اثر میدان مغناطیسی بر توان‌زیستی، بتولینیک اسید و خواص آنتی‌اکسیدان توس (Betula pendula Roth.)

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 دانشجوی دکتری ،گروه جنگل‌شناسی و اکولوژی جنگل، دانشکده علوم جنگل، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران.

2 دانشیار گروه جنگل‌شناسی و اکولوژی جنگل، دانشکده علوم جنگل، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران.

3 دانشیار ، گروه مکانیک بیوسیستم، دانشکده مهندسی آب و خاک، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، ایران.

چکیده

سابقه و هدف: تجمع متابولیت‌های ثانویه در گیاهان بخشی از پاسخ‌های دفاعی در برابر حملات پاتوژنی است که معمولاً به مقدار کم در سلول‌های گیاهی تجمع می‌یابند و محرک‌ها می­توانند مقدار آن‌ها را افزایش دهند. کشت‌ بافت و سلول گیاهی به‌عنوان یک روش پایدار برای بررسی متابولیت‌های ثانویه گیاهی به‌طور گسترده مورد استفاده قرار می­گیرد. بتولینیک اسید یکی از مهم‌ترین متابولیت‌های ضد سرطان و ضد ویروس ایدز است که اصلی‌ترین منبع اولیه تهیه آن درخت توس است. با توجه به در حال انقراض بودن این درختان در ایران، جایگزین نمودن روش‌های نوین کشت ‌سلول و بافت به‌جای استخراج از پوست این‌گونه امری ضروری است. پژوهش حاضر در همین راستا و باهدف افزایش مقدار ماده مؤثر تولیدی صورت گرفته است.
مواد و روش‌ها: ریزنمونه‌های پوست داخلی توس در محیط کشت NT با 3 درصد ساکارز، 8/0 درصد آگار، 5/2 میلی‌گرم بر لیتر 2-4,D و 5/0 میلی‌گرم بر لیتر BAP کشت شد و کالزایی صورت گرفت. بازکشت­ها به‌صورت ماهانه انجام شد و سلول‌های 8 ماهه تحت اثر میدان مغناطیسی قرار گرفتند. سلول‌های توس در کشت تعلیقی (حدود 1 گرم کالوس نرم و سفید در 30 میلی‌لیتر محیط کشت NT با ترکیبات هورمونی فوق‌الذکر بدون آگار) توسط میدان مغناطیسی ایستا با شدت 30 میلی تسلا و در روزهای 11-8 بعد از واکشت، روزی 4 ساعت تیمار شدند. سپس زنده‌مانی سلول‌ها، میزان رشد، بتولینیک اسید و خواص آنتی‌اکسیدانی در سلول‌های تیمار شده با میدان مغناطیس، اندازه‌گیری و با سلول‌های تیمار نشده مقایسه گردید.
یافته‌ها: نتایج نشان داد میزان رشد، بتولینیک اسید و خواص آنتی‌اکسیدانی در سلول‌های تیمار شده توسط میدان مغناطیس نسبت به شاهد افزایش یافت؛ اما زنده‌مانی سلول‌ها تحت‌تأثیر میدان مغناطیسی قرار نگرفت، بااین‌حال میزان مغناطیس واردشده باعث از بین رفتن و مرگ سلولی نشد. در کشت‌های تیمار شده تولید بتولینیک اسید کل (98/18 گرم بر لیتر) نسبت به کشت‌های شاهد (28/7 گرم بر لیتر) 5/2 برابر و نسبت خواص آنتی‌اکسیدانی در کشت‌های تیمار شده (09/40 درصد) نسبت به کشت‌های شاهد (2/14 درصد) 8/2 و نسبت رشد در کشت‌های تیمار شده (12/0 درصد) نسبت به کشت‌های شاهد (09/0 درصد) 2/1 برابر شد.
نتیجه‌گیری: در این پژوهش میدان مغناطیسی نیز مثل دیگر القاکننده‌ها یا محرک­ها با راه‌اندازی زنجیره انتقال پیام از طریق استرس اکسیداتیو ایجادشده، متابولیسم ثانویه را در سلول‌ها القاء نموده و باعث افزایش تولید متابولیت‌های ثانویه در سلول‌های توس نسبت به نمونه‌های شاهد شدند. هم‌چنین می‌توان گفت میدان مغناطیس به‌عنوان یک تنش مصنوعی باعث واکنش سلول‌ها و افزایش رشد و تولید مواد مؤثره شد. این محرک می‌تواند با افزایش میزان بتولینیک اسید و ظرفیت آنتی‌اکسیدان، در کشت­های این­ویترو این گیاه دارویی در حال انقراض به کار رود.

کلیدواژه‌ها


عنوان مقاله [English]

The effect of magnetic field on vitality, betulinic acid and antioxidant properties in birch (Betula pendula Roth.)

نویسندگان [English]

  • Mansooreh Aghasizadeh Shaarbaf 1
  • Vahideh Payamnoor 2
  • Abbas Rezaei Asl 3
1 Gorgan University of Agricultural Sciences and Natural Resources, Iran
2 Associate Prof., Dept. of Silviculture and Forest Ecology, Faculty of Forest Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
3 Gorgan University of Agricultural Sciences and Natural Resources, Iran
چکیده [English]

Background and Objectives: Accumulation of secondary metabolites in plants is part of the defensive responses to pathogenic attacks that induce and activate the inducers. Plant tissue or cell culture is widely used as a sustainable method for the study of plant secondary metabolites. Betulinic acid is one of the most important anti-cancer and anti-AIDS metabolites, which mainly comes from birch species (Betula pendula Roth). Due to the near extinction of these species in Iranian forests, there is need to develop modern cell and tissue culture techniques instead of traditional methods of skin extraction. The present study was carried out in the same direction and with the aim of increasing the amount of effective substance produced.
 
Materials and Methods: Birch inner skin explants were cultured in
NT+ (2.5 mlg/l) 2-4, D + (0.5 mlg/l) BAP with 3% sucrose and 0.8%
agar and callogenesis was conducted. Subcultures were performed once a month and the 8-month-old cells were exposed to a magnetic field.
Birch cells were treated in suspension culture (approx. 1 g of soft and
white callus in 30 ml of NT culture medium with the above-mentioned hormonal compounds without agar) by static magnetic field with
an intensity of 30 mT and on days 8-11 after sub culture, 4 hours a day. Next, cell viability, growth rate, betulinic acid and antioxidant properties in cells treated with a magnetic field were measured and compared to untreated cells.
 
Results: The results of this study showed that the growth rate, betulinic acid and antioxidant properties of treated cells increased compared to control cells. However, cell viability was not affected by the magnetic field. The amount of magnetic input did not cause cell death or death. In the treated cultures, the total betulinic acid production (18.51 g/l), the ratio of antioxidant properties (40.09%) and the ratio of growth (0.12%) were 2.5, 2.8 and 1.2 times higher than the control cultures of (7.28 g/l), (40.09%) and (0.09), respectively.
Conclusion: In this study, the magnetic field, like other inducers or elicitors, induces secondary metabolism in cells by triggering the message chain through oxidative stress, and increased the production of secondary metabolites in birch cells compared to control samples. It can also be said that the electromagnetic field as an artificial stress caused the cells to react and increase the growth and production of active substances. This elicitor can increase the medicinal properties of birch by increasing the amount of betulinic acid and antioxidant capacity.

کلیدواژه‌ها [English]

  • Cell culture
  • birch
  • betulinic acid
  • antioxidant
  • magnetic field
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