SCREENING OF ENDOPHYTIC BACTERIA PRODUCING ANTIFUNGAL ISOLATED FROM INDONESIA MEDICINAL PLANT, JAVA GINSENG (TALINUM TRIANGULARE) (JACQ.) WILLD

  • Alimuddin Ali Laboratory of Microbiology, Department of Biology, Faculty of Mathematic and Natural Sciences, Universitas Negeri Makassar, South Sulawesi, Indonesia 90233
  • Herlina Rante Laboratory of Microbiology, Faculty of Pharmacy, Hasanuddin University, South Sulawesi, Indonesia 90245

Abstract

Objective: The objective of this research was to isolate and characterize endophytic bacteria from Talinum triangulare having antifungal activity.

Methods: The endophytic bacteria were isolated from roots tissue of Talinum triangulare by surface sterilization method. The isolates were cultured in Trypticase Soybean agar and antagonist activities were evaluated by dual culture assay against Fusarium oxysporum, Trichoderma reesei and Candida albicans. For metabolite antifungal activities, bacterial isolates were grown for 4 d in TS broth at 35 °C under shaking condition. The antifungal activities of the supernatant extract were determined by using the disk agar diffusion. Polyketide synthase (PKS I) and NRPS genes fragments of all isolates were amplified.

Results: The result reveals that 4 of 23 endophytic bacterial isolates demonstrated great antifungal potentiality against many tested fungi. Polyketide synthase (PKS I) and NRPS genes amplification were showed 10 and 4 of endophytic isolates detected harboring PKS type I and NRPS genes, respectively. In general, high frequencies of positive PCR amplification were obtained for PKS I (43.47%). Phylogenetic analyses based on the 16S rRNA gene sequence, morphological, physiological and biochemical showed that the isolates were identified as a member of genus Bacillus and Brevibacillus.

Conclusion: These results indicated that the endophytic bacteria from java ginseng could be used as an alternative source of antifungal agents.

Keywords: Endophytic bacteria, Antifungal, Talinum triangulare, PKS I, NRPS, 16S rRNA

Downloads

Download data is not yet available.

Author Biography

Alimuddin Ali, Laboratory of Microbiology, Department of Biology, Faculty of Mathematic and Natural Sciences, Universitas Negeri Makassar, South Sulawesi, Indonesia 90233
Department of Biology

References

1. Pupo MT, Guimarães DO, Furtado NAJC, Borges WS. Microbial natural products: a promising source of bioactive compounds. In: Modern Biotechnology in Medicinal Chemistry and Industry. Taft, C. A. Ed. Kerala, India; Research Signpost; 2006. p. 51-78.
2. Hasegawa S, Meguro A, Shimizu M, Nishimura T, Kunoh H. Endophytic actinomycetes and their interactions with plants. Actinomycetol 2006;20:72–81.
3. Sun L, Qiu F, Zhang X, Dai X, Dong X, Song W. Endophytic bacterial diversity in rice (Oryza sativa L.) roots estimated by 16S rDNA sequence analysis. Microb Ecol 2008;55:415–24.
4. Braga RM, Dourado MN, Araũjo WL. Microbial interactions: ecology in a molecular perspective. Braz J Microbiol 2016;47S:86-98.
5. Kerry BR. Rhizosphere interactions and the exploitation of microbial agents for the biological control of plant-parasitic nematodes. Annu Rev Phytopathol 2000;38:423–41.
6. Sturz AV, Christie BR, Nowak J. Bacterial endophytes: potential role in developing sustainable systems of crop production. Crit Rev Plant Sci 2000;19:1–30.
7. Ping L, Boland W. Signals from the underground: bacterial volatiles promote growth in Arabidopsis. Trends Plant Sci 2004;9:263–66.
8. Berg G, Hallmann J. Control of plant pathogenic fungi with bacterial endophytes. Microbial Root Endophytes (Schulz BJE, Boyle CJC and Sieber TN, eds), Springer-Verlag, Berlin; 2006. p. 53–69.
9. Qin S, Wang HB, Chen HH, Zhang YQ, Jiang CL, Xu LH, et al. Glycomyces endophyticus sp. nov., an endophytic actinomycete isolated from the root of Carex baccans Nees. Int J Syst Evol Microbiol 2008;58:2525-8.
10. Clay K, Schardl CL. Evolutionary origins and ecological consequences of endophyte symbiosis with grasses; Am Midl Nat 2002;160:99–127.
11. Arnold AE, Maynard Z, Gilbert GS. Fungal endophytes in dicotyledonous neotropical trees: patterns of abundance and diversity. Mycol Res 2001;105:1502-7.
12. Ezra D, Castillo UF, Strobel GA, Hess WM, Porter H, Jensen JB, et al. Coronamycins, peptide antibiotics produced by a verticillate Streptomyces sp. (MSU-2110) endophytic on Monstera sp. Microbiology 2004;150:785–93.
13. Rodrigues KF, Petrini O. Biodiversity of endophytic fungi in tropical regions. In: Hyde KD. Ed. Biodiversity of Tropical Microfungi. Hong Kong, University of Hong Kong Press; 1997. p. 57-69.
14. Gasong BT, Tjandrawinata RR. Production of secondary metabolite E2.2 from Phaleria macrocarpa endophytic fungus. Asian Pac J Trop Biomed 2016;6:881-5.
15. Hawksworth DL. The magnitude of fungal diversity: the 1.5 million species estimated revised. Mycol Res 2001;105:1422-32.
16. Arnold AE, Meffa LC, Kyollo D, Rojas EI, Maynard Z, Robbins N, et al. Fungal endophytes limit pathogen damage in a tropical tree. Proc Natl Acad Sci USA 2003;100:15649–54.
17. Mayer KM, Ford J, Macpherson GR, Padgett D, Kohlmeyer BV, Kohlmeye J, et al. Exploring the diversity of marine-derived fungal polyketide synthases. Can J Microbiol 2007;53:291-302.
18. Cane DE, Walsh CT, Khosla C. Harnessing the biosynthetic code: combinations, permutations, and mutations. Science 1998;282:63–8.
19. Menzella HG, Reid R, Carney JR, Chandran SS, Resinger SJ, Patel KG, et al. Combinatorial polyketide biosynthesis by de novo design and rearrangement of modular polyketide synthase genes. Nat Biotechnol 2005;23:1171–6.
20. Nimnoi P, Pongsilp N, Lumyong S. Endophytic actinomycetes isolated from Aquilaria crassna Pierre ex Lec and screening of plant growth-promoters production. World J Microbiol Biotechnol 2010;26:193–203.
21. He RL, Wang GP, Liu XH, Zhang CL, Lin FC. Antagonistic bioactivity of an endophytic bacterium isolated from Epimedium brevicornu Maxim. Afr J Biotechnol 2009;8:191-5.
22. Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST. Bergey’s Manual of Determinative Bacteriology. 9th edition. 1994.
23. Barakate M, Ouhdouch Y, Oufdou KH, Beaulieu C. Characterization of rhizosperic soil Streptomyces from Moroccan habitats and their antimicrobial activities. World J Microb Biot 2002;18:49-54.
24. Ayuoso-Sacido A, Genilloud O. New PCR primers for the screening of NRPS and PKS-I systems in Actinomycetes: detection and distribution of these biosynthetic gene sequences in major taxonomic groups. Microb Ecol 2005;49:10-24.
25. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence allignment aided by quality analysis tools. Nucleic Acids Res 1997;25:4876-82.
26. Saitou N, Nei M. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406–25.
27. Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 1985;39:783–91.
28. Chun J. PHYDIT (The Phylogenetic Editor) Version 3.0. User’ Manual; 1999.
29. Posada F, Vega FE. Establishment of the fungal entomopathogen Beauveria bassiana (Ascomycota: Hypocreales) as an endophyte in cocoa seedlings (Theobroma cacao). Mycologia 2005;97:1195–200.
30. Sardi P, Saracchi M, Quaroni S, Petrolini B, Borgonovi GE, Merli S. Isolation of endophytic Streptomyces from surface-sterilized roots. Appl Environ Microbiol 1992;58:2691-3.
31. Shimizu M, Nakagawa Y, Sato Y, Furumai T, Igarashi Y, Onaka H, Yoshida R, et al. Studies on endophytic actinomycetes (I) Streptomyces sp. Isolated from Rhododendron and its antifungal activity. J Gen Plant Pathol 2000;66:360-6.
32. Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. Bacterial endophytes in agricultural crops. Can J Microbiol 1997;43:895–914.
33. Baek SH, Im WT, Oh HW, Lee JS, Oh HM, Lee ST. Brevibacillus ginsengisoli sp. nov., a denitrifying bacterium isolated from soil of a ginseng field. Int J Syst Evol Microbiol 2006;56:2665–9.
34. Das M, Prakash HS, Nalini MS. Antioxidative and antibacterial potentials of fungal endophytic from Justicia mynaadensis Heyne: An ethnomedicinal rainforest species of Western Ghats. Asian J Pharm Clin Res 2017;10:203-9.
35. Zinniel DK, Lambrecht P, Harris NB, Feng Z, Kuchzmarski D, Higley P, et al. Isolation and characterization of endophytic colonizing bacteria from agronomic crops and prairie plants. Appl Environ Microbiol 2002;68:2198–208.
36. Araùjo WL, Maccheroni WJr, Aguilar-Vildoso CI, Barroso PA, Saridakis HO, Azevedo JL. Variability and interactions between endophytic bacteria and fungi isolated from leaf tissues of citrus rootstocks. Can J Microbiol 2001;47:229-36.
37. Hori K, Yamamoto Y, Minetoki T, Kurotsu T, Kanda M, Miura S, et al. Molecular cloning and nucleotide sequence of the gramicidin S synthetase 1 gene. J Biochem 1989;106:639-45.
38. Chamberlain K, Crawford DL. In vitro and in vivo antagonism of pathogenic turfgrass fungi by Streptomyces hygroscopicus strains YCED9 and WYE53. J Ind Microbiol Biotechnol 1999;23:641-6.
39. Chathurdevi G, Gowrie SU. Endophytic fungi isolated from medicinal plant-a promising source of potential bioactive metabolites. Int J Curr Pharm Res 2016;8:50-60.
40. Parekh J, Chanda S. In vitro screening of antibacterial activity of aqueous and alcoholic extracts of various Indian plant species against selected pathogens from Enterobacteriaceae. Afr J Microbiol Res 2007;1:92–9.
41. Logan NA, Forsyth L, Lebbe L, 8 other authors. Polyphasic identification of Bacillus and Brevibacillus strains from clinical, dairy and industrial specimens and proposal of Brevibacillus invocatus sp. nov. Int J Syst Evol Microbiol 2002;52:953–66.
42. Goto K, Fujita R, Kato Y, Asahara M, Yokota A. Reclassification of Brevibacillus brevis strains NCIMB 13288 and DSM 6472 (NRRL NRS-887) as Aneurinibacillus danicus sp. nov. and Brevibacillus limnophilus sp. nov. Int J Syst Evol Microbiol 2004;54:419–27.
Statistics
354 Views | 738 Downloads
Citatons
How to Cite
Ali, A., and H. Rante. “SCREENING OF ENDOPHYTIC BACTERIA PRODUCING ANTIFUNGAL ISOLATED FROM INDONESIA MEDICINAL PLANT, JAVA GINSENG (TALINUM TRIANGULARE) (JACQ.) WILLD”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 6, June 2018, pp. 152-8, doi:10.22159/ijpps.2018v10i6.26037.
Section
Original Article(s)