Contamination control in Iranian seedless barberry micropropagation

Document Type : Research Paper


1 Department of Horticultural Science and Landscape, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

2 Khorasan Science and Technology Park (KSTP), Mashhad, Iran.

3 Department of Ornamental Plants, Research Center for Plant Sciences, Ferdowsi University of Mashhad, Mashhad, Iran.


Iranian seedless barberry is a species with high degree of contamination in in vitro culture. The current study was aimed to investigate the effects of mercuric chloride (HgCl2), sodium hypochlorite (NaOCl) + HgCl2, HgCl2 + 8-hydroxyquinoline, cefotaxime + streptomycin, HgCl2 + shoot type, HgCl2 + explant position and HgCl2 + shoot color on the reduction of contamination. The results showed that the combined use of NaOCl and HgCl2 was more effective than their single use, but the contamination rate was relatively high. Interestingly, the results showed that explants sterilized with 100 mg/L HgCl2 and cultured in the medium containing 150 mg/L 8-hydroxyquinoline exhibited the lowest fungal contamination (15%), while those treated with 50 mg/L HgCl2 and cultured in the 8-hydroxyquinoline-free medium showed the highest fungal contamination (96%). Furthermore, treatment of 75 mg/Lcefotaxime + 70 mg/L streptomycin exhibited no bacterial contamination compared to the control with 58% bacterial contamination. Explants prepared from suckers and treated with 150 mg/L HgCl2 showed the lowest rates of fungal (16%) and bacterial (13%) contamination. The results also showed that explants collected from upper position had the lowest rates of fungal (20.87%) and bacterial (18.4%) infection. Explants taken from pinkish-colored shoots showed surprisingly the lowest rates of fungal contamination (13.33%), bacterial contamination (10%) and bud mortality (36.47%). In general, choice of explants from upper position of pinkish-colored suckers, sterilization of them with 100 mg/L HgCl2 for 5 min and their culture in medium containing 150 mg/L 8-hydroxyquinoline can be recommended for reducing the contamination.


Article Title [Persian]

کنترل آلودگی در ریزازدیادی زرشک بیدانه ایرانی

Abstract [Persian]

زرشک بیدانه ایرانی گونه‌ای با درجه آلودگی بالا در کشت بافت می‌ باشد. مطالعه حاضر با هدف بررسی تأثیر کلرید جیوه، هیپوکلرید سدیم + کلرید جیوه، کلرید جیوه + 8 هیدروکسی کینولین، سفوتاکسیم + استرپتومایسین، کلرید جیوه + نوع شاخه، کلرید جیوه + موقعیت ریزنمونه و کلرید جیوه + رنگ شاخه بر کاهش آلودگی اجرا گردید. بر اساس نتایج حاصل، استفاده ترکیبی از هیپوکلرید سدیم و کلرید جیوه مؤثرتر از استفاده یگانه کلرید جیوه بود، اما میزان آلودگی نسبتاً بالا بود. نتایج به‌طور شگفت‌انگیزی نشان داد که ریزنمونه­ های ضدعفونی­شده با 100 میلی‌گرم بر لیتر کلرید جیوه و کشت آن‌ ها در محیط حاوی 150 میلی­گرم بر لیتر 8 هیدروکسی کینولین کمترین میزان آلودگی  قارچی (%15) را داشتند، درحالی‌که ریزنمونه ­های تیمار شده با 50 میلی‌گرم کلرید جیوه و کشت آن‌ ها در محیط فاقد 8-هیدروکسی کینولین بیشترین میزان آلودگی قارچی (%96) را دارا بودند. همچنین، در مقایسه با تیمار کنترل با 58% آلودگی باکتریایی، تیمار 75 میلی‌گرم بر لیتر سفوتاکسیم + 70 میلی‌گرم بر لیتر استرپتومایسین فاقد هر ‌گونه آلودگی باکتریایی بود. ریزنمونه ­های تهیه‌شده از پاجوش و تیمار شده با 150 میلی‌گرم بر لیتر کلرید جیوه کمترین میزان آلودگی قارچی (%16) و باکتریایی (%13) را نشان دادند. همچنین نتایج حاکی از آن بود که ریزنمونه­ های جمع‌آوری‌شده از موقعیت بالایی، از کمترین میزان آلودگی قارچی (87/%20) و باکتریایی (4/%18) برخوردار بودند. ریزنمونه ­های تهیه شده از شاخه‌های صورتی‌رنگ به‌طور شگفت‌انگیزی کمترین میزان آلودگی قارچی (33/%13)، آلودگی باکتریایی (%10) و مرگ جوانه (47/%36) را نشان دادند. به‌طورکلی، انتخاب ریزنمونه­ ها از موقعیت بالایی پاجوش با رنگ صورتی، ضدعفونی آن‌ها با 100 میلی‌گرم بر لیتر کلرید جیوه به مدت 5 دقیقه و کشت آن‌ها در محیط حاوی 150 میلی‌گرم بر لیتر 8-هیدروکسی کینولین برای کاهش آلودگی پیشنهاد می‌ گردد.

Keywords [Persian]

  • 8-هیدروکسی کینولین
  • آلودگی
  • جوانه
  • ریزازدیادی
  • زرشک بیدانه ایرانی
  • کلرید جیوه
  • نوع شاخه
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