بررسی تنوع ژنتیکی قارچ عامل بیماری آنتراکنوز گردو Ophiognomonia leptostyla (Fr.) Sogonov در منطقه ITS و IGS با استفاده از نشانگر CAPS در شمال غرب ایران
محورهای موضوعی :
مهدی میانجی
1
(کارشناس ارشد بیو تکنولوژی دانشگاه آزاد اسلامی واحد میانه)
حمید عبدالهی
2
(عضو هیأت علمی گروه بیوتکنولوژی کشاورزی دانشگاه آزاد اسلامی تهران. واحد علوم تحقیقات.)
سلیمان جمشیدی
3
(عضو هیأت علمی گروه گیاهپزشکی دانشگاه آزاد اسلامی واحد میانه.)
کلید واژه: PCR-RFLP, آنتراکنوز, لکه سیاه گردو, Marssoniella juglandis,
چکیده مقاله :
به منظور ارزیابی تنوع ژنتیکی قارچ عامل بیماری آنتراکنوز گردو Ophiognomonia leptostyla در شمال غرب ایران تعداد 25 جدایه از مناطق مختلف جمع آوری و با استفاده از روش تک اسپورسازی خالص شدند. استخراج DNA از میسلیوم قارچ با استفاده از روش لیو و همکاران انجام شد. تکثیر منطقه ITS-rDNA توسط جفت آغازگر اختصاصی، ITS1 و ITS4 انجام گردید. همچنین منطقه IGS-rDNA با استفاده از جفت آغازگر NS1R و LR13R تکثیر شد. نوارهایی به طول 600 جفت باز و 2320-2300 جفت باز از تکثیر منطقه ITS و IGS به دست آمد. قطعات تکثیر شده توسط پنج آنزیم برش دهنده EcoRI، HindIII، TagI، HinfI و BamHI مورد هضم قرار گرفتند. از مجموع آنزیم های برشی مورد استفاده EcorI، TagI و BamHI فاقد محل برش بر محصولات تکثیری منطقه ITS و HinfI و BamHI نیز فاقد محل برش بر محصولات تکثیری منطقه IGS بودند. گروه بندی جدایه های مورد مطالعه بر اساس الگوهای نواری هر دو منطقه در سطح شباهت 75 درصد، آنها را در چهار گروه طبقه بندی نمود.
Genetic diversity of Ophiognomonia leptostyla, walnut anthracnose casual agent in northwest of Iran was studied. 30 isolates were collected from different regions and purified as single spore cultures. DNA extraction from fungi mycelium was carried out. The ITS-rDNA region amplified with ITS1 and ITS4 primers. The IGS-rDNA region was amplified by NS1R and LR13R primers. Bands with 600 and 2300-2320 bp length were obtained in ITS and IGS amplification, respectively. Amplified fragments were digested by EcorI, HindIII, TagI, HinfI and BamHI restriction enzymes. EcorI, TagI and BamHI had no restriction site on amplified ITS region and HinfI and BamHI had no restriction site on IGS region. All isolates were grouped in four clusters based on ITS and IGS CAPS with 75% similarities.
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