مطالعه الگوی پروتئوم آنزیم‌های مؤثر در فرآیند مایکوپارازیتیسم جدایه جهش‌یافته قارچ Trichoderma harzianum

نوع مقاله: مقاله علمی-پژوهشی

نویسندگان

1 گروه گیاه‌پزشکی، دانشکده تولیدات گیاهی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، ایران

2 گروه‌گیاه‌پزشکی، پژوهشکده کشاورزی هسته‌ای، پژوهشگاه علوم و فنون هسته‌ای، سازمان انرژی اتمی ایران

چکیده

این پژوهش به‌منظور بررسی الگوی بیان آنزیم­های کیتیناز در جدایه­های جهش‌یافته قارچ Trichoderma harzianum، نسبت به جدایه وحشی انجام شد. ابتدا جدایه­های گونه T. harzianum با دوز بهینه250 گری پرتوگاما، در معرض جهش قرار گرفتند. توان آنتاگونیستی جدایه­های جهش­یافته و وحشی با آزمون کشت متقابل در حضور بیمارگر گیاهیSclerotinia sclerotiorum مورد بررسی قرار گرفت. تمامی آنالیزهای آماری با نرم­افزار SAS و آزمون حداقل اختلاف معنی­دار (LSD) انجام شد. پروفایل پروتئینی آنزیم­های تولیدشده کلیه جدایه­ها با آزمون الکتروفورز SDS-PAGE و الگوی پروتئوم گونه وحشی به‌همراه جدایه جهش یافته Th M8 با سوبسترای کیتین کلوئیدی و با استفاده از تکنیک الکتروفورز دو­بعدی و نرم­افزار ImageMaster 2D Platinum مورد بررسی قرار گرفت. نتایج نشان داد که اکثر جدایه­های جهش­یافته نسبت به جدایه وحشی در تولید آنزیم­های خارج سلولی کیتیناز، از نظر آماری در سطح پنج و یک درصد دارای اختلاف معنی­دار بودند. همچنین نتایج آزمون آنتاگونیستی نشان داد، پتانسیل کنترل جدایه­های جهش­یافته نسبت به جدایه وحشی در مقابله با قارچ S. sclerotiorum به‌طور معنی­داری افزایش یافته است. بیشترین و کمترین بازدارندگی به‌ترتیب مربوط به جدایه Th M8 با 72 درصد و Th M3با 43 درصد بازدارندگی بود. از طرفی الگوی پروتئوم بر اساس تکنیک الکتروفورز دوبعدی نشان داد که لکه­های پروتئینی جدایه‌ جهش­یافته در مقایسه با جدایه وحشی به‌صورت کمّی و کیفی متفاوت هستند. این موضوع نشان می­دهد که موتاسیون می­تواند منجر به تغییر در الگوی بیان پروتئین­های مترشحه شود. به‌طورکلی نتایج این مطالعه نشان داد که تکنیک پرتودهی گاما می­تواند در تولید آنزیم­های مؤثر در مایکوپارازیتیسم و بهبود پتانسیل کنترل بیولوژیک در مدیریت بیماری­های گیاهی مؤثر واقع شود.

کلیدواژه‌ها

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