ردیابی و توالی‌یابی ژن کُدکنندة پروتئین مرتبط با بیماریزایی PR10 در نخود (Cicer arietinum L.)

نوع مقاله : مقالات پژوهشی

نویسندگان

1 مجتمع اموزش عالی شیروان

2 مجتمع آموزش عالی شیروان

3 استادیار بیوتکنولوژی کشاورزی و عضو هیأت علمی پژوهشکده علوم گیاهی، دانشگاه فردوسی مشهد

چکیده

در بین حبوبات، نخود از جایگاه ویژه‌ای برخوردار است و از جمله عوامل محدودکننده عملکرد آن، می‌توان به برق‌زدگی نخود اشاره کرد. استفاده از پروتئین­های مرتبط با بیماریزایی یکی از راهکارهای مقابله گیاهان با عوامل بیماریزا به‌شمار می­رود. یکی از ژن‌های کُدکننده این نوع پروتئین­ها، ژن کدکننده PR-10 است. این پژوهش با هدف ردیابی ژنPR-10 در ژنوم گیاه نخود انجام شد. بدین منظور DNAی ژنومی شِش ژنوتیپ نخود با مقاومت متفاوت نسبت به بیماری شامل دو رقم مقاوم MCC142 و MDD528، دو رقم متحمل MCC150 و MCC20 و دو رقم حساس MCC507 و MCC506 به روشCTAB استخراج گردید. با استفاده از آغازگرهای PSH-91 و MN واکنش PCR طی دو مرحله انجام شد. مرحله اول با استفاده از DNAی ژنومی نخود و با آغازگرPSH91 و مرحله دوم با استفاده از محصولات مرحله اول و آغازگرMN انجام شد. سپس تخلیص باندها به­وسیله کیت صورت گرفت و جهت توالی‌یابی ارسال شد. حضور تک‌باند 1350جفت بازی در مرحله اول و حضور تک‌باند 1289بازی در مرحله دوم بیانگر حضور ژنPR-10 در ژنوم نخود می باشد. نتایج این آزمون حاکی از عدم تفاوت در توالی­های ژن کدکننده پروتئینPR-10 در ارقام نخود با سطوح مقاومتی متفاوت بود. با توجه به نتایج به‌دست‌آمده، می­توان نتیجه گرفت که تفاوت در واکنش ارقام نسبت به بیماری، به تفاوت در توالی این ژن مربوط نیست و دلایل دیگری مقاومت متفاوت را ایجاد می‌کنند.

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