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لیپوپپتیدهای خانوادۀ ایتورین به‌عنوان ترکیب‌های کلیدی در خاصیت آنتاگونیستی باکتری UTB96 Bacillus subtilis علیه Aspergillus flavus | ||
| کنترل بیولوژیک آفات و بیماری های گیاهی | ||
| مقاله 8، دوره 5، شماره 1، خرداد 1395، صفحه 79-95 اصل مقاله (1.26 M) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22059/jbioc.2016.58619 | ||
| نویسندگان | ||
| حمیده افشارمنش1؛ مسعود احمدزاده* 2 | ||
| 1استادیار، پژوهشکدۀ کشاورزی هسته‌ای، پژوهشگاه علوم و فنون هسته‌ای، صندوق پستی 498-3148، کرج | ||
| 2استاد، گروه گیاه‌پزشکی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، صندوق پستی 77871-31587، کرج | ||
| چکیده | ||
| 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. | ||
| کلیدواژه‌ها | ||
| Aspergillus؛ Bacillus؛ کنترل بیولوژیک؛ لیپوپپتید | ||
| عنوان مقاله [English] | ||
| The Iturin Lipopeptides as Key Compounds in Antagonism of Bacillus subtilis UTB96 toward Aspergillus flavus | ||
| نویسندگان [English] | ||
| Hamideh Afsharmanesh1؛ Masoud Ahmadzadeh2 | ||
| چکیده [English] | ||
| Members of the genus Bacillus are among the most promising candidates for biological control of plant pathogenic bacteria. In this study, cell free supernatants of three strains B. subtilis UTB96, B. subtilis UTB1 and B. subtilis UTB70 against Aspergillus flavus R5 were investigated using dual culture assay. Strain B. subtilis UTB96 showed the highest inhibition effect. Thin lyer chromatography in extracted lipopeptides of the bacterial cell free supernatants proved the presence of the three families of lipopeptides iturin, fengycin and surfactin. Bioautography analysis of the chromatogarams against A. flavus R5 revealed the antifungal potential of iturins in strain B. subtilis UTB96. Gene bymB (the third gene in iturin biosynthesis) was disrupted usisng targeted mutagenesis to confirm role of iturins in antifungal activity. Molecular study (PCR analysis and sequencing the amplified fragment) of the mutants revealed that the targeted gene (bmyB) was replaced by spectinomycin resistance gene, hence the gene was disrupted. The phenotypical comparison of the bmyB mutants with the parental strain demonstrated that the antifungal activity of cell free supernatants and extracted lipopeptides was decreased interestingly in a dual culture assay with A. flavus, bioautography analysis and also on pistachio nuts. To sum up, these findings allowed us to conclude that iturin-like lipopeptides are indispensable for the biocontrol ability of the strain UTB96 against A. flavus R5. | ||
| کلیدواژه‌ها [English] | ||
| Aspergillus, Bacillus, biological control, lipopeptide | ||
| مراجع | ||
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