بهینه‌سازی رشد گیاهچه و بررسی تولید تاکسان در کشت درون‌شیشه‌ای سرخدار (Taxus baccata L.)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری بیوتکنولوژی کشاورزی، پردیس دانشگاهی دانشگاه گیلان، گیلان، ایران

2 استاد گروه علوم باغبانی، دانشکده علوم کشاورزی دانشگاه گیلان، گیلان، ایران

3 استاد گروه زراعت و اصلاح نباتات، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، تهران، ایران

4 دانشیار گروه کشاورزی، پژوهشکده گیاهان و مواد اولیه دارویی دانشگاه شهید بهشتی، تهران، ایران

چکیده

سرخدار (Taxus baccata L.) به‌علت دارا بودن ترکیبات دی­ترپنوئیدی با اثرات ضد سرطان، از مهم­ترین گیاهان دارویی شناخته شده در جهان است که در معرض انقراض قرار دارد. در این تحقیق که در پژوهشکده گیاهان دارویی دانشگاه شهید بهشتی صورت پذیرفت، ابتدا اثر نوع محیط کشت (MS٬ MS۲/۱ و NN)، جاذب‌های فنولی (زغال فعال و PVP) و تنظیم کننده­های رشد گیاهی (IBA، GA3 و BA) در قالب طرح فاکتوریل با سه تکرار، با هدف بهبود جوانه‌زنی و رشد گیاهچه‌های سرخدار در کشت درون­شیشه‌ای بررسی گردید. در ادامه تولید تاکسان در گیاهچه‌هایی که طی ده ماه در محیط انتخابی رشد کرده بودند، اندازه‌گیری شد. محیط کشت NN به­دلیل غلظت نمک‌های پایه کم­تر در محیط کشت تأثیر معنی‌داری بر درصد جوانه‌زنی نداشت، ولی منجر به افزایش طول ریشه و درصد رویش عادی گیاهچه گردید. همچنین استفاده از جاذب‌های فنولی موجب افزایش بیش­تر طول ریشه و ساقه و بهبود نرخ رویش عادی گیاهچه گردید. به­علاوه ترکیب هورمونی IBA × GA3 (۲/۰ میلی‌گرم بر لیتر جیبرلیک اسید × ۰۱/۰ میلی‌گرم بر لیتر ایندول بوتیریک اسید) بیش­ترین تأثیر را بر میزان رویش عادی گیاهچه‌ها داشته و بیش­ترین طول ریشه با ۱/۰ میلی‌گرم بر لیتر ایندول بوتیریک اسید و حداکثر ارتفاع ساقه از ترکیب هورمونی BA × GA3 (۲/۰ میلی‌گرم بر لیتر جیبرلیک اسید × ۲ میلی‌گرم بر لیتر بنزیل آدنین) به‌دست آمد. همچنین میزان تاکسان‌ها در اندام هوایی بیش­تر از ریشه بوده و بیش­ترین تاکسان در اندام هوایی ۱۰- دی استیل باکاتین III و در ریشه تاکسول و ۱۰- دی استیل تاکسول بود که حداکثر مقدار آن‌ها در هفته چهلم به‌دست آمد. در کل نتایج این تحقیق حاکی از تأثیر مثبت استفاده از تنظیم‌کننده‌های رشد، زغال فعال و کاهش غلظت نمک‌های محیط کشت بر رشد ریشه و توسعه عادی گیاهچه به‌منظور بهینه‌سازی تکثیر گیاهچه‌های سرخدار در کشت درون­شیشه‌ای بود. همچنین نشان داده شد کشت‌ سلولی حاصل از اندام‌ هوایی نسبت به کشت ریشه‌های مویین گزینه بهتری در جهت تولید تاکسان‌ها در شرایط آزمایشگاهی است.

کلیدواژه‌ها


عنوان مقاله [English]

Seedling growth optimization and investigation of taxane production in Taxus baccata L. via in vitro culture

نویسندگان [English]

  • Mahsa Bamneshin 1
  • Abdolah Hatamzadeh 2
  • Mohammad Reza Naghavi 3
  • Mohammad Hossin Mirjalili 4
1 Ph.D. Student, Department of Biotechnology, Agricultural and Natural Resources College, Pardis of Guilan University, Guilan, Iran
2 Professor., Department of Horticultural Sciences, Agricultural College, Guilan University, Guilan, Iran
3 Professor., Department of Agronomy and Plant Breeding, Agricultural and Natural Resources College, Tehran University, Tehran, Iran
4 Associate Professor., Department of Agriculture, Medicinal Plant and Drugs Research Institute, Shahid Beheshti University, Tehran, Iran
چکیده [English]

Yew (Taxus baccata L.) has diterpenoid compounds with anti-cancer effects therefore it is one of the most well-known medicinal plants that is in danger of extinction. In this study, which was conducted at the Research Institute of Medicinal Plants of Shahid Beheshti University, the effect of culture medium (MS, 1/2MS and NN), phenolic absorbents (PVP and activated charcoal) and plant growth regulators (IBA, GA3 and BA) was investigated in a factorial design with three replications, with the aim of improving germination and growth of seedlings in in vitro culture. In the following, taxane production was measured in plantlets that were grown in select medium for ten months. According to our results, NN culture medium due to lower concentration of basal salts had no significant effect on germination percentage but increased root length and normal seedling growth percentage. Also supplementing the media with phenolic absorbent extended root length and shoot height as well as increasing normal seedling growth rate. In using PGRs, the highest normal seedling growth rate, shoot height and root length was obtained respectively from IBA × GA3 (0.2 mg/l gibberellic acid × 0.01 mg/l indole butyric acid), IBA × BA (0.2 mg/l gibberellic acid × 2 mg/l benzyl adenine) and 0.1 mg/l indole butyric acid, while PGRs had no significant differences in germination rate. Also, the amount of taxan in the aerial part was higher than the root and the highest taxan in the aerial part was 10-deacetylbaccatinIII  and in the root were taxol and 10-deacetyltaxol, the maximum of all was achieved at the end of the experiment. Overall, the results indicate the positive effect of activated charcoal, PGRs and reducing culture medium macronutrients on root length, shoot height and the normal growth process of Taxus baccata seedling, in order to increase the yeild of yew propagation under in vitro culture condition. The cell culture obtained from the aerial part is much better than the hairy root cell culture. 

کلیدواژه‌ها [English]

  • Culture medium
  • Taxan
  • Taxol
  • Yew
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