جداسازی و شناسایی باکتری‌های اندوفیت گیاه دارویی مرزه خوزستانی (Satureja khuzestanica) و ارزیابی متابولیت‌های ثانویه آن‌ها در فعالیت ضدمیکروبی علیه باکتری‌های بیماری‌زای گیاهی

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانشجوی دکتری بیماری شناسی گیاهی دانشکده کشاورزی دانشگاه بوعلی سینا همدان، همدان، ایران

2 استاد گروه گیاه‌پزشکی، دانشکده کشاورزی دانشگاه بوعلی سینا همدان، همدان، ایران

چکیده

مرزه خوزستانی (Satureja khuzestanica) گیاه معطری است که به دلیل محتوای متابولیت‌های ثانویه فعال زیستی که در طب سنتی قابل استفاده هستند، توجه بسیاری از محققین را به خود جلب کرده‌است. با این حال، اطلاعات در مورد اندوفیت‌های باکتریایی مرتبط با مرزه خوزستانی (S. khuzestanica) محدود است. در مطالعه حاضر، 17 اندوفیت باکتریایی از گیاه دارویی S. khuzestanica جداسازی شد. مطالعات آنتاگونیستی این جدایه‌ها برابر سه باکتری بیماری‌زای گیاه،Ralstonia solanacearum ،Pectobacterium carotovorum subsp. carotovora  و Clavibacter insidiosus در شرایط آزمایشگاهی روی محیط کشت Tryptic Soy Broth به روش کشت سه نقطه‌ای بر اساس آزمایش فاکتوریل در قالب طرح کامل تصادفی در سه تکرار انجام شد. داده‌های حاصل با استفاده از نرم‌افزار SAS تجزیه و تحلیل شدند و مقایسه میانگین‌ها از طریق آزمون چند دامنه‌ای دانکن انجام و نماینده‌ها انتخاب شدند. اندوفیت‌های باکتریایی با تعیین توالی ژن 16S rRNA شناسایی شدند و به سه جنس Bacillus, Pseudomonas و Streptomyces تعلق داشتند. متابولیت‌های این باکتری‌ها به وسیله اتیل استات استخراج و آنالیز گاز کروماتوگرافی- طیف‌سنجی جرمی عصاره (GC-MS) با روش‌های استاندارد انجام شد. نتایج آزمون حداقل غلظت مهاری (MIC) با استفاده از روش میکرودایلوشن، نشان دهنده فعالیت ضد میکروبی با غلظت مهار قابل توجهی از 5/2-312/0 میلی‌گرم در میلی‌لیتر بودند. یافته‌های ما بینش جدیدی را در مورد فعالیت‌های ضد میکروبی اندوفیت‌های باکتریایی از S. khuzestanica ارائه می‌دهد.

کلیدواژه‌ها

عنوان مقاله [English]

Isolation and identification of endophytic bacteria from Satureja Khuzestanica and evaluation of its secondary metabolites for antimicrobial activity against plant pathogenic bacteria

نویسندگان [English]

  • M. Omidi Nasab 1
  • G. Khodakaramian 2
1 PhD candidate Plant Pathology, Faculty of Agriculture, Bu Ali Sina University, Hamadan, Hamadan, Iran
2 Professor of Department of Plant Pathology, Faculty of Agriculture, Bu Ali Sina University, Hamadan, Hamadan, Iran
چکیده [English]

Background and Objectives
In many parts of the world, medicinal plants have been used as an alternative medicine to promote human health and longevity since ancient times. Microbes residing within plant tissues are known as endophytes. The compounds produced by these microbes have the potential to be employed in modern medicine, agriculture, and other industries. Endophytic bacteria isolated from medicinal plants are valuable sources of novel bioactive compounds with diverse activities, including antimicrobial, anticancer, and antiviral properties. This study aimed to isolate, identify, and screen endophyte bacteria with antimicrobial activity against plant pathogenic bacteria. Aromatic plants such as Satureja khuzestanica are utilized in traditional medicine due to their secondary metabolites, but data regarding its naturally occurring bacterial endophytes is limited.
Materials and Methods
In the current study, 17 strains of bacterial endophytes were isolated in a Tryptic Soy Agar medium from the medicinal plant Satureja khuzestanica. Based on sequencing the 16S rRNA encoding gene, researchers isolated bacterial strains from the Bacillus, Streptomyces, and Pseudomonas genera with the highest activity against plant pathogenic bacteria. Under standard conditions, these endophytic bacteria’s bioactive secondary metabolites were extracted with ethyl acetate and analyzed using Gas Chromatography-Mass Spectrometry (GC-MS). The mass spectra of the compounds were compared to the National Institute of Standards and Technology (NIST) library’s database. Moreover, microbroth dilution techniques were used to determine the minimum inhibitory concentration (MIC) values for five different bacteria species.
Results
The GC-MS analysis revealed the presence of a number of compounds, including dibutyl phthalate (DBP), eicosane, octadecanoic acid, hexadecanoic acid, and hexadecane 1,4- dicyclohexylbutane. Four of the selected bacterial endophytes exhibited antimicrobial activity against three plant pathogens: Ralstonia solanacearum, Pectobacterium carotovorum subsp., carotovorum, and Clavibacter insidiosus. Data analysis revealed significant differences in antimicrobial activity, with the minimum inhibitor concentration ranging from 0.312 mg/ml to 2. 5 mg/ml. Furthermore, we identified bioactive secondary metabolites with reported biological activities in antimicrobial, anti-inflammatory, and antioxidant properties with biotechnological applications in medicine, agriculture, and other industries based on an endophytic crude extract data analysis.
Conclusion
Our findings shed new light on the antimicrobial properties of naturally occurring bacterial endophytes in S. khuzestanica.
 

کلیدواژه‌ها [English]

  • antioxidant
  • crude extract
  • biological activity
  • pathogenic agents

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گیاه پزشکی
دوره 45، شماره 3
مهر 1401
صفحه 43-62

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