تحریک ریشه‌های موئین تراریخت در گیاه دارویی شقایق (Papaver somniferum L) با استفاده از آگروباکتریوم رایزوژنز (Agrobacterium rhizogenes)

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان

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

3 مربی، گروه زیست‌شناسی، دانشکده علوم پایه، دانشگاه صنعتی خاتم الانبیاء بهبهان

چکیده

گیاه شقایق (Papaver somniferum L.) یکی از قدیمی‌ترین گیاهان داروئی شناخته شده است. با توجه به این‌که پتانسیلمواد دارویی گیاهان دارویی درشرایططبیعیبسیارمحدودمی‌باشد، از این رو کشت بافت گیاهی بهعنوان یکی ازروشهای کاربردی نقش مهمی درراستایتولیدصنعتیمتابولیت‌هایثانویهدرگیاهان دارویی دارد. از میان سیستم‌های کشت بافت، سیستمریشه‌هایموئیندرشرایطکشتبدون تنظیمکننده رشد بسیار پایداراستو به دلیل ثبات ژنتیکی و بیوشیمیایی ذاتی، متابولیت‌های ثانویه را در طی یک دوره‌ی طولانی تولید می‌کنند. در این پژوهش به منظور بهینهسازی شرایط کشت ریشه‌های موئین از سویه‌های مختلف آگروباکتریوم رایزوژنز (ATCC15834،A4  و GMI9534)، محیط‌های کشت MS و ½MS، دو دمای 25 و 17 درجه سلسیوس در دوره هم‌کشتی و دو نوع ریزنمونه‌ی اندام هوایی و محور زیرلپه استفاده گردید. صفت‌های درصد تحریک ریشه موئین، تعداد ریشه موئین در ریزنمونه و تعداد انشعاب فرعی ریشه موئین در طول یک سانتی‌متر از ریزنمونه اندازه‌گیری شد. واکاوی نتایج نشان داد که سویه‌ی ATCC15834، محیط کشت ½ MS، دمای 25 درجه سلسیوس همکشتی و ریزنمونه اندام هوایی بهترین ترکیب تیماری جهت تحریک ریشه موئین در گیاه شقایق هستند. با استفاده از واکنش PCR تأیید تراریختی ریشه‌های موئین با آغازگرهای اختصاصی ژن rolB انجام گرفت.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Induction of Transgenic Hairy Roots in Medicinal Plant Poppy (Papaver Somniferum L.) by Agrobacterium Rhizogenes-Mediated Transformation

نویسندگان [English]

  • A Jalilian 1
  • A A 2
  • F N 2
  • S.Z H 3
چکیده [English]

Background and Objectives
Opium poppy plant (Papaver somniferum L.) is one of the oldest known medicinal plants. Since the potential of medicinal components production in medicinal plants is very limited in natural conditions, plant tissue culture, as a new technology, has an important role in industrial production of secondary metabolites in medicinal plants. Among different tissue culture systems, hairy root is very stable in hormone-free culture conditions and these roots stably produce secondary metabolites over a long period due to their inherent genetic and biochemical stability. Hence, the objective of this study was to evaluate the effect of some different parameters on optimization of hairy production in P. somniferum.
Materials and methods
In the present research, for the optimization of hairy root in papaver, a factorial experiment was conducted based on a completely randomized design with 4 replications. The factors included different Agrobacterium rhizogenes strains (ATCC15834, A4 and GMI9534), medium (MS and ½ MS medium), two temperatures (17 and 25 °C) at co-cultivation period, and two different explants (excised shoot and hypocotyl). Different traits were evaluated including percentage of induced hairy roots, number of lateral branches/1cm, number of hairy root per explants and days after inoculation (DAI).
Results
Hairy roots were induced at the wound site of explants after weeks of culture. Analysis of the results showed that strain ATCC15834, ½ MS medium, temperature 25 °C and excised shoot explant were the best treatment combination for hairy roots induction in the poppy plant. Transformed hairy roots were confirmed by polymerase chain reaction (PCR) using rolB gene specific primers.
Discussion
All of the A. rhizogenes strains led to hairy root induction, but there were no hairy roots formed from control explants. Differences in virulence, morphology and growth rate can be partially explained by the variety of plasmids harbored by the A. rhizogenesis strains. In the present study, excised shoot explants were highly and significantly susceptible to infection by each strain of A. rhizogenes and hypocotyl explants showed low rate of hairy root induction.

کلیدواژه‌ها [English]

  • Hairy roots
  • Agrobacterium rhizogenes
  • Explant
  • Opium poppy
  • Strain
  • Medium
  • Temperature
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