ارزیابی روش‌های ضدعفونی ریزوم و بذر زنبق مردابی در شرایط درون‌شیشه‌ای

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم باغبانی، دانشگاه محقق اردبیلی، اردبیل، ایران

2 پژوهشکده ژنتیک و زیست فناوری کشاورزی طبرستان، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ایران

چکیده

زنبق مردابی یکی از گونه‌های زینتی بومی و ارزشمند است. اصلی‌ترین مشکل در شروع کشت بافت این گیاه وجود آلودگی‌های قارچی و باکتریایی شدید است که باعث شکست روش‌های معمول سترون‌کردن می‌شود. به‌منظور بهینه‌سازی کم‌خطرترین روش ضدعفونی ریزوم و بذر زنبق مردابی در شرایط کشت درون‌شیشه‌ای انجام شد. دو آزمایش جداگانه در قالب طرح کاملا تصادفی با 5 تکرار انجام شد. به ترتیب 16 و 15 ‌نوع روش برای ضدعفونی بذر و ریزوم به‌کار برده شد. ریز‌نمونه‌های تیمار شده پس از طی مراحل ضدعفونی در محیط‌کشت موراشی اسکوگ بدون هورمون کشت شدند، دو هفته پس از کشت، شاخص‌های مختلف از قبیل درصد آلودگی، درصد جوانه‌زنی، میانگین طول برگ و میانگین طول ریشه اندازه‌گیری شد. پس از این مرحله نمونه‌های سالم در محیط کشت موراشی اسکوگ حاوی غلظت‌های مختلف تیدیازرون و نفتالین استیک اسید جهت پرآوری، کشت شدند. نتایج حاصل از این پژوهش نشان داد که در تیمار پانزدهم (ریز‌نمونه‌های ریزوم، 60 دقیقه حمام آبگرم و 60 دقیقه قارچ‌کش فلوکونازول و آنتی‌بیوتیک استرپتومایسین و یک دقیقه الکل‌70 درصد و 10 دقیقه هیپوکلریت سدیم 4 درصد)، کمترین درصد آلودگی (11درصد) و بیشترین جوانه‌زنی 88 درصد، برترین تیمار بود. رتبه دوم مربوط به تیمار سیزدهم (60 دقیقه حمام آبگرم دمای 42 درجه+60 دقیقه در قارچ‌کش فلوکونازول+جنتامایسین+کلیندامایسین‌+یک دقیقه الکل 70درصد+10دقیقه هیپوکلریت سدیم3درصد) با جوانه‌زنی 85درصد و آلودگی 15درصد بود. در ریز‌نمونه‌‌های بذری، تیمار دوم و تیمار پنجم بیشترین میزان جوانه‌زنی (74/71درصد) مشاهده شد. نتایج پرآوری نشان داد که غلظت 5 میکرومولار تیدیازرون +6میکرومولار از نفتالین استیک اسید بیشترین میزان پرآوری را به همراه داشت.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Exploring Disinfection Methods for Iris pseudacorus L. Rhizome and Seed Under In-Vitro Conditions

نویسندگان [English]

  • vajihe Abbasi ghadi 1
  • Esmaeil Chamani 1
  • Younes Pourbeyrami hir 1
  • Mehdi Mohebodini 1
  • Vali Ollah Ghasemi Omran 2
1 Department of Horticultural Sciences, Mohaghegh Ardebili University, Ardebil, Iran.
2 Genetic and Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
چکیده [English]

Iris pseudacorus L. is one of the native and prominent ornamental species in Iran. Its propagation by tissue culture is prone to sever bacterial and fungal contaminations of the explants, particularly in rhizomes., which causes the failure of conventional sterilization methods. In order to optimize the least dangerous method for sterilization of Iris pseudacorus rhizome and seed under in vitro culture condition, two separate experiments were performed each in a completely randomized design with five replications. 16 and 15 methods were applied for rhizome and seeds steriliztion, respectively. The treated specimens were cultured in a hormone-free MS medium and two weeks later, various indices such as infection percentage, germination percentage, mean leaf length and mean root length were measured. Then intact specimens were transferred to the MS media containing different concentrations of thidiazuron (TDZ) and naphthalen acetic acid (NAA) for proliferation.
 The results showed that A15 treatment (water bath (42 °C) for 60 mins, fluconazole fungicide and antibiotics streptamycin for 60 mins, alcohol 70% for one min. and NaoCl 4% for 10 mins) on rhizome explants with the lowest contamination percentage (11%) and a high germination rate of 88% was the best treatment. Also, A13 treatment with 85% germination and 15% contamination obtained the second place. Seeds-treated with treatment B2 using hot water and 3% sodium hypochlorite and treatment B5 using hydropriming with running water and hypochlorite 2% had the highest germination rate of 71.74% and contamination was almost zero. Meanwhile the highest proliferation was obtained in the culture medium containing MS and a combination of 5 μM TDZ and 6 μM NAA.

کلیدواژه‌ها [English]

  • Heat treatment
  • antibiotic
  • sterilization
  • benomyl
  • tissue culture

Extended Abstract

Introduction

Iris pseudacorus L. is one of the native and prominent ornamental species in Iran. Its propagation by tissue culture is prone to sever bacterial and fungal contaminations of the explants, particularly in rhizomes., which causes the failure of conventional sterilization methods. In addition, low speed propagation by seed and season-dependent propagation by rhizomes, altogether have reduced florists tendency to use this beautiful flower in the ornamental plants industry. The present research aimed to optimize the best and least dangerous disinfection method for the in vitro cultivation of rhizomes and seeds of this species of Iris.

 

Materials and Methods

This research was conducted as two separate trials based on a completely randomized design with 16 treatments on rhizomes and 15 treatments on seeds, in five replications to overcome the microbial infections under in vitro propagation. After disinfecting, the treated explants were cultured in a hormone-free MS medium. Two weeks after cultivation, some parameters, including infection percentage, germination percentage, average leaf length, and average root length, were recorded. After that, healthy samples were cultivated in MS culture medium containing different concentrations of thidiazuron and naphthaleneacetic acid for proliferation.

 

Results and Discussion

Based on the results, compounds such as sodium hydroxide, warm water, and hydro-priming performed the best for seed sterilization. The highest germination rate (71.74 %) and least infection rate (almost zero) were observed in the seed explants of treatments B2 in which warm water and 3% sodium hypochlorite were applied, and also B5 in which hydro-priming with running water and immersion in 2% sodium hypochlorite were used. The sterilization of the rhizomes is much more difficult than that seeds since underground organs are in contact with a wide range of fungal and bacterial infections. Also I. pseudacorus habitats on the margins of marshes, banks of rivers and streams are subjected to wider infections. Based on the findings, sodium hypochlorite does not alone sufficent to control the bacterial and fungal infections of I. pseudacorus L. It was found that the rhizome explants treated by A15 exhibited an infection rate of 11 percent and a germination rate of 88 percent. Also, treatment A13 with a germination rate of 85 percent and an infection rate of 15 percent was the second-best treatment. In general, the use of hot water bath at 42°C and 46°C and the use of fungicides benomyl and fluconazole and antibiotics streptomycin, gentamicin, and clindamycin outperformed antibiotics terbinafine and nystatin. Data analysis of sterilized samples showed that the concentration of 5 μM TDZ and 6 μM NAA hormones resulted in the highest amount of proliferation.

 

Conclusion

The fungicides benomyl and fluconazole along with clotrimazole and antibiotics streptomycin, gentamicin and clindamycin were more effective in controlling intensive fungal and bacterial infections of rhizomes. With the advantage of better effectiveness and greater safety, utilizing hot water bath, along with fungicides and antibiotics can be a more efficient method for providing the required growth conditions for  the rhizomes-derived explants of I. pseudacorus instead of the use of toxic, dangerous, and harmful compounds.

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علوم باغبانی ایران
دوره 55، شماره 1
فروردین 1403
صفحه 153-175

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  • تاریخ دریافت: 27 بهمن 1401
  • تاریخ بازنگری: 07 شهریور 1402
  • تاریخ پذیرش: 19 شهریور 1402

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