Determination of Biocontrol Potentials of Endophytic Bacteria in Biological Control of Citrus Sour Rot Disease Caused by Geotrichum citri-aurantii

Yıl 2022, Cilt: 19 Sayı: 1, 177 - 191, 26.01.2022

Soner Soylu , Merve Kara , Emine Mine Soylu , Aysun Uysal , Şener Kurt

https://doi.org/10.33462/jotaf.944704

Cited By: 8

Öz

Citrus fruits are susceptible to postharvest decays caused by various pathogenic fungal disease agents. Citrus sour rot, caused by Geotrichum citri-aurantii is one of the most important postharvest diseases of citrus fruits Although synthetic fungicides are commonly used to combat the disease, ineffectiveness of these applications as well as their harmful effects on human health and the environment have been reported in many studies recently. Biological control of postharvest diseases utilizing antagonistic bacteria has been explored in different host plant-pathogen interactions as a promising alternative to synthetic fungicides. In this study, biocontrol potentials of endophytic bacterial isolates, obtained from fruits, branch and leaves of healthy citrus trees, were investigated against Geotrichum citri-aurantii on petri plates (in vitro) and fruit tests (semi in vivo conditions). Among the 24 putative endophytic bacterial isolates isolated, sixteen different antagonist bacterial isolates were selected and assessed against mycelial growth inhibition of G. citri-aurantii in vitro studies. Among the bacterial isolates tested, Bacillus spp. isolates significantly inhibited mycelial growth of fungal agent by 59.5-78.6%. The highest mycelial growth inhibition was caused by B. subtilis CM8 (78.6%). Bacterial isolates of Acinetobacter johnsonii CT7, Erwinia herbicola CM5, Pseudomonas putida CL2, Pantoea agglomerans CM14, Acinetobacter lwoffii CP1 and Stenotrophomonas maltophilia CP3 failed to inhibit mycelial growth in dual culture assays. Antagonistic activities of bacterial isolates increased by the pre-incubation time before fungal inoculation. Bacterial isolates of Bacillus spp. caused considerably morphological changes such as vacuolation, shrivelling, swelling and lysis of hyphae close to inhibition zone. On artificially inoculated fruits, isolates of Bacillus spp. also found to suppressed disease incidence significantly by 56.7-83.3%. Due to strong antagonistic activities, isolates of Bacillus spp, tested in this study have the potential to be used as biofungicide for controlling post-harvest disease agent(s) of citrus fruits.

Anahtar Kelimeler

Citrus, Sour rot, Geotrichum citri-aurantii, Antagonist, Biological control, Endophyte bacteria

Geotrichum citri-aurantii’nin Sebep Olduğu Turunçgil Ekşi Çürüklük Hastalığının Biyolojik Mücadelesinde Endofit Bakterilerin Biyokontrol Potansiyellerinin Belirlenmesi

Öz

Turunçgil meyveleri, çeşitli fungal hastalık etmenleri tarafından neden olunan hasat sonrası hastalıklara karşı oldukça duyarlıdır. Geotrichum citri-aurantii'nin neden olduğu ekşi çürüklük, turunçgillerin en önemli hasat sonrası hastalıklarından biridir. Hastalıkla mücadelede genellikle sentetik fungisitler yaygın olarak kullanılsa da söz konusu uygulamaların son yıllarda etkisizliğinin yanı sıra insan sağlığı ve çevre üzerine olan zararlı etkileri birçok çalışmada bildirilmiştir. Hasat sonrası hastalıklara karşı antagonistik bakteri etmenlerinin kullanıldığı biyolojik mücadele, sentetik fungisitlere umut verici bir alternatif olduğu birçok farklı konukçu bitki patojen ilişkilerinin irdelendiği çalışmalarda araştırılmıştır. Bu çalışmada sağlıklı turunçgil ağaçlarının meyve, dal ve yapraklarından elde edilen endofitik bakteri izolatlarının biyokontrol potansiyelleri G. citri-aurantii'ye karşı petri denemeleri (in vitro) ve meyve testleri (yarı in vivo) ile araştırılmıştır. İzole edilen 24 aday bakteri izolatı arasından seçilen on altı farklı bakteri izolatı G. citri-aurantii'nin misel gelişiminin engellenmesi üzerine olan antagonistik potansiyeli in vitro çalışmalarda değerlendirilmiştir. Test edilen bakteriler arasında Bacillus spp. ait izolatlar fungal etmenin misel gelişimini % 59.5-78.6 oranında engellemiştir. En yüksek antagonistik etkinlik B. subtilis CM8 (% 78.6) izolatı tarafından neden olunmuştur. Acinetobacter johnsonii CT7, Erwinia herbicola CM5, Pseudomonas putida CL2, Pantoea agglomerans CM14, Acinetobacter lwoffii CP1 ve Stenotrophomonas maltophilia CP3 izolatları, ikili kültür denemelerinde misel gelişimini engellemede başarısız olmuştur. Bakteriyel izolatların antagonistik etkinlikleri, fungus inokulasyonundan önceki ön inkübasyon süresi ile artmıştır. Bacillus spp ait izolatlar engellenme bölgesine yakın noktalardaki hiflerde vakuolleşme, büzüşme, şişme ve erime gibi önemli morfolojik değişikliklere neden olmuştur. Yapay olarak enfekte edilen turunçgil meyveleri üzerinde hastalık çıkışı Bacillus spp. ait izolatlar tarafından % 56.7-83.3 gibi önemli oranlarda baskılanmıştır. Elde edilen sonuçlar, Bacillus spp. ait izolatların güçlü antagonistik aktivitelerinden dolayı, turunçgillerin hasat sonrası hastalık etmen(ler)i ile mücadelesinde biyofungisit olarak kullanılma potansiyeline sahip olduğunu göstermiştir.

Anahtar Kelimeler

Turunçgil, Ekşi çürüklük, Geotrichum citri-aurantii, Antagonist, Biyolojik mücadele, Endofit bakteri

Kaynakça

• Akgül, D. S., Mirik, M. (2008). Biocontrol of Phytophthora capsici on pepper plants by Bacillus megatarium strains. Journal of Plant Pathology, 90 (1): 29-34.
• Aktan, C., Soylu., S. (2020). Diyarbakır ilinde yetişen badem ağaçlarından endofit ve epifit bakteri türlerinin izolasyonu ve bitki gelişimini teşvik eden mekanizmalarının karakterizasyonu. KSÜ Tarım ve Doğa Dergisi 23 (3): 641-654.
• Amer, G.A., Aggarwal, R., Singh, D.V. and Srivastava, K. D. (1997). Interaction of Bacillus thuringiensis with Pythium ultimum and Fusarium oxysporum f. sp. lycopersici: Possible role in biological control. Currennt Sciences 3: 284–286
• Anonim (2020a). Yaş Meyve Sebze Sektörü Türkiye Geneli Değerlendirme Raporu. https://www.akib.org.tr/files/documents/2020/Rapor/Ekim%202020%20YMS%20%C4%B0hracat%20De%C4%9Ferlendirme%20Raporu.pdf, 38 sayfa.
• Anonim (2020b). TUİK Bitkisel Üretim İstatistikleri, https://biruni.tuik.gov.tr/medas/?kn=92&locale=tr, (Erişim tarihi: 20.04.2021)
• Anonymous (2019). Food and Agriculture Organization of the United Nations (FAO), http://www.fao.org/site, (Erişim tarihi: 20.04.2021)
• Barka, E.A., Gognies, S., Nowak, J., Audran, J.C., Belarbi, A. (2002). Inhibitory effect of endophyte bacteria on Botrytis cinerea and its influence to promote the grapevine growth. Biological Control 24: 135-142.
• Bazioli, J.M., Belinato, J.R., Costa, J.H., Akiyama, D.Y., Pontes, J.G.D., Kupper, K.C., Augusto, F., de Carvalho, J.E., Fill, T.P. (2019). Biological control of citrus postharvest phytopathogens. Toxins 11 (8): 460.
• Bora, P, Bora, L.C. (2020). Disease management in horticulture crops through microbial interventions: An overview. Indian Journal Of Agricultural Sciences 90 (8): 1389-1396
• Bozkurt, İ.A., Soylu, S. (2019). Elma kök uru hastalığı etmeni Rhizobium radiobacter’e karşı epifit ve endofit bakteri izolatlarının antagonistik potansiyellerinin belirlenmesi. Tekirdağ Ziraat Fakültesi Dergisi 16: 348-361.
• Carmona-Hernandez, S., Reyes-Perez, J.J., Chiquito-Contreras, R.G., Rincon-Enriquez, G., Cerdan-Cabrera, C.R., Hernandez-Montiel, L.G. (2019). Biocontrol of postharvest fruit fungal diseases by bacterial antagonists: a review. Agronomy-Basel 9 (3): 121.
• Chalupová, J., Raus, M., Sedlarova, M, Sebela, M. (2014). Identification of fungal microorganisms by MALDI-TOF mass spectrometry. Biotechnology Advances 32(1):230-41.
• Chaurasia, B., Pandey, A., Palni, L.M.S., Trivedi, P., Kumar, B., Colvin, N. (2005). Diffusible and volatile compounds produced by an antagonistic Bacillus subtilis strain cause structural deformations in pathogenic fungi in vitro. Microbiological Research 160: 75-81.
• Duman, K., Soylu, S. (2019). Characterization of plant growth-promoting traits and antagonistic potentials of endophytic bacteria from bean plants against Pseudomonas syringae pv. phaseolicola. Bitki Koruma Bülteni 59:59-69.
• Eckert J.W., Eaks I.L. (1989). Postharvest disorders and diseases of citrus fruits. In: Reuter W., Calavan E.C. & Carman G.E. (Eds), The citrus industry, vol. 5. Univ. Calif. Press, Berkeley, USA, pp. 179-260.
• Eljounaidi, K., Lee, S.K., Bae, H. (2016). Bacterial endophytes as potential biocontrol agents of vascular wilt diseases - Review and future prospects. Biological Control 103: 62-68.
• Feng, W., Chen, J., Zheng, X., Liu, Q. (2011). Thyme oil to control Alternaria alternate in vitro and in vivo as fumigant and contact treatments. Food Control 22: 78-81.
• Fravel, D.R. (2005). Commercialization and implementation of biocontrol. Annual Review of Phytopathology 43: 337-359.
• Gedik, M. (2019). Turunçgilde derim sonrası ekşi çürüklük hastalığı etmeni Geotrichum citri-aurantii’ye karşı antagonist bakteriler ve bor ürünlerinin antifungal etkileri. (Yüksek Lisans Tezi) Hatay Mustafa Kemal Üniversitesi, Bitki Koruma A.B.D., Hatay
• Ghazanfar, M.U., Hussain, M., Hamid, M.I., Ansari, S.U. (2016). Utilization of biological control agents for the management of postharvest pathogens of tomato. Pakistan Journal Of Botany 48 (5): 2093-2100.
• Gupta, C.P., Dubey, R.C., Kang, S.C., Maheshwari, D.K. (2001). Antibiosis-mediated necrotrophic effect of Pseudomonas GRC2 against two fungal plant pathogens. Current Science 81: 91-94.
• Han, T., You, C., Zhang, L., Feng, C., Zhang, C., Wang, J., Kong, F. (2016). Biocontrol potential of antagonist Bacillus subtilis Tpb55 against tobacco black shank. Biocontrol 61 (2): 195-205.
• Hao, W., Zhong, G., Hu, M., Luo, J., Weng, Q., Rizwan-ul-Haq, M. (2010). Control of citrus gren and blue mold and sour rot by tea saponin combined with imazalil and prochloraz. Postharvest Biology and Technology 56: 39-43.
• Hong, P., Hao, W., Luo, J., Chen, S., Hu, M., Zhong, G. (2014). Combination of hot water, Bacillus amyloliquefaciens HF-01 and sodium bicarbonate treatments to control postharvest decay of mandarin fruit. Postharvest Biology and Technology 88: 96-102.
• Horuz, S. (2009). Turunçgillerde ekşi çürüklük etmeni Geotrichum citri-aurantii (Ferraris) E. E. Butler'e bazı fungisitlerin etkililikleri üzerinde araştırmalar. (Yüksek Lisans Tezi) Ege Üniversitesi, Bitki Koruma A.B.D., İzmir.
• Horuz, S., Karut, Ş.T., Aysan, Y. (2019). Domates bakteriyel kanser ve solgunluk hastalığı etmeni Clavibacter michiganensis subsp. michiganensis’in tohumda aranması ve tohum uygulamalarının patojen gelişimine etkisinin belirlenmesi. Tekirdağ Ziraat Fakültesi Dergisi 16: 284-296
• Horuz, S., Kınay, P. (2010). The effects of some new postharvest fungicides and combination of hot water with sodium bicarbonate against Geotrichum citri-aurantii on citrus. Acta Horticulturae 877: 1551-1557.
• Kara, M., Soylu, E.M. (2020). Assessment of glucosinolate-derived isothiocyanates as potential natural antifungal compounds against citrus sour rot disease agent Geotrichum citri-aurantii. Journal of Phytopathology 168: 279-289
• Kara, M., Soylu, S., Kurt, Ş., Soylu, E. M., Uysal, A. (2020). Determination of antagonistic traits of bacterial isolates obtained from apricot against green fruit rot disease agent Sclerotinia sclerotiorum. Acta Horticulturae 1290:135-142.
• Kong, Q.J., Liang, Z., Xiong, J., Li, H.B., Ren, X.Y. (2016). Overexpression of the bivalent antibacterial peptide genes in Pichia pastoris delays sour rot in citrus fruit and induces Geotrichum citri-aurantii cell apoptosis. Food Biotechnology 30 (2): 79-97.
• Lelliot, R.A., Stead, D.E. (1987). Methods for the diagnosis of bacterial diseases of plants. (T.F. Preece, Editör). In: Methods in plant pathology. Vol 2, Blackwell Scientific Publications. pp. 176-177, Oxford.
• Liu, X., Wang, L.P, Li, Y.C., Li, H.Y., Yu, T., Zheng, X.D. (2009). Antifungal activity of thyme oil against Geotrichum citri-aurantii in vitro and in vivo. Journal of Applied Microbiology 107: 1450-1456.
• Liu, X., Fang, W., Liu, L., Yu, T., Lou, B., Zheng, X. (2010). Biological control of postharvest sour rot of citrus by two antagonistic yeasts. Letters in Applied Microbiology 51: 30-35.
• Liu, Y., Yao, S., Deng, L., Ming, J., Zeng, K. (2019). Different mechanisms of action of isolated epiphytic yeasts against Penicillium digitatum and Penicillium italicum on citrus fruit. Postharvest Biology and Technology 152: 100-110.
• Maldonado, M.C., Corona, J., Gordillo, M.A., Navarro, A.R. (2009). Isolation and partial characterization of antifungal metabolites produced by Bacillus sp. IBA 33. Current Microbiology 59: 646-650.
• McKay, A.H., Förster, H., Adaskaveg, J.E. (2012). Distinguishing Galactomyces citri-aurantii from G. geotrichum and characterizing population structure of the two postharvest sour rot pathogens of fruit crops in California. Phytopathology 102: 528-538.
• Mohammadi, P., Tozlu E., Kotan R., Kotan Ş.M. (2017). Potential of some bacteria for biological control of postharvest citrus green mould caused by Penicillium digitatum. Plant Protection Science 53: 134-143.
• Pane, C., Zaccardelli, M. (2015). Evaluation of Bacillus strains isolated from solanaceous phylloplane for biocontrol of Alternaria early blight of tomato. Biological Control 84: 11-18.
• Perez-Garcia, A., Romero, D., de Vicente, A. (2011). Plant protection and growth stimulation by microorganisms: biotechnological applications of Bacilli in agriculture. Current Opinion in Biotechnology 22 (2): 187-193.
• Soto, F., Tramon, C.P., Aqueveque, P.M., de Bruijn, J. (2018). Antagonist microorganisms that inhibit the development of post-harvest pathogens in lemons (Citrus limon L.). Chilean Journal of Agricultural and Animal Sciences 34 (2): 173-184.
• Soylu, S., Soylu, E.M., Kurt, Ş., Ekici, Ö.K. (2005). Antagonistic potentials of rhizosphere-associated bacterial isolates against soil-borne diseases of tomato and pepper caused by Sclerotinia sclerotiorum and Rhizoctonia solani. Pakistan Journal of Biological Sciences 8: 43-48.
• Soylu, E.M., Soylu, S., Kara, M., Kurt, Ş. (2020). Sebzelerde sorun olan önemli bitki fungal hastalık etmenlerine karşı vermikomposttan izole edilen mikrobiyomların in vitro antagonistik etkilerinin belirlenmesi. KSU Tarım ve Doğa Dergisi 23: 7-18.
• Stein, T. (2005). Bacillus subtilis antibiotics: structures, syntheses and specific functions. Molecular Microbiology 56 (4): 845-857.
• Sturz, A.V., Christie, B.R., Nowak, J. (2000). Bacterial endophytes: potential role in developing sustainable systems of crop production. Critical Reviews in Plant Sciences 19 (1): 1-30.
• Sülü, S.M., Bozkurt, İ.A., Soylu, S. (2016). Bitki büyüme düzenleyici ve biyolojik mücadele etmeni olarak bakteriyel endofitler. Mustafa Kemal Üniversitesi Ziraat Fakültesi Dergisi 21 (1): 103-111.
• Tan, S., Dong, Y., Liao, H., Huang, J., Song, S., Xu, Y., Shen, Q. (2012). Antagonistic bacterium Bacillus amyloliquefaciens induces resistance and controls the bacterial wilt of tomato. Pest Management Science 69: 1245-1252.
• Veliz, E.A., Martinez-Hidalgo, P., Hirsch, A.M. (2017). Chitinase-producing bacteria and their role in biocontrol. Aims Microbiology 3 (3): 689-705.
• Wang, Z.S., Sui, Y., Li, J.S., Tian, X.L., Wang, Q. (2020). Biological control of postharvest fungal decays in citrus: a review. Critical Reviews in Food Science and Nutrition: DOI: 10.1080/10408398.2020.1829542.
• Yoshida, S., Hiradate, S., Tsukamato, T., Hatakeda, K., Shirata, A. (2001). Antimicrobial activity of culture filtrate Bacillus amyloliquefaciens RC-2 isolated from mulberry leaves. Phytopathology 91: 181-187.