بررسی ویژگی‌های کمی و کیفی فرمولاسیون مبتنی بر آلژینات جدایه‌ی بومی نماتد بیمارگر حشرات (Rhabditida: Steinernematidae) Steinernema carpocapsae

نوع مقاله : علمی پژوهشی-فارسی

نویسنده

استادیار بخش تحقیقات کنترل بیولوژیک، موسسه تحقیقات گیاه‌پزشکی، سازمان تحقیقات، ترویج و آموزش کشاورزی، تهران، ایران.

چکیده

جدایه­‌ی بومیSteinernema carpocapsae  با رس‌شمارMF187616  در پایگاهNCBI  (مرکز بین‌­المللی اطلاعات بیوتکنولوژی) که از منطقه‌­­ی مغان جداسازی شده است، به­‌دلیل برخورداری از پتانسیل بالای بیماری­‌زایی و تحمل محیطی در بررسی‌­های آزمایشگاهی، برای فرمولاسیون مبتنی بر آلژینات مورد استفاده قرار گرفت. ویژگی‌­های فرمولاسیون، درصد زنده‌­مانی نماتدها در داخل فرمولاسیون، میزان آب خارج شده از کپسول­‌ها و بیماری‌­زایی نماتدهای فرموله شده روی لاروهای Galleria mellonella به عنوان میزبان آزمایشگاهی در فواصل زمانی چهار تا 90 روز مورد ارزیابی قرار گرفت. براساس نتایج به­‌دست آمده، بهترین غلظت آلژینات سدیم و کلرید کلسیم به‌­ترتیب دو درصد و نیم درصد بود که برای ادامه کار انتخاب شدند. قطر کپسول‌های تشکیل شده در دامنه­‌ی 5/3 تا 5/4 میلی­متر قرار داشت. تجزیه واریانس داده­‌های مربوط به­‌میزان آب خروجی از کپسول و زنده‌­مانی لاروهای آلوده‌­کننده­‌ی نماتد در داخل کپسول بیانگر تفاوتی معنی‌­دار بین نقاط زمانی مختلف بود (P ≤ 0.01). در نقاط زمانی چهار تا 90 روز پس از فرمولاسیون، نماتدهای فرموله شده درصد بقای بیشتری نسبت به نماتدهای فرموله نشده (در سوسپانسیون آبی) نشان دادند (P ≤ 0.01). نتایج تجزیه واریانس زیست­‌سنجی فرمولاسیون آلژینات جدایه بومی S. carpocapsae IRMoghan1 نشان‌­دهنده وجود تفاوت معنی­‌دار بین نقاط مختلف زمانی در درصد مرگ‌ومیر G. mellonella بود (P ≤ 0.01). نتایج مطالعه حاضر نشان دهنده­‌ی پتانسیل بالای جدایه‌­ی بومی S. carpocapsae IRMoghan1 برای فرمولاسیون آلژینات سدیم بود. فرموله کردن موفقیت‌­آمیز یک نماتد عامل کنترل بیولوژیک به منظور افزایش مدت زمان نگهداری بدون یخچال نیز از نتایج این پژوهش است که می‌­تواند مبنای بررسی‌های بعدی با هدف تجاری­‌سازی این جدایه باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Quantitative and qualitative characteristics of alginate-based formulation of a native isolate of the entomopathogenic nematode Steinernema carpocapsae (Rhabditida: Steinernematidae)

نویسنده [English]

  • L. Ebrahimi
Iranian Research Institute of Plant Protection, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran.
چکیده [English]

Background and Objectives
Entomopathogenic nematodes (EPNs), in the genera Steinernema and Heterorhabditis, are potent biocontrol agents that are widely used against various insect pests. The non-feeding and non-developing third stage infective juvenile (IJ), or the dauer juvenile, is the survival stage of EPNs that is able to survive outside the host and is used for formulation. In recent years, alginate has been recognized as the most promising biopolymer in the formulation of living cells and microorganisms. According to the results of scientific research, native isolates of each region have a greater impact than non-native and foreign isolates due to high ecological compatibility.
Materials and Methods
The native isolate of Steinrnema carpocapsae IRMoghan1 from the Moghan region, with accession number MF187616 in the NCBI database, was selected for use in the alginate-based formulation owing to its high potential of survival and pathogenicity. The best and most cost-effective capsule composition was selscted by evaluating 0.5%, 1%, 2%, and 3% alginate concentrations with 0.5%, 1%, and 2% concentrations of a calcium chloride solution. Formulation characteristics, the survival rate of the nematodes inside the formulation, exited water volume, escaped nematode rate, and pathogenicity of the formulated nematodes against Galleria mellonella larvae as the laboratory host were measured and recorded during 4 to 90 days post formulation. The analysis of variance (ANOVA) and comparison of means by Duncan’s test were performed on data using SAS software.
Results
Based on the results, the best concentrations for sodium alginate and calcium chloride were 2% and 0.5%, respectively, which were therefore selected for the next experiments. The diameter of the formed capsules ranged from 3.5 to 4.5 mm. The ANOVA of the amount of water leaving the capsules, the number of nematodes escaped from the capsules, and the survival rates of the nematodes inside the capsules were significantly different at various time intervals post formulation (P ≤ 0.01). Statistical analysis showed that the formulated nematodes had a higher survival rate than the unformulated congeners in all the time points from (4 to 90 days) after formulation. The results of the ANOVA for bioassay experiments showed significant differences between different time points in the percentage of mortality of G. mellonella (P ≤ 0.01).
Discussion
The present study showed the high potential of S. carpocapsae IRMoghan1 native isolate for sodium alginate formulation and, the successful formulation of an entomopathogenic nematode as a biocontrol agent, which is a valuable finding as it increases the shelf life without refrigeration. This finding should be used as the basis of further studies for the commercialization of this isolate.

کلیدواژه‌ها [English]

  • slow-release formulation
  • Sodium alginate
  • storage formulation
  • Biological control
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