تاثیر عوامل باکتریایی آنتاگونیست جدا شده از باغات پسته روی قارچ آسپرژیلوس فلاووس

محورهای موضوعی : قارچ شناسی

سعیده دهقانپورفراشاه ¹ (استادیار گروه کشاورزی دانشگاه پیام نور، تهران، ایران.)
مهرداد صالح زاده ² (شیراز- دانشگاه شیراز- دانشکده کشاورزی)

تاریخ دریافت : 1401/12/08 تاریخ پذیرش : 1402/06/10 تاریخ انتشار : 1402/06/15

کلید واژه: آفلاتوکسین, باسیلوس, آنتی بیوز, متابولیت, سیدروفور,

چکیده مقاله :

سابقه و هدف: آفلاتوکسین در پسته و انواع مواد غذایی سبب بروز مشکلات و بیماری در افراد می شود. گونه آسپرژیلوس فلاووس بهعنوان تولید کننده مهم آفلاتوکسین گزارش شده است. در این پژوهش عوامل باکتریایی آنتاگونیست از باغات پسته استان یزد جدا‌سازی و میزان خاصیت آنتاگونیستی آن ها روی قارچ آسپرژیلوس فلاووس بررسی شد. مواد و روش ها: جدایه های مختلف باسیلوس و آسپرژیلوس از باغات پسته استان یزد جداسازی و گونه های آسپرژیلوس فلاووس شناسایی شدند. خصوصیات مختلف بیوشیمیایی و مولکولی جدایه های باکتریایی مورد بررسی قرار گرفت. همچنین خاصیت آنتی‌ بیوزی و تولید آنتی بیوتیک و تاثیر عصاره خارجسلولی، مواد فرار ضدقارچی و سیدروفور جدایه ها روی آسپرژیلوس فلاووس ارزیابی شد. یافته ها: بر اساس نتایج آزمون های بیوشیمیایی و مولکولی، همه جدایه ها از جنس باسیلوس تشخیص داده شدند. سویه Pr 3 با 95 درصد بیشترین تأثیر و سویه‌ های Pr 5 وPr 7 با 50 درصد کمترین تأثیر را روی آسپرژیلوس فلاووس داشتند. نتایج حاصل از آزمون تولید آنتی‌ بیوتیک بهروش کلروفرم نشان داد که جدایه هایPr3 وPr7 بهترتیب با 84/35 درصد و 38 درصد بیشترین و کمترین میزان تولید آنتی بیوتیک را داشتند. در این پژوهش، جدایه‌ های Pr1، Pr2، Pr3 و Pr9 بهصورت معناداری مایع خارجسلولی، سیدروفور و مواد فرار ضد قارچی تولید کردند و سطوح متفاوتی از تأثیر را روی آسپرژیلوس فلاووس نشان دادند. نتیجه گیری: با توجه به نتایج به دست آمده از این پژوهش و یافته های سایر محققان، گونه های مختلف باسیلوس و متابولیت های آن‌ ها دارای خاصیت آنتاگونیستی بالایی روی قارچ آسپرژیلوس فلاووس پسته هستند که میتوانند به عنوان روشی با آینده روشن و موثر برای بازدارندگی رشد این قارچ و جلوگیری از تولید آفلاتوکسین بهکار روند.

چکیده انگلیسی:

Background &Objectives: Aflatoxin in pistachios and some other foods and feeeds causes diseases in human and animals. Aspergillus flavus has been reported as an important producer of aflatoxin. In this research, antagonistic bacterial agents were isolated from pistachio orchards in Yazd province, Iran, and their antagonistic properties were investigated on A. flavus isolates. Materials & Methods: Different isolates of Bacillus sp. and Aspergillus sp. were isolated from different pistachio orchards in Yazd. A. flavus species were identified and various biochemical and molecular characteristics of bacterial isolates were investigated. Also, the antibiosis properties and production of antibiotics and effect of extracellular secretion, antifungal volatile substances and siderophore of the isolates were evaluated on A. flavus. Results: According to the results of biochemical and molecular tests, all isolates were identified as Bacillus. Strain Pr3 had the most effect with 95% and strains Pr5 and Pr7 had the least effect on A. flavus with 50%. The results of the antibiotic production test using chloroform method showed that isolates Pr3 and Pr7 had the highest and lowest antibiotic production with 84.35 and 38%, respectively. In this study, isolates Pr1, Pr2, Pr3 and Pr9 significantly produced extracellular     secretion, siderophore and antifungal volatile substances and showed different amounts of effect on A. flavus. Conclusion: According to the results obtained from this research and the findings of other related researches, different Bacillus species and their metabolites have high antagonistic properties against A. flavus, which can be used as an effective and promising method to inhibit the growth of the fungus and to prevent the production of aflatoxin.  

منابع و مأخذ:

References

1. Bagheri Marzooni B, Jahed Khaniki G, Shariatifar N, Molaee-Aghaee E. Assessment of aflatoxins in some pistachio cultivars supplied in bulk in Tehran food stores. Int J Environ Anal Chem. 2022; 17:1-10.


2. Cho HJ, Son SH, Chen W, Son YE, Lee I, Yu JH, Park HS. Regulation of Conidiogenesis in Aspergillus flavus. Cells. 2022;11(18):2796.


3. Sayadi M, Tajik H. Detoxification of aflatoxin b1 by Lactobacillus rhamnosus in a simulated model of the human digestive system. JMW. 2019; 11(4): 380-391.


4. Mojtahedi H, Danesh D, Haghighi B, Barnett R. Postharvest pathology and mycotoxin contamination of Iranian pistachio nuts. Phytopathology. 1978; 68(12): 1800-1804.


5. Younesi S, Salehzadeh M, Soleymani Pari MJ. Control of strawberry gray mold fungus with combined application of different species of Trichoderma and salicylic acid. JMW. 2023;16(1): 72-88.


6. Moyne AL, Shelby R, Cleveland TE, Tuzun SA. Bacillomycin D: an iturin with antifungal activity against Aspergillus flavus. J appl microbiol. 2001; 90(4): 622-629.


7. Pakizeh M, Nouri L, Azizi MH. Probiotic-Based Optimization of Pistachio Paste Production and Detoxification of Aflatoxin B1 Using Bifidobacterium lactis. J Food Qual. 2022: 1-31.


8. Koumoutsi A, Chen XH, Henne A, Liesegang H, Hitzeroth G, Franke P, Vater J, Borriss R. Structural and functional characterization of gene clusters directing nonribosomal synthesis of bioactive cyclic lipopeptides in Bacillus amyloliquefaciens strain FZB42. J Bacteriol. 2004; 186(4): 1084-1096.


9. Alberts JF, Engelbrecht Y, Steyn PS, Holzapfel WH, Van Zyl WH. Biological degradation of aflatoxin B1 by Rhodococcus erythropolis cultures. Int j food microbiol. 2006; 109(1-2): 121-126.


10. Zhu Y, Hassan YI, Lepp D, Shao S, Zhou T. Strategies and methodologies for developing microbial detoxification systems to mitigate mycotoxins. Toxins. 2017; 9(4): 130.


11. Sommartya, T . 1997. Peanut disease. Bangkok: kasetsart University publishing.


12. Liang-bin HU, Fei WA, Luo-gen CH, Zhi-qi SH. Identification of B-FS06 and the antagonistic activity of its cultural productions against Aspergillus flavus. Chin J Biol Control. 2007; 23(2): 160.


13. ONo M, KIMURA N. Antifungal peptides produced by Bacillus subtilis for the biological control of aflatoxin contamination. JSM Mycotoxins. 1991; 1991(34): 23-28.


14. 14. Choudhary AK. Influence of microbial co‐inhabitants on aflatoxin synthesis of Aspergillus flavus on maize kernels. Lett appl microbiol. 1992; 14(4): 143-147.


15. Cotty PJ. Virulence and cultural characteristics of two Aspergillus flavus strains pathogenic on cotton. Phytopathology. 1989; 79(7):808-814.


16. Diener UL, Davis ND. Aflatoxin production by isolates of Aspcrgillus flavus. Phytopathology. 1966;56(12):1390-3.


17. Dorner JW, Cole RJ, Connick WJ, Daigle DJ, McGuire MR, Shasha BS. Evaluation of biological control formulations to reduce aflatoxin contamination in peanuts. Biol Control. 2003; 26(3): 318-324.


18. McClenny N. Laboratory detection and identification of Aspergillus species by microscopic observation and culture: the traditional approach. Med mycol J. 2005; 43(Supplement_1): S125-128.


19. Samson RA, Visagie CM, Houbraken J, Hong SB, Hubka V, Klaassen CH, Perrone G, Seifert KA, Susca A, Tanney JB, Varga J. Phylogeny, identification and nomenclature of the genus Aspergillus. Stud mycol. 2005; 53(1): 1-27.


20. Palu AP, Gomes LM, Miguel MA, Balassiano IT, Queiroz ML, Freitas-Almeida AC, de Oliveira SS. Antimicrobial resistance in food and clinical Aeromonas isolates. Food microbiol. 2006; 23(5): 504-509.


21. Kiu R, Caim S, Alcon-Giner C, Belteki G, Clarke P, Pickard D, Dougan G, Hall LJ. Preterm infant-associated Clostridium tertium, Clostridium cadaveris, and Clostridium paraputrificum strains: genomic and evolutionary insights. Genome Biol Evol. 2017; 9(10): 2707-2714.


22. Weller DM, Cook RJ. Suppression of take-all of wheat by seed treatment with Pseudomonas fluorescent. Phytopathology. 1983; 73: 463-469.


23. Singh V, Deverall BJ. Bacillus subtilis as a control agent against fungal pathogens of citrus fruit. Trans Br Mycolo Soc. 1984; 83(3): 487-490.


24. Fiddaman PJ, Rossall S. The production of antifungal volatiles by Bacillus subtilis. J Appl Bacteriol. 1993; 74(2): 119-126.


25. Geels FP, Schippers B. Selection of antagonistic fluorescent Pseudomonas spp. and their root colonization and persistence following treatment of seed potatoes. J Phytopathol. 1983; 108(3‐4): 193-206.


26. Emmert EA, Handelsman J. Biocontrol of plant disease: a (Gram-) positive perspective. FEMS Microbiol lett. 1999; 171(1): 1-9.


27. Broadbent P, Baker KF, Waterworth Y. Bacteria and actinomycetes antogonistic to fungal root pathogens in Australian soils. Aust J Biol Sci. 1971; 24(4): 925-944.


28. Munimbazi C, Bullerman LB. Inhibition of aflatoxin production of Aspergillus parasiticus NRRL 2999 by Bacillus pumilus. Mycopathologia. 1997; 140(3): 163-169.


29. Bottone EJ, Peluso RW. Production by Bacillus pumilus (MSH) of an antifungal compound that is active against Mucoraceae and Aspergillus species: preliminary report. J Med Microbiol. 2003; 52(1): 69-74.


30. Chang PK, Hua SS. Nonaflatoxigenic Aspergillus flavus TX9-8 competitively prevents aflatoxin accumulation by A. flavus isolates of large and small sclerotial morphotypes. Int J Food Microbiol. 2007; 114(3): 275-279.


31. Reddy KR, Reddy CS, Muralidharan K. Potential of botanicals and biocontrol agents on growth and aflatoxin production by Aspergillus flavus infecting rice grains. Food control. 2009; 20(2): 173-178.


32. Farzaneh M, Ahmadzadeh M, Ghasempour A, Mir-Abolfathi M, Javan Nikkhah M, Sharifi-Tehrani A. Mode of Action of Bacillus subtilis on Aflatoxin Control of Aspergillus flavus. Iran J Plant Prot Sci. 2011; 42(2): 191-198.


33. Farzaneh M, Shi ZQ, Ghassempour A, Sedaghat N, Ahmadzadeh M, Mirabolfathy M, Javan-Nikkhah M. Aflatoxin B1 degradation by Bacillus subtilis UTBSP1 isolated from        pistachio nuts of Iran. Food control. 2012; 23(1): 100-106.


34. Chen G, Liao Z, Xu C, Liang Z, Liu T, Zhong Q, Wang L, Fang X, Wang J. Detoxification of Aflatoxin B1 by a Potential Probiotic Bacillus amyloliquefaciens WF2020. Front microbiol. 2022; 13.


35. Mohammadi A, Akhavan SA, Hosseinidoost R. Evaluation of antifungal activity of iturin producing Bacillus subtilis strains. JMW. 2016; 8(4): 299-307.


36. Ashrafi A, Salehzadeh M, Khezrinezhad N. Detection and identification of tomato wilt disease in East Azerbaijan province and controlling it using antagonist bacteria. Genetic Engineering and Biosafety Journal. 2020; 10 9(1): 28-39.