بررسی عامل‌های مؤثر در القای ریشۀ موئین و افزایش میزان رزمارینیک‌اسید زرین‌گیاه (Dracocephalum kotschyi Boiss)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشیار، دانشکدۀ کشاورزی، دانشگاه ارومیه، ارومیه، ایران

2 دانشجوی سابق کارشناسی ارشد، دانشکدۀ کشاورزی، دانشگاه ارومیه، ارومیه، ایران

3 استادیار دانشکدۀ کشاورزی، دانشگاه ارومیه، ارومیه، ایران

چکیده

زرین­گیاه یا همان بادرنجبویۀ دنایی با نام علمی Dracocephalum kotschyi متعلق به تیرۀ نعناعیان و جنس بادرشبو (Dracocephalum) است که به علت پراکنش گستردۀ آن در ایران با نام­های چندی از آن یاد می­شود. ترکیب‌های فلاونوئید موجود در زرین­ گیاه، خاصیت ضدسرطانی دارد و استفاده از روش­های مختلف مانند تولید ریشه­های موئین می‌تواند به تولید ترکیب‌هایی با ارزش دارویی بالا کمک کند. در این بررسی تأثیر چهار نوع ریزنمونه (لپه یا کوتیلدون، محور ریزلپه یا هیپوکوتیل، برگ و میانگره( و سن ریزنمونه (برگ یک‌هفته‌ای، دوهفته‌ای و چهارهفته‌ای) بر میزان القای ریشۀ موئین گیاه زرین­گیاه بررسی شد. تأیید تراریختی با استفاده از آغازگر اختصاصی ژن rolB به روش PCR انجام شد. به‌منظور شناسایی محیط کشت مناسب برای رشد و افزونش ریشه­های موئین، تأثیر چهار نوع محیط کشت پایۀ مایع MS، 1/2MS، 1/4 MS وB5  و محرک UV-B در سه سطح: 3 دقیقه، 6 دقیقه و 9 دقیقه بر تغییر ویژگی‌های بیوشیمیایی و رزمارینیک اسید ریشه­های موئین بررسی شد. بیشترین درصد القاء ریشه در ریزنمونۀ برگ یک‌هفته‌ای (66/56 درصد) و لپه (66/36 درصد) مشاهده شد. بیشترین وزن­تر (34/4 گرم) و وزن خشک (31/0 گرم) در محیط کشت 1/4MS و کمترین وزن­تر (04/1 گرم) و وزن خشک (10/0 گرم) در محیط کشتB5  ثبت شد. بیشترین فعالیت پاداکسندگی (آنتی­اکسیدانی) در تیمار شش دقیقه (33/92 درصد) و کمترین میزان فعالیت پاداکسندگی در ریشه­های غیرتراریخت (63 درصد) به دست آمد.نتایج تجزیۀ HPLCنشان داد که تولید رزمارینیک اسید در ریشه­های تراریخت تیمارشده با UV-B با زمان شش دقیقه، 5/2 برابر بیشتر از ریشه­های غیر تراریخت (g /mg DW 9/34 در مقایسه با،µg /mg DW 8/13) است.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Study of factors affecting hairy roots induction and Rosmarinic Acid production in Dracocephalum kotschyi Boiss

نویسندگان [English]

  • Bahman Hosseini 1
  • Nasrin Ayyobi 2
  • Mohammad Fattahi 3
1 Associate Professor, Faculty of Agriculture, Urmia University, Iran
2 Former M.Sc. Student, Faculty of Agriculture, Urmia University, Iran
3 Assistant Professor, Faculty of Agriculture, Urmia University, Iran
چکیده [English]

Abstract: The flavonoid compounds in the Dracocephalum kotschyi has anti-cancer properties and using various methods such as hairy roots production can used to produce valuable secondary metabolites. In this study, the effect of explant type (cotyledon, hypocotyl, leaf and internode) and explants age (1 week leaf, 2 weeks leaf and 4 weeks old leaf) on hairy roots induction percent were surveyed. The transgenic status of hairy roots was confirmed by PCR using rolB genes-specific primers. In order to find optimum culture media for hairy roots growth and proliferation, four types of liquid basal media including, MS, 1/2MS, 1/4 MS and B5 and UV-B elicitor effects in three levels (3 min, 6 min and 9 min) on biochemical characteristics and Rosmarinic acid (RA) contents were investigated. The maximum hairy roots induction was obtained in 1 week leaves old (56.66 %); cotyledon explants (36.66 %), respectively. The highest fresh (4.34 gr) and dry weight (0.31 gr) and lowest fresh (1.04 gr) and dry weight (0.10 gr) was recorded in 1/4 MS and B5 media, respectively. The maximum and minimum antioxidant activities (92.33% and 63 %) were obtained in UV-B exposure time to 6 min radiation and non-transformed hairy roots. High performance liquid chromatography analysis revealed that the production of RA in UV-B (6 min) elicited hairy roots of D. kotschyi was almost 2.5-fold higher than that of the non-transformed roots (34.9 µg /mg DW compared to 13.8 µg /mg DW).

کلیدواژه‌ها [English]

  • Agrobacterium rhizogenes
  • antioxidant activitiy
  • Catalase
  • Total Flavonoid
  • Zarrin-giah

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  • تاریخ دریافت: 14 مهر 1395
  • تاریخ بازنگری: 25 اسفند 1395
  • تاریخ پذیرش: 26 اردیبهشت 1396

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