Growth Inhibition of Aspergillus spp. by Endophytic Bacteria

Dwi N. Susilowati, Untung Haryono


Aspergillus spp. (A. niger, A. flavus, and A. fumigatus) contaminate food commodities through production of secondary metabolites (mycotoxins) and aspergillosis, and thus pose severe hazard to human and animal health. Hence, the inhibition of mycotoxin-producing fungi on agricultural storage commodities needs to be considered. The aims of this study were to evaluate endophytic bacteria isolated from rice tissues that inhibit Aspergillus spp. growth, as well as to characterize the selected isolates morphologically and biochemically. Dual culture and disk diffusion method tests on 155 endophytic bacteria obtained three isolates, i.e. FB-Endo 65, FB-Endo 73, and FB-Endo 95, which showed inhibition zone from 13 to 17 mm against Aspergillus spp. growth. The inhibition zone and quantity of antifungal compounds increased positively with the length of incubation periods from 0 to 6 days. Antifungal compounds from the three isolates were insoluble in ethyl acetate, but soluble in methanol. The methanol soluble substance(s) from FB-Endo 73 showed higher inhibition zone than that of the other isolates. This result indicated that all three isolates produced strong antifungal activity. Morphological and biochemical identifications of the isolates revealed that all isolates belonged to the genus Bacillus sp. Further studies include identification and production methods of antifungal compounds of those endophytic bacteria and their application on stored seeds.


Endophytic bacteria; antifungal; Aspergillus niger; Aspergillus fumigatus; Aspergillus flavus

Full Text:



Afsharmanesh, H., Ahmadzadeh, M., Javan-Nikkhah, M. &

Behboudi, K. (2014) Improvement in biocontrol activity

of Bacillus subtilis UTB1 against Aspergillus flavus

using gamma irradiation. Crop Protection, 60, 83–92.

Bauer, A.W., Kirby, W.M.M., Sherris, J.C. & Turck, M.

(1966) Antibiotic susceptibility testing by a

standardized single disk method. American Journal of

Clinical Pathology, 45 (4), 493–496.

Beernaert, L.A., Pasmans, F., Van Waeyenberghe, L.,

Haesebrouck, F. & Martel, A. (2010) Aspergillus

infections in birds: A review. Avian Pathology, 39 (5),


Caldeira, A.T., Arteiro, J.M.S., Coelho, A.V. & Roseiro, J.C.

(2011) Combined use of LC–ESI-MS and antifungal

tests for rapid identification of bioactive lipopeptides

produced by Bacillus amyloliquefaciens CCMI 1051.

Process Biochemistry, 46 (9), 1738–1746.

Cho, S.J., Lee, S.K., Cha, B.J., Kim, Y.H. & Shin, K.S.

(2003) Detection and characterization of the

Gloeosporium gloeosporioides growth inhibitory

compound iturin A from Bacillus subtilis strain KS03.

FEMS Microbiology Letters, 223 (1), 47–51.

Cho, K.M., Math, R.K., Hong, S.Y., Islam, S.M.A.,

Mandanna, D.K., Cho, J.J., Yun, M.G., Kim, J.M. &

Yun, H.D. (2009) Iturin produced by Bacillus pumilus

HY1 from Korean soybean sauce (kanjang) inhibits

growth of aflatoxin producing fungi. Food Control, 20

(4), 402–406.

Das, P., Mukherjee, S. & Sen, R. (2008) Antimicrobial

potential of a lipopeptide biosurfactant derived from a

marine Bacillus circulans. Journal of Applied

Microbiology, 104 (6), 1675–1684.

de Melo, F.M.P., Fiore, M.F., de Moraes, L.A.B., Silva-

Stenico, M.G., Scramin, S., de Araujo-Teixeira, M. & de

Melo, I.S. (2009) Antifungal compounds produced by

cassava endophyte Bacillus pumilus MAIIIM4A.

Scientia Agricola, 66 (5), 583–592.

Gong, A.D., Li, H.P., Yuan, Q.S., Song, X.S., Yao, W., He,

W.J., Zhang, J.B. & Liao, Y.C. (2015) Antagonistic

mechanism of iturin A and plipastatin A from Bacillus

amyloliquefaciens S76-3 from wheat spikes against

Fusarium graminearum. PLOS One, 10 (2), e0116871.


Hidayatun, N., Susilowati, D.N. & Mulya, K. (2011)

Identifikasi 26 isolat bakteri endofitik dan filosfer padi

dengan analisis sekuen 16S rDNA. Berita Biologi, 10

(4), 455–461.

Hwang, B.K., Lim, S.W., Kim, B.S., Lee, J.Y. & Moon, S.S.

(2001) Isolation and in vivo and in vitro antifungal

activity of phenylacetic acid and sodium phenylacetate

from Streptomyces humidus. Applied and

Environmental Microbiology, 67 (8), 3739–3745.

Irkin, R. & Korukluoglu, M. (2007) Control of Aspergillus

niger with garlic, onion, and leek extracts. African

Journal of Biotechnology, 6 (4), 384–387.

Ji, S.H., Paul, N.C., Deng, J.X., Kim, Y.S., Yun, B.S. & Yu,

S.H. (2013) Biocontrol activity of Bacillus

amyloliquefaciens CNU114001 against fungal plant

diseases. Mycobiology, 41 (4), 234–242.

Kalemba, D. & Kunicka, A. (2003) Antibacterial and

antifungal properties of essential oils. Current

Medicinal Chemistry, 10 (10), 813–829.

Kowalska, A., Walkiewicz, K., Koziel, P. & Muc-Wierzqoǹ ,

M. (2017) Aflatoxins in animal and human health.

Postȩpy Higieny i Medycyny Doświadczalnej, 71 (0),


Liswara, N. (2000) Karakterisasi senyawa antibiotik yang

resisten terhadap β-laktamase tipe TEM-1, dari isolat

ICBB 1171 asal ekosistem air hitam, Kalimantan

Tengah. Tesis S2, Institut Pertanian Bogor.

Onishi, J., Meinz, M., Thompson, J., Curotto, J., Dreikorn,

S., Rosenbach, M., Douglas, C., Abruzzo, G., Flattery,

A. & Kong, L. (2000) Discovery of novel antifungal (1,

-β-D-glucan synthase inhibitors. Antimicrobial Agents

and Chemotherapy, 44 (2), 368–377.

Paster, N., Juven, B.J. & Harshemesh, H. (1988)

Antimicrobial activity and inhibition of aflatoxin B-1

formation by olive plant tissue constituenrs. Journal of

Applied Bacteriology, 64, 293–297.

Phister, T.G., O’Sullivan, D.J. & McKay, L.L. (2004)

Identification of bacilysin, chlorotetaine, and iturin A

produced by Bacillus sp. strain CS93 isolated from

pozol, a Mexican fermented maize dough. Applied and

Environmental Microbiology, 70 (1), 631–634.

Prakash, B., Singh, P., Kedia, A. & Dubey, N.K. (2012)

Assessment of some essential oils as food

preservatives based on antifungal, antiaflatoxin,

antioxidant activities and in vivo efficacy in food

system. Food Research International, 49 (1), 201–208.

Prapagdee, B., Kuekulvong C.H., & Mongkolsuk, S. (2008)

Antifungal potential of extracellular metabolites

produced by Streptomyces hygroscopicus against

phytopathogenic fungi. International Journal of

Biological Sciences, 4, 330–337.

Regnard, J.F., Icard, P., Nicolosi, M., Spagiarri, L.,

Magdeleinat, P., Jauffret, B. & Levasseur, P. (2000)

Aspergilloma: A series of 89 surgical cases. The

Annals of Thoracic Surgery, 69 (3), 898–903.

Romero, D., de Vicente, A., Olmos, J.L., Davila, J.C. &

Perez-Garcia, A. (2007) Effect of lipopeptides of

antagonistic strains of Bacillus subtilis on the

morphology and ultrastructure of the cucurbit fungal

pathogen Podosphaera fusca. Journal of Applied

Microbiology, 103, 969–976.

Ruiqian, L., Qian, Y., Thanaboripat, D. & Thansukon, P.

(2004) Biocontrol of Aspergillus flavus and aflatoxin

production. KMITL Science Journal, 4, 132–155.

Ryan, R.P., Germaine, K., Franks, A., Ryan, D.J. & Dowling,

D.N. (2008) Bacterial endophytes: Recent

developments and applications. FEMS Microbiology

Letters, 278 (1), 1–9. doi:10.1111/j.1574-


Satish, S., Mohana, D.C., Raghavendra, M.P. & Raveesha,

K.A. (2007) Antifungal activity of some plant extracts

against important seed borne pathogens of Aspergillus

sp. International Journal of Agricultural Technology, 3

(1), 109–119.

Selvi, A.T., Joseph, G.S. & Jayaprakasha, G.K. (2003)

Inhibition of growth and aflatoxin production in

Aspergillus flavus by Garcinia indica extract and its

antioxidant activuty. Food Microbiology, 20, 455–460.

Smirnov, V.V., Reznik, S.R. & Vasilievskaya, I.A. (1986)

Aerobe Endospore-Forming Bacteria. Budapest:

Medicina Konyvkiado.

Sneath, P.H.A. (1986) Endospore-forming Gram-positive

rods and cocci. In Sneath, P.H.A., Mair, N.S., Sharpe,

M.E. & Holt, J.G. (eds.) Bergey’s Manual of Systematic

Bacteriology, Vol. 2. Baltimore: William & Wilkins.


Susilowati, D.N., Hidayatun, N., Tasliah & Mulya, K. (2010)

Keragaman bakteri endofitik pada empat jenis varietas

padi dengan metode ARDRA (amplified ribosomal

DNA restriction analysis). Berita Biologi, 10 (2), 241–

Yoshida, S., Hiradate, S., Tsukamoto, T., Hatakeda, K. &

Shirata, A. (2001) Antimicrobial activity of culture

filtrate of Bacillus amyloliquefaciens RC-2 isolated

from mulberry leaves. Phytopathology, 91 (2), 181–

Yu, H., Zhang, L., Li, L., Zheng, C., Guo, L., Li, W., Sun, P.

& Qin, L. (2010) Recent development and future

prospects of antimicrobial metabolites produced by

endophytes. Microbiological Research, 165, 437–449.



  • There are currently no refbacks.

Copyright (c) 2018 Jurnal AgroBiogen

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

 Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.


P-ISSN : 1907-1094
E-ISSN : 2549-1547

Jurnal AgroBiogen

Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Jl. Tentara Pelajar 3A, Bogor 16111
Jawa Barat, Indonesia
Telp.: (0251) 8339793, 8337975
Faks.: (0251) 8338820

View My Stats