جداسازی و شناسایی ویژگیهای ضد میکروبی اکتینومیستهای بومی جدا شده از خاک های کشاورزی استان گیلان
محورهای موضوعی : Biotechnological Journal of Environmental Microorganisms
الهام امیری
1
(گروه میکروبیولوژی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد لاهیجان، لاهیجان، ایران)
میر ساسان میر پور
2
(گروه میکروبیولوژی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد لاهیجان، لاهیجان، ایران)
خسرو عیسی زاده
3
(گروه میکروبیولوژی ، دانشکده علوم پایه، واحد لاهیجان، دانشگاه آزاد اسلامی، لاهیجان ، ایران)
بهنام راستی
4
(گروه میکروبیولوژی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد لاهیجان، لاهیجان، ایران)
کلید واژه: اکتینومیست, فعالیت ضد میکروبی, 16S rRNA, آمیکولاتوپسیس, استرپتومایسس,
چکیده مقاله :
اکتینومیست ها به عنوان بزرگترین مخزن آنتی بیوتیک های طبیعی در جهان شناخته می شوند. هدف از این تحقیق جداسازی و شناسایی مولکولی اکتینومیست ها با خاصیت ضدمیکروبی ازخاکهای کشاورزی مناطق بومی استان گیلان می باشد. نمونه های خاک از مناطق کشاورزی جنوب غربی استان گیلان جمع آوری شدند. برای جداسازی اکتینومیست ها از رقیق سازی متوالی استفاده گردید. سپس به شناسایی مورفولوژی، فیزیولوژی و بیوشیمیایی نمونه ها پرداخته شد و در نهایت شناسایی مولکولی ایزوله ها با استفاده از توالی یابی 16S rRNA و تجزیه و تحلیل فیلوژنتیک انجام گرفت. بررسی فعالیت ضدمیکروبی در برابر میکروارگانیسم های پاتوژن انجام گردید. در مجموع 14 ایزوله شناسایی شد. 2 ایزوله با خاصیت ضد میکروبی بیشتر انتخاب شد. براساس نتایج مطالعات فیلوژنتیکی و توالی یابی 16S rRNA سویه Amycolatopsis roodepoortensis Strain EA7 با 63/99% اطمینان و سویه EA6 Streptomyces Microfluve Strain با 92/93 % اطمینان شناسایی شدند.باکتری های جدا سازی شده دارای فعالیت ضدمیکروبی بیشترعلیه میکروارگانیسم های پاتوژن گرم مثبت Staphylococcus aureus و نمونه استاندارد PTCC 1112 بودند. این تحقیق اولین گزارش از شناسایی اکتینومیست ها با خاصیت ضد میکروبی در خاکهای کشاورزی مناطق جنوب غربی استان گیلان واقع در کوههای البرز می باشد. شناسایی سویه نادرroodepoortensis Strain EA7 Amycolatopsis از مناطق شمالی ایران خاک های این مناطق را بسیار ارزشمند می سازد.
Actinomycetes are known as the largest reservoir of natural antibiotics in the world. For this reason, due to their ability to produce various antibiotics and other compounds of therapeutic importance, they are considered the golden microorganisms of the 21st century. The purpose of this research is the isolation and molecular identification of actinomycetes with antimicrobial properties from agricultural soils in the native areas of Guilan province. Soil samples were collected from the southwestern agricultural areas of Guilan province. Serial dilution was used to isolate actinomycetes. Then the morphological, physiological, and biochemical identification of the samples was done and finally, the molecular identification of the isolates was done using 16S rRNA sequencing and phylogenetic analysis. Antimicrobial activity was investigated against pathogenic microorganisms. A total of 14 isolates were identified.2 isolates with more antimicrobial properties were selected. Based on the results of phylogenetic studies and 16S rRNA sequencing, Amycolatopsis roodepoortensis strain EA7 with 99.63% confidence, and Streptomyces microflaveus strain EA6 with 93.92% confidence were identified. The isolated bacteria had more antimicrobial activity against Gram-positive pathogenic microorganisms Staphylococcus aureus and standard sample Staphylococcus aureus PTCC 1112. This research is the first report on the identification of actinomycetes with antimicrobial properties in the agricultural soils of the southwestern regions of Guilan province located in the Alborz mountains. The identification of the rare strain of Amycolatopsis roodepoortensis strain EA7 from the northern regions of Iran makes the soils of these regions very valuable.
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