphylococcus aureus strains. J Theor Biol 2000;205(2):231-40.
41. Liang H, Xing Y, Chen J, Zhang D, Guo S, Wang C. Antimicrobial activities of endophytic fungi isolated from Ophiopogon japonicus (Liliaceae). BMC Complement Altern Med 2012;12:238.
42. Borges A, Ferreira C, Saavedra MJ, Simoes M. The antibacterial activity and mode of action of ferulic and gallic acids against pathogenic bacteria. Microb Drug Resist 2013;19(4):256-65.
43. Coates J. In: Meyers RA, editor. Interpretation of Infrared Spectra, A Practical Approach in Encyclopedia of Analytical Chemistry. Chichester: John Wiley and Sons Ltd.; 2000. p. 10815-37.
44. Clarke JD, Volko SM, Ledford H, Ausubel FM, Dong X. Roles of salicylic acid, jasmonic acid, and ethylene in cpr-induced resistance in arabidopsis. Plant Cell 2000;12(11):2175-90.
45. Forchetti G, Masciarelli O, Izaguirre MJ, Alemano S, Alvarez D, Abdala G. Endophytic bacteria improve seedling growth of sunflower under water stress, produce salicylic acid, and inhibit growth of pathogenic fungi. Curr Microbiol 2010;61(6):485-93.
46. Li S, Yu Y, Chen J, Guo B, Yang L, Ding W. Evaluation of the antibacterial effects and mechanism of action of protocatechualdehyde against Ralstonia solanacearum. Molecules 2016;21(6). pii:E754.
47. Das B, Thirupathi P, Ravikanth B, Aravind Kumar R, Sarma AV,Basha SJ. Isolation, synthesis, and bioactivity of homoisoflavonoids from Caesalpinia pulcherrima. Chem Pharm Bull (Tokyo) 2009;57(10):1139-41.
48. Ramesh R, Dhanaraj TS. Identification of bioactive compounds in the ethyl acetate extract of Terminalia arjuna root by GC-MS analysis. Int J Res Sci Res 48. 2016;7(3):9747-51.
49. Bouker KB, Hilakivi-Clarke L. Genistein: Does it prevent or promote breast cancer? Environ Health Perspect 2000;108(8):701-8.
50. Rajeswari G, Murugan M, Mohan VR. GC-MS analysis of bioactive components of Hugonia mystax L. (Linaceae). RJPBCS 2012;3(4):301.
51. Duke JA, Beckstrom-Sternberg SM. Dr. Duke’s Phytochemical and Ethnobotanical Databases. Beltsville, Maryland: ARS/USDA; 1994.
528 Views | 149 Downloads
How to Cite
Bose, P., and U. Gowrie. “ASSESSMENT OF BIOACTIVE METABOLITES FROM THE ROOT ENDOPHYTE ISOLATED FROM CASUARINA JUNGHUHNIANA Miq.”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 10, no. 4, Apr. 2017, pp. 137-43, doi:10.22159/ajpcr.2017.v10i4.16382.
Original Article(s)