واکنش بیوشیمیایی شب‌پره مدیترانه‌ای آرد Ephestia kuehniella Zeller (Lep.: Pyralidae) به سمیت ترانس آنتول

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانش آموخته‌ی دکتری گروه گیاه‌پزشکی، دانشکده علوم کشاوری، دانشگاه گیلان، رشت، ایران

2 دانشیار گروه گیاه‌پزشکی، دانشکده کشاوری، دانشگاه زابل، زابل، ایران

3 دانشیار گیاه‌پزشکی، دانشکده علوم کشاوری، دانشگاه گیلان، رشت، ایران

چکیده

ترکیبات مونوترپنوئیدی از اجزای اصلی اسانس‌های گیاهی بوده و با ایجاد اختلال در عملکردهای فیزیولوژیکی و رفتاری حشرات، می‌­توانند در مدیریت آفات مورد استفاده قرار گیرند. شب‌پره مدیترانه‌ای آرد Ephestia kuehniella Zeller, 1879 (Lep.: Pyralidae) علاوه بر ایجاد خسارت روی محصولات انباری، به دلیل سهولت پرورش در شرایط آزمایشگاهی، به عنوان یک حشره‌ی مدل در آزمایش‌­های سم­‌شناسی و بیوشیمیایی مورد توجه محققین قرار دارد. در این پژوهش به منظور درک بهتر سازوکار عمل ترانس آنتول به­‌عنوان یکی از مهم‌ترین متابولیت‌های ثانوی گیاهان تیره چتریان، اثر غلظت کشنده 50 درصد (L g-1µ 03/7LC50=) این ترکیب بر فعالیت آنزیم‌های گوارشی، سم‌­زدا و مواد موثر در متابولیسم حد واسط لاروهای سن 4 شب‌پره مدیترانه‌­ای آرد مورد ارزیابی قرار گرفت. کاهش معنی‌داری در فعالیت آنزیم‌­های گوارشی (آلفا-آمیلاز، آلفا و بتاگلوکوزیداز، لیپاز) و پروتئازهای اختصاصی (تریپسین، کیموتریپسین، الاستاز، آمینو و کربوکسی پپتیداز) مشاهده شد در حالی­که فعالیت آنزیم­‌های سم‌زدا (استرازها و گلوتاتیون اس‌ترنسفرازها) در حشرات تیمارشده افزایش نشان داد. تغییرات افزایشی در فعالیت آمینو ترنسفرازها (آلانین، آسپارتات و گاما گلوتامیل) و کاهشی در فعالیت لاکتات دهیدروژناز، اسید و آلکالین فسفاتاز به­‌عنوان آنزیم‌های دخیل در متابولیسم حد واسط لاروهای شب‌پره مدیترانه‌ای آرد مشاهده شد. میزان ذخایر درشت­‌مولکول‌های ذخیره‌ای (پروتئین کل، گلیکوژن، تری‌گلیسرید) در حشرات تیمارشده کاهش معنی‌­داری در مقایسه با شاهد نشان داد. القاء و بازدارندگی فعالیت آنزیم‌­ها پس از تیمار با غلظت LC50 ترانس آنتول، نشان­‌دهنده کارایی استفاده از این ترکیب در ایجاد اختلالات فیزیولوژیکی در شب‌پره مدیترانه‌­ای آرد است.

کلیدواژه‌ها


عنوان مقاله [English]

Biochemical response of Mediterranean flour moth, Ephestia kuehniella Zeller (Lep.: Pyralidae) to the toxicity of trans-anethole

نویسندگان [English]

  • M. Shahriari 1
  • N. Sahebzadeh 2
  • A. Zibaee 3
1 PhD graduated, Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht. Iran
2 Associate Professor, Department of Plant Protection, Faculty of Agriculture, University of Zabol, Zabol, Iran
3 Associate Professor, Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht. Iran
چکیده [English]

Background and Objectives
Essential oils are a variety of compounds, and their monoterpenoids, as the main constituents may impair insects' physiological and behavioral functions and could thus be applied in pest management. In addition to causing damage to stored products, the Mediterranean flour moth, Ephestia kuehniella Zeller, 1879 (Lep.: Pyralidae), is actually an easy model to grow in the laboratory and is one of the suitable insects to determine toxicity and in vivo interactions of xenobiotics with physiological systems of insects. In the present study, in order to better understand the mechanism of action of trans-anethole as one of the most important secondary metabolites of the Apiaceae family, the oral toxicity of this compound was evaluated on the activities of digestive and detoxifying enzymes, and metabolites involving in intermediate metabolism of the fourth instar of E. kuehniella.
Materials and Methods
Fourth instar larvae of E. kuehniella were randomly selected and separately exposed to 1 g of the artificial diet containing 1, 2, 4, 8, and 16 μL/g of trans-anethole while the control larvae fed on the diet containing acetone. The bioassay test was performed in 3 replicates with 10 larvae per replication. Mortality was determined after 24 h and LC50 value was calculated by POLO-plus software. Ephestia kuehniella larvae were exposed to the LC50 value of trans-anethole to determine its effects on the enzymatic and non-enzymatic components. After 24 of exposure, the larvae hemolymph, midgut, and fat bodies were extracted. Hemolymph samples were immediately centrifuged at 15000 × g at 4°C for 20 min, while samples of midgut and fat bodies were initially homogenized by a glass pestle and then centrifuged under the same conditions. The hemolymph samples were used to assay the detoxifying enzymes and enzymes of intermediary metabolism, while the midgut samples were used to assess the digestive enzymes. Fat body samples and head capsules were also used for energy reserves.
Results
The LC50 value of trans-anethole against the larvae was 7.03 μL g-1. Significant decrease in the digestive enzyme activities (α-amylase, α and β-glucosidase, lipase) and specific proteases (trypsin, chymotrypsin, elastase, amino and carboxy peptidase) were observed. In contrast, the activity of the detoxifying enzymes (esterases, and glutathione S- transferase) increased in the treated insects. The activities of amino transferases (alanine, aspartate, and gamma-glutamyl) significantly increased in the treated larvae by trans-anethole. Significant decreases in lactate dehydrogenase and phosphatase (acid and alkaline) were observed after treatment. Moreover, the amount of storage macromolecules (total protein, glycogen, and triglyceride) in the treated insects was significantly decreased compared to the control.
Discussion
Our research revealed that trans-anethole is a toxic compound to E. kuehniella by reducing the survival and digestive efficiency of the larvae. Moreover, trans-anethole significantly enhanced the detoxifying enzymes activities. When the larvae were exposed to LC50 of trans-anethole, the digestive activity was reduced due to the cytotoxic effects of trans-anethole on epithelial cells of the larval alimentary canal. Our results demonstrated that trans-anethole might have a promising potential to develop as a safe compound to suppress E. kuehniella population. For the practical use of trans-anethole to control E. kuehniella, it is required to determine the accurate mode of action, and using appropriate formulations to increase its efficacy in long-term applications.

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