Research Article
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Year 2025, Volume: 12 Issue: 4, 1068 - 1074, 17.10.2025
https://doi.org/10.30910/turkjans.1673786

Abstract

Project Number

TUBITAK 2209A-Project number 1919B012214573

References

  • Albrigo, L.G., Stelinski, L.L., & Timmer, L. (2019). Citrus. CABI, Wallingford.
  • Anonymous, (2024). Food and Agriculture Organization of the United Nations (FAO), http://www.fao.org/site, (Accessed date: 28.03.2025)
  • Arora, N.K. (2018). Agricultural sustainability and food security. Environmental Sustainability, 1(3), 217–219.
  • Chand, K., Shah, S., Sharma, J., Paudel, M.R., & Pant, B. (2020). Isolation, characterization, and plant growth-promoting activities of endophytic fungi from a wild orchid Vanda cristata. Plant Signaling Behavior, 15(5), 1744294.
  • Chen, X., Luo, X., Fan, M., Zeng, W., Yang, C., Wu, J., Zhao, C., Zhang, Y., & Zhao, P. (2019). Endophytic fungi from the branches of Camellia taliensis (W.W. Smith) Melchior, a widely distributed wild tea plant. World Journal of Microbiology Biotechnology, 35, 1–15.
  • Clay, K., Shearin, Z.R.C., Bourke, K.A., Bickford, W.A., & Kowalski, K.P. (2016). Diversity of fungal endophytes in non-native Phragmites australis in the Great Lakes. Biological Invasions, 18, 2703–2716.
  • Das, S.K., & Mahapatra, S. (2019). Isolation and characterization of bioactive compound from endophytic fungus of spoiled fruits. International Journal of Research and Analytical Reviews, 7, 65–72.
  • De Silva, N.I., Brooks, S., Lumyong, S., & Hyde, K.D. (2019). Use of endophytes as biocontrol agents. Fungal Biology Reviews, 33, 133–148.
  • Dhayanithy, G., Subban, K., & Chelliah, J. (2019). Diversity and biological activities of endophytic fungi associated with Catharanthus roseus. BMC Microbiology, 19, 1–14.
  • Douanla-Meli, C., Langer, E., & Talontsi Mouafo, F. (2013). Fungal endophyte diversity and community patterns in healthy and yellowing leaves of Citrus limon. Fungal Ecology, 6(3), 212–222.
  • Du, T.Y., Karunarathna, S.C., Zhang, X., Dai, D.Q., Mapook, A., Suwannarach, N., & Tibpromma, S. (2022). Endophytic fungi associated with Aquilaria sinensis (Agarwood) from China show antagonism against bacterial and fungal pathogens. Journal of Fungi, 8(11), 1197.
  • Durán, E.L., Ploper, L.D., Ramallo, J.C., Grandi, R.A.P., Giancoli, A.C.H., & Azevedo, J.L. (2005). The foliar fungal endophytes of Citrus limon in Argentina. Canadian Journal of Botany, 83, 350–355.
  • Fu, J.X., Jiao, J., Gai, Q.Y., Fu, Y. J., Zhang, Z.Y., Gao, J., & Wang, X.Q. (2024). Enhanced accumulation of health-promoting cajaninstilbene acid in pigeon pea hairy root cultures cocultured with an endophytic fungus during early stages of colonization. Journal of Agricultural and Food Chemistry, 72(42), 23389–23400.
  • Garganese, F., Ippolito, A., Di Rienzo, V., Lotti, C., Montemurro, C., & Sanzani, S.M. (2018). A new high resolution melting assay for genotyping Alternaria species causing citrus brown spot. Journal Science Food Agriculture, 98(12), 4578–4583.
  • Giménez-Sanchis, A., Zhong, K., Pintor, A., Farina, V., & Besada, C. (2022). Understanding blood versus blond orange consumption: A cross-cultural study in four countries. Foods, 11(17), 2686.
  • Glienke-Blanco, C., Aguilar-Vildoso, C.I., Vieira, M.L.C., Barroso, P.A.V., & Azevedo, J.L. (2002). Genetic variability in the endophytic fungus Guignardia citricarpa isolated from citrus plants. Genetics Molecular Biology, 25, 251-255.
  • Juybari, H.Z., Tajick Ghanbary, M.A., Rahimian, H., Karimi, K., & Arzanlou, M. (2019). Seasonal, tissue and age infuences on frequency and biodiversity of endophytic fungi of Citrus sinensis in Iran. Forest Pathology, 49(6), e12559.
  • Korkom, Y. (2023). Drought stress is a global problem in sustainable agriculture: response of plant, role of fungal endophytes. In Türker HB, Gül A (ed) Architectural Sciences and Urban Agriculture II, Iksad, Türkiye, pp 223-254. https://doi.org/10.5281/zenodo.8385856
  • Liu, K.H., Ding, X.W., Deng, B.W., & Chen, W.Q. (2009). Isolation and characterization of endophytic taxol-producing fungi from Taxus chinensis. Journal Industrial Microbiology Biotechnology, 36, 1171–1177.
  • Li, P.Q., Wu, Z., Liu, T., & Wang, Y.N. (2016). Biodiversity, phylogeny, and antifungal functions of endophytic fungi associated with Zanthoxylum bungeanum. International Journal of Molecular Sciences, 17, 1541.
  • Lugtenberg, B.J.J., Caradus, J.R., & Johnson, L.J. (2016). Fungal endophytes for sustainable crop production. FEMS Microbiology Ecology, 92, fw94.
  • Mattoo, A.J., & Nonzom, S. (2021). Endophytic fungi: understanding complex cross-talks. Symbiosis, 83(3), 237–264.
  • Moghaddam, M.S.H., Safaie, N., Soltani, J., & Hagh-Doust, N. (2021). Desertadapted fungal endophytes induce salinity and drought stress resistance in model crops. Plant Physiology Biochemistry, 160, 225–238.
  • Moharram, A.M., Zohri, A.A., Omar, H.M., & Abd El-Ghani, O.A. (2017). In vitro assessment of antimicrobial and anti-inflammatory potential of endophytic fungal metabolites extracts. European Journal of Biological Research, 7(3), 234–244.
  • Moraes Bazioli, J., Belinato, J.R., & Costa, J.H. (2019). Biological control of citrus postharvest phytopathogens. Toxins (Basel) 11, 460.
  • Muñoz-Guerrero, J., Guerra-Sierra, B.E., & Alvarez, J.C. (2021). Fungal endophytes of Tahiti Lime (Citrus citrus × latifolia) and their potential for control of Colletotrichum acutatum JH simmonds causing anthracnose. Frontiers Bioengineering Biotechnology, 9, 650351.
  • Narayanasamy, P. (2011). Diagnosis of fungal diseases of plants. In: Microbial plant pathogens-detection and disease diagnosis. pp 273–284, Springer, Dordrecht.
  • Ortega, H.E., Torres-Mendoza, D., & Cubilla-Rios, L. (2020). Patents on endophytic fungi for agriculture and bio-and phytoremediation applications. Microorganisms, 8, 1237.
  • Qi, F., Jing, T., & Zhan, Y. (2012). Characterization of endophytic fungi from Acer ginnala maxim in an artificial plantation: media effect and tissue-dependent variation. PLoS One, 7, e46785.
  • Pazarlar, S., & Şimşek, E. (2024). The endophytic fungus Serendipita indica colonization protects chickpea plants against Fusarium wilt disease. Journal of Agriculture Faculty of Ege University, 61(4), 449–459.
  • Potshangbam, M., Devi, S. I., Sahoo, D., & Strobel, G.A. (2017). Functional characterization of endophytic fungal community associated with Oryza sativa L. and Zea mays L. Frontiers in Microbiology, 8, 325.
  • Priyashantha, A.K.H., Karunarathna, S.C., Lu, L., & Tibpromma, S. (2023). Fungal endophytes: An alternative biocontrol agent against phytopathogenic fungi. Encyclopedia, 3(2), 759–780.
  • Ramos, A.P., Talhinhas, P., Sreenivasaprasad, S., & Oliveira, H. (2016). Characterization of Colletotrichum gloeosporioides as the main causal agent of citrus anthracnose, and C. karstii as species preferentially associated with lemon twig dieback in Portugal. Phytoparasitica, 44, 549–561.
  • Sadeghi, F., Samsampour, D., Seyahooei, M.A., Bagheri, A., & Soltani, J. (2019). Diversity and spatiotemporal distribution of fungal endophytes associated with Citrus reticulata cv. siyahoo. Current Microbiology, 76, 279–289.
  • Saito, S., & Xiao, C.L. (2017). Prevalence of postharvest diseases of mandarin fruit in California. Plant Health Progress, 18, 204–210.
  • Sharma, D., Pramanik, A., & Agrawal, P.K. (2016). Evaluation of bioactive secondary metabolites from endophytic fungus Pestalotiopsis neglecta BAB-5510 isolated from leaves of Cupressus torulosa Don. 3 Biotech, 6(2), 210.
  • Soltani, J. (2017). Endophytism in Cupressoideae (Coniferae): a model in endophyte biology and biotechnology. In: Maheshwari D (ed) Endophytes: biology and biotechnology. Sustainable Development and Biodiversity, vol 15. Springer, Cham.
  • Syamsia, S., Idhan, A., Latifah, H., Noerfityani, N., & Akbar, A. (2021). Alternative medium for the growth of endophytic fungi. In IOP Conference Series: Earth and Environmental Science, 886(1), 012045.
  • Talukdar, R., & Tayung, K. (2021). Endophytic fungal assemblages of Zanthoxylum oxyphyllum Edgew. and their antimicrobial potential. Plant Science Today, 8, 132-139.
  • Wang, Y., Lai, Z., Li, X.X., Yan, R.M., Zhang, Z.B., Yang, H.L., & Zhu, D. (2016). Isolation, diversity and acetylcholinesterase inhibitory activity of the culturable endophytic fungi harboured in Huperzia serrata from Jinggang Mountain, China. World Journal Microbiology Biotechnology, 32, 1–23.
  • Wani, Z.A., Ashraf, N., Mohiuddin, T., & Riyaz-Ul-Hassan, S. (2015). Plant-endophyte symbiosis, an ecological perspective. Applied Microbiology Biotechnology, 99(7), 2955–2965.
  • Vega, F.E., Simpkins, A., Aime, M.C., Posada, F., Peterson, S.W., Rehner, S.A., Infante, F., Castillo, A., & Arnold, E. (2010). Fungal endophyte diversity in cofee plants from Colombia, Hawai’i, Mexico and Puerto Rico. Fungal Ecology, 3, 122–138.
  • Yan, L., Zhu, J., Zhao, X., Shi, J., Jiang, C., & Shao, D. (2019). Benefcial efects of endophytic fungi colonization on plants. Applied Microbiology Biotechnology, 103(8), 3327–3340.

Comparison of Different Media for Isolating Endophytic Fungi from Plant Tissues of Citrus Species

Year 2025, Volume: 12 Issue: 4, 1068 - 1074, 17.10.2025
https://doi.org/10.30910/turkjans.1673786

Abstract

Endophytic fungi play an ecological role in promoting plant growth and controlling diseases. This study compared the efficacy of different media for isolating fungal endophytes. We used potato dextrose agar (PDA), corn meal agar (CMA), water agar (WA), czapek dox agar (CDA), and sabouraud agar (SB). This study resulted in the isolation of 43 endophytic fungal isolates from healthy leaf and fruit tissues of mandarin and orange, with 69.7% of these isolates coming from leaf tissue. Morphological identification revealed these isolates belonged to Alternaria spp., Colletotrichum spp., Botryosphaeriaceae, and Trichoderma spp. Principal component analysis (PCA) showed that PDA and SB were effective for isolating culturable endophytic fungi. The unweighted pair-group method of arithmetic (UPGMA) dendrogram showed that PDA was in the same group as the SB, while CDA and CMA were separated. The WA was distinct from all other media, forming an outgroup. The results of this investigation demonstrated the influence of different media on the isolation of endophyte fungi from various plant tissues of Citrus species. Future research should focus on the efficacy of plant disease of the endophyte fungi and their potential for biocontrol under field conditions.

Ethical Statement

The authors declare no competing interests.

Supporting Institution

The Scientific and Technological Research Council of Türkiye (TUBITAK)

Project Number

TUBITAK 2209A-Project number 1919B012214573

Thanks

We acknowledge the Scientific and Technological Research Council of Türkiye (TUBITAK) for the financial support of this work.

References

  • Albrigo, L.G., Stelinski, L.L., & Timmer, L. (2019). Citrus. CABI, Wallingford.
  • Anonymous, (2024). Food and Agriculture Organization of the United Nations (FAO), http://www.fao.org/site, (Accessed date: 28.03.2025)
  • Arora, N.K. (2018). Agricultural sustainability and food security. Environmental Sustainability, 1(3), 217–219.
  • Chand, K., Shah, S., Sharma, J., Paudel, M.R., & Pant, B. (2020). Isolation, characterization, and plant growth-promoting activities of endophytic fungi from a wild orchid Vanda cristata. Plant Signaling Behavior, 15(5), 1744294.
  • Chen, X., Luo, X., Fan, M., Zeng, W., Yang, C., Wu, J., Zhao, C., Zhang, Y., & Zhao, P. (2019). Endophytic fungi from the branches of Camellia taliensis (W.W. Smith) Melchior, a widely distributed wild tea plant. World Journal of Microbiology Biotechnology, 35, 1–15.
  • Clay, K., Shearin, Z.R.C., Bourke, K.A., Bickford, W.A., & Kowalski, K.P. (2016). Diversity of fungal endophytes in non-native Phragmites australis in the Great Lakes. Biological Invasions, 18, 2703–2716.
  • Das, S.K., & Mahapatra, S. (2019). Isolation and characterization of bioactive compound from endophytic fungus of spoiled fruits. International Journal of Research and Analytical Reviews, 7, 65–72.
  • De Silva, N.I., Brooks, S., Lumyong, S., & Hyde, K.D. (2019). Use of endophytes as biocontrol agents. Fungal Biology Reviews, 33, 133–148.
  • Dhayanithy, G., Subban, K., & Chelliah, J. (2019). Diversity and biological activities of endophytic fungi associated with Catharanthus roseus. BMC Microbiology, 19, 1–14.
  • Douanla-Meli, C., Langer, E., & Talontsi Mouafo, F. (2013). Fungal endophyte diversity and community patterns in healthy and yellowing leaves of Citrus limon. Fungal Ecology, 6(3), 212–222.
  • Du, T.Y., Karunarathna, S.C., Zhang, X., Dai, D.Q., Mapook, A., Suwannarach, N., & Tibpromma, S. (2022). Endophytic fungi associated with Aquilaria sinensis (Agarwood) from China show antagonism against bacterial and fungal pathogens. Journal of Fungi, 8(11), 1197.
  • Durán, E.L., Ploper, L.D., Ramallo, J.C., Grandi, R.A.P., Giancoli, A.C.H., & Azevedo, J.L. (2005). The foliar fungal endophytes of Citrus limon in Argentina. Canadian Journal of Botany, 83, 350–355.
  • Fu, J.X., Jiao, J., Gai, Q.Y., Fu, Y. J., Zhang, Z.Y., Gao, J., & Wang, X.Q. (2024). Enhanced accumulation of health-promoting cajaninstilbene acid in pigeon pea hairy root cultures cocultured with an endophytic fungus during early stages of colonization. Journal of Agricultural and Food Chemistry, 72(42), 23389–23400.
  • Garganese, F., Ippolito, A., Di Rienzo, V., Lotti, C., Montemurro, C., & Sanzani, S.M. (2018). A new high resolution melting assay for genotyping Alternaria species causing citrus brown spot. Journal Science Food Agriculture, 98(12), 4578–4583.
  • Giménez-Sanchis, A., Zhong, K., Pintor, A., Farina, V., & Besada, C. (2022). Understanding blood versus blond orange consumption: A cross-cultural study in four countries. Foods, 11(17), 2686.
  • Glienke-Blanco, C., Aguilar-Vildoso, C.I., Vieira, M.L.C., Barroso, P.A.V., & Azevedo, J.L. (2002). Genetic variability in the endophytic fungus Guignardia citricarpa isolated from citrus plants. Genetics Molecular Biology, 25, 251-255.
  • Juybari, H.Z., Tajick Ghanbary, M.A., Rahimian, H., Karimi, K., & Arzanlou, M. (2019). Seasonal, tissue and age infuences on frequency and biodiversity of endophytic fungi of Citrus sinensis in Iran. Forest Pathology, 49(6), e12559.
  • Korkom, Y. (2023). Drought stress is a global problem in sustainable agriculture: response of plant, role of fungal endophytes. In Türker HB, Gül A (ed) Architectural Sciences and Urban Agriculture II, Iksad, Türkiye, pp 223-254. https://doi.org/10.5281/zenodo.8385856
  • Liu, K.H., Ding, X.W., Deng, B.W., & Chen, W.Q. (2009). Isolation and characterization of endophytic taxol-producing fungi from Taxus chinensis. Journal Industrial Microbiology Biotechnology, 36, 1171–1177.
  • Li, P.Q., Wu, Z., Liu, T., & Wang, Y.N. (2016). Biodiversity, phylogeny, and antifungal functions of endophytic fungi associated with Zanthoxylum bungeanum. International Journal of Molecular Sciences, 17, 1541.
  • Lugtenberg, B.J.J., Caradus, J.R., & Johnson, L.J. (2016). Fungal endophytes for sustainable crop production. FEMS Microbiology Ecology, 92, fw94.
  • Mattoo, A.J., & Nonzom, S. (2021). Endophytic fungi: understanding complex cross-talks. Symbiosis, 83(3), 237–264.
  • Moghaddam, M.S.H., Safaie, N., Soltani, J., & Hagh-Doust, N. (2021). Desertadapted fungal endophytes induce salinity and drought stress resistance in model crops. Plant Physiology Biochemistry, 160, 225–238.
  • Moharram, A.M., Zohri, A.A., Omar, H.M., & Abd El-Ghani, O.A. (2017). In vitro assessment of antimicrobial and anti-inflammatory potential of endophytic fungal metabolites extracts. European Journal of Biological Research, 7(3), 234–244.
  • Moraes Bazioli, J., Belinato, J.R., & Costa, J.H. (2019). Biological control of citrus postharvest phytopathogens. Toxins (Basel) 11, 460.
  • Muñoz-Guerrero, J., Guerra-Sierra, B.E., & Alvarez, J.C. (2021). Fungal endophytes of Tahiti Lime (Citrus citrus × latifolia) and their potential for control of Colletotrichum acutatum JH simmonds causing anthracnose. Frontiers Bioengineering Biotechnology, 9, 650351.
  • Narayanasamy, P. (2011). Diagnosis of fungal diseases of plants. In: Microbial plant pathogens-detection and disease diagnosis. pp 273–284, Springer, Dordrecht.
  • Ortega, H.E., Torres-Mendoza, D., & Cubilla-Rios, L. (2020). Patents on endophytic fungi for agriculture and bio-and phytoremediation applications. Microorganisms, 8, 1237.
  • Qi, F., Jing, T., & Zhan, Y. (2012). Characterization of endophytic fungi from Acer ginnala maxim in an artificial plantation: media effect and tissue-dependent variation. PLoS One, 7, e46785.
  • Pazarlar, S., & Şimşek, E. (2024). The endophytic fungus Serendipita indica colonization protects chickpea plants against Fusarium wilt disease. Journal of Agriculture Faculty of Ege University, 61(4), 449–459.
  • Potshangbam, M., Devi, S. I., Sahoo, D., & Strobel, G.A. (2017). Functional characterization of endophytic fungal community associated with Oryza sativa L. and Zea mays L. Frontiers in Microbiology, 8, 325.
  • Priyashantha, A.K.H., Karunarathna, S.C., Lu, L., & Tibpromma, S. (2023). Fungal endophytes: An alternative biocontrol agent against phytopathogenic fungi. Encyclopedia, 3(2), 759–780.
  • Ramos, A.P., Talhinhas, P., Sreenivasaprasad, S., & Oliveira, H. (2016). Characterization of Colletotrichum gloeosporioides as the main causal agent of citrus anthracnose, and C. karstii as species preferentially associated with lemon twig dieback in Portugal. Phytoparasitica, 44, 549–561.
  • Sadeghi, F., Samsampour, D., Seyahooei, M.A., Bagheri, A., & Soltani, J. (2019). Diversity and spatiotemporal distribution of fungal endophytes associated with Citrus reticulata cv. siyahoo. Current Microbiology, 76, 279–289.
  • Saito, S., & Xiao, C.L. (2017). Prevalence of postharvest diseases of mandarin fruit in California. Plant Health Progress, 18, 204–210.
  • Sharma, D., Pramanik, A., & Agrawal, P.K. (2016). Evaluation of bioactive secondary metabolites from endophytic fungus Pestalotiopsis neglecta BAB-5510 isolated from leaves of Cupressus torulosa Don. 3 Biotech, 6(2), 210.
  • Soltani, J. (2017). Endophytism in Cupressoideae (Coniferae): a model in endophyte biology and biotechnology. In: Maheshwari D (ed) Endophytes: biology and biotechnology. Sustainable Development and Biodiversity, vol 15. Springer, Cham.
  • Syamsia, S., Idhan, A., Latifah, H., Noerfityani, N., & Akbar, A. (2021). Alternative medium for the growth of endophytic fungi. In IOP Conference Series: Earth and Environmental Science, 886(1), 012045.
  • Talukdar, R., & Tayung, K. (2021). Endophytic fungal assemblages of Zanthoxylum oxyphyllum Edgew. and their antimicrobial potential. Plant Science Today, 8, 132-139.
  • Wang, Y., Lai, Z., Li, X.X., Yan, R.M., Zhang, Z.B., Yang, H.L., & Zhu, D. (2016). Isolation, diversity and acetylcholinesterase inhibitory activity of the culturable endophytic fungi harboured in Huperzia serrata from Jinggang Mountain, China. World Journal Microbiology Biotechnology, 32, 1–23.
  • Wani, Z.A., Ashraf, N., Mohiuddin, T., & Riyaz-Ul-Hassan, S. (2015). Plant-endophyte symbiosis, an ecological perspective. Applied Microbiology Biotechnology, 99(7), 2955–2965.
  • Vega, F.E., Simpkins, A., Aime, M.C., Posada, F., Peterson, S.W., Rehner, S.A., Infante, F., Castillo, A., & Arnold, E. (2010). Fungal endophyte diversity in cofee plants from Colombia, Hawai’i, Mexico and Puerto Rico. Fungal Ecology, 3, 122–138.
  • Yan, L., Zhu, J., Zhao, X., Shi, J., Jiang, C., & Shao, D. (2019). Benefcial efects of endophytic fungi colonization on plants. Applied Microbiology Biotechnology, 103(8), 3327–3340.
There are 43 citations in total.

Details

Primary Language English
Subjects Phytopathology, Plant Protection (Other)
Journal Section Research Articles
Authors

Yunus Korkom 0000-0001-5859-9026

Gizem Özgün 0000-0002-2262-8136

Ayhan Yıldız 0000-0001-9443-2362

Project Number TUBITAK 2209A-Project number 1919B012214573
Publication Date October 17, 2025
Submission Date April 11, 2025
Acceptance Date August 20, 2025
Published in Issue Year 2025 Volume: 12 Issue: 4

Cite

APA Korkom, Y., Özgün, G., & Yıldız, A. (2025). Comparison of Different Media for Isolating Endophytic Fungi from Plant Tissues of Citrus Species. Turkish Journal of Agricultural and Natural Sciences, 12(4), 1068-1074. https://doi.org/10.30910/turkjans.1673786