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اثر کیتوزان در بیان ژن و فعالیت آنزیم‌های مؤثر در القای مقاومت به بلایت فوزاریومی خوشۀ گندم

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد بیماری‌شناسی گیاهی، گیاه‌پزشکی، دانشکدۀ کشاورزی، دانشگاه زابل

2 استادیار بیماری‌شناسی گیاهی، گروه گیاه‌پزشکی، دانشکدۀ کشاورزی، دانشگاه زابل و پژوهشکدۀ زیست‌فناوری کشاورزی دانشگاه زابل

چکیده

در این تحقیق، نقش کیتوزان به عنوان یک محرک زیستی مکانیسم دفاعی در گندم آلوده به Fusarium graminearum عامل بلایت فوزاریومی خوشۀ گندم بررسی شد. به منظور بررسی بیان ژن‌های مرتبط با بیماری‌زایی و فعالیت آنزیمی، آزمایش گلخانه‌ای به‌صورت طرح فاکتوریل در قالب طرح کاملاً تصادفی در چهار تکرار انجام گرفت. در این مطالعه محلول کیتوزان با غلظت‌های مختلف (0، 100، 200 و 500 میلی‌گرم در لیتر) به‌صورت اسپری برگی استفاده شد. گیاهان تیمارشده پس از 24 ساعت با سوسپانسیون اسپور قارچ (با غلظت 106 ماکروکنیدی در میلی‌لیتر) به روش تزریق خوشه، مایه‌زنی شده و تحت شرایط گلخانه نگهداری شدند. نمونه‌برداری در بازه‌های زمانی مختلف بعد از آلودگی انجام گرفت و سپس سطوح بیان برخی ژن‌های دخیل در مقاومت و فعالیت آنزیم‌های مرتبط بررسی شد. تجزیه و تحلیل داده‌ها کاهش شدت بیماری را در گیاهان تیمارشده نسبت به گیاه شاهد نشان داد. همچنین افزایش معناداری در فعالیت آنزیم پراکسیداز و پلی فنل اکسیداز مشاهده شد. نتایج آنالیز مولکولی به روش qRT-PCR نشان از افزایش شایان توجه سطوح بیان mRNA ژن‌های بتا- 1 و 3 گلوکانازو اگزالات اکسیداز داشت. نتایج این پژوهش نشان می‌دهد که کیتوزان با القای مقاومت سیستمیک اکتسابی مقاومت گیاهان را علیه قارچ‌های بیماری‌زا تحت تأثیر قرار می‌دهد.
 
 
 

کلیدواژه‌ها

عنوان مقاله [English]

Effect of chitosan on gene expression and activity of enzymes involved in resistant induction to fusariuse of wheat

نویسندگان [English]

  • Vahid Ghazimohseni 1
  • Seyed Kazem Sabbagh 2
1 M.Sc. Student of Plant Pathology, University of Zabol
2 Assistant Professor, Department of Plant Protection, Faculty of Plant Pathology, University of Zabol and Institute of Plant Biotechnology, University of Zabol
چکیده [English]

In this research, the role of chitosan as a bio-inducer of defense mechanism in infected wheat by Fusarium graminearum, the causal agent of Fusarium head blight was investigated. To investigate the expression of genes associated with pathogenicity and enzymatic activity, a factorial experiment in a completely randomized design with four replications was done under greenhouse condition. The adjusted solution containing different concentrations of chitosan (0, 100, 200 and 500 mg l-1) were used via spray. Treated plants after 24 hours with a spore suspension (at a concentration of 106 Macro conidia ml-1) were inoculated by spikelet injection and were kept under greenhouse conditions. Sampling was performed at various time points after inoculation and then the expression level of some genes involved in resistance and activity of relative enzyme was studied. Data analysis showed that disease severity was reduced in treated plants compared with control plants. Enzyme measurement showed the greatest variation for peroxidase and polyphenol oxidase enzymes. The results of molecular analysis by qRT-PCR showed significantly increased mRNA expression levels of beta-1, 3-glucanase and oxalate oxidase genes. The results of this research indicate that chitosan could influence plant resistance against pathogenic fungi through induction of systemic acquire resistance.
 
 
 

کلیدواژه‌ها [English]

  • Chitosan
  • qRT-PCR
  • head blight
  • polyphenol oxidase
  • Peroxidase

مراجع

  1.  

    1. Arloria, M., Ludwing, A., Boller, T. & Bonfante, P. (1992) Inhibition of fungal growth by plant chitinases and β-1, 3-glucanases. Protoplasma, 177, 34-43.
    2. Ban, T., Kawad, N., Yanagisaway, A. & Takezaki, A. (2008) Progress and future prospects of resistance breeding to Fusarium head blight in Japan. Cereal Research Communications, 36, 23-29.
    3. Bernardo, A., Bai, G., Guo, P., Xiao, K., Guenzi, A. C. & Ayoubi, P. (2007) Fusarium graminearum-induced changes in gene expression between Fusarium head blight-resistant and susceptible wheat cultivars. Functional & Integrative Genomics, 7, 69-77.
    4. Benhamou, N., Kloepper, J. W. & Tuzun, S. (1998). Induction of resistance against Fusarium wilt of tomato by combination of chitosan with an endophytic bacterial strain: ultrastructure and cytochemistry of the host response. Planta, 204, 153-168.
    5. Ben- Shalom, N., Ardi, R., Pinto, R., Aki, C. & Fallik, E. (2003). Controlling gray mould caused by Botrytis Cinerea in cucumber plants by means of chitosan. Crop Protection, 22, 285-290.
    6. Bradford, M. M. (1976). A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254.
    7. Chen, C., Belanger, R., Benhamon, N. & Paulitz, T. (2000) Defense enzymes induced in cucumber roots by treatment with plant growth promoting rhizobacteria (PGPR) and Pythium aphanidermatum. Physiological and Molecular Plant Pathology, 56, 13- 23.
    8. Collinge D, Lyngs Jrgensen H. (2009). Effects of b-1, 3-glucan from Septoria tritici on structural defense responses in wheat. Journl of Exprimental Botny, 60(15), 4287-300.
    9. De Azevedo Neto, A. D., Prisco, J. T., Enéas-Filho, J., Abreu, C. E. & Gomes-Filho, E. (2006) Effect of salt stress on antioxidative enzymes and lipid peroxidation in leaves and roots of salt-tolerant and salt-sensitive maize genotypes. Environmental and Experimental Botany, 56, 87-94.
    10. Dordas, C. (2009). Role of Nutrients in Controlling Plant Diseases. In Lichtfouse, E., Navarrete, M., Debaeke, P., Véronique S, Alberola C. (Ed.), Sustainable Agriculture: A Review. (pp. 369-360). Springer Netherlands.
    11. El Ghaouth, A., Arul, J., Asselin, A. & Benhamou, N. (1992) Antifungal activity of chitosan on post-harvest pathogens: induction of morphological and cytological alterations in Rhizopus stolonifer. Mycological Research, 96, 769-779.
    12. Falcon-Rodriguez, A. B., Costales, D., Cabrera, J. C. & Martinez-Tellez, M. A. (2011) Chitosan physico–chemical properties modulate defense responses and resistance in tobacco plants against the Oomycete Phytophthora nicotianae. Pesticide Biochemistry and Physiology, 100, 221-228.
    13. Goswami, R. S. & Kistler, H. C. (2004). Heading for disaster: Fusarium graminearum on cereal crops. Molecular Plant Pathology, 5, 515-525.
    14. Jayaraj, J., Rahman, M., Wan, A. & Punja, Z. K. (2009). Enhanced resistance to foliar fungal pathogens in carrot by application of elicitors. Annals of Applied Biology, 155, 71-80.
    15. Koga, J., Kubota, H., Gomi, S., Umemura, K., Ohnishi, M. & Kono, T. (2006). Cholic acid a bile acid elicitor of hypersensitive cell death, pathogenesis- related protein synthesis, and phytoalexin accumulation in rice. Plant Physiology, 140, 1475-1483.
    16. Mackintosh, C. A., Lewis, J., Radmer, L. E., Sin, S., Heinen, S., J., Smith, L. A.,Wyckoff, M. & Muehlbauer, G. J. (2007). Over expression of defense response genes in transgenic wheat enhances resistance to Fusarium Head Blight. Plant Cell Report, 26, 479-488.
    17. Mayer, A. M. (2006). Polyphenol oxidases in plants and fungi. Phytochemistry, 67, 2318-2331.
    18. Mitchell, H., Hall, J. & Barber, M. (1994) Elicitor-induced cinnamyl alcohol dehydrogenasa activity in lignifying wheat (Triticum aestivum L.) leaves. Plant Physiology, 104, 551-556.
    19. Nandeeshkumar, P., Sudisha, J., Ramachandra, K., Prakash, H. S., Niranjana, S. R. & Shekar. S. (2008) Chitosan induced resistance to downy mildew in sunflower caused by Plasmoparahalstedii. Physiological and Molecular Plant Pathology, 72, 188-194.
    20. Nemati, M. & Navabpour, S. (2012) Study on quantitative expression pattern of oxalate oxidase and β-1, 3 glucanase genes under Fusarium graminearum treatment in wheat by quantitative Real time PCR. International Journal of Agriculture and Crop Sciences, 4, 443-447.
    21. Parry, D. W., Jenkinson, P. & Mcleod, L. (1995). Fusarium ear blight (scab) in small grain cereals – a review. Plant Pathology, 44, 207-238.
    22. Pfaffl, M. W. (2001) A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Research, 29, e45.
    23. Propagdee, B., Kotchdat, Kumsopa, A. & Visarathanonth, N. (2006). The role of chitosan in protection of soybean from suolden death syndrome caused by Fusarium solani f. sp. Glycines. Bioresource Technology, 98, 1353-1358.
    24. Schweigkofler, W., O'Donnell, K. & Garbelotto, M. (2004). Detection and quantification of airborne conidia of Fusarium circinatum, the causal agent of pine pitch canker, from two California sites by using a real-time PCR approach combined with a simple spore trapping method. Applied and Environmental Microbiology, 70, 3512-3520.
    25. Sharathchandra, R. G., Niranjan Raj, S., Shetty, N. P., Amruthesh, K.N. & ShekarShetty, H. (2004) A chitosan formulation Elexa induces downy mildew disease resistance and growth promotion in pearl millet. Crop Protection, 23, 881-888.
    26. Shetty, N.P., Mehrabi, R., Lütken, H., Haldrup, A., Kema, G.H., Collinge, D.B. & Jorgensen, H. (2007) Role of hydrogen peroxide during the interaction between the hemibiotrophic fungal pathogen Septoria tritici and wheat. New Phytologist 174, 637-647.
    27. Stack, R. W. & McMullen, M. P. (2010). A visual scale to estimate severity of Fusarium head blight in wheat. North Dakota State University. Reviewed November, p. 1095.
    28. Trotel-Aziz, P., Couderchet, M., Vernet, G. & Aziz, A. (2006) Chitosan stimulates defense reactions in grapevine leaves and inhibits development of Botrytis cinerea. Europian Journal Plant  Pathology, 114, 405-413.
    29. Wiese, M. (1977). Compendium of wheat diseases: American Phytopathological Society.

    Yoshizawa, T. (1997) Geographic difference in trichothecene occurrence in Japanese wheat and barley. Bulletin of the Institute for Comprehensive Agricultural Sciences, Kinki University,

دانش گیاهپزشکی ایران
دوره 46، شماره 2 - شماره پیاپی 2
پاییز و زمستان
مهر 1394
صفحه 363-371

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سابقه مقاله

  • تاریخ دریافت: 14 دی 1393
  • تاریخ بازنگری: 03 مهر 1394
  • تاریخ پذیرش: 12 آذر 1394

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