شناسایی ژن‌های احتمالی مرتبط با سیستم ضد احساس حد نصاب (توانایی کوئنچری) در باکتری‌های دارای قابلیت مهار زیستی

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 استادیار دانشگاه ایلام

2 استادیار، دانشگاه لرستان

چکیده

یکی از راه­ های جدید کنترل باکتری‌ های بیمارگر، استفاده از باکتری ­های دارای توانایی مهار زیستی مبتنی بر سیستم ضداحساس حد نصاب (توانایی کوئنچری) است. بر این اساس جدایه­ های باکتریایی دارای قدرت کوئنچری از ریزوسفر و فیلوسفر گیاهان مختلف جداسازی گردیدند. در بین جدایه ­های باکتریایی مورد بررسی، چهار جنس و گونه باکتریایی شامل Pseudomonas chlororaphis، Pseudomonas putida،Acinetobacter sp .  و Bacillus sp . به عنوان جدایه ­های باکتریایی برتر با توانایی کوئنچری و قابلیت مهار زیستی بیمارگرهای باکتریایی جنس Pectobacterium معرفی شدند. فراوان­ترین­ و قوی­ترین­ جدایه ­های دارای توانایی کوئنچری، فعالیت آنزیمی مرتبط با این پدیده از خود نشان دادند. سپس به منظور شناسایی ژن ­های آسیلازی و لاکتونازی این باکتری ­ها، آغازگرهایی طراحی و مورد آزمایش قرار گرفت. بر اساس نتایج حاصل از بررسی آغازگرها، در جنس باسیلوس ژن لاکتونازی aiia، در جدایه­­های  p . chlororaphis سه ژن آسیلازی همولوگ pvdQ، quip  و  hacB(یا فقط دو ژن اول)، درP. putida دو ژن همولوگ pvdQ  وquip  و در Acinetobacter یک ژن آسیلازی ردیابی گردید.  

کلیدواژه‌ها


عنوان مقاله [English]

Detection and identification of potentially anti-quorum sensing related genes in biocontrol bacteria

نویسندگان [English]

  • M. R. Alimanesh 1
  • H. Mirzaei Najafgholi 2
1 Assistant Professor, Ilam University
2 Assistant Professor, Lorestan University
چکیده [English]

Background and Objectives
Quorum sensing (QS) is a process of cell to cell communication that allows bacteria to be aware about cell density. In this phenomenon bacteria communicate with each other through signaling molecules such as N-acylhomoserine lactones (AHLs). Also, virulence gene expression in many pathogenic bacteria like Pectobacterium is under control of QS. Some bacteria can degrade AHLs molecules by a process called anti quorum sensing or quorum quenching (QQ); therefore, QQ can be used in biocontrol of plant pathogenic bacteria. The main purpose of this research was to detect QQ genes in strong QQ-based biocontrol bacteria.
Materials and Methods
In this investigation, QQ bacteria were isolated from rhizosphere and phyllosphere of some agricultural and non-agricultural plants using minimum media containing AHL. QQ bacteria were identified using biosensors CV026 and VIR07. Mechanism of QQ action was determined by heating, proteinase treatment and filtering methods. Afterwards, in order to find acylase and lactonase genes in these bacteria, homology searches were performed using BLASTn and BLASTp in NCBI. Then, several primers were designed by several softwares including CLC main workbench 5.5, Primer Premier 6, Primer3, Oligo7 and primer BLAST.
Results
The most abundant and strongest QQ isolates were shown enzymatic activity. Main twenty-seven isolates with high QQ activity were detected. Four genera and species including Pseudomonas chlororaphis, Pseudomonas putida, Acinetobacter sp < em>. and Bacillus sp < em>. were found as the bacteria with highest QQ and biocontrol ability against Pectobacterium. As a results of this study, in the Bacillus Aiia lactonase gene;in p < /em>. chlororaphis three acylase genes of pvdQ، quip < /em> and hacB; in P.putida two acylase genes of  pvdQ and quip < /em>  and in Acinetobacter one acylase genes were traced.
Discussion
According to the results of this investigation, Pseudomonas genus was determined as the frequent QQ bacteria. Also, p < /em>. chlororaphis species with the strongest QQ activity, were divided in to two groups; the first isolates with two acylase genes (pvdQ and quip < /em>) and the second isolates containing extra homologous acylase hacB. Isolates of the latter group with the most detected genes involved in QQ, were identified as the best QQ-based biocontrol bacteria against Pectobacterium.

کلیدواژه‌ها [English]

  • Cell communication
  • Pseudomonas
  • Pectobacterium
  • acylase and lactonase genes
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