جداسازی و شناسایی باکتری‌های اندوفیت از ریشه گیاه یونجه و بررسی اثرات ضد میکروبی آنها بر پاتوژن‌های انسانی
محورهای موضوعی :
میکروب شناسی
فاطمه منگلیان
1
(دانشجوی کارشناسی، گروه میکروبیولوژی، واحد سیرجان، دانشگاه آزاد اسلامی، سیرجان، ایران)
زهرا معصومعلی نژاد
2
(دکتری، گروه میکروبیولوژی، واحد سیرجان، دانشگاه آزاد اسلامی، سیرجان، ایران)
تاریخ دریافت : 1401/04/28
تاریخ پذیرش : 1401/06/17
تاریخ انتشار : 1401/07/01
کلید واژه:
اندوفیت,
باکتری‌های اندوفیت,
پاتوژن‌های انسانی,
خواص ضد باکتریایی,
گیاه یونجه,
چکیده مقاله :
هدف: یونجه با نام علمی Medicago sativa مهم‌ترین گیاه علوفه‌ای در ایران و بسیاری از نقاط جهان است. این گیاه به دلیل داشتن ارزش غذایی بالا و امکان کاشت در اقلیم‌های مختلف، به ملکه گیاهان علوفه‌ای مشهور است. هدف پژوهش حاضر جداسازی و شناسایی باکتری‌های اندوفیت از ریشه گیاه یونجه و بررسی اثرات آنتاگونیستی آنها بر باکتری‌های بیماری‌زای انسانی است.مواد و روش‌ها: قطعه‌های ریشه ضدعفونی شده گیاه یونجه پس از له شدن در محیط‌های کشت مایع تایوگلی کولات براث و مولر هینتون براث، جهت غنی‌سازی، کشت داده شدند. پس از کشت‌های مایع، بر روی محیط‌های کشت مولر هینتون آگار، ائوزین متیلن بلو و مک کانکی آگار کشت انجام شد. باکتری‌های اندوفیت با روش رنگ‌آمیزی گرم و تست‌های بیوشیمیایی، مورد شناسایی قرار گرفتند. فعالیت آنتاگونیستی باکتری‌های جدا شده به روش‌های استریک متقاطع و دولایه‌ای بررسی گردید. از باکتری‌های دارای اثرات آنتاگونیستی، استخراج DNA انجام شد و با کاربرد پرایمر یونیورسال، تکثیر ژن SrRNA16 صورت پذیرفت. محصول PCR تعیین توالی شده و درخت فیلوژنی رسم گردید.یافته‌ها: نتایج نشان داد باکتری‌های اندوفیت یونجه دارای فعالیت آنتاگونیستی بودند و در نهایت دو اندوفیت از لحاظ هویت مولکولی مورد بررسی قرار گرفتند که متعلق به جنس‌های انتروباکتر و سودوموناس بودند.نتیجه‌گیری: براساس نتایج حاصل، باکتری‌های اندوفیت دارای تنوع زیاد و اثرات قابل توجه ضد باکتریایی می‌باشند.
چکیده انگلیسی:
Objectives: alfalfa with the scientific name Medicago sativa is the most important fodder plant in Iran and many parts of the world. This plant is known as the queen of fodder plants due to its high nutritional value and the possibility of planting in different climates. The aim of the current research is to isolate and identify endophytic bacteria from alfalfa roots and investigate their antagonistic effects on human pathogenic bacteria.Materials and methods: The sterilized alfalfa root pieces, after being crushed, were cultured in Tyoglycollate Broth and Muller Hinton Broth liquid culture media for enrichment. After liquid cultures, culture was done on Muller Hinton agar, eosin methylene blue and McConkey agar. Endophytic bacteria were identified by gram staining method and biochemical tests. The antagonistic activity of isolated bacteria was investigated by steric crossing and bilayer methods. From the bacteria with antagonistic effects, DNA extraction was performed and with the use of universal primer, 16SrRNA gene was amplified. The PCR product was sequenced and a phylogeny tree was drawn.Findings: The results showed that alfalfa endophyte bacteria had antagonistic activity and finally, two endophytes were investigated in terms of molecular identity, which belonged to Enterobacter and Pseudomonas genera.Conclusions: Based on the results, endophytic bacteria have a great diversity and significant antibacterial effects.
منابع و مأخذ:
Tiwari P, Kang S & Bae H. Plant-endophyte associations: Rich yet under-explored sources of novel bioactive molecules and applications. Microbiological Research. 2022; Oct 17: 127241.
Strobel G. The emergence of endophytic microbes and their biological promise. Journal of Fungi. 2018; 4(2): 57.
Yadav G & Meena M. Bioprospecting of endophytes in medicinal plants of Thar Desert: An attractive resource for biopharmaceuticals. Biotechnology Reports. 2021; 30: e00629.
Del Carmen Orozco-Mosqueda M & Santoyo G. Plant-microbial endophytes interactions: scrutinizing their beneficial mechanisms from genomic explorations. Current Plant Biology. 2021; 25: 100189.
Harshitha K, Nair AR & Pillai PP. Overview of bioactive metabolite (s) produced by endophytes and future perspectives on epigenetic modification/regulation of cryptic biosynthetic pathways. Phytochemistry Letters. 2023; 53: 116-31.
Gao H, Li G & Lou HX. Structural diversity and biological activities of novel secondary metabolites from endophytes. Molecules. 2018; 23(3): 646.
Sharma P & Kumar S. Bioremediation of heavy metals from industrial effluents by endophytes and their metabolic activity: Recent advances. Bioresource Technology. 2021; 339: 125589.
Tanvir R, Javeed A & Bajwa AG. Endophyte bioprospecting in South Asian medicinal plants: an attractive resource for biopharmaceuticals. Applied microbiology and biotechnology. 2017; 101(5): 1831-44.
Pasrija P, Girdhar M, Kumar M, Arora S & Katyal A. Endophytes: an unexplored treasure to combat Multidrug resistance. Phytomedicine Plus. 2022: 100249.
Toghueo RM & Boyom FF. Endophytes from ethno-pharmacological plants: Sources of novel antioxidants-A systematic review. Biocatalysis and Agricultural Biotechnology. 2019; 22: 101430.
Omidi Nasab M & Khodakaramian G. Inhibitation of alfalfa endophytic bacteria against Clavibacter michiganensis subsp. insidiosus causal agent of wilt disease in in vitro and greenhouse conditions. BioControl in Plant Protection. 2017; 5(1): 1-3.
Hawkins C & Yu LX. Recent progress in alfalfa (Medicago sativa L.) genomics and genomic selection. The Crop Journal. 2018; 6(6): 565-75.
Attia EZ, Farouk HM, Abdelmohsen UR & Mo'men H. Antimicrobial and extracellular oxidative enzyme activities of endophytic fungi isolated from alfalfa (Medicago sativa) assisted by metabolic profiling. South African Journal of Botany. 2020; 134: 156-62.
El-Deeb B, Fayez K & Gherbawy Y. Isolation and characterization of endophytic bacteria from Plectranthus tenuiflorus medicinal plant in Saudi Arabia desert and their antimicrobial activities. Journal of plant interactions. 2013; 8(1): 56-64.
Chowdhary K & Kaushik N. Biodiversity and In vitro inhibition study of fungal endophytes of Chlorophytum borivilianum against selected phytopathogens. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2019; 89(1): 113-21.
Carrim A, Ribeiro ID, Araújo MV, Oliveira B & Vieira JD. Atividade antimicrobiana de bactérias endofíticas isoladas de Jacaranda decurrens Cham (carobinha-do-campo). Enciclopédia Biosfera. 2016; 13(23).
López JL, Alvarez F, Príncipe A, Salas ME, Lozano MJ, Draghi WO, Jofré E & Lagares A. Isolation, taxonomic analysis, and phenotypic characterization of bacterial endophytes present in alfalfa (Medicago sativa) seeds. Journal of biotechnology. 2018; 267: 55-62.
Hawkins C & Yu LX. Recent progress in alfalfa (Medicago sativa L.) genomics and genomic selection. The Crop Journal. 2018; 6(6): 565-75.
Ashitha A, Midhun SJ, Sunil MA, Nithin TU, Radhakrishnan EK & Mathew J. Bacterial endophytes from Artemisia nilagirica (Clarke) Pamp., with antibacterial efficacy against human pathogens. Microbial pathogenesis. 2019; 135: 103624.
Myo EM, Maung CE, Mya KM & Khai AA. Characterization of bacterial endophytes from Myanmar medicinal plants for antimicrobial activity against human and plant pathogens. Brazilian Journal of Pharmaceutical Sciences. 2020; 56.
Kaur N & Arora DS. Prospecting the antimicrobial and antibiofilm potential of Chaetomium globosum an endophytic fungus from Moringa oleifera. AMB Express. 2020; 10(1): 1-3.
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Tiwari P, Kang S & Bae H. Plant-endophyte associations: Rich yet under-explored sources of novel bioactive molecules and applications. Microbiological Research. 2022; Oct 17: 127241.
Strobel G. The emergence of endophytic microbes and their biological promise. Journal of Fungi. 2018; 4(2): 57.
Yadav G & Meena M. Bioprospecting of endophytes in medicinal plants of Thar Desert: An attractive resource for biopharmaceuticals. Biotechnology Reports. 2021; 30: e00629.
Del Carmen Orozco-Mosqueda M & Santoyo G. Plant-microbial endophytes interactions: scrutinizing their beneficial mechanisms from genomic explorations. Current Plant Biology. 2021; 25: 100189.
Harshitha K, Nair AR & Pillai PP. Overview of bioactive metabolite (s) produced by endophytes and future perspectives on epigenetic modification/regulation of cryptic biosynthetic pathways. Phytochemistry Letters. 2023; 53: 116-31.
Gao H, Li G & Lou HX. Structural diversity and biological activities of novel secondary metabolites from endophytes. Molecules. 2018; 23(3): 646.
Sharma P & Kumar S. Bioremediation of heavy metals from industrial effluents by endophytes and their metabolic activity: Recent advances. Bioresource Technology. 2021; 339: 125589.
Tanvir R, Javeed A & Bajwa AG. Endophyte bioprospecting in South Asian medicinal plants: an attractive resource for biopharmaceuticals. Applied microbiology and biotechnology. 2017; 101(5): 1831-44.
Pasrija P, Girdhar M, Kumar M, Arora S & Katyal A. Endophytes: an unexplored treasure to combat Multidrug resistance. Phytomedicine Plus. 2022: 100249.
Toghueo RM & Boyom FF. Endophytes from ethno-pharmacological plants: Sources of novel antioxidants-A systematic review. Biocatalysis and Agricultural Biotechnology. 2019; 22: 101430.
Omidi Nasab M & Khodakaramian G. Inhibitation of alfalfa endophytic bacteria against Clavibacter michiganensis subsp. insidiosus causal agent of wilt disease in in vitro and greenhouse conditions. BioControl in Plant Protection. 2017; 5(1): 1-3.
Hawkins C & Yu LX. Recent progress in alfalfa (Medicago sativa L.) genomics and genomic selection. The Crop Journal. 2018; 6(6): 565-75.
Attia EZ, Farouk HM, Abdelmohsen UR & Mo'men H. Antimicrobial and extracellular oxidative enzyme activities of endophytic fungi isolated from alfalfa (Medicago sativa) assisted by metabolic profiling. South African Journal of Botany. 2020; 134: 156-62.
El-Deeb B, Fayez K & Gherbawy Y. Isolation and characterization of endophytic bacteria from Plectranthus tenuiflorus medicinal plant in Saudi Arabia desert and their antimicrobial activities. Journal of plant interactions. 2013; 8(1): 56-64.
Chowdhary K & Kaushik N. Biodiversity and In vitro inhibition study of fungal endophytes of Chlorophytum borivilianum against selected phytopathogens. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences. 2019; 89(1): 113-21.
Carrim A, Ribeiro ID, Araújo MV, Oliveira B & Vieira JD. Atividade antimicrobiana de bactérias endofíticas isoladas de Jacaranda decurrens Cham (carobinha-do-campo). Enciclopédia Biosfera. 2016; 13(23).
López JL, Alvarez F, Príncipe A, Salas ME, Lozano MJ, Draghi WO, Jofré E & Lagares A. Isolation, taxonomic analysis, and phenotypic characterization of bacterial endophytes present in alfalfa (Medicago sativa) seeds. Journal of biotechnology. 2018; 267: 55-62.
Hawkins C & Yu LX. Recent progress in alfalfa (Medicago sativa L.) genomics and genomic selection. The Crop Journal. 2018; 6(6): 565-75.
Ashitha A, Midhun SJ, Sunil MA, Nithin TU, Radhakrishnan EK & Mathew J. Bacterial endophytes from Artemisia nilagirica (Clarke) Pamp., with antibacterial efficacy against human pathogens. Microbial pathogenesis. 2019; 135: 103624.
Myo EM, Maung CE, Mya KM & Khai AA. Characterization of bacterial endophytes from Myanmar medicinal plants for antimicrobial activity against human and plant pathogens. Brazilian Journal of Pharmaceutical Sciences. 2020; 56.
Kaur N & Arora DS. Prospecting the antimicrobial and antibiofilm potential of Chaetomium globosum an endophytic fungus from Moringa oleifera. AMB Express. 2020; 10(1): 1-3.
