ارزیابی اثر ضد قارچی اسانس آویشن و نعناع فلفلی و مونوترپن‌های عمده آنها در کنترل قارچ ورتیسیلیوم پسته

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 نویسنده مسئول، بخش تحقیقات گیاهپزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان سمنان (شاهرود)، سازمان تحقیقات، آموزش و ترویج کشاورزی، شاهرود، ایران

2 بخش تحقیقات صنایع غذایی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان سمنان (شاهرود)، سازمان تحقیقات، آموزش و ترویج کشاورزی، شاهرود، ایران

3 مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان سمنان (شاهرود)، سازمان تحقیقات، آموزش و ترویج کشاورزی، شاهرود، ایران.

4 بخش تحقیقات گیاهپزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان سمنان (شاهرود)، سازمان تحقیقات، آموزش و ترویج کشاورزی، شاهرود، ایران

چکیده

سابقه و هدف: بیماری پژمردگی ناشی از قارچ بیماریزای Verticillium dahliae، یکی از عوامل خطرناک تهدید کننده کِشت درختان پسته در سطح کشور می‌باشد که در دو دهه اخیر در مناطق مختلف کشور، از جمله استان سمنان گسترش پیدا کرده است. عدم وجود قارچکش مناسب جهت کنترل قارچ V. dahliae و استفاده مداوم از چند قارچکش عمومی در سطح کشور به گسترش قارچ کمک کرده است. از این‌رو در شرایط کنونی نیازمند استفاده از ترکیبات جدید با کارایی بالا جهت کنترل این قارچ بیماریزا هستیم. اسانس‌های گیاهی و ترکیبات مونوترپنی آنها با توجه به مزیت نسبی تولید گیاهان دارویی در کشور، به عنوان یکی از گزینه‌های مهم مطرح هستند.
مواد و روش‌ها: در این پژوهش از جدایه بومی V. dahliae بدست آمده از سطح باغات استان سمنان استفاده شد. با استفاده از روش‌های معمول فرآیند جداسازی و شناسایی قارچ انجام پذیرفت و با استفاده از پرایمر عمومی ITS شناسایی جدایه‌ها انجام شد. در ادامه از اسانس دو گیاه آویشن شیرازی (Zataria multiflora) و نعناع فلفلی (Mentha piperita) جهت بررسی اثر ضد قارچی استفاده شد. پس از تهیه مواد گیاهی، فرایند اسانس‌گیری با استفاده از دستگاه کلونجر انجام پذیرفت. بررسی اثر ضد قارچی با استفاده از روش اختلاط با محیط کشت در غلظت‌های 20، 40، 60، 80 و 100 میکرولیتر بر میلی‌لیتر صورت گرفت. همچنین حداقل غلظت بازدارندگی (MIC) و کُشندگی (MBC) برای دو اسانس تعیین گردید. در ادامه با بکارگیری دو ترکیب مونوترپنی تیمول و کارواکرول اثرات ضد قارچی با استفاده از روش اختلاط با محیط کشت در سه غلظت 15، 150 و 1500 میکرولیتر برمیلی لیتر ضد V. dahliae مورد آزمایش قرار گرفت.
یافته‌ها: آزمون‌های جداسازی و شناسایی، وجود قارچ V. dahliae را در مناطق نمونه برداری شده تائید نمود. نتایج نشان داد استفاده از پرایمر ITS تایید کننده گونه قارچی V. dahliae بود. نتایج اثر ضد قارچی نشان داد هر دو اسانس در جلوگیری از رشد قارچ موثر بودند، اما اسانس آویشن شیرازی قوی‌تر ارزیابی شد بطوری که در هفته سوم، رشد فقط در غلظت 20 میکرولیتر بر میلی‌لیتر مشاهده شد اما در مورد اسانس نعناع فلفلی در تمامی غلظت‌ها رشد قارچ مشاهده گردید. نتایج حداقل غلظت بازدارندگی و کُشندگی به ترتیب برای آویشن شیرازی 20 و 30 میکرولیتر بر میلی‌لیتر و برای اسانس نعناع فلفلی 100 و 150 میکرولیتر بر میلی‌لیتر بدست آمد. سایر نتایج نشان داد ترکیب تیمول در جلوگیری از رشد قارچ V. dahliae قوی تر از ترکیب کارواکرول عمل نمود.
نتیجه گیری: نتایج این پژوهش نشان داد اسانس‌های گیاهی و ترکیبات مونوترپنی تشکیل دهنده اسانس‌ها از توانایی بسیار بالایی در کنترل قارچ ورتیسیلیوم برخوردار بودند بطوری که در مقایسه با قارچکش‌های طیف وسیع رایج مورد استفاده در کشور، در غلطت‌های 10 تا 15 برابر کمتر از فعالیت قارچ جلوگیری نمودند.

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation of Antifungal Effect of Thyme and Peppermint Essential Oils and Their Major Monoterpenes in Controlling Verticillium Fungus on Pistachios

نویسندگان [English]

  • Mahdi Akhlaghi 1
  • Hamid reza Ziaolhagh 2
  • Esmaiel babakhanzade 3
  • Vahide Rafiee 4
1 Corresponding Author, Plant Protection Research Department, Agricultural and Natural Resources Research Center of Semnan Province (Shahrood), AREEO, Shahrood, Iran
2 Food Technology Research Department, Agricultural Engineering Research Department, Agricultural and Natural Resources Research and Education Center of Semnan Province (Shahrood), AREEO, Shahrood, Iran
3 Agricultural Engineering Research Department, Agricultural and Natural Resources Research and Education Center of Semnan Province (Shahrood), AREEO, Shahrood, Iran
4 . Plant Protection Research Department, Agricultural and Natural Resources Research and Education Center of Semnan Province (Shahrood), AREEO, Shahrood, Iran
چکیده [English]

Background and purpose: Wilt disease caused by the pathogenic fungus Verticillium dahliae is one of the dangerous factors threatening the cultivation of pistachio trees in the country, which has spread in different parts of the country in the last two decades, including Semnan province. Lack of suitable fungicides to control V. dahliae and continuous use of several common fungicides in the country has contributed to the spread of the fungus. Therefore, in the current situation, we need to use new compounds with high efficiency to control this pathogenic fungus. Plant essential oils and their monoterpene compounds are considered as one of the important options due to the comparative advantage of the production of medicinal plants in the country.
Materials and Methods: In this study, V. dahliae native isolates obtained from gardens of Semnan province were used. Isolation and identification process was performed using conventional methods and isolates were identified using general ITS primer. Then, the essential oils of thyme (Zataria multiflora) and Peppermint (Mentha piperita) were used to evaluate the antifungal effect. After preparing the herbal plants, the essential oil extraction process was performed using Clevenger method. The antifungal effect was investigated by mixing with culture medium at concentrations of 20, 40, 60, 80 and 100 μg/ml. Also, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MBC) were determined for the two essential oils. Then, using thymol and carvacrol monoterpenes, antifungal effects were tested by mixing with culture medium at three concentrations of 15, 150 and 1500 μg/ml against V. dahliae.
Results: Isolation and identification tests confirmed the presence of V. dahliae in the sampled areas. The use of ITS primer confirmed the fungal species V. dahliae. The results of antifungal effect showed that both essential oils were effective in preventing the growth of fungi, but Zataria multiflora essential oil was evaluated as stronger, so that in the third week, growth was observed only at a concentration of 20 μg/ml, but in the case of peppermint essential oil, fungus growth was observed in all concentrations. The results showed that the minimum inhibitory (MIC) and minimum fungicidal (MFC( concentrations were 20 and 30 μg/ml for thyme and 100 and 150 μg/ml for peppermint essential oils, respectively. In addition, the results showed that thymol was more potent than carvacrol in preventing the growth of V. dahliae.
Conclusion: The results of this study showed that plant essential oils and monoterpene compounds of essential oils had a very high ability to control Verticillium fungus, so that compared to common fungicides in the country, their inhibitory concentration was 10 to 15 times lower

کلیدواژه‌ها [English]

  • Carvacrol
  • Thymol
  • Verticillium dahliae
  • Verticillium wilt
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