تأثیر ترکیبات محیط‌ کشت بر القای ریشه‌های مویین در کاسنی (Cichorium intybus L.) و افزایش متابولیت‌های ثانویه

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشیار گروه باغبانی دانشگاه محقق اردبیلی

2 گروه علوم باغبانی، دانشکده‌ی کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، ایران،

چکیده

هدف: کاسنی گیاه دارویی مهمی از خانواده‌ی گل‌ستاره‌ای‌ها ( (Asteraceaeبوده و دارای ترکیبات دارویی ارزشمندی می‌باشد. القای ریشه‌های مویین با استفاده از rhizogenes Agrobacterium یکی از روش‌های کاربردی برای افزایش بیوسنتز متابولیت‌های ثانویه می‌باشد.
مواد و روش‌ها: در این تحقیق، در آزمایش اول، تأثیر غلظت‌های مختلف عناصر ماکرو محیط‌ کشت بر کارایی القای ریشه‌های مویین در کاسنی بررسی شد. تائید مولکولی ریشه‌های مویین به وسیله‌ی PCR با استفاده از آغازگر‌های اختصاصی ژن rolB انجام شد. در آزمایش دوم میزان رشد لاین‌های مختلف ریشه‌های مویین کاسنی بررسی شد و همچنین تاثیر الیسیتور هورمونی و منبع کربن و بر رشد ریشه‌های مویین بررسی شد.
نتایج: نتایج به‌دست آمده نشان داد که بیشترین درصد القای ریشه‌های مویین در غلظت x1 پتاسیم نیترات، x5/1 نیترات آمونیوم، x1 و x5/1 منیزیم سولفات، x5/1 کلسیم کلرید و x5/0 پتاسیم فسفات (به‌ترتیب 66/46، 33/53، 66/46، 33/53 و 66/66 درصد) مشاهده شد. بیشترین میزان فنول کل در غلظت x1 پتاسیم نیترات، x2 نیترات آمونیوم، x5/1 منیزیم سولفات، x5/1 کلسیم کلرید و x5/0 پتاسیم فسفات (به‌ترتیب 21/4، 33/4، 26/4، 58/4 و 84/4 میلی‌گرم بر گرم وزن خشک) به‌دست آمد. غلظت x5/1 پتاسیم نیترات، x2 نیترات آمونیوم، x1 منیزیم سولفات، x5/1 کلسیم کلرید و x5/0 پتاسیم فسفات بالاترین میزان فلاونوئید را داشتند (به‌ترتیب 25/17، 52/18، 22/17، 3/18 و 96/17 میلی‌گرم بر گرم وزن خشک). لاین C بیشترین افزایش زیست‌توده (36/0 گرم) را نشان دادند همچنین بیشترین وزن تر (96/1 و 72/1 گرم در فلاسک) به ترتیب در ریشه‌های رشد کرده در محیط کشت حاوی 5/1 میلی‌گرم در لیتر NAA و 3 درصد و 4 درصد ساکارز به‌دست آمد.
نتیجه‌گیری: کشت ریشه‌های مویین یک فرآیند نوین برای تولید متابولیت‌های ثانویه در سطح وسیع است که قابل استفاده در صنایع داروسازی و غذایی می‌باشد. نتایج این پژوهش نشان داد که استفاده از سویه‌ی ATCC15834 و محیط هم‌کشتی حاوی x 5/0 KH2PO4 باعث افزایش میزان القای ریشه‌‌های مویین‌ و متابولیت‌های گیاه کاسنی شد.

کلیدواژه‌ها


عنوان مقاله [English]

The effect of medium compounds on hairy root induction in chicory (cichorium intybus l.) and enhancement of secondary metabolites

نویسندگان [English]

  • Mehdi Mohebodini 1
  • Roghayeh Fathi 2
1 Assoc. Prof, plant breeding, University of Mohaghegh Ardabili
2 Department of Horticultural science, Faculty of Agriculture and Natural resources, University of Mohaghegh Ardabili, Iran
چکیده [English]

Objective
Chicory (Cichorium intybus L.) is an important medicinal plant from Asteraceae family; contain a number of valuable medicinal compounds. Hairy root induction by Agrobacterium rhizogenes are an effective method for production of secondary metabolites.
 
Materials and methods
In this study, through first experiment, different concentrations of macro elements (0.5x, 1x, 1.5x, 2x, 2.5x and 3x concentrations of MS base medium) were investigated. In second experiment inoculation time was examined to establish an efficient transformation system for chicory. Molecular confirmation of transgenic hairy roots was done with PCR using gene-specific primers for rolB gene and also growth rate of hairy root lines obtained from different explants were investigated. Also the effect of hormonal elicitor and carbon source were studied in hairy roots growth.
 
Results
The results show that maximum chicory hairy roots induction was observed by using 1x KNO3, 1.5x NH4NO3, 1x and 1.5x MgSO4, 1.5x CaCl2 and 0.5x KH2PO4 (45.66, 53.33, 46.66, 53.33 and 66.66). The highest phenolic content was obtained by using 1x KNO3, 2x NH4NO3, 1.5x MgSO4, 1.5x CaCl2 and 0.5x KH2PO4 (4.21, 4.33, 4.6, 4.58 and 4.84). Also 1.5x KNO3, 2x NH4NO3, 1x MgSO4, 1.5 CaCl2 and 0.5x KH2PO4showed maximum flavonoid content (17.25, 18.52, 17.22, 18.3, 17.96 mg g DW respectively).The observation confirmed that grow of hairy root lines were significantly different and Line C showed higher biomass (0.36 g). Also the results of experiments revealed that 1.5 mg l-1 NAA in combination with 3 and 4% sucrose were superior for highest fresh (1.96 and 1.72 g) weight.
 
Conclusions
Hairy roots culture was developed as the innovative path for bulky production of secondary metabolites which find relevance in the pharmaceutical, food and flavor industries. The results indicated significant increases in hairy root induction and total metabolites content by ATCC15834 strain and 0.5x KH2PO4 co-culture MS medium.

کلیدواژه‌ها [English]

  • Inoculation
  • KH2PO4
  • Medium
  • Phenolic
  • RolB gene
فتحی رقیه؛ محب الدینی مهدی؛ چمنی اسماعیل (1397) بهینه سازی القای ریشه‌های مویین در کاسنی (Cichorium intybus L.)  و تأثیر اکسین و منبع کربن بر رشد آن ها. علوم باغبانی ایران (94)3، 667-657.
کبیرنتاج سارا؛ ذوالعلی جعفر؛ نعمت زاده قربانعلی؛ شکری احسان (1391) بهینه سازی شرایط القا و تثبیت کشت ریشه‌های موئین گیاه کاسنی (cichorium intybus) حاصل از تلقیح اگروباکتریوم رایزوژنز. مجله‌ی بیوتکنولوژی کشاورزی (4)2، 75-61.
مرادی فاطمه؛ زارع مهرجردی محبوبه؛ وحدتی کوروش؛ حسنلو طاهره (1397) تأثیر عوامل مختلف بر القای ریشه‌های مویین در سیر ایرانی. تولیدات گیاهی )مجله علمی کشاورزی( (14)1، 54- 43.
 
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