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بررسی دخالت در تنظیم ژنی وابسته به حدنصاب احساس در Pectobacterium atrosepticum و تغییر فعالیت LuxI و LuxR توسط ریزوباکتریهای تجزیهکننده ان-اسیل‌هموسرین‌لاکتون | ||
| کنترل بیولوژیک آفات و بیماری های گیاهی | ||
| مقاله 2، دوره 6، شماره 1، خرداد 1396، صفحه 11-21 اصل مقاله (661.66 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22059/jbioc.2017.217827.172 | ||
| نویسندگان | ||
| عادله سبحانی پور1؛ کیوان بهبودی* 2؛ اسماعیل محمودی3 | ||
| 1استادیار، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه آزاد اسلامی واحد دامغان | ||
| 2دانشیار، گروه گیاهپزشکی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج | ||
| 3استادیار، گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه آزاد اسلامی واحد اصفهان و قطب علمی ترانسژنز، دانشگاه آزاد اسلامی واحد اصفهان (خوراسگان)، اصفهان | ||
| چکیده | ||
| سیستم حدنصاب احساس (Quorum sensing) یک سیستم ارتباطی است که به باکتریها اجازه میدهد تا تراکم جمعیتشان را از طریق تولید و ردیابی مولکولهای پیامرسان بسنجند. در باکتریهای گرم منفی این مولکولها شامل ان-اسیلهموسرینلاکتونها (AHLs) است که توسط پروتئینهای سازنده (LuxI) ساخته میشود و سپس به بیرون از سلول ترشح شده و به پروتئینهای گیرنده (LuxR) در باکتریهای مجاور متصل میشوند و رونویسی از ژنهای هدف را کم یا زیاد میکنند. تأثیر 66 ریزوباکتری تجزیهکننده AHL بر تغییر فعالیت و کارایی LuxI/LuxR و یا بازداری از سنتز AHL با استفاده از سیستم انتشار در آگار و Chromobacterium violaceum CV026 بهعنوان گزارشگر مورد ارزیابی قرار گرفت. جدایه Pectobacterium atrosepticum SRI1043 نیز بهعنوان مولد AHL استفاده شد. برای بررسی توانایی بازدارندگی از فعالیت LuxI، جدایه مولد AHL در نزدیکی باکتریهای خاموشکننده و باکتری گزارشگر دورتر از آن قرار گرفت. برای بررسی بازداری از فعالیت LuxR، جای استرینهای مولد AHL و گزارشگر عوض شد. هر دو خاصیت ممانعت از فعالیت LuxI و LuxR در 21 جدایه مشاهده شد. این جدایهها بر اساس تستهای بیوشیمیایی و بررسی توالی 16srDNA در جنسهای Bacillus، Brevundimonas، Citrobacter، Bordetella، Acinetobacter، Stenotrophomonas، Pseudomonas و Escherchia قرار گرفتند. برای بررسی دخالت در تنظیم ژنی وابسته به AHL در P. atrosepticum، باکتریهای آنتاگونیست با فاصله کمی از بیمارگر کشت شد و کاهش تولید آنزیم پروتئاز با مشاهده کاهش هاله شفاف اطراف این بیمارگر روی محیط کشت شیر خشک بدون چربی-آگار (SMA) مشخص شد. پنج جدایه تا حد زیادی تولید پروتئاز را کاهش دادند. | ||
| کلیدواژه‌ها | ||
| AHL؛ CV026؛ حدنصاب احساس | ||
| عنوان مقاله [English] | ||
| Bioassay for interference of AHL-dependent gene regulation in Pectobacterium atrosepticum and modulation of LuxI and LuxR by N-acylhomoserine lactone degrading rhizobacteria | ||
| نویسندگان [English] | ||
| Adeleh Sobhanipour1؛ Keivan Behboudi2؛ Esmaeil Mahmoudi3 | ||
| 1Assistant Professor, Department of Plant Protection, Damghan Branch, Islamic Azad University, Iran | ||
| 2Associate Professor, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran | ||
| 3Assistant Professor, Department of Plant Protection, Isfahan (Khorasgan) Branch, Islamic Azad University, Iran and Transgenesis Center of Excellence, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan | ||
| چکیده [English] | ||
| Quorum sensing is a communication system that allows bacteria to monitor their population density through the production and sensing of signal molecules. In Gram-negative bacteria, signal molecules include acyl-homoserine lactones (AHLs) which are synthesized by the protein synthases (LuxI). The synthesized signaling molecules are secreted out of the cell and bind with receptor proteins (LuxR) of neighboring bacterial and decrease or increase the rate of transcription in target genes. The effect of the 66 AHL-degrading rhizobacteria (quencher) on modulation of LuxI/LuxR activity and inhibition of AHL synthesis was determined using an agar diffusion double streak assay and the Chromobacterium violaceum CV026 biosensor system. The isolate Pectobacterium atrosepticum SRI1043 used as AHL-producer. To test for potential LuxI inhibition, the AHL–producer isolate was placed in close proximity to the test quencher bacteria and the AHL biosensor strain placed distantly. In order to test for LuxR inhibition, the location of the AHL–producer and biosensor strains was reversed. Both LuxI and LuxR modulation was observed in 21 isolates. These isolates were identified as genera Bacillus, Brevundimonas, Citrobacter, Bordetella, Acinetobacter, Stenotrophomonas, Pseudomonas and Escherichia using biochemical tests and 16s rDNA sequencing. Bioassay for interference of AHL-dependent gene regulation in P. atrosepticum performed on Skim Milk Agar medium and antagonistic bacteria cultured near the pathogen and the reduction of protease production by this pathogen was evaluated by the reduction of clear zone in the agar. Five isolates reduced the protease production significantly. | ||
| کلیدواژه‌ها [English] | ||
| AHL, CV026, Quorum Sensing | ||
| مراجع | ||
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