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 , میر ساسان میر پور ² , خسرو عیسی زاده ³ , بهنام راستی ⁴

1 - گروه میکروبیولوژی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد لاهیجان، لاهیجان، ایران
2 - گروه میکروبیولوژی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد لاهیجان، لاهیجان، ایران
3 - گروه میکروبیولوژی ، دانشکده علوم پایه، واحد لاهیجان، دانشگاه آزاد اسلامی، لاهیجان ، ایران
4 - گروه میکروبیولوژی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد لاهیجان، لاهیجان، ایران

تاریخ دریافت : 1402/01/20 تاریخ پذیرش : 1402/02/25 تاریخ انتشار : 1402/11/21

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.

چکیده انگلیسی:

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.

منابع و مأخذ:

Adamek M, Alanjary M, Sales-Ortells H, Goodfellow M, Bull A, Winkler A, Wibberg D, Kalinowski J, Ziemert N. Comparative genomics reveals phylogenetic distribution patterns of secondary metabolites in Amycolatopsis species.BMC Genomics .2018: 19(426):1-15.
Peng Q, Zhi-Xiang F, Jie-Wei T, Zu-Chao L, Lei W, Zhi-Gang Z, Yi-Wen C, Qiang T. Diversity, bioactivities, and metabolic potentials of endophytic actinomycetes isolated from traditional medicinal plants in Sichuan, China. Chinese Journal of Natural Medicines. 2015; 13(12): 0942- 0953.
Song Z, Xu T, Wang J, Hou Y, Liu C, Liu S, Wu S. Secondary Metabolites of the Genus Amycolatopsis: Structures, Bioactivities, and Biosynthesis.Molecules.2021;26(18):1-35.
Law JWF, Ser HL, Duangjai A, Sao Kaew S, Bukhari SI, Khan TM, et al. Streptomyces colonosanans sp. nov., a novel actinobacterium isolated from Malaysia mangrove soil exhibiting antioxidative activity and cytotoxic potential against human colon cancer cell lines. Frontiers in microbiology. 2017;8(1):877-891.
Takahashi Y, Nakashima T. Actinomycetes, an Inexhaustible Source of Naturally Occurring Antibiotics.Antibiotics Journal. 2018;7(45):1-17.
Ibnouf EO, Aldawsari MA, Waggiallah HA. Isolation and extraction of some compounds that act as antimicrobials from actinomycetes. Saudi Journal of Biological Sciences.2022; 29(8):1-7.
Tan LT-H, Chan K-G, Pusparajah P, Yin W-F, Khan TM, Lee L-H, et al. Mangrove-derived Streptomyces sp. MUM265 as a potential source of antioxidant and anti-cancer agents. BMC microbiology. 2019;19(1):1-16.
Kalaba MH, Moghannem SA, El-Hawary AS, Radwan AA, Sharaf MH, Shaban AS. Green synthesized ZnO nanoparticles mediated by Streptomyces plicatus: Characterizations, antimicrobial and nematicidal activities, and cytogenetic effects. Plants. 2021;10(9):1760-1786.
Ranjitha V, Ravishankar V. Extracellular synthesis of selenium nanoparticles from an actinomycetes Streptomyces griseoruber and evaluation of its cytotoxicity on HT-29 cell line. Pharmaceutical nanotechnology. 2018;6(1):61-68.
Fahmy NM, Abdel-Tawab AM. Isolation and characterization of marine sponge–associated Streptomyces sp. strain NMF6 producing secondary metabolite(s) possessing antimicrobial, antioxidant, anticancer, and antiviral activities. Journal of Genetic Engineering and Biotechnology. 2021;19(102):1-14.
Ansari MA, Alkubaisi N, Vijayaragavan P, Murugan P. Antimicrobial Potential of Streptomyces sp. To The Gram-Positive and Gram-Negative Pathogens. Journal of Infection and Public Health 2019; 12(1): 861-866.
Nabila MI, Kannabiran K. Biosynthesis, characterization and antibacterial activity of copper oxide nanoparticles (CuO NPs) from actinomycetes. Biocatalysis and agricultural biotechnology. 2018;15(1):56-62.
Oliveros KM,Rosana AR, Montecillo AD, Opulencia RB, Jacildo AJ, Zulaybar TO, Raymundo AK.Genomic Insights into the Antimicrobial and Anticancer Potential of Streptomyces sp. A1-08 Isolated from Volcanic Soils of Mount Mayon, Philippines. Philippine Journal of Science. 2021; 150 (6A): 1351-1377.
Osama N, Bakeer W, Raslan M, Soliman HA, Abdelmohsen UR, Sebak M. Anti-cancer and antimicrobial potential of five soil Streptomyces: a metabolomics-based study. Royal Society Open Science. 2022;9(2):1-17.
Li R, Wang M, Ren Z, Ji Y, Yin M, Zhou H, Tang SH-K. Amycolatopsis Aidingensis sp.nova., Halotolerant Actinobacterium, Produces New Secondary Metabolites. Frontiers in Microbiology.2021;12:1-14.
Kisil OV, Efimenko TA, Efremenkova OV. Looking Back to Amycolatopsis: History of the Antibiotic Discovery and Future Prospects. Antibiotics. 2021; 10(125): 1-25.
Kurnianto MA, Kusumaningrum HD, Lioe HN, Chasanah E. Antibacterial and antioxidant potential of ethyl acetate extract from Streptomyces AIA12 and AIA17 isolated from the gut of Chanos Chanos. BIODIVERSITAS .2021; 22(8):3196-3206.
Chen C, Ye Y, Wang R, Zhang Y, Wu C, Debnath S, Ma Z, Wang J, and Wu M. Streptomyces nigra sp. nov., is a novel actinobacterium isolated from mangrove soil and Exerts a Potent Antitumor Activity in Vitro. Frontiers in microbiology. 2018;9(1):1-14.
Alam M, Jha DK. Optimization of culture conditions for antimetabolite production by a rare tea garden actinobacterial isolate, Amycolatopsis sp. ST-28. African Journal of Clinical and Experimental Microbiology. 2019; 20 (3): 209-220.
Shaik M, Sankar G, Iswarya M, Rajitha P. Isolation and characterization of bioactivities of metabolites Producing marine Streptomyces parvulus Strain Sankarensis-A10. Journal Of Genetic Engineering and Biotechnology .2017;15,87-94.
Abd-Elnaby HM, Abo-Elala GM, Abdel-Raouf UM, Hamed MM. Antibacterial and anticancer activity of extracellular synthesized silver nanoparticles from marine Streptomyces rochei MHM13. The Egyptian Journal of Aquatic Research. 2016;42(3):301-312.
Nikbakht M, Omidi B, Amoozegar MA, Amini K. Cytotoxicity effect of secondary metabolites of Streptomyces Koyangensis and Streptomyces Tunisiensis isolated from saline soils of Garmsar City on human breast cancer cell line (MCF-7, IBRC C10082). Medical Science Journal of Islamic Azad Univesity-Tehran Medical Branch. 2021;31(4):367-376.
Sharma M, Manhas RK. Purification and characterization of actinomycins from Streptomyces strain M7 active against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus. BMC Microbiology. 2019;19(44):1-14.
Kawuri R, Darmayasa I. Bioactive Compound from Extract Filtrat Streptomyces sp. as Biocontrol of Vibriosis on Larvae of Macrobrachium Rosenbergii shrimps. Journal of Biosciences. 2019; 26(10):15-25.
Kumar R, Jadeja V. Isolation of Actinomycetes: A Complete Approach. International Journal of Current Microbiology and Applied Sciences. 2016;5(2): 606-618.
Gupta A, Singh D, Singh SK, Singh VK, Singh AV, Kumar A. Role of actinomycetes in bioactive and nanoparticle synthesis. Role of Plant Growth Promoting Microorganisms in Sustainable Agriculture and Nanotechnology: Elsevier. 2019; 4(1): 163-182.