Limon’da (Citrus limon (L.) Burm. f.) Hasat Sonrası Hastalıklara Karşı Antifungal Aktiviteye Sahip Antagonist Bakterilerin Taranması

Öz

Bu çalışma, limon meyvesinde hasat sonrası kayıplara neden olan farklı fungal etmenlere karşı biyolojik mücadele etmeni olabilecek bakterileri belirlemek amacıyla in vitro koşullarda yürütülmüştür. Bu amaçla, hastalıklı limon meyvelerinden izolasyonlar yapılmış, izolasyonlar sonucu üç farklı cinse ait fungus türü izole edilmiş, patojenite testleri yapılmış ve moleküler tanı sonuçlarına göre bu izolatların Alternaria alternata (ET 119), Colletotrichum gleosporoides (ET 120) ve Penicillium chrysogenum (ET 121) olduğu tespit edilmiştir. Patojen fungus izolatlarına karşı sekiz farklı cinse ait otuz altı adet antagonist bakteri izolatının yüzde engelleme oranları ikili kültür testi ile tesadüf parselleri deneme desenine göre 3 tekerrürlü olarak belirlenmiştir. Elde edilen sonuçlara göre üç patojen izolata karşı en yüksek etki TV 53D (%67.46: Brevibacillus choshinensis) izolatında saptanırken, bu izolatı sırasıyla TV 16F (%66.27: Bacillus subtilis) ve FDG 37 (%62.30: Pseudomonas fluorescens) izolatları takip etmiştir. En etkili sonuç alınan antagonist bakteri izolatları ile gelecekte kontrollü şartlarda daha detaylı çalışmaların yürütülmesi gerekmektedir.

Anahtar Kelimeler

• Antifungal aktivite
• Bacillus subtilis
• antagonist bakteri
• biyolojik mücadele
• Pseudomonas fluorescens

Screening of Antagonist Bacteria with Antifungal Activity Against Postharvest Diseases on Lemon (Citrus limon (L.) Burm.f.)

Öz

This study was carried out in vitro conditions to determine the bacteria biological control agents of against different fungal agents causing postharvest losses in lemon fruit. For this purpose, isolations were made from diseased lemon fruits, as a result of isolations three different fungal species were isolated, pathogenicity tests were carried out and according to the results of molecular diagnosis, these isolates were found to be Alternaria alternata (ET 119), Colletotrichum gleosporoides (ET 120) and Penicillium chrysogenum (ET 121). Percent inhibition rate of thirty-six antagonist bacterial isolates belonging to eight different genera against pathogenic fungus isolates was determined by dual culture test according to a randomized plot experiment design with 3 replications. According to the results obtained, the highest effect against the three pathogen isolates was detected in TV 53D (67.46%: Brevibacillus choshinensis), this isolate was followed by TV 16F (66.27%: Bacillus subtilis) and FDG 37 (62.30%: Pseudomonas fluorescens) isolates, respectively. More detailed studies should be carried out in the future under controlled conditions with the most effective antagonist bacterial isolates.

Anahtar Kelimeler

• Bacillus subtilis
• Pseudomonas fluorescens
• Antifungal activity
• antagonist bacteria
• biological control

Kaynakça

1. Ab Rahman, S. F. S., Singh, E., Pieterse, C. M., & Schenk, P. M. (2018). Emerging microbial biocontrol strategies for plant pathogens. Plant Science, 267, 102-111.
2. Akgün, C. (2006). Turunçgiller sektör profili. Dış Ticaret Servisi Uygulama Şubesi, Türkiye. Erişim adresi http:///20684676Turuncgillersektorprofili.html.
3. Akimitsu, K., Peever, T. L., & Timmer, L. W. (2003). Molecular, ecological and evolutionary approaches to understanding Alternaria diseases of citrus. Molecular Plant Pathology, 4(6), 435-446.
4. Aktaş, S. (2015). Domates öz nekrozuna neden olan etmenlere karşı PGPR ve biyoajan bakterileri kullanılarak kontrollü koşullarda biyolojik mücadele imkânlarının araştırılması. Yüksek Lisans Tezi, Atatürk Üniversitesi Fen Bilimleri Enstitüsü.
5. Anonim, (2020). Tarım ve Orman Bakanlığı, Limon. Erişim adresi https://arastirma.tarimorman.gov.tr/tepge.
6. Barkai-Golan, R. (2001). Postharvest diseases of fruits and vegetables: Development and control. Elsevier, Amsterdam, Netherlands.
7. Benli, M. (2003). Hasat sonrası fungal hastalıklarla kimyasal ve biyolojik mücadele. Orlab On Line Mikrobiyoloji Dergisi, 01(08), 1-25.
8. Bonaterra, A., Badosa, E., Cabrefiga, J., Francés, J., & Montesinos, E. (2012). Prospects and limitations of microbial pesticides for control of bacterial and fungal pomefruit tree diseases. Trees, 26(1), 215-226.

9. Brown, G. E., & Eckert, J. W. (2000). Postharvest Fungal Diseases, in: Timmer LW, Garnsey SM, Graham JH (eds.), Compendium of Citrus Diseases (2nd) (pp. 37-45). APS Press, Saint Paul, Minnesota.
10. Chandel, S., Allan, E. J., & Woodward, S. (2010). Biological control of Fusarium oxysporum f. sp. lycopersici on tomato by Brevibacillus brevis. Journal of phytopathology, 158(7‐8), 470-478.
11. Couillerot, O., Prigent-Combaret, C., Caballero-Mellado, J., & Moënne-Loccoz, Y. (2009). Pseudomonas fluorescens and closely-related fluorescent Pseudomonads as biocontrol agents of soil-borne phytopathogens. Letters in Applied Microbiology, 48(5), 505-512.
12. Dadaşoğlu, F., & Şahin, F. (2010). Bakterilerin yüzük kelebeği Malacosoma neustria L. (Lepidoptera: Lasiocampidae)’nın biyolojik mücadelesinde kullanımı. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 41(2), 97-104.
13. Droby, S., Wisniewski, M., Teixidó, N., Spadaro, D., & Jijakli, M. H. (2016). The science, development, and commercialization of postharvest biocontrol products. Postharvest Biology and Technology, 122, 22-29.
14. Dukare, A. S., Prasanna, R., Dubey, S. C., Chaudhary, V., Nain, L., Singh, R., & Saxena, A. K. (2011). Evaluating novel microbe amended composts as biocontrol agents in tomato. Crop Protection, 30, 436-442.
15. Dukare, A. S., Paul, S., Nambi, V. E., Gupta, R. K., Singh, R., Sharma, K. & Vishwakarma, R. K. (2019). Exploitation of microbial antagonists for the control of postharvest diseases of fruits: A review. Critical Reviews in Food Science and Nutrition, 59(9), 1498-513.
16. Eckert, J. (1989). Recent developments in the chemical control of postharvest diseases. Sympos Tropical Fruit Int. Trade, 269, 477-494.
17. Eckert, J. W., & Eaks, I. L. (1989). Postharvest disorders and diseases of citrus fruit, in: Reuter W, Calavan EC, Carman GE, Berkeley CA (Eds.), The Citrus Industry, (pp. 179-260). University of California Press, USA.
18. Edwards S. G., McKay, T., & Seddon, B. (1994). Interaction of Bacillus species with phytopathogenic fungi. Methods of analysis and manipulation for biocontrol purposes. in: Blakeman JP, Williamson B (eds) Ecology of Plant Pathogens (pp. 101-118). Wallingford, Great Britain, British Society for Plant Pathology.
19. Edwards, S. G., & Seddon, B. (2001). Mode of antagonism of Brevibacillus brevis against Botrytis cinerea in vitro. Journal of Applied Microbiology, 91, 652-659.
20. Ekinci, M., Turan, M., Yıldırım, E., Güneş, A., Kotan, R., & Dursun A. (2014). Effect of plant growth promoting rhizobacteria on growth, nutrient, organic acid, amino acid and hormone content of cauliflower (Brassica oleracea L. var. botrytis) transplants. Acta Scientiarum Polonorum, 13(6), 71-85.
21. Ekinci, M., Yıldırım, E., & Kotan, R. (2015). Effects of different plant growth promoting rhizobacteria on growth and quality of broccoli (Brassica oleraceae L. var. italica) seedling. Akdeniz University Journal of Agriculture, 28(2), 53-59.
22. El-Mabrok, A., Hassan, Mokhtar, A. M., Hussain, K., & Aween, M. (2012). Isolation and identification of lactic acid bacteria with antifungal activity against anthracnose disease. Research Journal of Biological Sciences, 7(9-12), 335-339.
23. Elshafie, H., Camele, I., Racioppi, R., Scrano, L., Iacobellis, N., & Bufo, S. (2012). In vitro antifungal activity of Burkholderia gladioli pv. agaricicola against some phytopathogenic fungi. International Journal of Molecular Sciences, 13, 16291-16302.
24. Elsherif, M., & Grossmann, F. (1994). Comparative investigations on the antagonistic activity of fluorescent pseudomonads against Gaeumannomyces graminis var. tritici in vitro and in vivo. Microbiological Research, 149(4), 371-377.
25. Erdoğan, O., & Benlioğlu, K. (2010). Biological control of Verticillium wilt on cotton by the use of fluorescent Pseudomonas spp. Biological Control, 53(1), 39-45.
26. Erman, M., Kotan, R., Çakmakçı, R., Çığ, F., Karagöz, K., & Sezen, M. (2010). Effect of nitrogen fixing and phosphatesolubilizing rhizobacteria isolated from Van Lake Basin on the growth and quality properties in wheat and sugar beet. Turkey IV. Organic Farming Symposium, 28 June - 1 July 2010, Erzurum, Turkey, 325-329.
27. Forner, C., Bettiol, W., Nascimento, L. M. D., & Terao, D. (2013). Postharvest control of Penicillium digitatum in pera orange trees with microorganisms and heat treatment. Revista Brasileira de Fruticultura, 35(1), 23-31.
28. Gökçe, A. Y., & Kotan, R. (2016). Buğday kök çürüklüğüne neden olan Bipolaris sorokiniana (Sacc.)’ya karşı PGPR ve antagonistbakterileri kullanılarak kontrollü koşullarda biyolojik mücadele imkânlarının araştırılması. Bitki Koruma Bülteni, 56(1), 49-75.
29. Güneş, A., Karagöz, K., Turan, M., Kotan, R., Yıldırım, E., Çakmakçı, R., & Şahin, F. (2015). Fertilizer effiency of some plant growth promoting rhizobacteria for plant growth. Research Journal of Soil Biology, 7(2), 28-45.
30. Heimpel, G. E., & Mills, N. (2017). Biological Control-Ecology and Applications. Cambridge University Press, Cambridge, England.
31. Ismail, M., & Zhang, J. (2004). Post-harvest citrus diseases and their control. Outlooks on Pest Management, 15, 29.
32. Janisiewicz, W. J., & Korsten, L. (2002). Biological control of postharvest diseases and fruits. Annual Review Phytopathology, 40, 411-441.
33. Karabulut, O. A., Tezcan, H., Daus, A., Cohen, L., Wiess, B., & Droby, S. (2004). Control of preharvest and postharvest fruit rot in strawberry by Metschnikowia fructicola. Biocontrol Science and Technology, 14, 513-521.
34. Karagöz, K., & Kotan, R. (2010). Bitki gelişimini teşvik eden bazı bakterilerin marulun gelişimi ve bakteriyel yaprak lekesi hastalığı üzerine etkileri. Türkiye Biyolojik Mücadele Dergisi, 1(2), 165-179.
35. Karahocagil, P., Tunalıoglu, R., Taskaya, B., & Anac, H. (2003).Turunçgiller Durum ve Tahmin: 2003/2004. Tarımsal Ekonomi Araştırma Enstitüsü, 111, 74.
36. Karakurt, H., Kotan, R., Dadaşoğlu, F., Aslantaş, R., & Sahin F. (2011). Effects of plant growth promoting rhizobacteria on fruit set pomological and chemical characteristics color values and vegetative growth of sour cherry Prunus cerasus cv. Kutahya. Turkish Journal of Biology, 35, 283-291.
37. Ketabchi, S., Taghipour, M., & Sharzei, A. (2012). Identification of lime fruit surface colonizing bacteria antagonistic against the green mold (Penicillium digitatum) and comparison of biological control with heat treatment and chemical control. Asian Journal of Experimental Biological Sciences, 3, 287-292.
38. Kinay, P., Mansour, M. F., Gabler, F. M., Margosan, D. A., & Smilanick, J. L. (2007). Characterization of fungicideresistant isolates of Penicillium digitatum collected in California. Crop Protection, 26, 647-656.
39. Kotan, R., Dikbaş, N., & Bostan, H. (2009). Biological control of postharvest disease caused by Aspergillus flavus on stored lemon fruits. African Journal of Biotechnology, 8(2), 209-214.
40. Kotan, R., Şahin, F., Demirci, E., & Eken, C. (2011). Biological control of the potato tubers dry rot caused by Fusarium species using PGPR strains. Biological Control, 59(3), 194-198.
41. Köhl, J., Kolnaar, R., & Ravensberg, W. J. (2019). Mode of action of microbial biological control agents against plant diseases: relevance beyond efficacy. Frontiers in Plant Science, 10, 845. https://doi.org/10.3389/fpls.2019.00845.
42. Kumar, P., Dubey, R. C., & Maheshwari, D. K. (2012). Bacillus strains isolated from rhizosphere showed plant growth promoting and antagonistic activity against phytopathogens. Microbiological Research, 167, 493-499.
43. Leelasuphakul, W., Sivanunsakul, P., & Phongpaichit, S. (2006). Purification, characterization and synergistic activity of β-1, 3-glucanase and antibiotic extract from an antagonistic Bacillus subtilis NSRS 89-24 against rice blast and sheath blight. Enzyme and Microbial Technology, 38(7), 990-997.
44. Liu, J., Wisniewski, M., Droby, S., Tian, S., Hershkovitz, V., & Tworkoski, T. (2011). Effect of heat shock treatment on stress tolerance and biocontrol efficacy of Metschnikowia fructicola. FEMS Microbiology and Ecology, 76, 145-155.
45. Liu, Y., Heying, E., & Tanumihardjo, S. A. (2012). History, global distribution, and nutritional importance of citrus fruits. Comprehensive Reviews in Food Science and Food Safety, 11(6), 530-538. doi: 10.1111/j.1541-4337.2012.00201.x.
46. Liu, J., Sui, Y., Wisniewski, M., Droby, S., & Liu, Y. (2013). Utilization of antagonistic yeasts to manage postharvest fungal diseases of fruit. International Journal of Food Microbiology, 167, 153-160.
47. Mohammadi, P. (2018). Domates bakteriyel solgunluk ve kanser hastalığı etmeni (Clavibacter michiganensis subsp. michiganensis (Smith) Davis vd.)’nin antagonistbakteriler kullanılarak mücadele imkânlarının araştırılması. Doktora Tezi, Atatürk Üniversitesi Fen Bilimleri Enstitüsü.
48. Mohammadi, P., Tozlu, E., & Kotan, R. (2014). Screening of antagonistic bacteria for biological control of green mold caused by Penicillium digitatum Sacc. of citrus fruits. 21th Iranian Plant Protection Congress, January 21-22, Urmia, Iran, 189.
49. Mohammadi, P., Tozlu, E., Kotan, R., & Senol Kotan, M. (2017). Potential of some bacteria for biological control of postharvest citrus green mould caused by Penicillium digitatum. Plant Protection Science, 53(3), 134-143.
50. Morales-Cedeño, L. R., del Carmen Orozco-Mosqueda, M., Loeza-Lara, P. D., Parra-Cota, F. I., de Los Santos-Villalobos, S., & Santoyo, G. (2021). Plant growth-promoting bacterial endophytes as biocontrol agents of pre-and post-harvest diseases: Fundamentals, methods of application and future perspectives. Microbiological Research, 242, 126612.
51. Ongena, M., & Jacques, P. (2008). Bacillus lipopeptides: versatile weapons for plant disease biocontrol. Trends in Microbiology, 16(3), 115-125.
52. Özaktan, H., Aysan, Y., Yıldız, F., & Kınay, P. (2010). Fitopatolojide biyolojik mücadele. Türkiye Biyolojik Mücadele Dergisi, 1(1), 61-78.
53. Palou, L., Smilanick, J. L., & Droby, S. (2008). Alternatives to conventional fungicides for the control of citrus postharvest green and blue moulds. Stewart Postharvest Review, 2, 1-16.
54. Palou, L. (2014). Penicillium digitatum, Penicillium italicum ( green mold, blue mold), in Bautista-Baños S. (ed.), Postharvest Decay (pp. 45–102). Academic Press, London, England.
55. Panda, A. K., Bisht, S. S., DeMondal, S., Senthil Kumar, N., Gurusubramanian, G., & Panigrahi, A. K. (2014). Brevibacillus as a biological tool: a short review. Antonie Van Leeuwenhoek, 105(4), 623-639.
56. Papoutsis, K., Mathioudakis, M. M., Hasperue, J. H., & Ziogas, V. (2019). Non-chemical treatments for preventing the postharvest fungal rotting of citrus caused by Penicillium digitatum (green mold) and Penicillium italicum (blue mold). Trends in Food Science and Technology, 86, 479-491.
57. Romanazzi, G., Feliziani, E., Baños, S. B., & Sivakumar, D. (2017). Shelf life extension of fresh fruit and vegetables by chitosan treatment. Critical reviews in food science and nutrition, 57(3), 579-601.
58. Ruffo Roberto, S., Youssef, K., Hashim, A. F., & Ippolito, A. (2019). Nanomaterials as alternative control means against postharvest diseases in fruit crops. Nanomaterials, 9(12), 1752.
59. Schirra, M., D’Aquino, S., Cabras, P., & Angioni, A. (2011). Control of postharvest diseases of fruit by heat and fungicides: efficacy, residue levels, and residue persistence. A review. Journal of Agricultural and Food Chemistry, 59(16), 8531-8542.
60. Schmitt, A., & Seddon, B. (2005). Biocontrol of plant pathogens with microbial BCAs and plant extracts advantages and disadvantages of single and combined use, in: Dehne HW, Gisi U, Kuck KH, Russell PE, Lyr H (eds), Modern Fungicides and Antifungal Compounds (pp. 205-225). Alton, England.
61. Seddon, B., McHugh, R. C., & Schmitt, A. (2000). Brevibacillus brevis– a novel candidate biocontrol agent with broad-spectrum antifungal activity. Proc Brit Crop Protec Con - Pests and Diseases, 563-572.
62. Senthilkumar, M., Swarnlakshmi, K., Govindasamy, V., Lee, Y. K., & Annapurna, K. (2009). Biocontrol potential of soybean bacterial endophytes against charcoal rot fungus Rhizoctonia bataticola. Current Microbiology, 58, 288-293.
63. Smilanick, J. L., Brown, G. E., Eckert, J. W. (2006). The biology and control of postharvest diseases, in: Wardowski WF, Miller WM, Hall DJ, Grierson W (Eds.), Fresh Citrus Fruits (2nd) (pp. 339-396). Florida Science Source, Inc.
64. Sunita, C., Eunice, J. A., & Steve, W. (2010). Biological control of Fusarium oxysporum f.sp. lycopersici on tomato by Brevibacillus brevis. Journal of Phytopathology, 158, 470-478.
65. Şahinoglu, E. (2019). Kapya biberde Fusarium proliferatum (Matsush.) Nirenberg ex Gerlach & Nirenberg, (1976) ile biyolojik mücadele imkânlarının araştırılması. Yüksek Lisans Tezi, Atatürk Üniversitesi Fen Bilimleri Enstitüsü.
66. Tekiner, N., Tozlu, E., & Kotan, R. (2019a). Domateste Alternaria alternata (Fr.) Keissl’nın bazı bakteriler ile biyolojik mücadelesi. Plant Protection Bulletin, 59(4), 57-68.
67. Tekiner, N., Kotan, R., Tozlu, E., & Dadasoglu, F. (2019b). Determination of some biological control agents against Alternaria fruit rot in quince. Alınteri Journal of Agricultural Sciences, 34(1), 25-31.
68. Tekiner, N., Tozlu, E., & Kotan, R. (2019c). Portakalda antraknoz hastalığı etmeni Colletotrichum gloeosporioides’in biyolojik mücadele imkânlarının araştırılması. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 50(3), 282-291.
69. Tekiner, N., Tozlu, E., & Guarnaccia, V. (2020). First report of Diaporthe foeniculina causing fruit rot of lemon in Turkey. Journal of Plant Pathology, 102, 277. https://doi.org/10.1007/s42161-019-00413-4.
70. Tozlu, E. (2016). Biological Control of Carrot Sour Rot (Geotrichum candidum Link) by some bacterial biocontrol agents. Atatürk University Journal of Agricultural Faculty, 47(1), 1-9.
71. Tozlu, E., Dadasoglu, F., Kotan, R., & Tozlu, G. (2011). Insecticidal effect of some bacteria on Bruchus dentipes Baudi (Coleoptera: Bruchidae). Fresenius Environmental Bulletein, 20(4), 918-923. Tozlu, E., Tekiner, N., Kotan, R., & Örtücü, S. (2018). Investigation on the biological control of Alternaria alternata. Indian Journal of Agricultural Sciences, 88(8), 1241-1247.
72. Wang, H., Yan, Y., Wang, J., Zhang, H., & Qi, W. (2012). Production and characterization of antifungal compounds produced by Lactobacillus plantarum IMAU10014. Plos One, 7(1), e29452.
73. Wang, X. Q., Zhao, D. L., Shen, L. L., Jing, C. L., & Zhang, C. S. (2018). Application and mechanisms of Bacillus subtilis in biological control of plant disease, in: Role of Rhizospheric Microbes in Soil (pp. 225-250). Springer, Singapore.
74. White, T. J., Brauns, T., Leeand, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, in: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds), PCR Protocols: A Guide to Methods and Applications (pp. 315-322). Academic Press, San Diego, California.
75. Zhen, S., Kaiqi, L., Changxu, L., Jian, Y., Ruicheng, J., & Xunli, L. (2011). Isolation and characterization of a potential biocontrol Brevibacillus laterosporus. African Journal of Microbiology Research, 5(18), 2675-2681.