تعیین دز مناسب مواد جهش‌زای فیزیکی و شیمیایی در بافت کالوس برگ کیوی‌فروت

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانشجوی دکتری میوه‌کاری، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران

2 استادیار، گروه باغبانی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران

3 سرپرست گروه تحقیقاتی، مرکز تحقیقات گیاه و غذا، مرکز پالمراستون نورث، نیوزلند

چکیده

چکیده
استفاده از مواد جهش‌زای فیزیکی و شیمیایی باعث ایجاد واریته‌های مقاوم به انواع بیماری، مقاوم به شرایط نامطلوب محیطی و حتی سبب بروز خصوصیات جدید در تولیدات کشاورزی در نقاط مختلف جهان شده‌ است. با توجه به اثرات کشنده خیلی از عوامل جهش‌زا در دزهای بالا، تعیین دزهای مناسب با توجه به نوع بافت، شرایط درون سلول و شرایط محیط رشد، گام اولیه در استفاده از این تکنیک‌ها است. استفاده از بافت کالوس به‌عنوان بافت هدف به‌جای جوانه یا بذر روش سریع‌تری جهت تولید گیاهچه‌های جدید جهش‌یافته بدون ظهور بافت شیمر را ممکن می‌سازد. در پژوهش حاضر با توجه به اهمیت اقتصادی کیوی و نقش ایران در تولید منطقه‌ای این محصول، از بافت کالوس آن به‌عنوان بافت هدف برای تعیین دزهای مناسب از اشعه گاما با شدت تابش صفر، 10، 20، 30، 40 و 50 گری به‌عنوان جهش‌زای فیزیکی و اتیل‌متان‌سولفونات (EMS) به‌عنوان جهش‌زای شیمیایی با غلظت‌های صفر، 4/0، 6/0، 8/0، 1 و 2/1 درصد در مرکز تحقیقات گیاه و غذا، نیوزلند در سال 2016انجام شد. شاخص‌های رشد کالوس و همچنین نشانگرهای مولکولی SSR در مرحله گیاهچه، جهت تعیین دزهای مناسب و تغییرات احتمالی ژنتیکی استفاده شد. نتایج نشان داد که با افزایش دز اشعه گاما و غلظت EMS، اندازه کالوس مرده نیز افزایش یافت اما بیشترین تعداد شاخه تولیدشده در تیمار اشعه گاما در دزهای 40 و 50 گری و در تیمار EMS در غلظت 1 درصد به‌دست آمد. لذا دز 50‌ گری اشعه گاما و غلظت 1 درصد EMS به‌عنوان مقادیر مناسب جهت استفاده در بافت کالوس دوماهه کیوی و ایجاد جهش به ‌منظور تولید خصوصیات متفاوت و جدید در جمعیت موتانت تشخیص داده‌ شدند.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Determination of the Optimal Dosage of Physical and Chemical Mutagenesis Agents in Callus Tissue of Kiwifruit Leaf

نویسندگان [English]

  • Hassan Saeiahagh 1
  • Mousa Mousavi 2
  • Ranjith Pathirana 3
1 Ph.D. Student of Pomology, Faculty of Agriculture, Shahid Chamran Unversity of Ahvaz, Ahvaz, Iran
2 Assistant Professor, Department of Horticultural Science, Faculty of Agriculture, Shahid Chamran Unversity of Ahvaz, Ahvaz, Iran
3 Science Team Leader, The New Zealand Institue for Plant and Food Research, Batchelar Road Palmerston North, New Zealand
چکیده [English]

Abstract
 
Background and Objectives
Induced mutations have been used in a wide range of horticultural crops for more than half a century. This approach has opened up great opportunities for biotechnologists and plant breeders to produce new varieties with several new traits including resistance to disease, and tolerance to adverse environmental conditions or even produce new horticultural characters in some crops. The optimization of dosage of the physical and chemical mutagenic agents for inducing mutations is the first step in using this technique in plant breeding. This study was thus conducted to determine the optimum dosage of two mutagenic agents including gamma radiation as a physical agent and ethyl methanesulphonate (EMS) as a chemical agent in kiwifruit callus in in-vitro conditions. To that end, simple sequence repeat (SSR) markers were used to detect the mutations in the regenerated plantlets.
 
Materials and Methods
Having induced the callus in tissue culture media containing naphthaleneacetic acid, 6-benzylamino purine and meta-Topolin, the produced calli were exposed to different dosage of gamma radiation (0, 10, 20, 30, 40, 50 Gy) or different EMS concentrations (0, 0.4, 0.6, 0.8, 1, 1.2 % for 60 min). Then, the callis were transferred to the shoot regeneration media. The number of dead calli, and the size of live calli as well as the number of shoots generated in live calli were then recorded. Afterwards, the regenerated shoots were transferred to elongation media, and the DNA was extracted. The SSR markers analysis was followed on the randomly selected plantlets to detect the SSR markers using a Genetic Analyzer.
 
Results
As expected, with increasing the dosage of gamma radiation, the number of dead calli increased and the callus growth was thus retarded. The lowest number of dead calli was, indeed, detected in the control and the 50 Gy dose resulted in the highest number of dead calli. The morphology of shoots generated in the treated calli and the SSR marker analysis showed that 50 Gy dose could be used as the optimum radiation dose to produce an acceptable mutation rate in kiwifruit callus. In turn, EMS as a chemical mutagenic agent showed a different pattern in increasing the number of shoots regenerated with increasing dosage, however the same trend happened with gamma radiation in the number of dead calli and live callus size. Considering the effect of EMS on callus and the number of shoots regenerated per callus, a treatment with 1% EMS for 60 min can be used to produce a high density mutant population in kiwifruit. The highest mutation rate (12.5 %) was observed in the 50 Gy gamma ray treatment with changes in 12 pairs of SSR markers among 25 loci. For EMS, this value was 2.6 % in 1% concentration treatment.
 
Discussion
To detect the induced mutations using Gamma ray and EMS, the SSR markers can be applied. However, for the EMS mutation detection, more studies are needed to evaluate the efficiency of other techniques such as single nucleotide polymorphism (SNPs). 

کلیدواژه‌ها [English]

  • Ethyl methanesulphonate
  • Gamma radiation
  • Mutagenesis
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 © 2019 by the authors. Licensee SCU, Ahvaz, Iran. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution-Non Commercial 4.0 International (CC BY-NC 4.0 license) (http://creativecommons.org/licenses/by-nc/4.0/)