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بررسی اثرات بازدارندگی برخی پپتیدهای حشرات علیه باکتری Pseudomonas tolaasii عامل بیماری لکه قهوه‌ای قارچ سفید دکمه ای | ||
| مهار زیستی در گیاه‌پزشکی | ||
| دوره 11، شماره 1، شهریور 1402، صفحه 109-123 اصل مقاله (740.21 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/bcpp.2024.365978.368 | ||
| نویسندگان | ||
| افسانه مسلمی مهنی1؛ محمد رهنمائیان* 2؛ نادر حسن زاده3؛ حمید فروتن فر4 | ||
| 1-دانشجوی دکتری رشته بیماری شناسی گیاهی، دانشکده کشاورزی و صنایع غذایی، واحد علوم و تحقیقات دانشگاه آزاد اسلامی، تهران، ایران | ||
| 2بخش شیمی دارویی، دانشکده داروسازی، دانشگاه فیلیپس، ماربورگ، آلمان. | ||
| 3دانشیار، گروه گیاه‌پزشکی، دانشکده کشاورزی و صنایع غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران. | ||
| 4استاد، مرکز تحقیقات علوم دارویی و فراورده های آرایشی، دانشگاه علوم پزشکی کرمان، کرمان، ایران. | ||
| چکیده | ||
| باکتری Pseudomonas tolaasii عامل بیماری لکه قهوه‌ای قارچ خوراکی از مهمترین عوامل محدود‌کننده تولید این محصول می‌‌باشد که تاثیر مستقیمی بر کاهش کیفیت محصول دارد. . باتوجه به عدم وجود راه حل مناسب در مدیریت این بیماری با کاربرد سموم شیمایی و آنتی بیوتیک‌ها ،که اثرات ناخواسته سمیت و مقاومت میکروبی را در پی دارند، تحقیقات گسترده‌ای در زمینه شناسایی ترکیبات ضد‌باکتری طبیعی و سازگار با سلامت محصول، درحال انجام است. در تحقیق حاضر نیز بدین منظور، پس از جداسازی و شناسایی گونه بیمارگر باکتری Pseudomonas tolaasii با روشهای معمول آزمایشگاهی، اثرات ضد‌باکتریایی ده پپتید حاصل از گونه‌های مختلف حشرات، نظیر پروانه سکروپیا، مورچه مرمیسیا، موریانه، زنبور عسل اروپایی، مگس میوه مدیترانه‌ای، مگس اسطبل و مگس گوشت، علیه این بیماری مورد بررسی قرار گرفتند. مقادیر حداقل غلظت بازدارندگی از رشد به میزان 50% (MIC) مقادیر حداقل غلظت کشندگی (MBC)پپتیدهای مورد نظر علیه باکتری بیمارگر به روش تست الایزا به‌دست آمد. برای تایید نتایج MBC محتوی میکروپلیت‌ها مجدد کشت شدند. جهت ارزیابی شدت و کیفیت علایم بیماری در محیط آزمایشگاهی، سطح بلوک‌های قارچ با غلظت1mM از پپتیدهای موثر پوشانیده شد و سپس با سوسپانسیون باکتری حاوی 108 cfu/ml مایه زنی شدند. پپتیدها با توجه به نوع بروز علایم در شش گروه متفاوت (B0-B1-B2-B3-B--B5) قرار گرفتند. سطوح مختلف شدت بروز علایم بیماری، از شاخص بدون علامت (B0) تا شاخص علایم پوسیدگی قهوه‌ای همراه با نقاط فرورفته(B5) مورد ارزیابی قرار گرفت. مقادیر MIC نشان داد که پپتیدهای Cecropin A(پروانه سکروپیا)، Scal-stomoxyn(مگس اسطبل)، Lser-Cecropin1(مگس گوشت) در غلظت‌های متفاوتی سبب کاهش رشد حداقل 50% جمعیت باکتری P. tolaasii شدند. کمترین مقدار MBC نیز در پپتید Cecropin A در غلظت µM 6.25 مشاهده شد. در نتایج بررسی کیفیت و شدت بروز علایم، پپتید Cecropin A در سطح B0، بطور کامل کلاهک قارچ را در شرایط آزمایشگاهی علیه باکتری P. tolaasiiمحافظت نمود و از بروز علایم بیماری جلوگیری کرد. پس از آن پپتید Scal-stomoxyn با ایجاد لکه‌های مختصر زرد رنگ در سطح شاخص B1 قرار گرفت و تا حدودی مانع پیشرفت علایم بیماری شد. پپتیدهای Spinigerin(موریانه)، Ceratotoxin (مگس میوه مدیترانه‌ای) ، Formaecin-1 (مورچه مرمسیا)،Metchnikowin1(مگس سرکه)، Apidaecin Ia(زنبور عسل اروپایی) و Metchnikowin- 2(مگس سرکه) نیز با ایجاد لکه‌های قهوه‌ای پوسیده در سطحB4 قرار گرفتند. شدیدترین سطح بروز علایم نیز در شاخص (B5) پوسیدگی قهوه‌ای همراه با نقاط فرورفته بر روی کلاهک قارچ بود که تنها در نمونه شاهد مثبت حاوی محیط کشت و باکتری مشاهده شد. تمامی آزمایشات در 3 تکرار انجام شد و آنالیز آماری با روش تجزیه واریانس و مقایسه میانگین‌های مقادیر MICو MBC به روش دانکن انجام شد. وفق نتایج این مطالعه، می‌توان از پپتیدهای ضد میکروبی با منشا حشرات به‌ویژه Cecropin ها‌در کاهش بیماری لکه قهوه‌ای قارچ سفید دکمه‌ای بهره برد. | ||
| کلیدواژه‌ها | ||
| Pseudomonas tolaasii؛ قارچ خوراکی؛ پپتیدهای ضد‌میکروبی؛ Cecropin | ||
| عنوان مقاله [English] | ||
| Effect of some Antibacterial Peptides Against pseudomonas tolaasii the causal agent of brown blotch in White button mushroom | ||
| نویسندگان [English] | ||
| Afsaneh Moslemi Mehni1؛ Mohammad Rahnamaeian2؛ Nader Hassanzadeh3؛ Hamid Forootan Far4 | ||
| 1Department of Plant Protection, Faculty of Agricultural Sciences and Food Industries, Science and Research Branch, Islamic Azad University, Tehran, Iran | ||
| 2School of Pharmacy, Philipps University of Marburg, Marburg, Germany. | ||
| 3Associate Professor, Department of Plant Protection, Faculty of Agricultural Sciences and Food Industries, Science and Research Branch, Islamic Azad University, Tehran, Iran. | ||
| 4Professor, Pharmaceutical Sciences and Cosmetic Products Research Center, Kerman University of Medical Sciences, Kerman, Iran. | ||
| چکیده [English] | ||
| One of the most important factors limiting the production of edible mushrooms is the brown spot disease caused by the bacteria Pseudomonas tolaasii, which has a direct effect on reducing the quality and commercial effect of product. Due to the lack of an effective solution to control of this disease and the impossibility of using chemical poisons and antibiotics because of their unvialling effect on product health and bacterial resistant, new research is being done to identify antibacterial compounds of natural origin. With this approach, in this research, the antibacterial effects of ten peptides obtained from insects like Cecropia moth, Mediterranean fruit fly, Stable fly, Termites, Western honey bee, Red bull ant, Common fruit fly, Common green bottle fly were investigated against this disease. The values of the minimum inhibitory concentration (MIC) 50% and the minimum lethal concentration (MBC) of the desired peptides against pathogenic bacteria were obtained by ELISA method. The MBC values of peptides were also determined by re–treating the contents of the microplates. Also, to determine of disease severity, the caps of mushrooms were covered by peptides(1Mm) befor inoculating with pathgenic bacteria (108 cfu/ml). Then, the severity of disease symptoms was evaluated with six standard scale levels ( B0–B1–B2–B3–B4–B5), where B0 indicates no infection and B5 showing brown and sunken symptoms (B5). MIC values of Cecropin A (Cecropia moth), Scal–stomoxyn (Stable fly), Lser–Cecropin1(Common green bottle fly) peptides in different concentrations were effective on the growth of P. tolaasii. The lowest amount of MBC was observed in Cecropin A peptide equal to 6.25 µM. Also, the results showed that Cecropin A peptide completely protected the mushroom cap at B0 level, and then Scal–stomoxyn peptide was placed at B1 index level with no occurrence or formation of pale–yellow spots. Peptides Spinigerin, Ceratotoxin (Mediterranean fruit fly), Formaecin–1(Red bull ant), Metchnikowin–1(Common fruit fly), Apidaecin Ia (Western honey bee) and Metchnikowin–2 (Common fruit fly) was also placed in B4 level infection by causing brown spots and decay. Brown rot symptoms with sunken spots on the mushroom cap were observed only in the positive control sample, which had the most severe quality level of symptoms (B5). The antimicrobial peptide Cecropin A completely inhibited P. tolaasii and the use of Scal–stomoxyn on the mushroom cap partially prevented the progress of the disease All experiments were performed in 3 replicates and the accuracy of MIC and MBC values was evaluated by comparing the means through analysis of variance. According to the results of this study, the capacity of insect antimicrobial peptides, especially Cecropins, can be used to control the brown spot of edible button mushrooms. . | ||
| کلیدواژه‌ها [English] | ||
| Antibacterial peptides, Mushroom, Pseudomonas tolaasii, Cecropin | ||
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