ISOLATION OF ENDOPHYTIC FUNGI FROM ARTEMISIA ANNUA, L AND IDENTIFICATION OF THEIR ANTIMICROBIAL COMPOUND USING BIOAUTOGRAPHY METHOD

Authors

  • Indah Purwantini Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia
  • Wahyono . Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia
  • Mustofa . Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia
  • Ratna Asmah Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia

Keywords:

Endophytic fungi, Artemisia annua, L, antibacterial, antifungal, bioautography

Abstract

Objective: The aims of this study were (1) to isolate endophytic fungi from A. annua, (2) to find out their activity in inhibiting microorganisms' growth and (3) to investigate the active compound of the selected fungus.

Methods: A. annua herb was cleaned by putting it in running water and then the leaves, stems, and roots were separated. They were washed using chlorinated water, 70% ethanol, and distilled sterile water successively. Each part of the plant was cut into small pieces, grown in potato dextrose agar (PDA) medium, and incubated at room temperature. The endophytic fungi obtained were tested for their antimicroorganism activity against bacteria Staphylococcus aureus, Streptococcus mutans, Salmonella typhi, Bacillus subtilis, and the fungi Malassezia furfur and Candida albicans. Identification of active compound was done using bio-autography method.

Results: This study obtained 7 endophytic fungi successfully and they were encoded IP-1, IP-2, IP-3, IP-4, IP-5, IP-6, and IP-7. All the fungi demonstrated antibacterial or antifungal activity, with IP-4 had the greatest activity. IP-4 showed septate brown hyphae, elliptical to cylindrical conidia, pale to dark brown in colony color, terminal and intercalary swellings conidiophores. Extract of endophytic fungi (IP-4) had only one antimicrobial compound indicated by the presence of clear zone at Rf value of 0.38 which could not be detected by using spray reagents.

Conclusion: Endophytic fungi were isolated successfully from the leaves of A. annua demonstrated antibacterial and antifungal activities. From 7endophytic fungi obtained, fungi encoded IP-4 had the greatest activity. It was classified into Cladosporium sp. By using bioautography method, the active compound was assumed as dimethyl cladosporine.

 

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Author Biographies

Indah Purwantini, Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia

Department of Pharmaceutical Biology

Faculty of Pharmacy

Wahyono ., Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia

Department of Pharmaceutical Biology

Faculty of Pharmacy

Mustofa ., Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia

Department of Pharmacology

Faculty of Medicine

Ratna Asmah, Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia

Department of Pharmaceutical Chemistry

Faculty of Pharmacy

References

WHO. World Health Statistic. Geneva; 2009.

Hundley NJ. Structure elucidation of bioactive compounds isolated from endophytes of Alstoniascholaris and Acmenagraveolens. Thesis, Brigham Young University; 2005.

Tan RX, Zou WX. Endophytes: a rich source of functional metabolites. Nat Prod Rep 2001;18:448-59.

Radji M. The role of biotechnology and microbial endophyte in the development of herbal medicine. PSR 2005;2:113-26.

Strobel G, Daisy B, Castillo U, Harper J. Natural products from endophytic microorganisms. J Nat Prod 2004;67:257-68.

Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 2003;67:491-502.

Suffness M, Wall ME. Discovery and development of Taxol. In: Suffness M. Ed. Taxol® science and applications. CRC Press: Boca Raton; 1995. p. 3–25.

Dreyfuss MM, Chapela IH. The discovery of natural products with therapeutic potential. Butterworth-Heinemann, Boston; 1994. p. 49-79.

Miller RV, Miller CM, Garton-Kinney D, Redgrave B, Sears J, Condron M, et al. Ecomycins, unique antimycotics from Pseudomonas viridiflava. J Appl Microbiol 1998;84:937-44.

Harrison L, Teplow D, Rinaldi M, Strobel GA. Pseudomycins, a family of novel peptides from Pseudomonas syringae, possessing broad spectrum antifungal activity. J Gen Microbiol 1998;137:2857-65.

Castillo UJ, Harper K, Strobel GA, Sears J, Alesi K, Ford E, et al. Kakadumycins, novel antibiotics from Streptomyces sp. NRRL 30566, an endophyte of Grevilleapteridifolia. FEMS Lett 2003;24:183-90.

Castillo UF, Strobel GA, Ford EJ, Hess WM, Poter H, Jenson JB. et al. Munumbicins, wide spectrum antibiotics produced by Streptomyces NRRL 30562, endophytic on Kennedia nisgricans. Microbiol 2002;148:2675-85.

Verdian-rizi MR, Sadat-Ebrahimi E, Hadjiakhoondi A, Fazeli MR, Hamedani M, Pirali. Chemical composition and antimicrobial activity of Artemisia annua, L. essential oil from Iran. Pharmacogn Res 2009;1 Suppl 1:21-4.

Bhakuni RS, Jain DC, Sharma RP, Kumar S. Review article: secondary metabolites of Artemisia annua and their biological activity. Curr Sci 2001;80:35-48.

Anonymous. WHO Monograph on Good Agricultural and Collection Practices (GACP) for Artemisia annua L. New York; 2006. p. 7-10.

Wright CW. Artemisia. Taylor and Francis Inc. New York; 2002.

Potawale SE, Waseem Md, Sadiq Md, Mehta UK, Dhalawat HJ, Luniya KP, Mantri RA, et al. Research and medicinal potential of Artemisia annua: a review. Pharmacologyonline 2008;2:220-35.

Pratiwi ST. Pharmaceutical microbiology. Erlangga Press. Jakarta; 2008.

Ellis D, Davis S, Alexiou H, Handke R, Bartley R. Description of medical fungi. Nexus Print Solution. Adelaide; 2007.

Guo L, Hyde K, Liew E. A method to promote sporulation in palm endophytic fungi. Fungal Divers 1998;1:109–13.

Lacap DC, Hyde K, Liew ECY. An evaluation of the fungal morphotype†concepts based on ribosomal DNA sequence. Fungal Divers 2003;12:53–66.

Scott PM, Walbeek WV, Maclean WM. Cladosporin, a new antifungal metabolite from Cladosporium cladosporioides. J Antibiot 1971;24:747-55.

Wagman GH, Weinstein MJ. Chromatography of antibiotics. 2nd Completely Revised Edition, Elsevier Science Pub. Company Inc., New York; 1984.

Published

01-12-2015

How to Cite

Purwantini, I., W. ., M. ., and R. Asmah. “ISOLATION OF ENDOPHYTIC FUNGI FROM ARTEMISIA ANNUA, L AND IDENTIFICATION OF THEIR ANTIMICROBIAL COMPOUND USING BIOAUTOGRAPHY METHOD”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 7, no. 12, Dec. 2015, pp. 95-99, https://journals.innovareacademics.in/index.php/ijpps/article/view/8374.

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