همسانه سازی و بیان پپتید ضدمیکروبی امیگانان پنتاهیدروکلراید (MBI226) در توتون (Nicotiana tabacum) به منظور ایجاد مقاومت به باکتری‌های بیماری‌زا

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه لرستان، خرم آباد، ایران

2 استاد گروه زراعت و اصلاح‌نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم آباد، ایران

3 دانشیار گروه زراعت و اصلاح‌نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم آباد، ایران

4 استادیار گروه بیوتکنولوژی، دانشکده کشاورزی، دانشگاه بوعلی‌سینا، همدان، ایران

چکیده

پپتیدهای ضدمیکروبی، مولکول‌های قدیمی و حفاظت شده‌ هستند که در مکانیسم‌های دفاعی موجودات زنده مانند باکتری‌ها، جانوران و گیاهان دیده می‌شوند. شناسایی و معرفی پپتیدهای ضدمیکروبی جدید، روشی مقرون به صرفه برای مقابله با میکروب‌های بیماری‌زا و همچنین بهبود مقاومت گونه‌های گیاهان زراعی با استفاده از تکنولوژی DNA نوترکیب است. به این منظور یک سازه ژنی حاوی توالی ژن کد کننده پپتید ضدمیکروبی امیگانان (Omiganan) نوتروفیل گاو پس از همسانه‌سازی به کمک اگروباکتریویوم تومفاشینز به دیسک‌های برگی توتون انتقال داده شد. با روش PCR حضور ژن کد کننده پپتید ضدمیکروبی در ژنوم گیاهان تراریخت اثبات و تعداد 6 گیاه تراریخت به همراه شاهد انتخاب شدند. پروتئین کل استخراج و از آن برای کنترل رشد برخی باکتری‌های مهم انسانی مثل؛ E.coli ، Staphylococcus aureus و Bacillus cereus و همچنین برخی باکتری‌های گیاهی مانند Xanthomonas campestris و Pseudomonas aeruginosa به روش دیسک و تشکیل هاله مهار کننده استفاده شد. نتایج نشان داد که پروتئین کل گیاهان تراریخت در مقایسه با گیاه غیرتراریخت، به‌طور معنی‌داری (05/0P

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Cloning and expression of an omiganan pentahydrochloride (MBI226) antimicrobial peptide in tobacco (Nicotiana tabacum) confers resist to bacterial pathogens

نویسندگان [English]

  • Azam Badr Hadad 1
  • Farhad Nazarian Firouzabadi 2
  • Ahmad Ismaili 3
  • Hedayat Bagheri 4
1 Ph.D. Student, Department of Biotechnology, Agricultural Faculty, Lorestan University, Khorramabad, Iran
2 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran
3 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran
4 Assistant Professor, Department of Biotechnology Bu-Ali Sina University, Hamedan, Iran
چکیده [English]

Antimicrobial peptides are ancient and conserved molecules which are found in defense mechanisms of almost all living organisms from bacteria to animal and plant species. Identification and introduction of novel antimicrobial peptides, is a cost-effective way to improve crop plants resistance to pathogens by using recombinant DNA technology. Therefore, an expression construct containing omiganan antimicrobial encoding gene from the cytoplasmic granules of bovine neutrophils, was cloned and transferred to the tobacco leaf disk using Agrobacterium tumefaciens mediated-transformation. The presence of the antimicrobial peptide encoding gene in the genome of transgenic plants was confirmed by PCR analysis. Six putative transgenic lines and a non-transgenic control plant were selected for further molecular analysis. Total protein was extract from transgenic and non-transgenic control plants and used for antimicrobial activity assay against some human; E. coli, Salmonella, Staphylococcus, Bacillus, and plant: Xanthomonas and Pseudomonas pathogens by disc diffusion method. Results of this experiment showed that total protein extract from transgenic lines, as compared to non-transgenic plant, was significantly (P

کلیدواژه‌ها [English]

  • Antimicrobial peptides
  • Agrobacterium tumefaciens
  • pathogen
  • recombinant protein
  • Tobacco

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