غربال و ارزیابی تنوع فنوتیپکی جدایه‌های غیرتوکسین‌زای Aspergillus flavus در باغات پسته ایران

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه گیاهپزشکی، دانشگاه ولیعصر رفسنجان (عج)، رفسنجان

2 بخش تحقیقات گیاه‌پزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان یزد، سازمان تحقیقات، آموزش و ترویج کشاورزی، یزد، ایران

3 پژوهشکده پسته، مؤسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، رفسنجان، ایران.

4 پژوهشکده پسته، مؤسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، رفسنجان، ایران

چکیده

استفاده از سویه‌های غیرتوکسین‌زای قارچ Aspergillus flavus به عنوان یک روش بیولوژیک مؤثر برای کاهش آفلاتوکسین در کشورهای مختلف در حال توسعه است. سویه‌های غیرتوکسین‌زا قادر به رقابت با سویه‌های توکسین‌زا و اشغال بستر هستند و این امر باعث کاهش جمعیت توکسین‌زا و میزان آفلاتوکسین در بستر غذایی می‌شود. هدف از این مطالعه، بررسی تنوع در جمعیت غیرتوکسین‌زای A. flavus از نظر مورفولوژیکی بود. تعداد 225 نمونه میوه و خاک پسته از استان‌های مختلف جمع‌آوری و برای جداسازی قارچ از روش سری‌های رقت و محیط کشت AFPA استفاده شد. برای غربال اولیه جدایه‌های غیرتوکسین‌زا از روش بخار آمونیاک در محیط کشت نارگیل آگار و برای تأیید نتایج از روش کروماتوگرافی لایه نازک (TLC) استفاده شد. تعیین گونه جدایه‌های حاصله به روش مولکولی با استفاده از آغازگرهای اختصاصی گونه FLAVIQ1/FlaQ2 انجام شد. ویژگی‌های ماکرومورفولوژیکی و میکرومورفولوژیکی 56 سویه غیرتوکسین­زای A. flavus به دست آمده، با استفاده از چهار محیط کشت CYA (czapek yeast agar)، MEA (malt extract agar)،CY20S  (czapek yeast 20% sucrose) و CZ (czapek) مورد مقایسه قرار گرفت. بررسی این خصوصیات، جدایه­ های غیرتوکسین‌زایA. flavus را در پنج گروه قرار داد. الگوی رشد پرگنه در محیط‌ کشت CYA25 قابلیت تفکیک خوبی در بین جدایه‌ها از خود نشان داد. نتایج نشان داد در میان جدایه‌های غیرتوکسین‌زا A. flavus به دست آمده از مکان‌های جغرافیایی مختلف تنوع زیادی وجود دارد که دلیل آن می‌تواند به وجود گونه‌های مختلف در کمپلکس A. flavus مربوط باشد.

کلیدواژه‌ها

عنوان مقاله [English]

Screening and assessment of phenotypic diversity in nontoxigenic strains of Aspergillus flavus in pistachio orchards of Iran

نویسندگان [English]

  • Razieh Pourhosseini 1
  • Ebrahim Sedaghati 1
  • Seyed Reza Fani 2
  • Marieh Nadi 3
  • Mohammad Moradi 4
1 Department of Plant Protection, Vali-e-Asr University of Rafsanjan, Rafsanjan
2 Plant Protection Research Department, Yazd Agricultural and Natural Resources Research and Education Center, AREEO, Yazd, Iran
3 Pistachio Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rafsanjan, Iran.
4 Pistachio Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rafsanjan, Iran
چکیده [English]

The use of nontoxigenic strains of Aspergillus flavus as an effective biological method to reduce aflatoxin is being developed in different countries. Nontoxigenic strains are able to compete with toxigenic strains by substrate occupation, which reduces the toxigenic population and the amount of aflatoxin in the substrate. The aim of study was to investigate the morphological diversity in the nontoxigenic population of A. flavus. To isolate nontoxigenic strains from 225 fruit and pistachio soil samples obtained from different provinces, AFPA culture medium and dilution series method were used. Primary nontoxigenicity screening was performed by ammonia vapor method in coconut agar medium and thin layer chromatography was used for result confirmation. Isolates were identified molecularly using FLAVIQ1/FlaQ2 species-specific primers. In order to identify the macromorphological and micromorphological features of 56 obtained strains, four culture media including CYA (Czapek yeast agar), MEA (malt extract agar), CY20S (Czapek yeast 20% sucrose) and CZ (Czapek) were used. Investigation of macro-micromorphological features, classified A. flavus strains into five groups. Colony growth pattern on CYA25 cultural media has shown good differentiation between isolates. The results showed that there is a great variety among nontoxigenic A. flavus isolates obtained from different agro ecological zones due to the presence of different species in the A. flavus complex.

کلیدواژه‌ها [English]

  • Biological Control
  • Food safety
  • Pistachio
  • Mycotoxin
  • Secondary metabolite

مراجع

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پژوهش های کاربردی در گیاهپزشکی
دوره 10، شماره 3
مهر 1400
صفحه 109-120

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  • تاریخ دریافت: 06 مرداد 1400
  • تاریخ پذیرش: 06 مرداد 1400

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