بهینه‌سازی کالوس زایی و بررسی میزان بیان ژن‌های فلاونول سنتاز و چالکون ایزومراز در مسیر تولید کوئرستین تحت تأثیر اشعۀ UV-B در کشت کالوس خیار تلخ ( .Momordica charantia L )
محورهای موضوعی : گیاهان دارویی
زیبا فولادوند
1
(دانشجوی دکتری ژنتیک مولکولی دانشگاه تربیت مدرس)
بهمن فاضلی نسب
2
(عضو هیات علمی دانشگاه)
کلید واژه: توفوردی, کوئرستین, فلاونوئید, کارلا, فنیل‌آمونیالیاز,
چکیده مقاله :
هدف از تحقیق حاضر بهینه‌سازی کالوس زایی و بررسی میزان بیان ژن‌های فلاونول سنتاز و چالکون ایزومراز در مسیر تولید کوئرستین تحت تأثیر اشعۀ UV-B در کشت کالوس خیار تلخ می باشد. غلظت‌های مختلف تنظیم‌کننده‌های رشدBAP و 2-4-D و ریز نمونه جهت تولید کالوس در گیاه خیار تلخ در طرح کاملاً تصادفی با سه تکرار ارزیابی شد. تاثیر اشعۀ UV-B (پنج گروه (شاهد (نور طبیعی)، 5 دقیقه در معرض اشعه (بررسی بیان ژن در 24 و 48 بعد از اعمال تیمار) و 10 دقیقه در معرض اشعه (بررسی بیان ژن در 24 و 48 بعد از اعمال تیمار)) بر بیان ژن‌های مسیر بیوسنتزی کوئرستین از جمله فلاونول‌سنتاز و چالکون‌ایزومراز، میزان فعالیت آنزیم آنتی‌اکسیدانی فنیل‌آمونیالیاز و فلاونوئید در بهترین کالوس های انتخاب شده بررسی شد. مقایسه میانگین داده‌ها نشان داد که بیشترین وزن خشک کالوس در غلظت 5/0 میلی‌گرم در لیتر 2-4-D و 1 میلی‌گرم در لیتر BAP برای ریز نمونه ساقه به دست آمد. بیشترین بیان ژن فلاونول سنتاز و چالکون ایزومراز در تیمار اعمال 10 دقیقه اشعۀ UV-B در زمان 24 ساعت بعد از شروع تیماردهی بدست آمده اما این بیان در 48 ساعت بعد از اعمال تیمار کاهش یافته است. بیشترین بیان فنیل‌آمونیالیاز، کوئرستین و فلاونوئید در زمان 5 دقیقه اعمال تیمار اشعه UV-B در 48 ساعت بعد از اعمال تیمار بدست آمده است. در کل نتایج این تحقیق نشان داد که اشعۀ UV-B در سطح کالوس تاثیر مثبتی بر بیان ژن‌های مسیر بیوسنتز فلاونوئیدها داشته است.
The present study aimed to optimize the callus induction and to determine the expression of flavonol synthase and chalcone isomerase genes in the pathway of quercetin production under UV-B rays in bitter cucumber callus cultivation. In this research was used concentration and combine of plant hormone 2, 4-D and BAP and explant to callus induction in Momordica charantia using complete random design with three replication. Genes expression study of Quercetin biosynthesis pathway such as FLS, CHI (were analyzed by 2-∆∆CT) and antioxidant activity assay of phenylalanine ammonia-lyase (Produced by concentration of cinnamic acid) and flavonoid using best callus were selected and affected by UV-B ray (five group(Control(natural light), 5 minute under UV-B ray(studying of gene expression after 24 and 48 hour) and 10 minute under UV-B (studying of gene expression after 24 and 48 hour)). Analysis of variance was shown that kind and concentration of plant hormone was significant on callus induction. Mean square of data was shown that the most callus dry weight was obtained from 0.5 mg/L 2,4-D and 1 mg/L BAP and stem explant. The highest expression of flavonol synthase and chalcone isomerase (chi) was observed in the 10-min UV-B treatment at 24 h after treatment but decreased at 48 hours after treatment. The highest expression of PAL, quercetin and flavonoid were obtained at 5 minutes UV-B treatment at 48 hours after treatment. Overall, the results showed that UV-B radiation at the callus level had a positive effect on the expression of flavonoids biosynthetic pathway genes.
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