بهینه‌سازی القا و تثبیت ریشه‌های مویین هویج به‌منظور کشت درون شیشه‌ای قارچ‌های میکوریز آربوسکولار

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشگاه ولی عصر رفسنجان

2 دانشگاه ولی عصر (عج) رفسنجان

3 هیات علمی دانشگاه ولی عصر رفسنجان

چکیده

هدف از این پژوهش، بهینه‌سازی شرایط القای ریشه مویین هویج با استفاده از ریشه و دمبرگ درون‌شیشه‌ای و همچنین دمبرگ و ساقه گلخانه‌ای به منظور کشت درون شیشه‌ای قارچ‌های میکوریز آربوسکولار می‌باشد. بذرهای هویج پس از ضدعفونی، در محیط کشت MS و گلدان کشت شدند. ریشه و دمبرگ درون‌شیشه‌ای چهار هفته‌ای و دمبرگ و ساقه گلخانه‌ای سه ماهه با استرین‌هایMSU، 15834 و A4 باکتری Rhizobium rhizogenes مایه‌زنی و در محیط کشت MS کشت شدند. تاثیر استرین باکتری و نوع ریزنمونه بر القای ریشه مویین بررسی شد. پس از حذف باکتری، میزان رشد ریشه‌های مویین حاصل از هر ریزنمونه پبه‌صورت فاکتوریل در قالب طرح کاملاً تصادفی با سه تکرار اندازه‌گیری گردید. تایید مولکولی تراریختگی ریشه‌های مویین و استرین‌های باکتری با استفاده از واکنش PCR برای تکثیر اختصاصی ژن rolC به اثبات رسید. نتایج نشان داد هر سه استرین توانایی القای تولید ریشه مویین را دارند. ریشه‌های تراریخته نسبت به ریشه‌های معمولی ضخامت، انشعاب و سرعت رشد بیشتری داشتند. استرین 15834 و ریزنمونه‌های دمبرگ گلخانه‌ای و ریشه درون‌‌شیشه‌ای بیشترین توانایی تولید ریشه مویین را نشان دادند. بالاترین میزان رشد ریشه‌های مویین در ریزنمونه ساقه گلخانه‌ای با استرین MSU مشاهده شد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Optimizing of carrot hairy root induction and stabilization for monoxenic culture of arbuscular mycorrhizal fungi

چکیده [English]

This study was conducted to optimizing of in vitro culture of Arbuscular Mycorrhizal Fungi (AMF) on in vitro root and petiole and greenhouse petiole and stem of carrot. Carrot seeds were disinfested and cultured on MS medium and pot. Four weeks old of in vitro root and petiole and 3 months old of greenhouse petiole and stem were inoculated with Rhizobium rhizogenes MSU, 15834 and A4 strains and cultured on MS medium. The effects of bacterial strain and explants type on hairy root production were studied. After the elimination of bacteria, growth rate of hairy roots obtained from each explants were measured in factorial experiment with completely randomized design with three replications and three explants in each replicate. Molecular confirmation of hairy roots induction and bacterial strains were performed using specific amplification of rolC gene in PCR reactions. The results showed that all bacterial strains could initiate hairy root production. Transgenic roots were thicker, more branches and higher growth rate compared to normal roots. 15834 bacterial strain and greenhouse petiole and in vitro root explants demonstrate the highest ability in hairy root production. The highest growth of hairy roots were observed in greenhouse stem explant with MSU bacterial strain.

کلیدواژه‌ها [English]

  • Agrobacterium
  • Arbuscular mycorrhizal propagation
  • Carrot
  • Root organ culture
  • Symbiosis
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