آنالیز بیوشیمیایی و آنتی اکسیدانی کافیئک اسید ریشه‌های مویین گیاه خرفه (Portulaca oleracea L.) تراریخته با آگروباکتریوم ریزوژنز
محورهای موضوعی : گیاهی
فرح فراهانی
1
*
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معصومه انصاری بیدار
2
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محسن زرگر
3
1 - استاد، گروه زیست‌شناسی، واحد قم، دانشگاه آزاد اسلامی، قم، ایران
2 - گروه زیست‌شناسی، واحد قم، دانشگاه آزاد اسلامی، قم، ایران
3 - گروه زیست‌شناسی، واحد قم، دانشگاه آزاد اسلامی، قم، ایران
کلید واژه: انتقال ژن, خرفه, ریشه مویین, آنتی اکسیدان.,
چکیده مقاله :
هدف: خرفه در طب سنتی بسیاری از کشورها استفاده می‌شود. ریشه خرفه حاوی ترکیبات ثانویه مفید و به مقدار بسیار کم هستند. هدف تحقیق حاضر بررسی کاربرد یکی از روش‌های زیست فناوری، به منظور افزایش تولید ترکیبات موثر دارویی با تولید ریشه‌های مویین می‌باشد.
مواد و روش‌ها: دانه‌های خرفه ضدعفونی شده کشت شدند، محور زیر لپه دانه رست‌ها با آگروباکتریوم ریزوژنز (جدایه‌های A15 و A4) تلقیح شدند. آزمایش‌های PCR (Polymerase Chain Reaction) برای تایید انتقال ژن انجام شدند. محور زیر لپه گیاهان تلقیح شده و شاهد (تلقیح نشده با باکتری) بر روی محیط کشت (Murashig & Skoog, 1962) MS جامد کشت داده شدند. وجود کافیئک اسید حاصل از گیاهان تراریخته و کنترل با آنالیزهای HPLC (High Performance Liquid Chromatography) و فعالیت آنتی اکسیدانی با آزمون DPPH (2,2-diphenyl-1-picrylhydrazyl) مورد ارزیابی قرار گرفتند.
یافته‌ها: آزمایش‌های PCR حضور ژن rolB از جدایه‌های A15 و A4 را در خرفه تراریخته نسبت به کنترل (فاقد ژن rol) نشان دادند. پس از تشکیل کالوس، ریشه‌های مویین القاء شدند. وجود ریشه‌های مویین خرفه تراریخته در سطح احتمال %5 معنی‌دار بودند. آنالیز HPLC بر وجود مقدار بیشتر کافئیک اسید در خرفه تراریخته را نسبت به شاهد تایید دارد، که 34 درصد بالاتر بودند. فعالیت آنتی اکسیدانی خرفه تراریخته با جدایه A4 737/86 بودند که درصد بازدارندگی در غلظت پایین انجام شده است.
نتیجه‌گیری: ریشه‌های مویین خرفه تراریخته می‌توانند برای افزایش بازده تولید آلکالوییدهای کافئیک اسید در صنایع داروسازی قابل استفاده باشند.
Objective: Purslane is used in traditional medicine in many countries. Its roots contain beneficial secondary metabolites but in very low quantities. The aim of this study was to evaluate the application of a biotechnological method to enhance the production of pharmaceutically active compounds through hairy root induction.
Materials and Methods: Disinfected purslane seeds were cultivated. Hypocotyl segments of seedlings were inoculated with Agrobacterium rhizogenes (strains A15 and A4). PCR (Polymerase Chain Reaction) was performed to confirm gene transfer. Inoculated hypocotyls and controls (non-inoculated) were cultured on solid Murashige and Skoog (MS, 1962) medium. Caffeic acid content in transgenic and control plants
was analyzed via HPLC (High Performance Liquid Chromatography), and antioxidant activity was assessed using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay.
Findings: PCR confirmed the presence of the rolB gene from strains A15 and A4 in transgenic purslane compared to controls (lacking the rol gene). Callus formation was followed by hairy root induction. Transgenic hairy roots showed significant induction at the 5% probability level. HPLC analysis revealed 34% higher caffeic acid content in transgenic plants compared to controls. The antioxidant activity of transgenic purslane with strain A4 exhibited 86.737% inhibition at a low concentration.
Conclusion: Transgenic hairy roots of purslane can be utilized to enhance the production yield of caffeic acid alkaloids for pharmaceutical industries.
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