شناسایی مولکولی قارچ‌های تولید کننده 8-کتو تریکوتسین همراه با بذر روناس و ردیابی ژن‌های سنتزکننده دی‌اکسی نیوالنول در آنها با استفاده از PCR

نوع مقاله : مقالات پژوهشی

نویسندگان

• سید علیرضا اسمعیل زاده حسینی ¹
• محمود دهقانی اشکذری ¹
• مصطفی عابدی تیزکی ²

¹ مرکز تحقیقات زیست فناوری پزشکی، واحد اشکذر، دانشگاه آزاد اسلامی، اشکذر، یزد

² دانشگاه بوعلی سینا همدان

10.22067/jpp.v31i3.58227

چکیده

روناس (Rubia tinctorum) یکی از محصولات مهم زراعی با مصارف دارویی و صنعتی در استان یزد می‌باشد. به منظور شناسایی قارچ‌های همراه با بذور روناس و تولید کننده توکسین، طی سال‌های زراعی 91 لغایت 93 از بذور روناس در مناطق مهم کاشت آن در استان یزد شامل اردکان و بافق نمونه‌برداری به عمل آمد. پس از کشت و خالص‌سازی جدایه‌های قارچی در محیط‌های کشت‌های سیب زمینی دکستروز آگار (PDA) و برگ میخک آگار (CLA)، شناسایی تکمیلی با آغازگرهای اختصاصی هر گونه با استفاده از واکنش زنجیره‌ای پلیمراز صورت گرفت. ردیابی قارچ‌های دارای پتانسیل تولید مایکوتوکسین 8-کتوتریکوتسین از جمله دی اکسی نیوالنول (DON) از طریق آغازگرهای اختصاصی ژن Tri13 انجام شد. جهت تأیید پتانسیل تولید مایکوتوکسین DON در جدایه‌های فوزاریوم از کروماتوگرافی مایع با کارایی بالا (HPLC) استفاده شد. در مجموع 249 جدایه قارچی از بذور روناس جداسازی شد که متعلق به شش جنس قارچی شامل فوزاریوم، آسپرژیلوس، پنیسلیوم، آلترناریا، رایزوکتونیا و رایزوپوس بودند که بیشترین فراوانی قارچ‌های جداسازی شده مربوط به فوزاریوم با 71 درصد کل جدایه‌ها بود. در بین قارچ‌های فوزاریوم، گونه‌های F.solani (55 جدایه) و F.oxysporum (41 جدایه) بیشترین فراوانی را داشتند. گونه‌های F. poae، F. semitectum و F. equiseti توانایی تولید دی اکسی نیوالنول را دارا بودند. ژن Tri13مؤثر در تولید دی اکسی نیوالنول نیز در این سه گونه ردیابی گردید و در آزمایش HPLCهمه قارچ‌های فوزاریوم مورد بررسی پتانسیل تولید مایکوتوکسین مورد نظر را داشتند.

کلیدواژه‌ها

• Rubia tinctorum
• مایکوتوکسین
• یزد

20.1001.1.20084749.1396.31.3.12.3

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عنوان مقاله [English]

Molecular identification of fungi trichothecens producing in seeds madder and detection of their nivalenol gene synthesis using PCR

نویسندگان [English]

• Seyyed Alireza Esmailzadeh Hosseini ¹
• Mahmod Dehghany Ashkezary ¹
• Mostafa Abedi Tizaki ²

¹ Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran

² Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran

چکیده [English]

Madder is one of the most important crops that used for medical and industrial applications and is widely cultivated in Yazd province. During 2012, sampling was done form seeds madder in important areas planted in Yazd province, including Bafq and Ardakan. After culturing and purification of fungal isolates in PDA and CLA media, additional identification was performed by PCR with specific primers for each species. Detection of fungi mycotoxins producing potential such as Nivalenol (NIV) using Tri13 primers was done. High-performance liquid chromatography (HPLC) was used to confirm the produce NIV mycotoxins potential in Fusarium species. 249 fungal strains were isolated from madder seed belonging to 6 genera of fungi including Fusarium spp., Aspergillus spp., Penicillium spp., Alternaria spp., Rhizoctonia solani and Rhizpous spp., that Fusarium isolates with 71 percent was the most frequency among fungi isolated. Among Fusarium fungi isolated, F. solani (55 isolates) and F. oxysporum (41 isolates) were the most frequency. F. poae, F. semitectum and F. equiseti ability to produce mycotoxins such as Nivalenol (NIV) that are harmful to human health and animals as well as effect on the quantity and quality of madder color production. Tri13 gene involved in production NIV was detected in three Fusarium species that all isolates produce NIV. The results of HPLC showed that all studied Fusarium fungi, have the potential to produce NIV mycotoxins. The results of this study showed that fungi associated with seeds madder are able to produce trichothecene mycotoxins that they can be dangerous for consumers. Given that, this is the first report of fungi mycotoxins producing on seeds madder in Yazd province, thus should be measures to control and reduce fungal agents in these products.

کلیدواژه‌ها [English]

• Mycotoxin
• Rubia tinctorum
• Yazd

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