اثرات اسانس های هل، سقز و میخک بر کنترل و بیان برخی از ژن‌های مایکوتوکسین داکسی‌نیوالنول فوزاریوم گرامینه‌آروم
محورهای موضوعی : میکروب شناسی پزشکی
1 - استادیار، گروه زراعت دانشکده کشاورزی، دانشگاه آزاد اسلامی واحد زاهدان، زاهدان، ایران
2 - مربی، پژوهشکده کشاورزی، دانشگاه زابل، زابل، ایران
کلید واژه: گیاهان دارویی, فعالیت ضد‌قارچی, داکسی‌نیوالنول, فوزاریوم گرامینه‌آروم,
چکیده مقاله :
سابقه و هدف: قارچ فوزاریوم گرامینه‌آروم عامل بیماری اسکب در گندم و سایر غلات دانه ریز است. زیرالنون (Zearalenone) و داکسی‌نیوالنول (Deoxynivalenol) مهمترین مایکوتوکسین های این قارچ هستند. این پژوهش با هدف ارزیابی تاثیر اسانس‌های هل، سقز و میخک بر مهار رشد فوزاریوم گرامینه‌آروم و بیان برخی از ژن‌های موثر در زیست‌ساخت داکسی‌یوالنول انجام شد.مواد و روش‌ها: پس از کشت فوزاریوم گرامینه‌آروم در محیط Potato Dextrose Broth، حداقل غلظت بازدارندگی (MIC) رشد قارچ تحت اثر اسانس‌ها با استفاده از روش میکروتیتر پلیت اندازه‌گیری شد. همچنین بیان ژن‌های TRI5، TRI6 و TRI14 با تکنیک Real-time PCR ارزیابی شد.یافته‌ها: اسانس هل کمترین مقدار MIC (100 میکرولیتر بر میلی‌لیتر) و اسانس‌های سقز و میخک بیشترین مقدار MIC 200 میکرولیتر بر میلی‌لیتر) را داشتند. اسانس هل با کمترین مقدار غلظت قارچ‌کشی (MFC) (200 میکرولیتر بر میلی‌لیتر) بالاترین خاصیت قارچ‌کشی روی فوزاریوم گرامینه‌آروم را داشت و اسانس‌های سقز و میخک بیشترین مقدار MFC (400 میکرولیتر بر میلی‌لیتر) را داشتند. میزان بیان ژن‌های TRI5، TRI6 و TRI14 به طور معنی‌داری توسط اسانس هل کاهش یافت.نتیجه گیری: نتایج این تحقیق نشان داد که اسانس هل دارای خاصیت قارچ‌کشی و مهارکنندگی بر رشد قارچ فوزاریوم گرامینه‌آروم است و موجب کاهش میزان بیان ژن‌های TRI5، TRI6 و TRI14 موثر در تولید داکسی‌نیوالنول می گردند.
Background & Objectives: Fusarium graminearum is the causal agent of scab disease in wheat and other small grains. Zearalenone and deoxynivalenol are the main toxins produced by F. graminearum. In this study, the effects of essential oils of Elettaria cardamomum, Pistacia atlantica, and Eugenia caryophillata on F. graminearum growth inhibition and the expression of some genes in deoxynivalenol biosynthetic pathway were investigated. Materials & Methods: Minimal inhibitory concentration (MIC) of the fungal growth was measured through the microtiter plate method after growing F. graminearum on Potato Dextrose Broth. In addition, the expression of TRI5, TRI6, and TRI14 genes were evaluated using real-time PCR (qRT-PCR) technique. Results: Elettaria cardamomum essential oil had the lowest MIC (100 µl/ml) and the essential oils of P. atlantica and E. caryophillata had the highest MIC (200 µl/ml). Elettaria cardamomum essential oil had the lowest MFC (200 µl/ml) and the highest fungicidal property against F. graminearum, and the essential oils of P. atlantica and E. caryophillata had the highest MFC value (400 µl/ml). The expression of TRI5, TRI6, and TRI14 genes was significantly decreased by E. cardamomum essential oil. Conclusion: The results of the study showed that the E. cardamomum essential oil has fungicidal and inhibitory effects against F. graminearum and leads to reduce the expression of TRI5, TRI6, and TRI14 genes relating to deoxynivalenol production.
devastating impact. Plant Dis. 1997; 81(12): 1340e1348.
2. Bottalico A, Perrone G. Toxigenic Fusarium species and mycotoxins associated with head
blight in smallcereals in Europe. Eur J Plant Pathol. 2002; 108(7): 611-624.
3. Luo Y, Yoshizawa T, Katayama T. Comparative study on the natural occurrence of Fusarium
mycotoxins (trichothecenes and zearalenone) in corn and wheat from high- and low-risk areas
for human esophageal cancer in China. Appl Environ Microbiol. 1990; 56(12):3723-3726.
4. Alexander NJ, Proctor RH, McCormick SP. Genes, gene clusters, and biosynthesis of
trichothecenes and fumonisinsin Fusarium. Toxin Rev. 2009; 28(2-3): 198-215.
5. Lee J, Jurgenson JE, Leslie JF, Bowden RL. Alignment of genetic and physical maps of
Gibberella zeae. Appl Environ Microbiol. 2008; 74(8): 2349-2359.
6. Harris CA, Renfrew MJ, Woolridge MW. Assessing the risk of pesticide residues to consumers:
recent and future developments. Food Addit Contam. 2001; 18(12): 1124-1129.
7. Pagnussatt FA, Del Ponte EM, Garda-Buffon J, Badiale-Furlong E. Inhibition of Fusarium
graminearum growth and mycotoxin production by phenolic extract from Spirulina sp. Pestic
Biochem Physiol. 2014; 108: 21-26.
8. Marín S, Velluti A, Ramos AJ. Sanchis V. Effect of essential oils on zearalenone and
deoxynivalenol production by Fusarium graminearum in non-sterilized maize grain. Food
Microbiol. 2004; 21(3): 313-318.
9. Velluti A, Sanchis V, Ramos AJ, Turon C, Marín S. Impact of essential oils on growth rate,
zearalenone and deoxynivalenol production by Fusarium graminearum under different
temperature and water activity conditions in maize grain. J Appl Microbiol. 2004; 96(4):
716-724.
10. Lahooji A, Mirabolfathy M, Karami-Osboo R. Effect of Zataria multiflora and Satureja
hortensis essential oils, thymol and carvacrol on growth of Fusarium gramineum isolates and
deoxynivalenol production. Iran J Plant Pathol. 2010; 46(1): 37-50.
11. Khatib A, Mahmoudi H, Derakhshan A. 2016. Assessment of antifungal activity potential
several plant essential oils against Ceratocystis paradoxa. Proceedings of 22nd Iranian plant
protection congress. 27–30 Aug, Karaj, Iran. 319.
12. Vijayalakshmi P, Thenmozhi S, Rajeswari P. The Evaluation of the virulence factors of
clinical Candida isolates and the anti-biofilm activity of Elettaria cardamomum against
multi-drug resistant Candida albicans. Curr Med Mycol. 2016; 2(2): 8-15.
13. Shialy Z, Zarrin M, Sadeghi Nejad B, Yusef Naanaie S. In vitro antifungal properties of
Pistacia atlantica and olive extracts on different fungal species. Curr Med Mycol. 2015; 1(4):
40-45.
14. Vesaltalab Z, Gholami M. The effect of clove buds and rosemary extracts and essences on
control of Botrytis cinerea growth. Plant Prod Technol. 2010; 11(2): 1-11.
15. Sanchulli N, Ghaffari M, Qaraee A. Comparison of antifungal effects of essential oils of
Zataria multiflora boiss, Cuminum cyminum and Eugenia caryophyllata with formalin on
aflatoxin-producing fungus Aspergillus parasiticus. J Comp Pathol. 2015; 12(3): 1691-1698.
16. Ćosić J, Vrandečić K, Postić J, Jurković D, Ravlić M. In vitro antifungal activity of essential
oils on growth of phytopathogenic fungi. Agriculture. 2010; 16(2): 25-28.
17. Moosavian SM, Darvishnia M, Derikvand N. 2016. Effect of clove (Eugenia caryophyllata)
essential oil in inhibiting growth of Botrytis cinerea, Fusarium solani and Aspergillus niger.
Proceedings of 22nd Iranian plant protection congress. 27–30 Aug, Karaj, Iran. 310.
18. Eloff JN. A sensitive and quick microplate method to determine the minimal inhibitory
concentration of plant extracts for bacteria. Planta Medica. 1998; 64(8): 711-713.
19. Abril M, Curry KJ, Smith BJ, Wedge DE. Improved microassays used to test natural
product based and conventional fungicides on plant pathogenic fungi. Plant Dis. 2008; 92:
106-112.
20. Espinel-Ingroff A, Fothergill A, Peter J, Rinaldi MG, Walsh TJ. Testing conditions for
determination of minimum fungicidal concentrations of new and established antifungal agents
for Aspergillus spp.: NCCLS Collaborative Study. J Clin Microbiol. 2002; 40(2): 3204-3208.
21. Jahanshiri Z, Shams-Ghahfarokhi M, Allameh A, Razzaghi-Abyaneh M. Inhibitory effect of
eugenol on aflatoxin B1 production in Aspergillus parasiticus by down-regulating the
expression of major genes in the toxin biosynthetic pathway. World J Microbiol Biotechnol.
2015; 31(7): 1071-1078.
22. Kim HK, Yun SH. Evaluation of potential reference genes for quantitative RT-PCR analysis in
Fusarium graminearum under different culture conditions. Plant Pathol J. 2011; 27(4): 301-309.
23. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (RESTª) for group-wise
comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids
Res. 2002; 30(9): 1-10.
24. AlTaweel Kh, Amarasinghe ChC, Brûlé-Babel AL, Dilantha Fernando WG. Gene expression
analysis of host–pathogen interaction between wheat and Fusarium graminearum. Eur J Plant
Pathol. 2017; 148(3): 617-629.
25. Aneja KR, Sharma C. Antimicrobial potential of fruit extracts of Elettaria cardamomum maton
(chhoti elaichi) against the pathogens Causing ear infection. Pharmacol Online. 2010; 3:
750-756.
26. Trapp SC, Hohn TM, McCormick S, Jarvis BB. Characterization of the gene cluster for
biosynthesis of macrocyclic trichothecenes in Myrothecium roridum. Mol Gen Genet. 1998; 257
(4): 421-432.
27. Rynkiewicz MJ, Cane DE, Christianson DW. Structure of trichodiene synthase from Fusarium
sporotrichioides provides mechanistic inferences on the terpene cyclization cascade. Proc Natl
Acad Sci USA. 2001; 98(24): 13543-13548.
28. Dyer RB, Plattner RD, Kendra DF, Brown DW. Fusarium graminearum TRI14 is required for
high virulence and DON production on wheat but not for DON synthesis in vitro. J Agric Food
Chem. 2005; 53(23): 9281-9287.
29. Pinson-Gadais L, Richard-Forget F, Frasse P, Barreau C, Cahagnier B, Richard-Molard D,
Bakan B. Magnesium represses trichothecene biosynthesis and modulates Tri5, Tri6, and Tri12
genes expression in Fusarium graminearum. Mycopathologia. 2008; 165(1): 51-59
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devastating impact. Plant Dis. 1997; 81(12): 1340e1348.
2. Bottalico A, Perrone G. Toxigenic Fusarium species and mycotoxins associated with head
blight in smallcereals in Europe. Eur J Plant Pathol. 2002; 108(7): 611-624.
3. Luo Y, Yoshizawa T, Katayama T. Comparative study on the natural occurrence of Fusarium
mycotoxins (trichothecenes and zearalenone) in corn and wheat from high- and low-risk areas
for human esophageal cancer in China. Appl Environ Microbiol. 1990; 56(12):3723-3726.
4. Alexander NJ, Proctor RH, McCormick SP. Genes, gene clusters, and biosynthesis of
trichothecenes and fumonisinsin Fusarium. Toxin Rev. 2009; 28(2-3): 198-215.
5. Lee J, Jurgenson JE, Leslie JF, Bowden RL. Alignment of genetic and physical maps of
Gibberella zeae. Appl Environ Microbiol. 2008; 74(8): 2349-2359.
6. Harris CA, Renfrew MJ, Woolridge MW. Assessing the risk of pesticide residues to consumers:
recent and future developments. Food Addit Contam. 2001; 18(12): 1124-1129.
7. Pagnussatt FA, Del Ponte EM, Garda-Buffon J, Badiale-Furlong E. Inhibition of Fusarium
graminearum growth and mycotoxin production by phenolic extract from Spirulina sp. Pestic
Biochem Physiol. 2014; 108: 21-26.
8. Marín S, Velluti A, Ramos AJ. Sanchis V. Effect of essential oils on zearalenone and
deoxynivalenol production by Fusarium graminearum in non-sterilized maize grain. Food
Microbiol. 2004; 21(3): 313-318.
9. Velluti A, Sanchis V, Ramos AJ, Turon C, Marín S. Impact of essential oils on growth rate,
zearalenone and deoxynivalenol production by Fusarium graminearum under different
temperature and water activity conditions in maize grain. J Appl Microbiol. 2004; 96(4):
716-724.
10. Lahooji A, Mirabolfathy M, Karami-Osboo R. Effect of Zataria multiflora and Satureja
hortensis essential oils, thymol and carvacrol on growth of Fusarium gramineum isolates and
deoxynivalenol production. Iran J Plant Pathol. 2010; 46(1): 37-50.
11. Khatib A, Mahmoudi H, Derakhshan A. 2016. Assessment of antifungal activity potential
several plant essential oils against Ceratocystis paradoxa. Proceedings of 22nd Iranian plant
protection congress. 27–30 Aug, Karaj, Iran. 319.
12. Vijayalakshmi P, Thenmozhi S, Rajeswari P. The Evaluation of the virulence factors of
clinical Candida isolates and the anti-biofilm activity of Elettaria cardamomum against
multi-drug resistant Candida albicans. Curr Med Mycol. 2016; 2(2): 8-15.
13. Shialy Z, Zarrin M, Sadeghi Nejad B, Yusef Naanaie S. In vitro antifungal properties of
Pistacia atlantica and olive extracts on different fungal species. Curr Med Mycol. 2015; 1(4):
40-45.
14. Vesaltalab Z, Gholami M. The effect of clove buds and rosemary extracts and essences on
control of Botrytis cinerea growth. Plant Prod Technol. 2010; 11(2): 1-11.
15. Sanchulli N, Ghaffari M, Qaraee A. Comparison of antifungal effects of essential oils of
Zataria multiflora boiss, Cuminum cyminum and Eugenia caryophyllata with formalin on
aflatoxin-producing fungus Aspergillus parasiticus. J Comp Pathol. 2015; 12(3): 1691-1698.
16. Ćosić J, Vrandečić K, Postić J, Jurković D, Ravlić M. In vitro antifungal activity of essential
oils on growth of phytopathogenic fungi. Agriculture. 2010; 16(2): 25-28.
17. Moosavian SM, Darvishnia M, Derikvand N. 2016. Effect of clove (Eugenia caryophyllata)
essential oil in inhibiting growth of Botrytis cinerea, Fusarium solani and Aspergillus niger.
Proceedings of 22nd Iranian plant protection congress. 27–30 Aug, Karaj, Iran. 310.
18. Eloff JN. A sensitive and quick microplate method to determine the minimal inhibitory
concentration of plant extracts for bacteria. Planta Medica. 1998; 64(8): 711-713.
19. Abril M, Curry KJ, Smith BJ, Wedge DE. Improved microassays used to test natural
product based and conventional fungicides on plant pathogenic fungi. Plant Dis. 2008; 92:
106-112.
20. Espinel-Ingroff A, Fothergill A, Peter J, Rinaldi MG, Walsh TJ. Testing conditions for
determination of minimum fungicidal concentrations of new and established antifungal agents
for Aspergillus spp.: NCCLS Collaborative Study. J Clin Microbiol. 2002; 40(2): 3204-3208.
21. Jahanshiri Z, Shams-Ghahfarokhi M, Allameh A, Razzaghi-Abyaneh M. Inhibitory effect of
eugenol on aflatoxin B1 production in Aspergillus parasiticus by down-regulating the
expression of major genes in the toxin biosynthetic pathway. World J Microbiol Biotechnol.
2015; 31(7): 1071-1078.
22. Kim HK, Yun SH. Evaluation of potential reference genes for quantitative RT-PCR analysis in
Fusarium graminearum under different culture conditions. Plant Pathol J. 2011; 27(4): 301-309.
23. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (RESTª) for group-wise
comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids
Res. 2002; 30(9): 1-10.
24. AlTaweel Kh, Amarasinghe ChC, Brûlé-Babel AL, Dilantha Fernando WG. Gene expression
analysis of host–pathogen interaction between wheat and Fusarium graminearum. Eur J Plant
Pathol. 2017; 148(3): 617-629.
25. Aneja KR, Sharma C. Antimicrobial potential of fruit extracts of Elettaria cardamomum maton
(chhoti elaichi) against the pathogens Causing ear infection. Pharmacol Online. 2010; 3:
750-756.
26. Trapp SC, Hohn TM, McCormick S, Jarvis BB. Characterization of the gene cluster for
biosynthesis of macrocyclic trichothecenes in Myrothecium roridum. Mol Gen Genet. 1998; 257
(4): 421-432.
27. Rynkiewicz MJ, Cane DE, Christianson DW. Structure of trichodiene synthase from Fusarium
sporotrichioides provides mechanistic inferences on the terpene cyclization cascade. Proc Natl
Acad Sci USA. 2001; 98(24): 13543-13548.
28. Dyer RB, Plattner RD, Kendra DF, Brown DW. Fusarium graminearum TRI14 is required for
high virulence and DON production on wheat but not for DON synthesis in vitro. J Agric Food
Chem. 2005; 53(23): 9281-9287.
29. Pinson-Gadais L, Richard-Forget F, Frasse P, Barreau C, Cahagnier B, Richard-Molard D,
Bakan B. Magnesium represses trichothecene biosynthesis and modulates Tri5, Tri6, and Tri12
genes expression in Fusarium graminearum. Mycopathologia. 2008; 165(1): 51-59