لیپوپپتیدهای خانوادۀ ایتورین به‌عنوان ترکیب‌های کلیدی در خاصیت آنتاگونیستی باکتری UTB96 Bacillus subtilis علیه Aspergillus flavus

نوع مقاله: مقاله کامل


1 استادیار، پژوهشکدۀ کشاورزی هسته‌ای، پژوهشگاه علوم و فنون هسته‌ای، صندوق پستی 498-3148، کرج

2 استاد، گروه گیاه‌پزشکی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، صندوق پستی 77871-31587، کرج


اعضای جنس باسیلوس به‌عنوان باکتری­های آنتاگونیست امیدبخش برای کنترل زیستی (بیولوژیک) عامل‌های بیماریزای گیاهی شناخته شده­اند. در این تحقیق عصارۀ بدون یاخته‌ای سه سویه Bacillus subtilis UTB96، B. subtilis UTB1 وB. subtilis UTB70 علیه قارچ Aspergillus flavus R5 به روش کشت متقابل بررسی شد که سویۀ B. subtilis UTB96  بیشترین اثر بازدارندگی (05/0p≤) را نشان داد. رنگ‌نگاری (کروماتوگرافی) لایۀ ­­نازک با لیپوپپتیدهای استخراج‌شده از باکتری­ها، نشان‌دهندۀ حضور هر سه خانواده از لیپوپپتیدهای ایتورین، فنجایسین و سورفکتین در هر سه سویه است. تجزیه‌وتحلیل بیواتوگرافی کروماتوگرام­ها علیه قارچ A. flavus R5، نقش بالقوۀ ایتورین­ها را در فعالیت ضدقارچی سویۀ B. subtilis UTB96 مشخص کرد. به‌منظور تأیید نقش ایتورین­ها در فعالیت ضدقارچی سویۀ B. subtilis UTB96، ژن bmyB (سومین ژن در مسیر زیست‌ساخت یا بیوسنتز ایتورین­ها) به کمک جهش­زایی هدفمند تخریب شد. بررسی مولکولی شامل تجزیه‌وتحلیل PCR و توالی­یابی قطعۀ افزایش‌یافته در جهش‌یافته‌ها مشخص کرد که ژن مقاومت به آنتی‌بیوتیک اسپکتینومایسین جایگزین ژن هدف (bmyB) شده و به‌این‌ترتیب این ژن تخریب شده است. مقایسۀ فنوتیپی جهش­یافته­های bmyB با سویۀ مادری نشان داد که فعالیت ضدقارچی عصارۀ بدون یاخته‌ای و لیپوپپتیدهای استخراج‌شدۀ جهش­یافته­ها  در کشت متقابل باA. flavus ، تجزیه‌وتحلیل اتوبیوگرافی و نیز روی میوۀ پسته کاهش چشمگیری یافته است. بنابراین، یافته­های این پژوهش گویای نقش اصلی لیپوپپتیدهای خانوادۀ ایتورین در کنترل زیستی سویۀ B. subtilis UTB96  علیه A. flavus R5 است.



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