Sains Malaysiana 47(8)(2018): 1685–1692

http://dx.doi.org/10.17576/jsm-2018-4708-07

Antimicrobial Activity of Fungal Endophytes from Vaccinium dunalianum var. urophyllum

(Aktiviti Antimikrob Kulat Endofit daripada Vaccinium dunalianum var. urophyllum)

XIN TONG, XIAO-YE SHEN* & CHENG-LIN HOU

College of Life Science, Capital Normal University, Beijing, PR China

Received: 20 March 2017/Accepted: 22 March 2018

ABSTRACT

Fungi associated with Vaccinium species play important roles in plant growth and disease control, especially in the final blueberry production. Vaccinium dunalianum var. urophyllum (Ericaceae) is a well-known medicinal plant in Southern China used to treat inflammation and microbial infections. The endophytic fungi from these plants are therefore anticipated as potential new sources of antimicrobials. In this report, the inhibitory effects of endophytes against clinical bacteria and yeast were comprehensively screened and 11 isolates indicated high bioactivity by the agar diffusion method. The corresponding crude extracts of these fungi under submerged fermentation also demonstrated distinct differences and n-butyl alcohol displayed the lowest extraction efficiency among the extracts. The ethyl acetate and dichloromethane extracts of filtrates from the Colletotrichum sp. VD001, Epicoccum nigrum VD021 and E. nigrum VD022 strains displayed good properties against pathogenic microorganisms according to disc diffusion assays and minimal inhibitory concentration (MIC). This study is the first indicating that cultivable endophytic fungi associated with blueberry plants produce potential compounds against clinical pathogens.

Keywords: Antimicrobial activity; fungal endophytes; minimum inhibitory concentration; Vaccinium species

ABSTRAK

Kulat yang dikaitkan dengan spesies Vaccinium memainkan peranan penting dalam pertumbuhan tumbuhan dan kawalan penyakit, terutamanya dalam pengeluaran beri biru akhir. Vaccinium dunalianum var. urophyllum (Ericaceae) ialah tumbuhan ubatan yang terkenal di Selatan China yang digunakan untuk merawat jangkitan keradangan dan mikrob. Oleh itu, kulat endofit daripada tumbuhan ini dijangka sebagai sumber baru antimikrob yang berpotensi. Dalam laporan ini, kesan perencatan endofit terhadap bakteria klinikal dan suatu ragi telah ditayangkan secara komprehensif dan 11 pencilan menunjukkan bioaktiviti tinggi melalui kaedah penyebaran agar. Ekstrak mentah yang sepadan dengan kulat ini di bawah penapaian terendam juga menunjukkan perbezaan yang ketara serta n-butil alkohol menunjukkan kecekapan pengekstrakan terendah dalam kalangan ekstrak. Ekstrak etil asetat dan diklorometana filtrat daripada Colletotrichum sp. VD001, Epicoccum nigrum VD021 dan E. nigrum VD022 menunjukkan sifat-sifat yang baik terhadap mikroorganisma patogen berdasarkan ujian penyebaran cakera dan kepekatan perencatan minimum (MIC). Kajian ini adalah kajian yang pertama menunjukkan bahawa kulat endofit yang boleh dikaitkan dengan tumbuhan beri biru menghasilkan sebatian yang berpotensi terhadap patogen klinikal.

Kata kunci: Aktiviti antimikrob; kepekatan perencatan minimum; kulat endofit; spesies Vaccinium

REFERENCES

Ali, S.M. & Olivo, M. 2002. Efficacy of hypocrellin pharmacokinetics in phototherapy. International Journal of Oncology 21: 1229-1237.

Bamford, P.C., Norris, G.L.F. & Ward, G. 1961. Flavipin production by Epicoccum spp. Transactions of the British Mycological Society 44(3): 354-356.

Baute, M.A., Deffieux, G., Baute, R. & Neveu, A. 1978. New antibiotics from the fungus Epicoccum nigrum. I. Fermentation, isolation and antibacterial properties. The Journal of Antibiotics 31(11): 1099-1101.

Bell, P.J.L. & Karuso, P. 2003. Epicocconone, a novel fluorescent compound from the fungus Epicoccum nigrum. Journal of the American Chemical Society 125(31): 9304-9305.

Cannon, P.F., Damm, U., Johnston, P.R. & Weir, B.S. 2012. Colletotrichum-current status and future directions. Studies in Mycology 73(1): 181-213.

Dos Santos, I.P., Da, S.L., Da, S.M., de Araújo, J.M., Cavalcanti, M.S. & Lima, V. L. 2015. Antibacterial activity of endophytic fungi from leaves of Indigofera suffruticosa Miller. (Fabaceae). Frontiers in Microbiology 6: 350.

Elfar, K., Torres, R., Díaz, G.A. & Latorre, B.A. 2013. Characterization of Diaporthe australafricana and Diaporthe spp. associated with stem canker of blueberry in Chile. Plant Disease Journal 97: 1042-1050.

Eloff, J.N. 1998. A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria. Planta Medica 64: 711-713.

Ermis, E., Hertel, C., Schneider, C., Carle, R., Stintzing, F. & Schmidt, H. 2015. Characterization of in vitro antifungal activities of small and American cranberry (Vaccinium oxycoccos L. and V. macrocarpon Aiton) and lingonberry (Vaccinium vitis-idaea L.) concentrates in sugar reduced fruit spreads. International Journal of Food Microbiology 204: 111-117.

Faeth, S.H. & Fagan, W.F. 2002. Fungal endophytes: Common host plant symbionts but uncommon mutualists. Integrative and Comparative Biology 42: 360-368.

Farr, D.F., Castlebury, L.A. & Rossman, A.Y. 2002. Morphological and molecular characterization of Phomopsis vaccinii and additional isolates of Phomopsis from blueberry and cranberry in the eastern United States. Mycologia 94: 494- 504.

Garcíapajón, C.M. & Collado, I.G. 2003. Secondary metabolites isolated from Colletotrichum species. Natural Product Reports 20(4): 426-431.

Girardot, M., Guerineau, A., Boudesocque, L., Costa, D., Bazinet, L., Enguehard-Gueiffier, C. & Imbert, C. 2014. Promising results of cranberry in the prevention of oral Candida biofilms. Pathogens and Disease 70: 432-439.

Hyde, K.D., Cai, L., Cannon, P.F., Crouch, J.A., Crous, P.W., Damm, U., Goodwin, P.H., Chen, H., Johnston, P.R., Jones, E.B.G., Liu, Z.Y., McKenzie, E.H.C., Moriwaki, J., Noireung, P., Pennycook, S.R., Pfenning, L.H., Prihastuti, H., Sato, T., Shivas, R.G., Tan, Y.P., Taylor, P.W.J., Weir, B.S., Yang, Y.L. & Zhang, J.Z. 2009. Colletotrichum - names in current use. Fungal Diversity 39(1): 147-182.

Huang, X. & Xie, Z. 1988. Beijing Medical College: Dictionary of Traditional Chinese Medicine. Hong Kong: Commercial Press.

Ikawa, M., Mcgratten, C.J., Burge, W.R., Iannitelli, R.C., Uebel, J.J. & Noguchi, T. 1978. Epirodin, a polyene antibiotic from the mold Eipoccum nigrum. The Journal of Antibiotics 31(2): 159-161.

Jeffers, S.N. 1991. Seasonal incidence of fungi in symptomless cranberry leaves and fruit treated with fungicides during bloom. Phytopathology 81: 636-644.

Kalt, W., Joseph, J.A. & Shukitt-Hale, B. 2007. Blueberries and human health: A review of current reseach. Journal- American Pomological Society 61: 151-160.

Kusari, P., Kusari, S., Spiteller, M. & Kayser, O. 2013. Endophytic fungi harbored in Cannabis sativa L.: Diversity and potential as biocontrol agents against host plant-specific phytopathogens. Fungal Diversity 60: 137-151.

Li, S.N., Li, Y.D. & Wang, Q. 2009. Bolting flavonoid-producing endophytic fungi from Vaccinium. Journal of Jilin Agricultural University 31: 587-591.

Li, Z.J., Shen, X.Y. & Hou, C.L. 2016. Fungal endophytes of South China blueberry (Vaccinium dunalianum var. urophyllum). Letters in Applied Microbiology 63: 482-487.

Ma, G., Khan, S.I., Jacob, M.R., Tekwani, B.L., Li, Z., Pasco, D.S., Walker, L.A. & Khan, I.A. 2004. Antimicrobial and antileishmanial activities of hypocrellins A and B. Antimicrobial Agents and Chemotherapy 48: 4450-4452.

Miao, Y., Wang, H., Li, G., Zhang, Z. & Tang, L. 2013. Morphology and structure of mycorrhiza and distribution of endophytic fungus in Vaccinium uliginosum L. in Changbai Mountain. Journal of Northeast Agricultural University 44: 81-85.

Mousa, W.K. & Raizada, M.N. 2013. The diversity of anti-microbial secondary metabolites produced by fungal endophytes: An interdisciplinary perspective. Frontiers in Microbiology 4: 65.

Perveen, I., Raza, M.A., Iqbal, T., Naz, I., Sehar, S. & Ahmed, S. 2017. Isolation of anticancer and antimicrobial metabolites from Epicoccum nigrum; endophyte of Ferula sumbul. Microbial Pathogenesis 110: 214-224.

Puišo, J., Jonkuvienė, D., Mačionienė, I., Šalomskienė, J., Jasutienė, I. & Kondrotas, R. 2014. Biosynthesis of silver nanoparticles using lingonberry and cranberry juices and their antimicrobial activity. Colloids and Surfaces B: Biointerfaces 121: 214-221.

Pusztahelyi, T., Holb, I.J. & Pócsi, I. 2015. Secondary metabolites in fungus-plant interactions. Frontiers in Plant Science 6: 573.

Qadri, M., Rajput, R., Abdin, M.Z., Vishwakarma, R.A. & Riyaz- Ul-Hassan, S. 2014. Diversity, molecular phylogeny, and bioactive potential of fungal endophytes associated with the Himalayan blue pine (Pinus wallichiana). Microbial Ecology 67: 877-887.

Reeh, K.W. & Cutler, G.C. 2013. Laboratory efficacy and fungicide compatibility of Clonostachys rosea against Botrytis blight on lowbush blueberry. Canadian Journal of Plant Science 93: 639-642.

Richardson, S.N., Walker, A.K., Nsiama, T.K., McFarlane, J., Sumarah, M.W., Ibrahim, A. & Miller, J.D. 2014. Griseofulvin-producing Xylaria endophytes of Pinus strobus and Vaccinium angustifolium: Evidence for a conifer-understory species endophyte ecology. Fungal Ecology 11: 107-113.

Sauer, M., Lu, P., Sangari, R., Kennedy, S., Polishook, J., Bills, G. & An, Z. 2002. Estimating polyketide metabolic potential among non-sporulating fungal endophytes of Vaccinium macrocarpon. Mycological Research 106: 460-470.

Shen, X.Y., Cheng, L.Y., Cai, C.J., Fan, L., Gao, J. & Hou, C.L. 2014. Diversity and antimicrobial activity of culturable endophytic fungi isolated from moso bamboo seeds. PLoS One 9: e95838.

Shu, Y.Z., Ye, Q., Li, H., Kadow, K.F., Hussain, R.A., Huang, S., Gustavson, D.R., Lowe, S.E., Chang, L.P., Pirnik, D.M. & Kodukula, K. 1997. Orevactaene, a novel binding inhibitor of HIV-1 rev protein to Rev response element (RRE) from Epicoccum nigrum WC47880. Bioorganic & Medicinal Chemistry Letters 7(17): 2295-2298.

Silva Araújo, F.D.D., Araújo, W.L., Aparicio, R. & Marsaioli, A.J. 2012. Epicolactone - natural product isolated from the sugarcane endophytic fungus Epicoccum nigrum. European Journal of Organic Chemistry 2012(27): 5225-5230.

Silva, S., Costa, E.M., Pereira, M.F., Costa, M.R. & Pintado, M.E. 2013. Evaluation of the antimicrobial activity of aqueous extracts from dry Vaccinium corymbosum extracts upon food microorganism. Food Control 34: 645-650.

Stewart, J.E., Brooks, K., Brannen, P.M., Cline, W.O. & Brewer, M.T. 2015. Elevated genetic diversity in the emerging blueberry pathogen Exobasidium maculosum. PLoS One 10: e0132545.

Stierle, A., Strobel, G. & Stierle, D. 1993. Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew. Science 260: 214-216.

Tadych, M., Bergen, M.S., Johnson-Cicalese, J., Polashock, J.J., Vorsa, N. & White, J.F. 2012. Endophytic and pathogenic fungi of developing cranberry ovaries from flower to mature fruit: diversity and succession. Fungal Diversity 54: 101-116.

Vohnik, M., Sadowsky, J.J., Lukesova, T., Albrechtova, J. & Vosatka, M. 2012. Inoculation with a ligninolytic basidiomycete, but not root symbiotic ascomycetes, positively affects growth of highbush blueberry (Ericaceae) grown in a pine litter substrate. Plant and Soil 355: 341-352.

Zhang, X.Y., Bao, J., Wang, G.H., He, F., Xu, X.Y. & Qi, S.H. 2012. Diversity and antimicrobial activity of culturable fungi isolated from six species of the South China Sea gorgonians. Microbial Ecology 64: 617-627.

Zhang, Y., Liu, S., Che, Y. & Liu, X. 2007. Epicoccins A-D, epipolythiodioxopiperazines from a cordyceps-colonizing isolate of Epicoccum nigrum. Journal of Natural Products 70(9): 1522-1525.

*Corresponding author; email: shenxiaoye2009@cnu.edu.cn