کنترل زیستی قارچ فوزاریوم اکسیسپوروم فرم اختصاصی لیکوپرسیسی و القای آنزیم دفاعی فنیل‌آلانین‌آمونیالیاز در گوجه‌فرنگی توسط جدایه‌های آنتاگونیست تریکودرما و باسیلوس
محورهای موضوعی : میکروب شناسی گیاهی
شهناز آل آقائی
1
,
سعید رضائى
2
,
مصطفی عبادی
3
,
حمیدرضا زمانی زاده
4
1 - دانشجوی دکتری، گروه گیاهپزشکی، دانشکده علوم کشاورزی و صنایع‌غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
2 - استادیار، گروه گیاهپزشکی، دانشکده علوم کشاورزی و صنایع‌غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
3 - استادیار، گروه زیست‌شناسی، دانشکده علوم‌پایه، واحد دامغان، دانشگاه آزاد اسلامی، دامغان، ایران.
4 - استاد، گروه گیاهپزشکی، دانشکده علوم کشاورزی و صنایع‌غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران
کلید واژه: تریکودرما هارزیانوم, کلونیزاسیون, مقاومت القایی, تریکودرما اتروویریده, واکنش دفاعی,
چکیده مقاله :
سابقه و هدف: در اغلب کشورها به ویژه ایران، بیماری پژمردگی آوندی فوزاریومی از عوامل اصلی محدود کننده تولید گوجه‌فرنگی است. باتوجه به تأثیر نامناسب کاربرد ترکیبات شیمیایی بر سلامت، مطالعه راهکارهای جایگزین برای کنترل بیماری حائز اهمیت است. این پژوهش با هدف ارزیابی قابلیت برخی جدایه‌های بومی قارچ تریکودرما و باکتری باسیلوس در کنترل زیستی قارچ فوزاریوم اکسیسپوروم فرم اختصاصی لیکوپرسیسی انجام شد.مواد و روش‌ها: بازدارندگی از رشد میسلیوم‌های قارچ بیمارگر فوزاریوم توسط جدایه‌های تریکودرما و باسیلوس در محیط کشت سیب‌زمینی دکستروز آگار مورد ارزیابی قرار گرفت. همچنین قابلیت کلونیزاسیون این جدایه‌ها نیز بررسی شد. تأثیر کاربرد مجزا و توأم جدایه‌های آنتاگونیست بر شاخص شدت بیماری پژمردگی آوندی فوزاریومی وبرخی عوامل رشدی گوجه‌فرنگی تعیین گردید. همچنین پتانسیل جدایه‌های یاد شده در القای آنزیم دفاعی فنیل‌آلانین‌آمونیالیاز در گیاهچه‌های آلوده به بیمارگر فوزاریوم بررسی شد.یافته‌ها: عوامل بیوکنترلی درجات مختلفی از بازدارندگی رشد فوزاریوم را نشان دادند. جدایه‌های تریکودرما با بازدارندگی قابل‌توجه (72.06% و 69.16%)، در مهار بیمارگر موفق‌‌تر عمل کردند. این جدایه‌ها کلونیزه کننده‌های قدرتمند بیمارگر نیز بودند. آنتاگونیست‌ها با سطوح متفاوت بر شاخص شدت بیماری تأثیرگذار بودند. تیمارهای مجزا و ترکیبی جدایه‌های تریکودرما در مقایسه با باسیلوس، در کاهش شدت بیماری و بهبود عوامل رشد مؤثرتر بودند. سطح فعالیت آنزیم فنیل‌آلانین‌آمونیالیاز نیز در تیمارهای تریکودرما در مقایسه با شاهد (فوزاریوم) افزایش یافت. کاربرد هم زمان این جدایه‌ها در مقایسه با تیمار مجزا در القای آنزیم موثرتر بود.نتیجه گیری: در این پژوهش جدایه‌های آنتاگونیست تریکودرما عملکرد موفقی در مهار بیمارگر داشتند، از این رو کاربرد این جدایه‌ها به عنوان عواملی با تأثیرات چند جانبه می‌تواند در کنترل زیستی بیماری پژمردگی آوندی فوزاریومی گوجه‌فرنگی کارآمد باشد.
Background & Objectives: Fusarium wilt disease is one of the limiting factors of tomato production in most countries, especially in Iran. Considering the malicious effect of chemicals on health, the study on the alternatives for disease controlling is important. This study aimed to evaluate the ability of Trichoderma and Bacillus isolates on biological control of Fusariumoxysporum f. sp. lycopersici.Materials & Methods: The inhibitory effects of Trichoderma and Bacillus isolates against Fusarium were evaluated on PDA. The ability of the colonization of these antagonists was also examined. The effect of separate and combined application of the antagonists isolates against Fusarium vascular wilt disease severity index and some growth factors of tomato were determined. Also, the potential of these isolates on the induction of phenylalanine ammonlyase was measured in seedlings infected with Fusarium.Results: The biocontrol agents showed different levels of inhibition of Fusarium growth. The Trichoderma isolates were more successful with a high inhibitory effect (72.06% and 69.16%). These isolates were also powerful colonizers of the pathogen. The disease severity was affected by the antagonists at different levels. Separate and combined treatments of Trichoderma isolates were more effective in the reduction of the disease severity and improving the growth factors. The activity of phenylalanine ammonialyas increased in the plants treated with Trichoderma isolates compared to control treatment (Fusarium). The combined application of these isolates was more effective than the separate using of them on the induction of the enzyme.Conclusion: The investigated Trichoderma isolates were successful in controlling the pathogen. Therefore, the use of these isolates as agents with multiplier effects can be effective in biocontrol of tomato Fusarium wilt.
managing a vascular wilt of tomato caused by Fusarium oxysporum f. sp. lycopersici. J Biol
Agric Healthcare. 2014; 4(27): 335-343.
2. Rajik M, Biswas SK, Shakti S. Biochemical basis of defense response in plant against Fusarium
wilt through bio-agents as an inducer. Afr J Agric Res. 2012; 7(43): 5849-5857.
3. Mwangi MV, Monda EO, Okoth SA, Jefwa JM. Inoculation of tomato seedlings with
Trichodermaharzianum and arbuscular mycorrhyzal fungi and their effect on growth and
control of wilt in tomato seedlings. Braz J Microbiol. 2011; 42(2): 508-513.
4. Sundaramoorthy S, Balabaskar P. Biocontrol efficacy of Trichoderma spp. against wilt of
tomato caused by Fusarium oxysporum f. sp. Lycopersici. J ApplBiol Biotechnol. 2013; 1(3):
36- 40.
5. Amini J. Physiological race of and Fusarium oxysporum f. sp. Lycopersici in Kurdistan
province of Iran and reaction of some tomato cultivars to race 1 of pathogen. J Plant Pathol.
2009; 8(2): 68-73.
6. Pourjam E, Kamali N, Sahebani N. Elicitation of defense responses in tomato against
Meloidogyne javanica and Fusarium oxysporum f. sp. Lycopersici wilt complex. J Crop
Protect. 2015; 4(1): 29-38.
7. Naing KW, NguyenXH, Anees M, Lee YS, Kim YC, Kim SJ, Kim MH, Kim YH, Kim KY.
Biocontrol of Fusarium wilt disease in tomato by Paenibacillus ehimensis KWN38.World J
Microbiol Biotechnol. 2015; 31(1): 165-174.
8. Vos CM, Yang Y, De-Coninck B, Cammue BPA. Fungal (-like) biocontrol organisms in tomato
disease control. Biol Control. 2014; 74(7): 65-81.
9. Amer MA, El-Samra IA, Abou-El-Seoud II, El-Abd SM, Shawertamin NK. Induced systemic
resistance in tomato plants against Fusarium wilt disease using biotic inducers. Middle East J
Agric Res. 2014; 3(4): 1090-1103.
10. Farhangniya S, Naraghi L, Ommati F, Pirnia M. Evaluation of the efficacy of the biological
compound affected by Talaromyces flavus in controlling tomato Fusarium wilt disease in the
field conditions. Int J Agric Sci Res. 2015; 5(2): 153-164.
11. Harman GE, Herrera-Estrella AH, Horwitz BA, Lorito M. Trichoderma- from basic biology to
biotechnology. Microbiol. 2012; 158(1): 1-2.
12. Nawrocka J, Malolepza V. Diversity in plant systemic resistance induced by Trichoderma. Biol
Control.2013; 67(2):149-156.
13. Verma N, Ahmed M, Upadhyay RS. Induction of resistance in tomato against Fusarium
oxysporum f. sp. lycopersici. Persian Gulf Crop Protect. 2014; 3(4): 25-36.
14. Ojha S, Chatterjee NC. Induction of resistance in tomato against Fusarium oxysporum f. sp.
Lycopersici mediated through salicylic acid and Trichoderma harzianum. J Plant Protect Res.
2012; 52(2): 220-225.
15. Leslie JF, Summerell BA. The Fusarium laboratory manual. Manhattan. Blackwell Press; 2006.
16. NelsonPE, Toussoun TA, Marascs WFO. Fusarium species, an illustrated manual for
identification. Pennsylvania.University Press; 1983.
17. Cheng AA, Wu FR. Fusarium wilt of sesame in Taiwan. Rep TAINAN Station Council Agric.
1991; 26: 53-60.
18. Menzies JG, Koch C, Seywerd F. Additions to the host range of Fusarium oxysporum f. sp.
radicis-lycopersici. Plant Dis. 1990; 74(8): 569-572.
19. Aleaghaee Sh, Shahriari D, Torabi M. 2012. Evaluation of the effect of Trichoderma spp.
isolates on biological control of Pythiumaphanidermatum, the causal agent of cucumber
damping-off and seed rot in laboratory andgreenhouse. Proceeding of the 20th Iranian Plant
Protection Congress.25-28 Aug, Shiraz, Iran. 252.[In Persian]
20. Hagedron C, Gould WD, Bardinelli TR. Rhizobacteria of cotton and their repression of
seedling disease pathogens. Appl Environ Microbiol. 1989; 55(11): 2743-2797.
21. Boyhan GE, Lagston DB, Lewis PM, Linton MO. Use of insulin syringe for Fusarium wilt
inoculation of watermelon germplasm. Cucurbit Genet Coop Rep. 2001; 24: 49-51.
22. Altinok HH. Activation of systemic resistance by acibenzolar-s-methyl and a non-pathogenic
Fusarium oxysporummelonis (FOM) strain against Fusarium wilt disease in eggplant seedlings.
J Turk Phytopathol. 2009; 38(1-3): 21-32.
23. El Khaldi R, Daami-Remadi M, Cherif M. Biological control of stem canker and black scurf on
potato by date palm compost and its associated fungi. J Phytopathol. 2016; 164(1): 40-51.
24. Chowdappa P, Mohan Kumar SP, Lakshmi MJ, Upreti KK. Growth stimulation and induction
of systemic resistance in tomato against early and late blight by Bacillussubtilis OTPB1 or
Trichoderma harzianum OTPB3. Biol Control. 2013; 65(1): 109-117.
25. Madhaiyan M, Poonguzhali S, Senthilkumar M, Seshadri S. Chung H, Yang J, Sundarm S, Sa,
T. Growth promotion and induction of systemic resistance in rice cultivar co-47 by
methylobacterium spp. Bot Bull Acad Sinica. 2004; 45: 315-324.
26. Wang JW, Zhang LP, Wu JY, Tan R. X. Involvement of nitric oxide in oxidative burst,
phenylalanine ammonia-lyase activation and taxol production induced by low-energy
ultrasound in Taxus Yunnanensis cell suspension culture. Nitric Oxide. 2006; 15(4): 351-358.
27. Bradford M. A rapid and sensitive method for the quantification of quantities of protein
utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248-252.
28. Raza W, Ling N, Zhang R, Huang Q, Xu Y, Shen Q. Success evaluation of the biological
control of Fusarium wilts of cucumber, banana, and tomato since 2000 and future research
strategies. Crit Rev Biotechnol. 2017; 37(2): 202-212.
29. Chandge A, Lim MH, Lee SW, Robb EJ, Nazar RN. Tomato PAL gene family: highly
redundant but strongly underutilized. J Biol Chem. 2008; 283(48): 33591-33601.
30. Elsharkawy MM, Shivanna M, Hyakumach M. Mechanism of induced systemic resistance
against anthracnose disease in cucumber by plant growth-promoting fungi. Acta Agric Scand,
Sec B-Soil Plant Sci. 2015; 65(4): 287-299.
31. Małolepsza U, Nawrocka J, Szczech M.Trichoderma virens 106 inoculation stimulates defense
enzyme activities and enhances phenolic levels in tomato plants leading to lowered Rhizoctonia
solani infection. Biocontrol Sci Technol. 2017. 27(2): 180-199.
32. Aleaghee Sh, Rezaee S, Ebadi M, Zamanizadeh HR. The efficacy of some native Trichoderma
isolates in induction of resistance in tomato against Fusarium oxyporum f. sp. lycoresici, the
causal agent of Fusarium wilt disease. Appl Entomol phytopathol. 2017; 85(2): 219-234.
33. Pieterse CMJ, Zamioudis Ch, Berendsen RL, Weller DM, Van Wees SCM, Bakker PAHM.
Induced systemic resistance by beneficial microbes. Annu Rev Phytopathol. 2014; 52:
347-375.
34. Zamioudis C, Pieterse CMJ. Modulation of host immunity by beneficial microbes. Mol
Plant-Microbe Interact. 2012; 25(2): 139-150.
35. Holand MA. Occams razor applied to hormonology. Arc cytokinins produced by plants?. Plant
Physiol. 1997; 115(3): 865-868.
36. Vos IA, Pieterse CMJ, Van Wees SCM. Costs and benefits of hormone-regulated plant
defences. Plant Pathol. 2013; 62:43-55.
37. Fu J, Huang B. Involvement of antioxidant and lipid peroxidation in the adaptation of two cool
-season grasses to localized drought stress. Environ Exper Bot. 2001; 45(2): 105-114.
38. Silva HSA, da Silva Romeiro R, Macagnan D, de Almeida Halfeld-Vieira B, Pereira MCB,
Mounteer A. Rhizobacterial induction of systemic resistancein tomato plants: non-specific
protection and increase in enzyme activities. Biol Control. 2004; 29(2): 288-295.
39. Mathys J, De-Cremer K, Timmermans P, VanKerckhove S, Lievens B, Vanhaecke M,
Cammue BP, De-Coninck B. Genome-wide characterization of ISR induced in Arabidopsis
thaliana by Trichoderma hamatum T382 against Botrytiscinerea infection. Front Plant Sci.
2012; 3(108)
_||_
managing a vascular wilt of tomato caused by Fusarium oxysporum f. sp. lycopersici. J Biol
Agric Healthcare. 2014; 4(27): 335-343.
2. Rajik M, Biswas SK, Shakti S. Biochemical basis of defense response in plant against Fusarium
wilt through bio-agents as an inducer. Afr J Agric Res. 2012; 7(43): 5849-5857.
3. Mwangi MV, Monda EO, Okoth SA, Jefwa JM. Inoculation of tomato seedlings with
Trichodermaharzianum and arbuscular mycorrhyzal fungi and their effect on growth and
control of wilt in tomato seedlings. Braz J Microbiol. 2011; 42(2): 508-513.
4. Sundaramoorthy S, Balabaskar P. Biocontrol efficacy of Trichoderma spp. against wilt of
tomato caused by Fusarium oxysporum f. sp. Lycopersici. J ApplBiol Biotechnol. 2013; 1(3):
36- 40.
5. Amini J. Physiological race of and Fusarium oxysporum f. sp. Lycopersici in Kurdistan
province of Iran and reaction of some tomato cultivars to race 1 of pathogen. J Plant Pathol.
2009; 8(2): 68-73.
6. Pourjam E, Kamali N, Sahebani N. Elicitation of defense responses in tomato against
Meloidogyne javanica and Fusarium oxysporum f. sp. Lycopersici wilt complex. J Crop
Protect. 2015; 4(1): 29-38.
7. Naing KW, NguyenXH, Anees M, Lee YS, Kim YC, Kim SJ, Kim MH, Kim YH, Kim KY.
Biocontrol of Fusarium wilt disease in tomato by Paenibacillus ehimensis KWN38.World J
Microbiol Biotechnol. 2015; 31(1): 165-174.
8. Vos CM, Yang Y, De-Coninck B, Cammue BPA. Fungal (-like) biocontrol organisms in tomato
disease control. Biol Control. 2014; 74(7): 65-81.
9. Amer MA, El-Samra IA, Abou-El-Seoud II, El-Abd SM, Shawertamin NK. Induced systemic
resistance in tomato plants against Fusarium wilt disease using biotic inducers. Middle East J
Agric Res. 2014; 3(4): 1090-1103.
10. Farhangniya S, Naraghi L, Ommati F, Pirnia M. Evaluation of the efficacy of the biological
compound affected by Talaromyces flavus in controlling tomato Fusarium wilt disease in the
field conditions. Int J Agric Sci Res. 2015; 5(2): 153-164.
11. Harman GE, Herrera-Estrella AH, Horwitz BA, Lorito M. Trichoderma- from basic biology to
biotechnology. Microbiol. 2012; 158(1): 1-2.
12. Nawrocka J, Malolepza V. Diversity in plant systemic resistance induced by Trichoderma. Biol
Control.2013; 67(2):149-156.
13. Verma N, Ahmed M, Upadhyay RS. Induction of resistance in tomato against Fusarium
oxysporum f. sp. lycopersici. Persian Gulf Crop Protect. 2014; 3(4): 25-36.
14. Ojha S, Chatterjee NC. Induction of resistance in tomato against Fusarium oxysporum f. sp.
Lycopersici mediated through salicylic acid and Trichoderma harzianum. J Plant Protect Res.
2012; 52(2): 220-225.
15. Leslie JF, Summerell BA. The Fusarium laboratory manual. Manhattan. Blackwell Press; 2006.
16. NelsonPE, Toussoun TA, Marascs WFO. Fusarium species, an illustrated manual for
identification. Pennsylvania.University Press; 1983.
17. Cheng AA, Wu FR. Fusarium wilt of sesame in Taiwan. Rep TAINAN Station Council Agric.
1991; 26: 53-60.
18. Menzies JG, Koch C, Seywerd F. Additions to the host range of Fusarium oxysporum f. sp.
radicis-lycopersici. Plant Dis. 1990; 74(8): 569-572.
19. Aleaghaee Sh, Shahriari D, Torabi M. 2012. Evaluation of the effect of Trichoderma spp.
isolates on biological control of Pythiumaphanidermatum, the causal agent of cucumber
damping-off and seed rot in laboratory andgreenhouse. Proceeding of the 20th Iranian Plant
Protection Congress.25-28 Aug, Shiraz, Iran. 252.[In Persian]
20. Hagedron C, Gould WD, Bardinelli TR. Rhizobacteria of cotton and their repression of
seedling disease pathogens. Appl Environ Microbiol. 1989; 55(11): 2743-2797.
21. Boyhan GE, Lagston DB, Lewis PM, Linton MO. Use of insulin syringe for Fusarium wilt
inoculation of watermelon germplasm. Cucurbit Genet Coop Rep. 2001; 24: 49-51.
22. Altinok HH. Activation of systemic resistance by acibenzolar-s-methyl and a non-pathogenic
Fusarium oxysporummelonis (FOM) strain against Fusarium wilt disease in eggplant seedlings.
J Turk Phytopathol. 2009; 38(1-3): 21-32.
23. El Khaldi R, Daami-Remadi M, Cherif M. Biological control of stem canker and black scurf on
potato by date palm compost and its associated fungi. J Phytopathol. 2016; 164(1): 40-51.
24. Chowdappa P, Mohan Kumar SP, Lakshmi MJ, Upreti KK. Growth stimulation and induction
of systemic resistance in tomato against early and late blight by Bacillussubtilis OTPB1 or
Trichoderma harzianum OTPB3. Biol Control. 2013; 65(1): 109-117.
25. Madhaiyan M, Poonguzhali S, Senthilkumar M, Seshadri S. Chung H, Yang J, Sundarm S, Sa,
T. Growth promotion and induction of systemic resistance in rice cultivar co-47 by
methylobacterium spp. Bot Bull Acad Sinica. 2004; 45: 315-324.
26. Wang JW, Zhang LP, Wu JY, Tan R. X. Involvement of nitric oxide in oxidative burst,
phenylalanine ammonia-lyase activation and taxol production induced by low-energy
ultrasound in Taxus Yunnanensis cell suspension culture. Nitric Oxide. 2006; 15(4): 351-358.
27. Bradford M. A rapid and sensitive method for the quantification of quantities of protein
utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248-252.
28. Raza W, Ling N, Zhang R, Huang Q, Xu Y, Shen Q. Success evaluation of the biological
control of Fusarium wilts of cucumber, banana, and tomato since 2000 and future research
strategies. Crit Rev Biotechnol. 2017; 37(2): 202-212.
29. Chandge A, Lim MH, Lee SW, Robb EJ, Nazar RN. Tomato PAL gene family: highly
redundant but strongly underutilized. J Biol Chem. 2008; 283(48): 33591-33601.
30. Elsharkawy MM, Shivanna M, Hyakumach M. Mechanism of induced systemic resistance
against anthracnose disease in cucumber by plant growth-promoting fungi. Acta Agric Scand,
Sec B-Soil Plant Sci. 2015; 65(4): 287-299.
31. Małolepsza U, Nawrocka J, Szczech M.Trichoderma virens 106 inoculation stimulates defense
enzyme activities and enhances phenolic levels in tomato plants leading to lowered Rhizoctonia
solani infection. Biocontrol Sci Technol. 2017. 27(2): 180-199.
32. Aleaghee Sh, Rezaee S, Ebadi M, Zamanizadeh HR. The efficacy of some native Trichoderma
isolates in induction of resistance in tomato against Fusarium oxyporum f. sp. lycoresici, the
causal agent of Fusarium wilt disease. Appl Entomol phytopathol. 2017; 85(2): 219-234.
33. Pieterse CMJ, Zamioudis Ch, Berendsen RL, Weller DM, Van Wees SCM, Bakker PAHM.
Induced systemic resistance by beneficial microbes. Annu Rev Phytopathol. 2014; 52:
347-375.
34. Zamioudis C, Pieterse CMJ. Modulation of host immunity by beneficial microbes. Mol
Plant-Microbe Interact. 2012; 25(2): 139-150.
35. Holand MA. Occams razor applied to hormonology. Arc cytokinins produced by plants?. Plant
Physiol. 1997; 115(3): 865-868.
36. Vos IA, Pieterse CMJ, Van Wees SCM. Costs and benefits of hormone-regulated plant
defences. Plant Pathol. 2013; 62:43-55.
37. Fu J, Huang B. Involvement of antioxidant and lipid peroxidation in the adaptation of two cool
-season grasses to localized drought stress. Environ Exper Bot. 2001; 45(2): 105-114.
38. Silva HSA, da Silva Romeiro R, Macagnan D, de Almeida Halfeld-Vieira B, Pereira MCB,
Mounteer A. Rhizobacterial induction of systemic resistancein tomato plants: non-specific
protection and increase in enzyme activities. Biol Control. 2004; 29(2): 288-295.
39. Mathys J, De-Cremer K, Timmermans P, VanKerckhove S, Lievens B, Vanhaecke M,
Cammue BP, De-Coninck B. Genome-wide characterization of ISR induced in Arabidopsis
thaliana by Trichoderma hamatum T382 against Botrytiscinerea infection. Front Plant Sci.
2012; 3(108)