نوع مقاله : علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناس ارشد بیوتکنولوژی، دانشکده‌ کشاورزی، ‌دانشگاه ‌لرستان، خرم‌آباد، ایران

2 دانشیار، گروه زراعت ‌و اصلاح ‌نباتات، دانشکده‌ کشاورزی، ‌دانشگاه ‌لرستان، خرم‌آباد، ایران

3 استاد، گروه زراعت ‌و اصلاح ‌نباتات، دانشکده‌ کشاورزی، ‌دانشگاه ‌لرستان، خرم‌آباد، ایران

چکیده

گیاه خشخاش (Papaver somniferum L.)، از مهم‌ترین گیاهان در صنعت دارویی جهان و منشأ تولید آلکالوئیدها می‌باشد. یکی از تکنیک‌های تولید آلکالوئید، استفاده از کشت ریشه مویین می‌باشد؛ امّا در بیشتر مواقع تولید آلکالوئیدها در مقیاس تجاری کم است و برای افزایش آن‌ها، نیاز به تحریک تولید با روش‌های مختلفی از جمله استفاده از محرک‌های زیستی و غیرزیستی است. در این پژوهش، به‌ منظور افزایش زیست‌توده ریشه‌های مویین، اثر 4 نوع محرک (سالیسیلیک اسید، نیترات نقره، سولفات مس و عصاره مخمّر) به‌صورت آزمایش‌های جداگانه و در قالب طرح کاملاً تصادفی با سه تکرار مورد مطالعه قرار گرفت. جهت انجام این آزمایش‌ها، ریشه‌های مویین اولیه تولید شده در نهایت درون ارلن‌های حاوی 30 میلی‌لیتر محیط کشت  ½MSاضافه شدند و هر ارلن به عنوان یک تکرار در نظر گرفته شد. تأیید تراریختگی ریشه‌های مویین با تکثیر اختصاصی ژن‌های rolC، انجام شد و باندی با اندازه‌ای مطابق انتظار، روی ژل الکتروفورز مشاهده گردید. نتایج نشان داد که غلظت 5/0 میلی‌گرم در میلی‌لیتر از محرک عصاره مخمّر بیش‌ترین تأثیر را بر صفات مورفولوژیکی داشت. تجزیه و تحلیل داده‌ها نشان داد که محرک‌های زیستی، نقش مؤثرتری نسبت به تیمارهای غیرزیستی دارند و غلظت‌های بالای محرک‌ها، اعم از زیستی و غیرزیستی، بر زیست‌توده ریشه‌های مویین، تأثیر منفی دارند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Effect of Different Elicitor Treatments on Hairy Root of Medicinal Plant Poppies (Papaver somniferum L.)

نویسندگان [English]

  • Sh. Siahmansour 1
  • A. Ismaili 2
  • F. Nazarian Firouzabadi 3

1 M.Sc. Student of Biotechnology, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

2 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

3 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran

چکیده [English]

Abstract
Background and Objectives
As a source of alkaloids, Poppy (Papaver somniferum L.), is one of the most important plants in the field of pharmaceutical industry. Various methods including use of elicitors, adding precursor, optimizing culture, culture of hairy roots and its metabolites engineering are used to increase the production of secondary metabolites in plant cell cultures. In recent years, cultivation of hairy roots, to the production of valuable metabolites in a number of species of medicinal plants, has taken on commercial scale; but in most cases, this technique produces low amount of alkaloid in commercial scale. Hence, it is needed to optimize the production rate using biological and non-biological elicitors.
Materials and Methods
This experiment was conducted in 2015 in biotechnology laboratory of Lorestan University, Khorramabad, Iran. Sterile explants were prepared from seedling and were then inoculated with Agrobacterium. Inoculated explants were kept in a dark condition for 2 days and then washed and transferred to glasses containing½ MS medium complemented with cefotaxime. Finally, the glasses were kept in tissue culture room until production of hairy roots. The produced hairy roots were used as explants for elicitors' experiments in flask containers. In this study, in order to increase biomass of hairy roots, 4 types of elicitor treatments (salicylic acid, silver nitrate, copper sulphate and yeast extract) were evaluated in separate experiments based on a completely randomized design with 3 replicates.
Results
Confirmation of transgenic hairy roots was tested by PCR using the rolC gene primers to ensure the removal of bacteria from the produced roots. Results of PCR products were separated by electrophoresis and expected fragments were observed in gel. The results showed that the concentration of 5.0 mg/mL of yeast extract elicitor had the most impact on morphological traits. Totally, results showed that biotic elicitors had better effects in comparison to abiotic elicitors. Also, high concentrations of elicitors in all biotic and abiotic cases had a negative impact on biomass of hairy roots.
Discussion
It was shown that biotic elicitors had better effects than abiotic elicitors. The remarkable result of this study was that the high concentration of elicitors had negative effects on biomass hairy root production in poppies plants. It seems that bio-active compounds elicitors induce the plant responses but high concentration of biotic and abiotic elicitors may cause cell death and browning of hairy root tissues.

کلیدواژه‌ها [English]

  • Abiotic
  • Agrobacterium rhizogenesis
  • Biomass
  • Biotic
  • Transgenic
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