شناسایی مولکولی و تشخیص ژن کدکننده دی اکسی نیوالنول در جدایه های Fusarium graminearum عامل بلایت فوزاریومی سنبله گندم در ایران
محورهای موضوعی : بوم شناسی گیاهان زراعیرویا رضائیان دلوئی 1 , سعید رضایی 2 , منصوره میرابوالفتحی 3 , حمید رضا زمانی زاده 4 , محمد رضوی 5
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2 - اعضای هیت علمی گروه بیماری شناسی گیاهی، دانشکدهکشاورزی و منابع طبیعی دانشگاه آزاد اسلامی تهران واحد علوم و تحقیقات
3 - اعضای هیت علمی بخش تحقیقات بیماری های گیاهی، موسسه تحقیقات گیاه پزشکی کشور، تهران
4 - اعضای هیت علمی گروه بیماری شناسی گیاهی، دانشکدهکشاورزی و منابع طبیعی دانشگاه آزاد اسلامی تهران واحد علوم و تحقیقات
5 - اعضای هیت علمی بخش تحقیقات بیماری های گیاهی، موسسه تحقیقات گیاه پزشکی کشور، تهران
کلید واژه: DON, FHB, آغازگر اختصاصی گونه, زهرابه, توکسین,
چکیده مقاله :
بیماری بلایت فوزاریومی سنبله یکی از مهم ترین بیماری های قارچی گندم در ایران و سایر نقاط جهان می باشد. عامل اصلی این بیماری Fusarium graminearum شناخته شده که گندم را در طی دوره گل دهی آلوده کرده و نه تنها باعث کاهش محصول می گردد، بلکه با تولید زهرابه قارچی، موجب مسمومیت در انسان و دام نیز می شود. در این مطالعه 60 جدایه F. graminearum جداشده از مزارع گندم آلوده از استان های مختلف ایران مورد بررسی قرار گرفتند. تشخیص مولکولی جدایه ها بر اساس روش PCR با استفاده از آغازگرهای اختصاصی Fg16F/Fg16R برای این گونه انجام و تعلق کلیه جدایه ها به گونه F. graminearum مورد تأیید قرار گرفت. در کلیه جدایه ها یک باند 420 جفت بازی تکثیر شد که در سایر گونه های نزدیک از جمله F. culmorum مشاهده نگردید. جدایه ها برای تشخیص ژن کدکننده زهرابه دی اکسی نیوالنول با استفاده از آغازگر اختصاصی Tri13F/Tri13DONR مورد آزمایش قرار گرفتند. تنها در 36 جدایه یک قطعه 228 جفت بازی تکثیر گردید. با توجه به این که روش های مرسوم بسیار وقتگیر و غیراختصاصی هستند، استفاده از آغازگرهای ویژه گونه برای تشخیص سریع کشت های مشکوک به F. graminearum برای تعیین گونه قارچ عامل بیماری به طور مستقیم دربافت آلوده و تعیین ترکیب زهرابه های تولیدی آن از اهمیت خاصی برخوردار است.
Fusarium Head Blight (FHB) is one of the most important fungal diseases of wheat in Iran and throughout the world. Fusarium species infect wheat during the flowering period. In addition to losses of yield, these fungi can also synthesize mycotoxins under suitable environmental conditions, thus threatening animal and human health. In the present study, 60 isolates of F. graminearum collected from infested wheat fields of some provinces of Iran and were characterized morphologically and genetically. Molecular characterization of isolates was performed using an optimized simple and low cost method for isolation of DNA from F. graminearum using polymerase chain reaction with species specific primers. All of the isolates were confirmed as F. graminearum using species specific primers Fg16F/Fg16R through PCR assays. The results indicated that a specific band of 420 bp was amplified, when species specific primers of F. graminearum was used. All of the isolates were molecularly identified as F. graminearum. The primers failed to amplify any band in negative controls including F. culmorum. All of F. graminearum isolates were tested for DON chemotype producing gene by specific Tri13F/Tri13DONR primer by PCR. Our results showed that a specific band of 282 bp was amplified only in 36 isolates (60%). Consequently, using of species specific primers for detection of suspected cultures of F. graminearum in infected tissues and toxin profile of isolates are very interesting when comparing with the morphological trait which are time consuming and non-specific.
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