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اثر تغذیه ساکارز بر افزایش رشد و تولید ترکیبات فنولی در کشت ریشه نابجا سرخارگل (Echinacea purpurea (L.) Monech) | ||
| تحقیقات گیاهان دارویی و معطر ایران | ||
| مقاله 8، دوره 40، شماره 3 - شماره پیاپی 121، مهر 1403، صفحه 521-543 اصل مقاله (1.27 M) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/ijmapr.2024.363372.3364 | ||
| نویسندگان | ||
| سیده محدثه حسینی1؛ سید علی اندی* 2؛ امیر صحرارو1؛ محمد باقر فرهنگی1 | ||
| 1دانشگاه گیلان | ||
| 2دانشگاه تخصصی فناوری های نوین آمل | ||
| چکیده | ||
| سابقه و هدف: ریشه نابجا در گیاهان دارویی منبع غنی از متابولیت‌های ثانویه باارزش هستند. سرخارگل (Echinacea purpurea (L.) Monech) یک گیاه دارویی مهم است و به‌طور گسترده در صنعت دارویی استفاده می‌شود. ریشه‌ها به‌طور سنتی در داروهای گیاهی و مکمل‌های غذایی به عنوان یک محرک سیستم ایمنی در درمان بیماری‌های التهابی، ویروسی و تنفسی استفاده می‌شوند. ناهمگنی سنتز ترکیبات مؤثره در محیط طبیعی، عملکرد پایین تولید گیاه در کنار عوامل دیگر ازجمله علل جست‌وجو برای رویکردهای جایگزین تولید مواد و اندام مؤثره گیاه هستند. در میان تکنیک‌های مختلف کشت‌های درون شیشه‌ای، کشت بافت‌های تمایز یافته همانند ریشه، معمولاً مقادیر بالاتری از متابولیت‌ها را نسبت به بافت‌های تمایز نیافته مانند کالوس و سلول‌های کشت مایع سلولی تولید می‌کنند. مواد و روش‌ها: در این مطالعه، هدف اولیه انتخاب بهترین ریزنمونه گیاهی و شناسایی دقیق مؤثرترین ترکیب از تنظیم‌کننده‌های رشد گیاهی بود که بتواند استقرار موفقیت‌آمیز کشت مایع ریشه نابجا را تسهیل کند. این بررسی شامل ارزیابی سه ریزنمونه گیاهی متمایز برگ، دمبرگ و ریشه بود. این ریزنمونه‌ها تحت تأثیر غلظت‌های مختلف 0، 0.5، 1، 1.5 و 2 میلی‌گرم در لیتر از دو نوع تنظیم‌کننده رشد گیاهی ایندول-3 - بوتیریک اسید (IBA) و 1- نفتالن استیک اسید (NAA) قرار گرفتند. تعداد ریشه‌های نابجا در ریزنمونه‌ها در غلظت‌های مختلف از تنظیم‌کننده‌های رشد گیاهی پس از شش هفته اندازه‌گیری شد. در آزمایش مجزای دیگری، اثر غلظت‌های مختلف ساکارز شامل 10، 20، 30، 40، 50 و 60 گرم در لیتر محیط کشت بر روی ویژگی‌های دینامیکی رشد (وزن تر و خشک، حجم محیط باقیمانده، هدایت الکتریکی (EC) و pH) و فیتوشیمیایی (فنول کل، فلاونوئید کل و فعالیت آنتی‌اکسیدانی) ریشه در طول چهار هفته کشت، بررسی شد. برای اعمال تیمارهای ساکارز، مقدار 0.35 گرم وزن تر از ریشه‌های نابجا در داخل ارلن مایرهای 250 میلی‌لیتری حاوی 50 میلی‌لیتر نصف محیط کشت MS، یک میلی‌گرم در لیتر IBA به همراه غلظت‌های مختلف ساکارز با pH= 5.8 قرار داده شدند. محتوای سه فلاسک هر تیمار در هر هفته برای اندازه‌گیری استفاده شد. مطالعات در قالب آزمایش فاکتوریل به صورت طرح پایه کاملاً تصادفی انجام شدند و مورد تجزیه‌وتحلیل آماری با استفاده از نرم‌افزار SPSS قرار گرفتند. نتایج: ریشه‌های نابجا در ریزنمونه‌های برگ بسیار مؤثرتر از ریزنمونه‌های ریشه و دمبرگ بودند. میانگین تعداد ریشه‌های نابجا در ریزنمونه برگ 93.5% بود، در حالی که ریزنمونه‌های دمبرگ و ریشه به ترتیب 6.5 و صفر درصد ریشه نابجا تولید کردند. محیط MS حاوی یک میلی‌گرم در لیتر IBA حداکثر تعداد ریشه را با میانگین 33.3% ایجاد کرد، در حالی که غلظت‌های صفر و دو میلی‌گرم در لیتر از هر دو هورمون استفاده شده ریشه‌ای تولید نکردند. بیشترین وزن تر با میزان 29.8 گرم در لیتر در هفته چهارم کشت مشاهده شد که این میزان 4.3 برابر وزن تلقیح اولیه بود. با پیشرفت مدت زمان کشت، میزان EC و pH محیط کشت کاهش یافت. البته افزایش زمان کشت، تأثیر قابل‌توجهی بر تولید فلاونوئید کل و فعالیت آنتی‌اکسیدانی مرتبط در ریشه‌های نابجا نداشته است. با وجود این، فنول کل با میزان 56.6 میلی‌گرم گالیک اسید در هر گرم وزن خشک در هفته چهارم تحت تأثیر افزایش معنی‌دار مدت زمان کشت قرار گرفت. به‌طورکلی، نتایج نشان داد که غلظت‌های کمتر ساکارز برای تولید زیست‌توده و محتوای ترکیبات مؤثره و فعالیت آنتی‌اکسیدانی کارآمدتر هستند. بالاترین تجمع زیست‌توده و محتوای فنول کل، بیشترین مقدار از محتوای فلاونوئید کل و بالاترین درصد مهار رادیکال آزاد به ترتیب با به‌کارگیری غلظت‌های سه، دو و 1% ساکارز در هفته چهارم کشت اندازه‌گیری گردید. در کل، همبستگی معکوسی بین میزان EC محیط کشت باقیمانده و زیست‌توده ریشه‌ها در غلظت‌های پایین‌تر ساکارز با افزایش مدت زمان کشت مشاهده شد. نتیجه‌گیری: از آنجایی که کشت مایع ریشه نابجا E. purpurea به عنوان یک منبع جایگزین بالقوه برای تولید متابولیت‌های ثانویه به‌ویژه مشتقات کافئیک اسید (Caffeic acid) شناسایی می‌شوند، یافته‌های این پژوهش ممکن است به مطالعات تولید انبوه اندام مؤثره ریشه و به دنبال آن، این ترکیب‌های خیلی با ارزش با استفاده از سیستم‌های بیوراکتور مناسب آزمایشگاهی کمک کند. | ||
| کلیدواژه‌ها | ||
| سرخارگل (E. purpurea (L.) Monech)؛ تغذیه ساکارز؛ کشت ریشه نابجا؛ زیست‌توده؛ ترکیبات فنولی | ||
| عنوان مقاله [English] | ||
| The impact of sucrose feeding on augmenting growth and phenolic compounds bio-production in adventitious root culture of Echinacea purpurea (L.) Monech | ||
| نویسندگان [English] | ||
| Seyedeh Mohaddeseh Hosseini1؛ Seyed Ali Andi2؛ Amir Sahraroo1؛ Mohammad Bagher Farhangi1 | ||
| 1Guilan University | ||
| 2Amol University of Special Modern Technologies | ||
| چکیده [English] | ||
| Background and objectives: Adventitious root cultures of medicinal plants represent a bountiful reservoir of valuable secondary metabolites. The esteemed medicinal plant Echinacea purpurea (L.) Monech is highly sought after for its phytochemical properties, with a traditional use of its roots in herbal medicines and dietary supplements as an immune system stimulant for treating inflammatory, viral, and respiratory diseases. However, the heterogeneous synthesis of bioactive compounds in natural environments and the limitations of conventional plant production methods necessitate exploring alternative approaches for obtaining active plant substances and organs. Among various in vitro cultivation techniques, the cultivation of differentiated tissues, specifically roots, consistently demonstrates the ability to yield higher quantities of metabolites than undifferentiated tissues such as callus and suspension cells. Methodology: The primary aim of this study was to accurately determine the ideal combination of plant explants and plant growth regulators that would facilitate the successful establishment of adventitious root suspension cultures. Three distinct plant explants, namely leaf, petiole, and root, were carefully evaluated in this study. These explants were subjected to varying concentrations (0, 0.5, 1, 1.5, and 2 mg.L-1) of two types of plant growth regulators: indole-3-butyric acid (IBA) and 1-naphthalene acetic acid (NAA). After six weeks, the number of adventitious roots developed by the explants under different concentrations of plant growth regulators was measured. The impact of different sucrose concentrations (10, 20, 30, 40, 50, and 60 grams per liter) in the culture medium was investigated over a four-week cultivation period. Various dynamic characteristics of root growth, including fresh weight, dry weight, volume of the remaining medium, electrical conductivity, and pH, were closely examined. Furthermore, phytochemical properties such as total phenol, total flavonoid, and antioxidant activity were assessed. To administer the sucrose treatments, 0.35 grams of adventitious roots were placed inside 250 ml Erlenmeyer flasks containing 50 ml of half MS culture medium, 1 mg.L-1IBA, and varying concentrations of sucrose, maintaining a pH of 5.8. Measurements were taken from three flasks of each treatment weekly. The experiments followed a Completely Randomized Factorial Design, with statistical analysis conducted using SPSS software. Results: Leaf explants exhibited superior root formation to root and petiole explants. The average number of adventitious roots in leaf explants reached 3.37, while petiole and root explants produced only 0.23 and no adventitious roots, respectively. The most effective root formation occurred in MS medium with 1 mg.L-1 IBA, resulting in an average of 4 roots per explant. Hormone concentrations of 0 and 2 mg.L-1 did not induce root formation. In the fourth week of cultivation, the highest fresh weight recorded was 29.8 grams per liter, marking a remarkable 3.4-fold increase from the initial inoculation weight. The culture medium's electrical conductivity and pH decreased as the cultivation progressed. Cultivation weeks did not significantly impact total flavonoid production and related antioxidant activity in adventitious roots. However, the cultivation period significantly affected the total phenol content at 57.56 mg of gallic acid per gram of dry weight. Altogether, lower sucrose concentrations proved more efficient for biomass production, bioactive compound content, and antioxidant activity. The highest biomass accumulation and total phenol content occurred at 3% sucrose. In comparison, in the fourth week of cultivation, the highest total flavonoid content and the highest free radical inhibition percentage were observed at 2% and 1% sucrose, respectively. Notably, a negative correlation was observed between the electrical conductivity of the remaining culture medium and root biomass in lower sucrose concentrations as the culture duration increased. Conclusion: In light of the immense potential harbored by adventitious root suspension cultures of E. purpurea as an alternative source for the production of secondary metabolites, particularly caffeic acid derivatives, the discoveries made in this research have the potential to advance the field of mass production for active root organs, thereby facilitating the synthesis of these precious compounds through the implementation of appropriate laboratory bioreactor systems. | ||
| کلیدواژه‌ها [English] | ||
| Purple coneflower (Echinacea purpurea (L.) Monech), sucrose feeding, adventitious root culture, biomass, phenolic compounds | ||
| مراجع | ||
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